University of Pennsylvania Cardiovascular Institute T32 Research Training Program in Cardiovascular Biology and Medicine This is an exciting and dynamic time in which molecular biology and genomic technologies have transformed cardiovascular research. The primary purpose of this training grant is to provide rigorous research training for cardiology fellows and PhD scientists in cardiovascular biology and medicine with the ultimate goal of preparing trainees as independent clinician-scientists and investigators. The specific rationale underlying the training program is that state-of-the-art cardiovascular research (basic, translational and patient-oriented) requires a strong foundation in molecular and cellular biology and multi-departmental collaborations. In recognition of this principle, the University of Pennsylvania has recently established the Penn Cardiovascular Institute to promote multi-disciplinary cardiovascular research programs and education across schools, institutes and departments. The Penn Cardiovascular Institute is directed by Dr. Michael Parmacek, Program Director and Chief of the Division of Cardiovascular Medicine. Professor H. Lee Sweeney, Chairman of Physiology, will continue to serve as Co-P.I. of the training program. Together, they coordinate a multi-disciplinary group of training faculty that include well-funded and established investigators from the Departments of Medicine, Surgery, Physiology, Pharmacology, Cell and Developmental Biology, Genetics, the Institute for Medicine and Engineering and the Center for Clinical Epidemiology and Biostatistics. Pre- and postdoctoral Trainees for this program will be selected from the Penn Cardiology Fellowship Training program, the Penn Combined Degree Program (MD/PhD) and the Biomedical Graduate Studies (BGS) program. In addition, a strategy to attract individuals from under-represented minorities has been implemented. Trainees will have full access to the newly established University of Pennsylvania Molecular Cardiology Center and Laboratories for Experimental Electrophysiology and Interventional Cardiology. The core training will be a well-supervised research preceptorship lasting a minimum of two years. This training will be supplemented by graduate school classes, lectures, seminars, and specific courses in the ethical conduct of research. The training program has implemented a formal mentoring program and established an Oversight Committee to monitor trainee progress. The inter-departmental structure of this training program provides potential for an unparalleled training environment in cardiovascular research and is aligned directly with the NIH Roadmap. Resources and Environment

Institution Founded by Benjamin Franklin, the University of Pennsylvania is home of the oldest medical school in the country. It is a major research institution with a distinguished history in biomedical research and education. It has an externally funded research budget in excess of $487 million and currently ranks third in the nation in NIH support. Practically all of its components are located in contiguous space on a single urban campus. The School of Art and Sciences, in which the Biology, Chemistry, Physics and Mathematics departments are located, the Schools of Medicine, Veterinary Medicine, Dental Medicine, and Engineering are all present, facilitating interaction among these schools. In addition, the Wistar Institute, an affiliated institution with strong research programs in virology, immunology, molecular biology, molecular oncology, and vascular biology is located on the University campus. During the last decade, the University of Pennsylvania Medical School dramatically expanded facilities devoted to basic science and molecular medicine. Over 1,000,000 new square feet of state-of-the-art basic and translational research facilities were built on the Penn medical campus including the Clinical Research Building (CRB), Stellar-Chance Building, and the Biomedical Research Building (BRB) II/III which currently houses the Molecular Cardiology Center. In addition to traditional departments, the University houses Centers and Institutes dedicated to certain research areas, whose members hold primary appointments in established departments. This structure fosters collaborations that broach traditional departmental barriers. These include the newly established University of Pennsylvania Cardiovascular Institute (CVI) whose mission is to promote cardiovascular research across Schools, Departments and Divisions. This newly created Penn Cardiovascular Institute is directed by Dr. Michael S. Parmacek, P.I. of this training grant. The Pennsylvania Muscle Institute, supported by a NIH Program Project that spans several Universities and two continents; its emphasis is on advanced molecular/biophysical research in muscle structure and function. Dr. Lee Sweeney is Associate Director of the Muscle Institute and serves as Co-Director of this grant. The Institute for Medicine and Engineering (IME), directed by Professor Peter Davies, brings together biomedical scientists and engineers in a collaborative environment focused upon understanding and treatment of various disease states. The Institute for Translational Medicine and Therapeutics and the GCRC, directed by Dr. Garret FitzGerald, a trainer on this grant, were created to translate advances in basic science into novel therapies. The Center for Clinical Epidemiology and Biostatistics is internationally recognized for studies in health services research including outcomes, bio-informatics, pharmaco-epidemiology and health economics. The University of Pennsylvania Cardiovascular Institute The Penn CVI was established to promote multi-disciplinary translational cardiovascular research across schools, institutes, centers, departments and divisions at the University of Pennsylvania. The Penn CVI is an umbrella structure over all cardiovascular research and clinical programs on the Penn medical campus. In FY03, NIH support to cardiovascular research programs at Penn exceeded $54,000,000 including three SCCORs and thirteen PO1s focused on cardiovascular research. The CVI will leverage this strong foundation in cardiovascular research by promoting collaboration across

entities, providing additional infrastructure for translational research, and perhaps most importantly, identifying and breaking down barriers between basic scientists, translational researchers, clinical investigators and faculty members involved primarily in delivery of patient care. The CVI has been organized into multi-disciplinary working groups in strategic areas including: atherosclerosis/interventional cardiology, heart failure/myocyte biology, cardiovascular development/congenital heart disease, cardiac electrophysiology/channel biology and molecular imaging/bioengineering. All trainers that participate in this program are members of the Penn CVI. A critical component of the newly established Penn CVI is that faculty members performing cardiovascular research will be geographically aggregated (whenever possible) into adjacent spaces that will serve as cardiovascular centers-of-excellence on the Penn medical campus. As described under Background, faculty members performing molecular and translational cardiovascular and metabolic research and core laboratories supporting these functions (see below) will occupy approximately 75% (4 floors) of the CRB Building. Cardiovascular research programs and collaborations located outside these spaces also fall under the Penn CVI umbrella including on-going collaborations with investigators in the Departments of Genetics and Cell and Developmental Biology, the Institute for Medicine and Engineering and the Abramson Cancer Institute. Each of these institutes and departments has a strong focus in basic and/or translational cardiovascular research and will provide trainers for this program. Key components available to the trainees on this grant include: The Molecular Cardiology Center Over the past five years, many trainees have chosen mentors housed in the Penn Molecular Cardiology Center. This new modular space is devoted to molecular and cellular Cardiology research. Dr. Jonathan A. Epstein, is the Director of the Molecular Cardiology program and serves as a trainer on this grant. This 16,000 square foot facility is home to eight clinician-scientists within the Division of Cardiovascular Medicine and has bench space for up to 60 trainees. The Molecular Cardiology Center includes integrated Transgenic/ES cell, Molecular Biology and Molecular Imaging and Cell Culture Core Facilities with dedicated support personnel. The laboratories are all well equipped with state-of-the-art molecular biology equipment. Of note, in addition to performing technical service functions for investigators and trainees in the core laboratories, the Core Laboratories also offer quarterly technical training courses in areas related to trainees research. As such, trainees generating transgenic mice and/or genetically engineered mice are taught the technical aspects of micro-injection and blastocyst implantation by experts in these areas. Specialized equipment in the Molecular Cardiology Center includes: two micro-injection stations for oocyte injection and production of aggregation chimeras, two cryostat, an automated slide processing center, a Molecular Dynamics PhosphorImager, a Zeiss Axiophot and a Nikon microscope with darkfield, fluorescent and Numarski optics, an ELISA reader, beta and gamma counters, multiple light and dissecting microscopes with teaching heads, ultra-centrifuges, low and high speed centrifuges, PCR machines, a digital image acquisition center, several

spectrophotometers, multiple tissue culture hoods and incubators, a dark room and X-ray developer, a cold room and several autoclaves. In addition to the office space and secretarial support space, the Molecular Cardiology Center contains a central conference room/library (seating capacity approximately 50) that is used for lab meetings, as well as the weekly research seminar series, journal club and outside speaker series. A separate space has been set aside for students and post-doctoral fellows. One of the major purposes of this room is to foster collaborative interactions between students and fellows from different laboratories. Molecular Cardiology Center activities relevant to this training program include a weekly Molecular Cardiology seminars presented by our faculty, post-doctoral fellows and pre-doctoral students, a weekly journal club, and a monthly post-doctoral fellow research symposium. Thus, the Molecular Cardiology Center provides an interdepartmental and interdivisional structure for coordinating molecular and cellular research in cardiovascular biology at the University of Pennsylvania. B. School of Medicine Resources Cell Center: The primary services of the Cell Center include consultation regarding establishment and use of cell lines and strategies for production of monoclonal antibodies; training in basic sterile and tissue culture techniques, hybridoma and monoclonal antibody production, immunological assays, preparation of special media; mycoplasma and endotoxin testing; liquid nitrogen storage; and stockroom services. The latter includes over 850 discounted molecular biology and tissue culture products from 28 vendors. DNA Sequencing Facility: The DNA Sequencing Facility uses state-of-the-art instrumentation (ABI 377), techniques and data analysis tools to analyze the nucleotide sequence of DNA fragments. Extended sequences are determined by the method of primer walking routinely yielding sequences of 500 nt/run. Flow Cytometry and Cell Sorting Facility: The Flow Cytometry and Cell Sorting Facility provides a diverse, technically sophisticated cell sorting (Becton-Dickinson FACStarPLUS, Becton-Dickinson FACSort and Becton-Dickinson FACSCAN) and analysis service to investigators, including operator-dependent services, access to state-of-the-art instruments on a 24 hour/day basis and technical advice. Protein Chemistry Facility: The protein chemistry laboratory offers services that include peptide synthesis, peptide purification, peptide coupling to proteins, protein purification, tryptic peptic preparation, peptide modification, capillary electrophoresis, protein/peptide microsequencing (ABI 473A gas phase sequencer) and mass spectrometric analysis (Fisons Instruments VG Quattro tandem quadrupole and VG Tof Spec laser systems). Genomics/Microarray Core Facility: The Genomics/Microarray Core Facility offers technical services including preparation, hybridization and interpretation of Affymetrix microarray chips and biostatistical support related to experimental

design and interpretation. Software is available including the SAM package to users of the facility C. Translational Research Facilities The Institute for Translational Medicine and Therapeutics (ITMAT): This ITMAT is located immediately adjacent to the Molecular Cardiology Center facilitating collaborations between the Molecular Cardiology and ITMAT faculty members. Trainees in the basic/translational research track will have access to both faculties and shared core laboratories and move easily between these laboratories. The Center fosters translational and clinical research designed to identify new targets for therapeutic intervention by the study of human pathophysiology, to elucidate the mechanisms of drug action and the factors that contribute to differences among individuals in their responses to drugs. Current research themes include signal transduction mechanisms, the molecular and cellular biology of lipid mediators, and oxidant injury to the cardiovascular system. Center resources include a mass spectrometry and bioanalytical facility, a clinical investigational unit, a DNA Genomics Unit. The ITMAT is home to Genetics/Genomics and Proteomics Core Facilities. The Institute for Medicine and Engineering (IME): The IME is a supra-departmental interschool Institute established in 1996 with resource allocation from both schools. Its mission is to stimulate fundamental research at the interface between medicine and the engineering/computational sciences that will lead to innovative applications in biomedical research and clinical practice. The IME has recruited 11 faculty investigators skilled in biomedical engineering into departments of both schools, provided specialized space and facilities for the intellectual pursuit of interdisciplinary research, and attracted outstanding predoctoral and postdoctoral trainees into molecular and cellular biomedical engineering. In FY04, these faculty members received $11,200,000 in total NIH support. Membership in the IME currently stands at 91 faculty extending throughout the university. The Institute is centered in the Vagelos Research laboratories (13,000 nsf), with additional laboratory space in the Medical and Engineering complexes adjacent (approx 6,000 nsf). IME members represent a true interdisciplinary thrust by virtue of expertise in quantitative biology, biological aspects of engineering and clinical medicine. Research thrusts are Cardiovascular, Cell and Tissue Engineering, and Neuroengineering. Penns Center for Bioinformatics and a Center for Bioactive Materials in Tissue Engineering are within the administrative structure of the IME. Heart Failure and Transplantation Research Center: The Heart Failure Translational Research Center is currently located in approximately 7,000 square feet of space that will be relocated adjacent to the Molecular Cardiology Center in the Penn CVI later this year. This space includes a Cardiac Myocyte Function Core Laboratory and a Mouse Cardiovascular Physiology Core Laboratory. It also includes Heart Failure and Transplantation database and human sample repository. Specialized equipment includes a primary cardiac myocyte isolation facility, a working-heart preparation, a muscle-strip biophysical assessment station, two Nikon fluorescent microscopes, and a robotic microarray facility. Experimental Interventional Cardiology and Electrophysiology Laboratories: In order to promote translation of basic cardiovascular research to the clinical arena, the Division of

Cardiovascular Medicine has recently received approval to build an Experimental Interventional Cardiology Laboratory. This GLP-certified large animal catheterization laboratory is located adjacent to the large animal veterinary facility in the BRBII/III building. It is anticipated that some basic research that occurs in the Molecular Cardiology center will translate into pre-clinical testing in the Experimental Interventional Cardiology Laboratory. This laboratory will also benefit cardiology fellows who wish to pursue careers in academic medicine. In addition, the Division recently opened an Experimental Electrophysiology Laboratory in the Medical Science Research Laboratory building. This 7,000 square foot laboratory houses two electrophysiology suites and includes electroanatomic mapping systems, radiofrequency catheter ablation systems and intracardiac ultrasound equipment. D. Patient-Oriented Research Facilities The General Clinical Research Center (GCRC): The GCRC is a multi-disciplinary research facility funded by the NIH under the direction of Dr. Garret FitzGerald (trainer and member of the Oversight Committee). It is centrally located within the University of Pennsylvania Medical Center on the first floor of the Dulles Building in the Hospital of the University of Pennsylvania. The GCRC occupies newly renovated quarters that include technologies for the treatment of critically ill patients, as well as for the precise conduct of research. The GCRC has an experienced nursing staff dedicated to providing the clinical service and to making assessments for clinical research protocols. Included within the 5000 square foot GCRC are the following resources: 8 inpatient beds with both negative and positive air-flow options, separate outpatient facilities with 5 beds, HP cardiac monitoring including continuous oxygen saturation capability, computing facilities. Trainees have full access to the GCRC. The Center for Clinical Epidemiology and Biostatistics (CCEB): The Center for Epidemiology and Biostatistics was established in 1993 on the University of Pennsylvania Medical campus. Dr. Brian L. Strom, MD, MPH, Professor of Biostatistics and Epidemiology, Professor of Medicine and Pharmacology is the Center Director. Dr. Steven Kimmel, Associate Professor Department of Medicine (Cardiology) is Director of Cardiovascular Research at the CCEB. The CCEB has 33 primary faculty members and an additional 28 faculty members have secondary appointments. The annual research budget of the CCEB exceeds $31 million. The CCEB is an inter-disciplinary and interdepartmental program. Its mission is to improve the health of the public by linking epidemiology, biostatistics, and clinical medicine, bringing epidemiologic research methodology to clinical medicine. The educational programs offered by the Center for Clinical Epidemiology and Biostatistics (CCEB) are designed for health care professionals and respond to the individual needs of the trainees. These programs include a Master of Science in Clinical Epidemiology degree program (M.S.C.E.), a PhD degree program in Epidemiology, M.S. and PhD programs in Biostatistics, and combined M.D./M.S.C.E. and M.D./PhD degree programs. Cardiology at The Hospital of the University of Pennsylvania The clinical facilities of the Cardiovascular Division are located at the Hospital of the University of Pennsylvania. The outpatient practices and the Noninvasive Cardiac Imaging center, that includes echocardiography, exercise testing, and nuclear cardiology, is housed in the Rhoads Pavilion. The Heart Failure and Transplantation Ambulatory Care Center, included Cardiac Biopsy Suite, is

currently located in the Penn Tower Building. This facility was designed to meet the needs of heart failure and transplantation patients and includes patient exam rooms, on-site V02 exercise testing, and laboratory services. It is also home to Heart Failure research which includes dedicated research nurses, computer databases and a DNA bank. The outpatient Cardiology practices including the Heart Failure Center will be relocated into the Cardiovascular Institute in the Center for Advanced Medicine (CAM) which is currently under construction. The Cardiac Magnetic Resonance Imaging facility is located immediately adjacent to the noninvasive cardiology imaging center. Newly constructed state-of-the-art digital cardiac catheterization and electrophysiology laboratories are located on the 8th floor of the Founders Pavilion, immediately adjacent to the Coronary Care Unit. Each EP lab contains integrated equipment and software for electroanatomic/NOGA mapping and intra-cardiac echocardiography. Penn is a beta testing site for Siemens Inc. which provides us with pre-release versions of imaging equipment including Cath Labs, EP Labs, cardiac CT and cardiac MRI machines. A 30-bed inpatient Heart Failure Unit and Intermediate Cardiac Care Unit opened in July of 2004. This state-of-the-art facility includes dedicated spaces for clinical research nurses and access to all hospital databases. In addition, Penn cardiology faculty members staff the Philadelphia VA Hospital and Presbyterian Hospital. University of Pennsylvania Health System-wide computer databases have been developed to facilitate clinical research at each site. Faculty Approximately, 40% of the over 1295 faculty in the School of Medicine hold appointments in Graduate Groups. In addition, many faculty members in the School of Arts and Sciences and in the School of Engineering and Applied Science are engaged in biomedical education and research. The graduate programs in biology and bioengineering are closely affiliated with the biomedical science training programs in the health professional schools. Over 125 faculty members at the University of Pennsylvania describe their primary research area in the areas of cardiovascular biology or medicine. There are 65 full-time faculty members in the Division of Cardiovascular Medicine and there are an additional 12 full-time faculty members who hold joint appointments in basic science departments. The Cardiovascular Medicine Division is divided into nine Subsections: Heart Failure/Transplantation, Interventional Cardiology, Cardiac Electrophysiology, Cardiovascular Imaging (including MRI, echocardiography, PET imaging, nuclear cardiology), Vascular Medicine, Adult Congenital Heart Disease, Preventive and Clinical Cardiology, Molecular Cardiology and Health Services Research. An integral component of each clinical section is an active program of translational and patient-oriented research. Several of the clinical faculty members hold joint appointments in the Center for Epidemiology and Biostatistics. The Molecular Cardiology section consists of eight faculty members whose research interests include development of the heart and vascular system, vascular

biology/atherosclerosis, thrombosis and hemostasis, cardiac muscle function, receptor biology, and cellular electrophysiology. Faculty members with interests in basic, translational and patient-oriented research have served as mentors on this training grant. A weekly series of state-of-the-art lectures on basic and clinical cardiovascular research, serves as a vehicle to promote collaborations between the basic and clinical faculty members. Research Space The proposed training faculty includes scientists from several parts of the University of Pennsylvania and two distinguished research institutions that have close affiliations with the University. Faculty from the schools of Medicine, Dental Medicine, Veterinary Medicine, Engineering and Applied Science and Arts and Sciences play active roles in the presentation of course work and supervision of research in the laboratory. Total research space in the biomedical sciences includes over 875,000 nsf in the Medical School, in addition to large areas elsewhere in the Medical Center and University. As discussed above, the Molecular Cardiology Center is housed in 16,000 nsf of laboratory space and includes integrated Transgenic/ES cell, Molecular Biology and Molecular Imaging Core Laboratories. The translational heart failure program and mouse electrophysiology laboratory are located in approximately 7,000 nsf of laboratory space. The Experimental Interventional Cardiology and Electrophysiology laboratories are located in an additional 8,000 nsf of space. G. Special Facilities The University as well as the Schools of Medicine and Veterinary Medicine have excellent research libraries. There is on-line access to the internet, MEDLINE, OVID and all major biological databases throughout the University including the laboratories within the new Molecular Cardiology Center. Essential facilities also include several machine and electronic shops (Departments of Biochemistry and Biophysics and Physiology); a Molecular Biology/Cell Center that provides molecular biology reagents, cell culture media, monoclonal antibody facilities, DNA sequencing and plasmid preparation. The Cancer Center houses a cell sorter, genomics core laboratory. The Center for Biostatistics and Epidemiology provides assistance in study design, statistics and bio-informatics. There are also core facilities in computing, magnetic resonance imaging, bio-imaging, high energy laser spectroscopy, mass spectroscopy, NMR and a protein sequencing facility. All of these facilities are available to trainers and trainees of this program. H. Financial Commitment The Cardiovascular Medicine fellowship training program is a four to six year training program, with the first two years devoted to ACC-specified core clinical training in Cardiology. Some fellows elect to spend one additional year obtaining subspecialty training in an area such as Cardiac Electrophysiology, Interventional Cardiology or Noninvasive Imaging. This clinical subspecialty training is also supported by the hospital. As described in the training program, fellows entering the basic research track generally enter the training program at the beginning of their third year of fellowship training and remain in the basic science training program for an additional 2-3 years. This training is supported either through this T32 grant, an NRSA award, a K08 award or restricted funds available to the Chief of the Division of Cardiovascular Medicine that have been set aside for fellows in transition to faculty. Fellows, preparing for careers in translational research, may elect to

spend one year of subspecialty clinical training which is supported by hospital funds prior to formally entering the research training program. As described above for those fellows entering the basic research track, fellows entering the translational/POR track are supported either through this T32 award, an NRSA or a K23 grant. Some fellows interested in translational or patient oriented research spend an additional one or two years obtaining advances training through formalized programs offered through the Center for Epidemiology and Biostatistics. The Center for Clinical Epidemiology has a separate training grant that funds fellows entering the MSCE program. This training grant does not support fellows entering the MSCE program. All fellows receive stipends consistent with the PGY scale of the NIH.The University of Pennsylvania Medical School also has a large M.D./Ph.D combined degree program. The purpose of this program is to provide extensive training in the basic sciences for future clinician scientists. Combined degree students typically spend two years in Medical School which is complemented with intensive training in basic science. Following a series of three laboratory rotations, M.D./PhD students identify a mentor and propose a thesis. The typical M.D./PhD student spends 3-4 years in the laboratory fulfilling the requirements for a PhD. Of note, institutional support for MD/PhD students after their second year of laboratory experience is no longer available for MD/PhD students and the research mentor must provide funds to cover the additional one to two years of training usually required to complete a research thesis. This is followed by an additional one to two years of clinical training required for M.D. certification. These students are supported by a training grant and the sponsoring laboratory throughout their thesis studies. Research Training Program Background The recent revolution in cardiovascular science and medicine has emphasized the importance of providing young MD and PhD cardiovascular scientists with state-of-the-art training in molecular and cellular biology and genetics. The purpose of this application is to renew and expand a formal training program for pre- and post-doctoral MD and PhD fellows in the area of modern cardiovascular biology. Over the last decade, the application of modern molecular and genetic approaches to problems in cardiovascular science has begun to yield novel and important insights into cardiovascular development, biochemistry and physiology. Identification of new genes and signaling pathways and the application of genomic technologies has provided important insights into the molecular basis of many cardiovascular diseases. Because of this, emphasis has been placed on structuring our training program to provide training and exposure in molecular and cellular biology as a foundation for academic careers in basic, translational and patient-oriented research. Our experience over the past five years has validated this central tenet as evidenced by the large percentage of trainees who have launched successful careers as physician-scientists or who are in the process of completing their training in preparation for faculty positions at leading medical schools across the country.

Since the original submission of this application, it has become increasingly clear that areas traditionally not considered relevant to cardiovascular biology are in fact critical to understanding normal and abnormal cardiovascular physiology. For example, identification of novel cardiac myocyte-specific transcription factors led to fundamental insights into cardiac development and the molecular basis of some forms of congenital heart disease. The recent dramatic increase in the scope of cardiovascular science has underscored the importance of providing young cardiovascular scientists with training in excellent basic science laboratories with wide-ranging interests. In fact the multi-disciplinary nature of training physician scientists that was incorporated in this training program four years ago is entirely consistent with the principles underlying the NIH Roadmap. This principle has been retained and expanded in this application by the inclusion of MD and PhD preceptors from multiple disciplines and departments who share broadly defined collaborative interests in problems relevant to cardiovascular biology. Moreover, as described below, since the recent establishment of the University of Pennsylvania Cardiovascular Institute (CVI), directed by Michael S. Parmacek, P.I., in many cases scientists in different departments will be located in geographically contiguous spaces and they will share expanded core facilities. Thus, the trainees will be exposed to multiple preceptors functioning in a collaborative environment; a model system for integrating previously unrelated molecular and cellular approaches to problems in cardiovascular biology. One of the most difficult problems faced by all cardiovascular sciences training programs is devising methods for training outstanding MD investigators who choose to follow academic research careers following the completion of their training program. This difficulty stems in part from the long period of training required for clinical cardiology, and in part from the salary differential between clinical and research careers in cardiology. The training program proposed in this application uses three approaches that address this problem. First, because we believe that it is much easier to interest MDs in basic research early in their careers, we are requesting two pre-doctoral training positions. These positions are especially important because they represent a mechanism for attracting MD/PhD students to the cardiovascular sciences, an area that has not been traditionally attractive to these students. Secondly, as described in detail below, we will continue to identify and recruit fellows into the Penn cardiology fellowship training program that have demonstrated previous interests and skills in basic research. Finally, we will train MD and PhD fellows together in MD and PhD run laboratories in the newly established Penn Cardiovascular Institute. This approach has the advantage of exposing relatively inexperienced MD fellows to the research expertise and high research standards of their PhD colleagues. In addition, it is felt that the PhD fellows will benefit from exposure to MD fellows with an understanding of human cardiovascular physiology.

It should be emphasized that training grant support will not be limited to MDs in the adult cardiology training program. MD fellows from other sections and departments are currently pursuing basic cardiovascular research projects in the laboratories of several of the preceptors of this application. Such fellows would be excellent candidates for funding by this training grant and such funding would facilitate collaborative interactions involving investigators with interests and expertise from multiple disciplines with relevance to cardiovascular biology. Given the challenges and opportunities that are rapidly evolving in molecular medicine, there will be an increasing need for clinician scientists and clinical investigators with extensive training in the basic sciences as well as PhDs interested in applying basic science to clinical problems. This training program is designed to meet these needs. We have learned that to prepare physician-scientists for careers in translational research trainees must have an extensive foundation in basic research. In recognition of this the proposed training program includes a basic research track and a translational/patient-oriented research track. The basic research track will involve 3-4 years of intensive training and basic/molecular research. In general, this pathway will involve training of MDs, or MD/PhDs who have competed two years of a clinical cardiology fellowship program, or PhDs interested in applying molecular biology to problems in molecular cardiology. Over the course of the past five years, all trainees in this track have been mentored by physician-scientists in the Molecular Cardiology Center, the Department of Physiology and/or the Department of Pharmacology. The translational/patient-oriented research track is designed to provide a strong foundation in molecular and cell biology upon which a career in patient-oriented cardiovascular research can be established. In addition, this track offers the option of training in health services research for those interested in genetic epidemiology, bio-informatics, outcomes and health economics. Because of the variety of pathways open to translational cardiovascular research each training program will be tailored to the individual needs of the trainee (within the general requirements described under Training Program). However, a fundamental tenet guiding this training is that success in translational research requires a foundation in a focused area of basic science that may then be applied. The proposed program differs from the existing program in several aspects. First, as discussed above, we believe it is important to attract MD/PhD or PhD graduate students into basic cardiovascular research at an early stage in their career. Accordingly, we are now requesting support to fund two graduate students per year (MD/PhD or PhD) to support their thesis studies. Second, we have expanded the multi-disciplinary nature of the training available to the trainees by expanding the list of trainers in other departments. In this regard we are particularly excited about incorporating trainers working in the important area of metabolic disease/diabetes in this application. Table II is a listing of the number of faculty members in each unit and department and total numbers of current pre-doctoral students and postdoctoral trainees in each unit or department is shown in Table III. Third, this proposal incorporates the expanded infrastructure for cardiovascular research as part of the newly established University of Pennsylvania Cardiovascular Institute (CVI).

The University of Pennsylvania Cardiovascular Institute (CVI) was established in January 2005 to promote multi-disciplinary cardiovascular research across schools, institutes, centers, departments and divisions at the University of Pennsylvania. It is directed by Dr. Michael S. Parmacek, P.I. of this training program. The Penn CVI is an umbrella structure over all cardiovascular research and clinical programs on the Penn medical campus. In FY03, NIH support to cardiovascular research programs at Penn exceeded $54,000,000 including two SCCORs and thirteen PO1s focused primarily on cardiovascular science and medicine. The CVI will leverage this strong foundation in cardiovascular research by promoting collaboration across entities, providing additional infrastructure for translational research, and perhaps most importantly, identifying and breaking down barriers between basic scientists, translational researchers, clinical investigators and faculty members involved primarily in delivery of patient care. The CVI has been organized into multi-disciplinary working groups including basic, translational and patient-oriented research faculty in strategic areas including: atherosclerosis/interventional cardiology, heart failure/myocyte biology, cardiovascular development/congenital heart disease, cardiac electrophysiology/channel biology and molecular imaging/bioengineering. A critical component of the newly established Penn CVI is that faculty members performing cardiovascular research will be geographically aggregated (whenever possible) into adjacent spaces that will serve as cardiovascular centers-of-excellence on the Penn medical campus. As described below, faculty members performing molecular and translational cardiovascular and metabolic research and core laboratories supporting these functions will be relocated into adjacent spaces in the Clinical Research Building (CRB). Cardiovascular research programs and collaborations located outside these spaces also fall under the Penn CVI umbrella including on-going collaborations with investigators in the Departments of Genetics and Cell and Developmental Biology, the Institute for Medicine and Engineering and the Abramson Cancer Institute. Each of these institutes and departments has a strong focus in basic and/or translational cardiovascular research and will provide trainers for this program. Some of the key participating units in the CVI and cardiovascular research programs that will be made available for trainees are described below: The Molecular Cardiology Center (Jonathan A. Epstein, M.D., Director) The Penn Molecular Cardiology Center was established in 1999 to promote interdepartmental collaborations and to facilitate scientific interactions among investigators with interests in the molecular and cellular biology of the cardiovascular system. The program is currently directed by Dr. Jonathan A. Epstein who is internationally recognized for his research in cardiovascular development. The Molecular Cardiology Center is currently located in the Biomedical Research Building (BRB) II/III building and will be relocated into the Penn Cardiovascular Institute in the CRB Building later this year.

The Molecular Cardiology Center is home to eight faculty members in the Cardiovascular Medicine and CT Surgery Divisions. The Molecular Cardiology Center contains integrated Transgenic/ES cell, Molecular Biology and Molecular Imaging and Mouse Physiology Core Laboratories that are partially subsidized by the Penn Cardiovascular Institute and a restricted endowment for cardiovascular research. In addition to the office space and secretarial support space, the Molecular Cardiology Center contains a central conference room/library (seating capacity approximately 50) that is used for lab meetings, as well as the weekly research seminar series, journal club and outside speaker series. A separate space has been set aside for students and post-doctoral fellows. One of the major purposes of this room is to foster collaborative interactions between students and fellows from different laboratories. Molecular Cardiology Center activities relevant to this training program include a weekly Molecular Cardiology seminar series for participating factulty, students and fellows, a weekly journal club, and a monthly post-doctoral fellow research symposium. The Molecular Cardiology Center and faculty members will serve as mentors for many trainees in the basic research track (see below). Over the past five years, eight trainees funded by this training grant have been mentored by physician scientists in the Molecular Cardiology program. As shown in Table IX, all of the graduates them have obtained faculty positions at leading academic institutions across the country and/or have gone on to obtain NIH and other extramural support. The Pennsylvania Muscle Institute (PMI) (Yale E. Goldman, M.D., Ph.D, Director) The Pennsylvania Muscle Institute was formed in 1973 as a cross‑disciplinary research institute to promote collaborative research among muscle and cell motility researchers with common interests and methodologies. Many important discoveries and development of significant new technologies followed. H. Lee Sweeney, Ph.D., Chairman of the Department of Physiology serves as the Co-P.I. of this training program. Today, the University of Pennsylvania is one of the outstanding centers for muscle and motility research in the world. As the range of interests and expertise of the trainers listed on this grant will attest, virtually all of the important areas of current research in muscle are covered by PMI faculty. Graduate and postdoctoral research opportunities are provided in: 1) Cell Migration and Intracellular Transport, 2) Mechanism and Regulation of Muscle Contraction, 3) Structure/Function Studies of Molecular Motors, 4) Chromosome Segregation and Cell Division, 5) Gene Therapy and Neuromuscular Disease, 6) Muscle Cell Development and Myofibril Assembly, and 6) Advanced Technological Development. This provides the potential for an unparalleled training environment in the area of cardiac muscle biology. Five trainees funded by this grant have been mentored by faculty members based in the PMI and the Department of Physiology (Table IX and X). The Institute for Medicine and Engineering (IME) (Peter Davies, Ph.D., Director) The IME is a supra-departmental interschool Institute established in 1996 with resource allocation from both schools. Its mission is to stimulate fundamental research at the interface between medicine

and the engineering/computational sciences that will lead to innovative applications in biomedical research and clinical practice. Dr. Peter Davies, an internationally recognized vascular biologist with a longstanding interest on the effect of shear stress (physical forces) on the vascular endothelium, serves as Director of the Institute. The IME has recruited 11 faculty investigators skilled in biomedical engineering into departments of both schools, provided specialized space and facilities for the intellectual pursuit of interdisciplinary research, and attracted outstanding predoctoral and postdoctoral trainees into molecular and cellular biomedical engineering. In FY04, these faculty members received $11,200,000 in total NIH support. Membership in the IME currently stands at 91 faculty extending throughout the university. The Institute is centered in the Vagelos Research laboratories (13,000 nsf), with additional laboratory space in the Medical and Engineering complexes adjacent (approx 6,000 nsf). IME members represent a true interdisciplinary thrust by virtue of expertise in quantitative biology, biological aspects of engineering and clinical medicine. Research thrusts are Cardiovascular, Cell and Tissue Engineering, and Neuroengineering. Penns Center for Bioinformatics and a Center for Bioactive Materials in Tissue Engineering are within the administrative structure of the IME. The IME Seminar Series hosts about 40 seminars each year by guest and local speakers on a variety of topics in cutting-edge interdisciplinary research (see Appendix). Heart Failure and Transplantation Research Program (Ken Margulies, M.D., Director) Over the past five years, the University of Pennsylvania has established one of the five busiest heart failure and transplantation programs in the United States. In parallel with establishing this large clinical program and database, we have successfully recruited three leading translational researchers in the area of heart failure and transplantation. The Heart Failure and Transplantation Translational Research Program is directed by Dr. Ken Margulies who is nationally recognized for his translational heart failure research focused on human cardiac myocyte physiology. The Heart Failure Translational Research Center is currently located in approximately 7,000 square feet of space that will be relocated to the Penn CVI. This space includes a Cardiac Myocyte Function Core Laboratory and a Mouse Cardiovascular Physiology Core Laboratory. Drs. Tom Cappola and Dan Dries who were recently recruited to Penn to establish a program in the Genetics and Genomics of Heart Failure and Transplantation are also located in this space. The Heart Failure and Transplantation program will provide a relatively unique environment for physician-scientists interested in obtaining postdoctoral training in translational heart failure and transplantation research. Cardiac Electrophysiology Research Program (Francis Marchlinski, Director) The University of Pennsylvania is internationally recognized for the pioneering research programs in the area of cardiac electrophysiology. The program established by Dr. Mark Josephson in the late seventies is now directed by Dr. Francis Marchlinski. Dr. Marchlinski is internationally recognized for his studies demonstrating the efficacy of radiofrequency catheter ablation and devices to treat and in some cases cure ventricular and atrial tachyarrhythmias. Penn EP faculty members (9 full-time clinical investigators and three basic/translational scientists) have a longstanding record of academic

productivity and of training investigators for careers in academic electrophysiology. Vickas Patel, M.D., Ph.D., a past-trainee on this grant, was recently recruited to Penn to establish a translational program in channel biology/mouse electrophysiology. He will oversee the training of physician-scientists interested in academic careers in basic cardiac electrophysiology resarch. Trainees interested in channel biology/cardiac electrophysiology have access to the Mouse Cardiac Electrophysiology Core laboratory and the large animal Experimental Electrophysiology Laboratory. Three Cardiology fellows have elected to obtain advanced research training in this laboratory under the mentorship of Drs. Marchlinski and Patel, MD (Tables IX and X). GLP-Certified Experimental Interventional Cardiology Laboratory (Robert Wilensky, Director) To facilitate preclinical research, a GLP-certified Experimental Interventional Cardiology Laboratory was established in the basement of the BRBII/III building. This laboratory is directed by Dr. Robert Wilensky, an interventional cardiologist with expertise in intra-coronary drug and gene delivery. The focus of the laboratory is to translate discoveries in vascular biology into new therapies for atherosclerosis. Through this program, trainees have the opportunity to obtain expertise in molecular biology, vascular biology and biochemistry. Two trainees sponsored by this grant have trained in this program (see Tables IX and X). The Abramson Family Cancer Research Institute/Angiogenesis Program (M. Celeste Simon, Director) The Angiogenesis and Vascular Development research program was established in 1999 under the direction of Professor M. Celeste Simon, Investigator HHMI. Dr. Simon is recognized for her research elucidating the role of specific transcription factors in regulating angiogenesis and hypoxic response. In addition, Dr. Gary Koretzky, who is internationally recognized for his expertise in intracellular signaling, is a member of the Abramson Institute. Dr. Koretzky has established collaborations with Dr. Mark Kahn (who is based in the Molecular Cardiology Center) studying the function of novel G-protein coupled receptors in platelets. The Abramson Family Cancer Research Institute hosts a weekly seminar series that is attended by many of the graduate students and postdoctoral fellows in the Molecular Cardiology program. Drs. Simon, Koretzky and Kahn serve as trainers on this training program. Dr. John Lee, a current trainee on this grant who is mentored by Dr. Mark Kahn and Gary Koretzky, is examining the function of lung Kruppel-like factor (LKLF) in the developing vasculature (Table IX and X). Patient-Oriented Cardiovascular Research As described below, the formalized training program in translational/patient-oriented research was structured to prepare MDs for careers in academic cardiology and related sub-specialties. We believe a critical component to this training is formalized training including training in study design, biostatistics and molecular and cellular biology. Formalized training programs tailored to the specific needs of trainees on this grant are available through the Center for Clinical Epidemiology and Biostatistics, the newly established Institute for Translational Medicine and Therapeutics and the NIH-sponsored Penn General Clinical Research Center. Accordingly, trainees have the option of entering

a degree granting program or entering a core curriculum that is individually designed to meet the future needs of the trainee. For example, trainees interested in interventional cardiology will be required to take specific courses in vascular biology and molecular medicine. Some of the participated units in this training grant that contain programs relevant to patient-oriented research are described below: The University of Pennsylvania General Clinical Research Center (Garret Fitzgerald, Director) Founded in 1962 with a NIH grant, the Penn General Clinical Research Center (GCRC) is a multidisciplinary research facility which is located in a recently constructed unit of the Hospital of the University of Pennsylvania. The GCRC is a core facility for patient related clinical research at the Penn Medical Center. It is directed by Garret Fitzgerald and the Associate Director is Dan Rader, both of whom serve as trainers for this program. The GCRC houses eight in-patient beds and five out-patient bays and includes a metabolic kitchen staffed by a dietician and a highly sophisticated cardiac monitoring system. The GCRC has state-of-the-art software and databases designed to store, protect and analyze research data. The GCRC is utilized by faculty members in the Center for Clinical Epidemiology and Biostatistics and the Institute for Translational Medicine and Therapeutics to provide practical training for students and postdoctoral fellows enrolled in degree granting programs. Trainees, under the mentorship of a trainer, have the opportunity to perform clinical protocols in the Penn GCRC. The Center for Clinical Epidemiology and Biostatistics (Brian Strom, Director) The Center for Epidemiology and Biostatistics was established in 1993 on the University of Pennsylvania medical campus. Brian L. Strom, MD, MPH, Professor of Biostatistics and Epidemiology, Professor of Medicine and Pharmacology is the Center Director. Dr. Stephen Kimmel, Associate Professor of Medicine (Cardiology) is Director of Cardiovascular Research at the CCEB. The CCEB has 33 primary faculty members and an additional 28 faculty members have secondary appointments. The annual research budget of the CCEB exceeds $31 million. The CCEB is an inter-disciplinary and inter-departmental program. Its mission is to improve the health of the public by linking epidemiology, biostatistics, and clinical medicine, bringing epidemiologic research methodology to clinical medicine. The educational programs offered by the Center for Clinical Epidemiology and Biostatistics (CCEB) are designed for health care professionals and respond to the individual needs of the trainees. These programs include a Master of Science in Clinical Epidemiology degree program (M.S.C.E.), a PhD degree program in Epidemiology, M.S. and PhD programs in Biostatistics, and combined M.D./M.S.C.E. and M.D./PhD degree programs. In addition, a Clinical Research Certificate program is available to those trainees who envision careers as clinician investigators. This program provides didactic course work, but does not require preparation of a thesis project. The curriculum for this program is described under section B3. Proposed Training (below).

The Institute for Translational Medicine and Therapeutics (Garret Fitzgerald, Director) Trainees in the translational/patient-oriented research track will have access to faculty members and core laboratories located in the ITMAT. Dr. Garret FitzGerald, M.D., Ph.D., a cardiologist who chairs the Dept. of Pharmacology, serves as director of the ITMAT. The Institute fosters translational and clinical research designed to identify new targets for therapeutic intervention by the study of human pathophysiology, to elucidate the mechanisms of drug action and the factors that contribute to differences among individuals in their responses to drugs. Center resources include a mass spectrometry and bioanalytical facility, a clinical investigational unit, a DNA Genomics Unit. The ITMAT is home to Genetics/Genomics and Proteomics Core Facilities. In addition to providing core resources for translational research, the ITMAT provides an internal framework for students and fellows wishing to purse careers in patient-oriented research. Translational research and patient-oriented trainees sponsored by this grant may enroll in the Masters of Science in Translational Research degree program, but at a minimum most complete the Translational Medicine Core Curriculum or CCEB sponsored certificate program. Each of these programs are described further under section B3. Proposed Training (below). B. Program Plan1. Program Direction Principal Investigator The Program will be directed by Dr. Michael S. Parmacek, the Herbert C. Rorer Professor of Medical Sciences, Chief of the Division of Cardiovascular Medicine and director of the newly established University of Pennsylvania Cardiovascular Institute (see above). He is an NIH/NHLBI-funded scientist who for the past 17 years has demonstrated interest and commitment to training medical and graduate students and MD, PhD and MD/PhD postdoctoral fellows. After completing Cardiology fellowship training, Dr. Parmacek performed a postdoctoral research fellowship in the Howard Hughes Medical Institute at the University of Michigan where he later joined the faculty. Subsequently he has been a faculty member at the University of Chicago (1992-1998) serving as Co-Director of the Cardiology Fellowship Training Program and a member of the Executive Committee of the University of Chicago Training Grant in Cardiovascular Sciences. He joined the faculty of the University of Pennsylvania in 1998 as Chief of the Division of Cardiovascular Medicine and established the Penn Molecular Cardiology Center in 1999. Recently, Parmacek was named director of the newly established University of Pennsylvania Cardiovascular Institute. Dr. Parmacek's research interest lies in the examining the molecular and genetic programs regulating cardiovascular development.He is currently the P.I. of two NIH-funded research grants and serves as an ad-hoc member of several different NHLBI study sections. He is an elected member of the ASCI and AAP and a former Established Investigator of the AHA. He is active in medical and graduate education in a number of ways, including by providing didactic lectures for undergraduate and graduate medical education, lecturing in courses in the Departments of Physiology and Pharmacology, providing lectures to the Cardiology fellows, and serving as attending physician in the CCU and Cardiology Consultation services where he trains medical

students, residents and Cardiology fellows. His duties as Director of this training program will include advising all trainees and overseeing and coordinating formal training exercises that constitute the program. He also serves as Chair of the training grant Advisory Committee (see below). This will involve 10% of Dr. Parmaceks total effort. Dr. Parmacek has served as the primary trainer for graduate and medical students and postdoctoral fellows including four trainees sponsored by this grant (Tables IX and X). In this position, he works closely with Dr. Martin St. John Sutton (Director of the Cardiology Fellowship Training Program) advising cardiovascular fellows and MD/PhD students in the choice of research areas, and in following their progress. Co-Director of the Training Program Dr. H. Lee Sweeney, Professor and Chairman of Physiology will serve as Co-Director of the Training Program. He is also Associate Director of the Pennsylvania Muscle Institute. He joined the University of Pennsylvania faculty in 1989, and has been an active participant in graduate teaching and curriculum development. Dr. Sweeney spearheaded the development of the Training Program in Muscle and Motility. Dr. Sweeney received formal training in biochemistry (S.B.) and in physiology and biophysics (PhD). His major research is examining muscle function and physiology at a molecular level. He also studies the molecular mechanisms regulating cardiac myocyte survival following ischemic injury and has utilized gene transfer to attenuate or abolish myocardial injury following cardiac ischemia. Moreover, he has pioneered novel methods to regulate virus-mediated gene transfer. His laboratory routinely utilizes techniques of molecular biology, biochemistry and biophysics. Over the last nine years, he has mentored five fellows on this training grant (Tables IX and X), all of whom were in clinical departments including Cardiology, Thoracic Surgery and Vascular Surgery. His duties as Co-Director of this grant will include advising all basic science trainees and overseeing and coordinating formal training exercises (i.e. journal clubs and retreats) that constitute the program. This will involve 5% of Dr. Sweeney's total effort. Fiscal administration of this grant will be handled by the Business Office of the Penn Cardiovascular Institute. Ms. Barbara Barras, the Grants Administrator for the Division or Cardiovascular Medicine and Penn CVI, administers all awards for the Division of Cardiovascular Medicine and will assume responsibility for the grant, and will provide a monthly financial summary. Ms. Barras will act as the liason between the University, the Training Program and the National Heart Lung and Blood Institute. An Oversight Committee, which consists of trainers in a variety of disciplines spanning basic, translational and patient-oriented research, will advise the Program Directors on all major policy matters pertaining to this training program and grant. This committee is responsible for the selection of all trainees, mentors for these trainees, and will decide the duration of the trainees support on the grant.

Thereafter, the committee will assess the performance of trainees on a quarterly basis. The committee also will help in the coordination of components of the training program, including courses and seminar programs. In addition, the program directors plan to invite two prominent researchers in academic cardiology as visiting professors each year to provide critical evaluation of the training program as well as to serve as guest lecturers. The Oversight Committee consists of: 1) Dr. Michael S. Parmacek, Program Director, Herbert C. Rorer Professor of Medical Sciences, Chief Division of Cardiovascular Medicine, Director of the Penn Cardiovascular Institute 2) Dr. H. Lee Sweeney, Program Co-Director, Professor of Physiology and Medicine, Chairman of the Department of Physiology 3) Dr. Jonathan Epstein, W.W. Smith Professor of Medicine, Director of the Molecular Cardiology Program, Associate Director of the Penn Cardiovascular Institute 4) Dr. Garret FitzGerald, Professor and Chairman of the Department of Pharmacology and Director of the ITMAT and GCRC 5) Dr. Dan Rader, Associate Professor of Medicine, Director of the Penn Preventive Cardiology program, Associate Director of the Penn GCRC 6) Dr. Martin St. John Sutton, Professor of Medicine, Director Cardiology Fellowship Training Program 7) Dr. Lawrence Brass, Professor of Medicine and Pharmacology, Assoc. Dean and Director Combined Degree and Physician Scholar Program (MD/PhD Program) 2. Program Faculty There are currently 36 faculty trainers associated with this grant. Table II is a list of program faculty including their departmental affiliation, role and percent effort. A brief description of their research interests is provided below. All of them are full-time members at the University of Pennsylvania and devote most of their time to basic, translational or patient-oriented research. Table III is a listing of all current and pending training support available to participating faculty members and departments including the funding source, complete id number, title of training program, name of program director, project period, number of training positions (pre and post doc) and the amount of award. Of note, none of these grants supports training in basic or translational cardiovascular research. Table IV lists all past and current students for whom program faculty are serving as thesis advisor or sponsor. Those trainees who were, or are, supported by this training grant are denoted with an asterisk. Trainer Research Descriptions Michael S. Parmacek, MD (Director) The Parmacek laboratory examines the molecular and genetic programs regulating cardiovascular development. In the Parmacek laboratory transgenic and gene knockout technologies are utilized to characterize the function of genes and proteins mediating critical functions in the developing cardiovascular system. The Parmacek laboratory cloned and characterized the function of the GATA-4/5/6 subfamily of zinc finger transcription factors in the cardiovascular system. The Parmacek laboratory also studies the transcriptional regulatory programs regulating vascular smooth muscle cell

differentiation and modulation of SMC phenotype. While primarily basic/molecular in nature, these fundamental studies have provided insights into the pathogenesis of common forms of congenital heart disease and vascular proliferative syndromes including atherosclerosis. In addition, Dr. Parmacek is involved in several on-going collaborations in translational cardiovascular research in the areas of metabolic/diabetic vascular disease and heart failure. H. Lee Sweeney, PhD (Co-Director) Dr. Sweeney’s laboratory integrates biochemical and molecular biological tools and techniques with cellular physiology. Dr. Sweeneys basic studies are focused primarily on elucidating muscle function at a molecular level. This work involves assessing protein structure/function relationships of the heavy chain and the light chains of myosin. Wild type and mutant cardiac myosin heavy chain (and other contractile proteins) are introduced into myogenic cells using a variety of techniques including transfection, micro-injection and adenovirus-mediated gene transfer. Among the many questions currently under study is examination of the molecular mechanisms that lead to hypertrophic cardiomyopathy. In addition, the Sweeney laboratory is actively investigating novel gene therapies that may be applicable in the future to the treatment of muscle dystrophies and cardiomyopathy. Morris Birnbaum, MD, PhD Dr. Birnbaum's laboratory studies the processes that coordinately regulate growth and metabolism in a number of contexts, including the regulation of glucose metabolism by insulin in vertebrates, and the determination of cell size in Drosophila melanogaster. A critical regulator of such processes is the serine/threonine protein kinase known as Akt or PKB. This enzyme has been implicated as a mediator of insulin action, a prime regulator of cellular growth, and as a potent anti-apoptotic factor. Dr. Birnbaum's laboratory is utilizing cell culture and mouse models to investigate the role and mechanism of Akt/PKB in such processes as vasculogenesis, glucose utilization, and hypertrophy of vascular smooth muscle, as well as cardiac and skeletal muscle. In the latter two organs, AMP-activated protein appears to have a major role in switching from an anabolic to a catabolic program. Lawrence F. Brass, MD, PhD Dr. Brass research focuses on understanding the molecular mechanisms underlying atherosclerosis and thrombosis. Specifically, they study the signal transduction pathways that allow platelets and endothelial cells to respond to extracellular events. Topics currently under investigation include: 1) Protease-activated G protein coupled receptors, including the thrombin receptor. Their work focuses on the identification of additional family members, the signaling pathways that are initiated when these receptors are activated, the trafficking of the receptors within the cell as used receptors are replaced, and the development of receptor antagonists and clinically-useful assays for detecting receptor activation in vivo. 2) The role of the G protein, Gz. Gz is one of the G proteins that are not universally expressed, its presence being limited mainly to some neural cells and to some hematopoietic cells, particularly megakaryocytes and platelets. 3) The role in platelet activation and endothelial cell biology of Eph tyrosine kinase receptors and their ligands, which are known as ephrins. David J. Callans, MD

Dr. Callans research interest is in defining the molecular and anatomic basis of ventricular tachyarrhythmias (VT) he serves as the Director of the Experimental Electrophysiology Laboratory (EEPL). In the EEPL a swine model of post-infarct VT is used to test the hypothesis that detailed mapping performed during VT superimposed on the voltage map obtained in sinus rhythm will: 1. Facilitate the development of a minimalistic linear lesion strategy, 2. Obviate the need for future mapping during VT, and 3. Allow for the development of prophylactic ablation strategies. Dr. Callans also performs patient-oriented research examining whether overlapping linear catheter ablation lesions of the electroanatomic VT substrate, performed in sinus rhythm guided by the CARTO mapping system, is feasible and will result in curative treatment for ventricular arrhythmias associated with healed myocardial infarction. Thomas P. Cappola, MD, MPH Dr. Cappolas primary research interest is examining the molecular basis of heart failure. He directs the genomics of heart failure program and has established a heart failure/ genomics database at Penn. Consenting heart failure patients are entered into a database that includes over 300 variables including clinical profile, hemodynamic indices, family history (detailed), standard laboratory data and neurohormonal profile. In addition, Dr. Cappola has established a DNA bank on all patients entered in the genetics and genomics of heart failure program. The focus of Dr. Cappolas current research is on identifying critical transcription factors that modulate heart failure with particular focus on the role of PPARg signaling mechanisms. Peter F. Davies, PhD The Davies lab is a cell and molecular facility focused on mechanism of cardiovascular diseases, particularly the biomechanics (mechanotransduction) of arterial biology and pathology. Of major interest are the mechanisms of interaction of hemodynamic forces with the arterial endothelium. The fluid dynamics which define mass transport characteristics at the endothelial surface are also of interest. Experimental approaches range from model flow systems to transgenic animals. Characterization of endothelial surfaces using atomic force microscopy and finite element analyses are linked to single cell and regional endothelial gene expression. A powerful technique in use is a combination of single cell mRNA amplification combined with high throughput microarray hybridizations to develop transcriptional profiles that are precisely defined spatially. A second thrust is in the area of cell-cell communication, specifically through analyses of endothelial gap junction biology. A third approach to cytomechanics is through the development and use of GFP-cytoskeletal constructs that facilitate real-time imaging of intermediate filament and actin dynamics during changes in the mechanical environment in living cell geometry arising both spontaneously and as a function of applied hemodynamic forces. Jonathan A. Epstein, MD The Epstein laboratory studies transcriptional programs controlling heart and outflow tract development with particular focus on defining the function(s) of the cardiac neural crest. This research has provided understanding of the molecular and genetic basis of congenital heart disease and cardiomyopathy. The Epstein laboratory has focused on examining the role of Pax3, a gene required for proper neural crest function, during cardiovascular morphogenesis. They are also studying the role

of the Neurofibromatosis Type I gene, Nf1, in neural crest cells and in many other tissues. Inactivation of Nf1 in mice leads to congenital heart defects that may or may not be related to neural crest defects. In collaboration with investigators at Albert Einstein the Epstein lab demonstrated that mutations in TBX1 are responsible for many of the defects observed in DiGeorge Syndrome. In addition, the Epstein laboratory cloned and characterized the cardiac-restricted homeobox gene, HOP, which regulates cardiac myocyte growth and differentiation. In a series of translational research studies, the Epstein laboratory has been examining the role that histone deacetylasaes (HDACS) plan in regulating cardiac hypertrophy. Garret A. FitzGerald, MD, PhD Dr. FitzGerald has a long-standing research interest in understanding the mechanical forces and chemical mediators that cause endothelial cell dysfunction and eventually atherosclerosis. He is internationally recognized for his studies demonstrating that oxidant stress plays an important role in the pathogenesis of atherosclerosis. His laboratory has used transgenic mouse and gene knock-out mouse models of atherosclerosis to examine the role that eicosanoids and iso-eicosanoids play in mediating atherosclerosis. PGI2 is a potent vasodilator and platelet antagonist. It may also modulate cellular proliferation, adhesive interactions of monocytes and platelets with endothelial cells, and cholesterol export, properties that may limit atherogenesis. Cyclooxygenase (COX)-2 is a major source of PGI2 in humans and COX-2 inhibitors suppress PGI2 formation without concomitant platelet inhibition. In a series of translational studies, Dr. FitzGerald is testing the mechanisms underlying the pro-thrombotic (and potentially pro-atherosclerotic) effects of selective inhibitors of COX-2 in humans, in conventional and novel mouse models of atherosclerosis and in genetic crosses of these models with mice lacking COX-2 or the PGI2 receptor. Yale Goldman, MD, PhD Muscle and myocardium are prototype biological energy transducers that can be understood at a particularly fine level of detail. In the Goldman lab, the reactions of the contractile proteins and their signaling and trigger molecules are initiated rapidly and abruptly by laser photolysis of "caged" precursors. This powerful method enables resolution of kinetic events in organized biological systems to the millisecond time scale, using intense laser pulses to cleave "caged" molecules within muscle preparations. This leads to rapid release of the parent molecules, such as nucleotides, Ca2+ and phosphoinositides. The laser photolysis technique is combined with detection methods such as advanced single-molecule optical spectroscopy, time-resolved electron microscopy, high time resolution mechanical recording from single fibers, and isotope exchange. Chromophores (fluorescent or phosphorescent probes) are covalently attached to specific sites on actin or myosin with known orientation in the protein crystal coordinates. Polarization of the absorption or emission transition moments of these probes during reactions initiated by photolysis of caged molecules is a powerful technique to detect motions of actomyosin and many other protein structural changes. All of these techniques are applicable in myocardiocytes and vascular smooth muscle cells toward understanding normal and pathophysiological processes in the cardiovascular system. Keith J. Gooch, PhD Blood vessels can remodel either physiologically or pathologically when exposed to altered

mechanical environments. The complex interdependence between components of the mechanical environment (e.g., pressure, shear, and strain) in vivo has hindered the identification of the specific mechanical stimuli responsible for remodeling. To help identify the mechanical stimuli responsible for vascular remodeling, a system for exposing viable, excised, blood vessels to precisely controlled flow and pressure regimes has been built. Using this system, Dr. Gooch is investigating the effects of transmural pressure, cyclic strain, fluid shear stress, and longitudinal tension on remodeling of excised porcine arteries and veins. In addition to improving our knowledge of physiological and pathological events, Dr. Gooch believes that an understanding of mechanically induced remodeling could be used to apply appropriate mechanical conditions to direct the remodeling of human vessels - vessels that could potentially be used as autologous vascular grafts after appropriate ex vivo conditioning. Joseph Gorman, MD The Gorman laboratory has two related primary areas of investigation. First, they are interested in understanding the remodeling process following myocardial infarction at a mechanical, cellular, biochemical and genetic level in order to develop novel medical and surgical interventions which are designed to prevent rather than reverse the remodeling process. MRI, 2D and 3D echocardiography as well as sonomicrometry are used to assess regional myocardial strains which are then correlated with regional biologic markers that are associated with adverse remodeling. Second, they are interested in improving mitral valve repair surgery for both degenerative and ischemic etiologies we are studying the dynamic anatomy of the valve using MRI, 2D and 3D echocardiography as well as sonomicrometry. Several new annuloplasty and replacement devices are currently being designed by members of the laboratory. Techniques for performing virtual surgery on 3D valve reconstruction are also under development. Peter J. Gruber, MD Dr. Grubers research interest is in the molecular and genetic basis of congenital heart disease. The Gruber lab takes a broad variety of molecular genetic and physiological approaches to understand the heart in development and disease. One portion of the lab examines the functional consequences of the targeted deletion of transcription factors that may play an important role in cardiogenesis. A second group is dedicated towards the examination of epigenetic modulation of cardiac function, both developmentally, and as therapy for ischemic heart disease. Third, they are interested in cardiac regeneration through the purification and characterization of a population of self-renewing, heart progenitor cells. A common theme running through these projects is the intersection of the rich clinical material available at the Children’s Hospital of Philadelphia (CHOP) and the use of molecular techniques in animal models to dissect pathways important in the diagnosis and therapy of congenital heart disease. Howard C. Herrmann, MD Dr. Herrmann performs patient-oriented research focused in both medication and device areas related to interventional cardiology. Dr. Herrmann's primary research interest is in novel percutaneous treatments for valvular heart disease, including balloon valvuloplasty, stented-aortic valves, and coronary sinus devices for mitral regurgitation. He is participating in a phase 1 trial of percutaneous edge-to-edge repair for mitral regurgitation and is designing his own percutaneous replacement mitral

valve. Dr. Herrmann is also participating in and serves on the steering committee of other device trials for acute myocardial infarction (FLAME trial), for PFO and ASD closure (CLOSURE-1), distal protection during saphenous vein graft intervention (FIRE), new drug-eluting stent trials (ATLAS), a randomized trial comparing CABG and PCI (FREEDOM), and several multi-center trials examining new adjunctive anti-platelet and anti-thrombotic therapies for PCI (TENACIY, ACUITY). Katherine A. High, MD The focus of the High laboratory is on the genetics, molecular biology and biochemistry of blood coagulation factors. Major areas of investigation include: 1) Structure-function analysis of Factors VII, IX and X. 2) Study of the regulation of expression of the genes encoding the vitamin K dependent clotting factors. These genes manifest tissue-specific expression; they are expressed only in the liver. They have isolated the 5' flanking sequences of all three genes, characterized promoter activity and determined, using DNase footprinting, the location of protein binding sites within these promoters. In the case of Factor X, they have determined through a variety of approaches the identity of the transcription factors binding to the promoter and are in the process of carrying out similar studies on the promoters of VII and IX. 3) Studies designed to establish an experimental and clinical basis for gene therapy of hemophilia, a bleeding disorder that results from a deficiency of functional Factor IX. They are currently using both viral and non-viral vectors to introduce the Factor IX cDNA into target cells of interest. Mark L. Kahn, MD The Kahn laboratory investigates vascular receptor signaling involved in thrombosis and angiogenesis. They are presently focused on platelet signaling responses to G protein coupled and non-G protein coupled receptors (GPCR). Specific areas of investigation include: 1) Non-GPCR activation of platelets: characterization of novel platelet receptors. 2) G protein activation of platelets: regulation of G protein signaling in vivo. Dr. Kahn in collaboration with Dr. Gary Koretzky have demonstrated that Slp-76 and Syk- signaling plays a critical role in embryonic angiogenesis and lymphogenesis. Ongoing studies are examining the molecular basis of Slp- and Syk- signaling in the vasculature. Dr. Kahn is engineering conditional mutations in mice to better elucidate the cell autonomous role of Slp-76- and Syk- signaling during embryonic cardiovascular development and there role in platelet signaling. Stephen E. Kimmel, MD, MSCE Dr. Kimmels research is in cardiovascular epidemiology with a focus on the effects of non-cardiac medications on the heart and on angioplasty outcomes. His work includes one of the largest cohort studies ever performed to examine protamine adverse drug reactions after cardiopulmonary bypass; an almost 4,000 person case-control study, involving 68 sites, of the relationship between myocardial infarction (MI) and nicotine patches; and an over 8,500 person case-control study examining the effects of non-steroidal anti-inflammatory drugs on MI. He also has performed numerous studies of angioplasty outcomes, including studies of the association between laboratory volume and complications. He is the PI of an NIH R01-funded prospective pharmacogentic epidemiology cohort study examining genetic predictors of anticoagulation control on warfarin. Dr. Kimmel also is the recipient of an NIH K24 Midcareer Investigator Award in Patient-Oriented Research to study genetic influences on the response to anticoagulation drugs. He recently received an NIH P20 Roadmap Grant for development of interdisciplinary research in the field of Human Pharmacogenomic

Epidemiology. Gary Koretzky, MD, PhD A major focus of research in the Koretzky laboratory is on the integrationof signal transduction pathways leading to cellular activation or programmed cell death. Recently his group has identified several novel adapter proteins, molecules with no intrinsic enzymatic activity but which function by mediating protein-protein interactions. One of these proteins (designated SLP-76) appears to play a critical role in T cell differentiation and activation as well as in platelet function. Over-expression of SLP-76 in T cell lines leads to a dramatic augmentation of T cell antigen receptor induced interleukin 2 production. Generation of mice deficient in SLP-76 leads to a complete loss of peripheral T cells with an arrest early in thymocyte development. SLP-76 deficient mice additionally exhibit severe perinatal hemorrhage, presumably due to defects in platelet signal transduction. His laboratory is now performing a detailed structure function analysis of SLP-76 in both T cells and megakaryocyte/platelets. They are utilizing biochemical, imaging, and genetic techniques to determine what inter-molecular interactions directed by SLP-76 are critical for its function as a regulator of signal transduction events. Mitchell A. Lazar, MD, PhD Dr. Lazar's research focuses on nuclear hormone receptors and their actions. In the absence of hormone, receptors interact with corepressor molecules to repress transcription; hormone ligand leads to coactivator recruitment and activation of transcription. The receptor complexes contain histone modifying activities that may be drug targets. Specifically, Dr. Lazar is studying the molecular mechanisms of nuclear receptors called PPARs. PPAR ligands actively promote fat cell differentiation. Excitement has been generated by the observation that PPAR ligands include a promising new class of antidiabetic drugs called thiazolidinediones (TZDs), including troglitazone (RezulinR), pioglitazone (ActosR), and rosiglitazone (AvandiaR). These drugs enhance the actions of insulin, which is critical because diabetes is a major cause of morbidity and mortality in the U.S, and more than 95% of diabetics have type 2 diabetes, associated with obesity and severe resistance to the action of insulin. The mechanism whereby PPAR activation leads to increased insulin sensitivity is not known. In a series of related studies, the Lazar laboratory has also identified and is further characterizing resistin, a hormone that in murine models is responsible in part for insulin resistance. Francis E. Marchlinski, MD Dr. Marchlinski's research focuses on elucidating the anatomic and electrophysiologic substrate for atrial and ventricular arrhythmias in man. On-going research studies involve a detailed characterization of the electroanatomy associated with atrial fibrillation and ventricular tachycardia. Pharmacologic manipulation and stimulation techniques are used to alter the substrate to enhance understanding of the conditions that promote arrhythmogenesis. State-of-the art magnetic mapping and intracardiac echo recording techniques are used during ablation procedures to characterize the creation and location of all lesions and their effect on the electroanatomic substrate. They also use radiofrequency ablation techniques as a powerful tool for further delineating the electroanatomic substrate. This detailed physiologically oriented approach has permitted the design of original strategies and technology for curing arrhythmias in man.

Kenneth Margulies, MD Dr. Margulie's research examines load-induced myocardial remodeling, myocardial failure and myocardial recovery. Mechanistic studies focus on cardiac mechanics, cardiac myocyte calcium homeostasis, and regulation of the myocardial extracellular matrix. To better understand these areas, his laboratory performs research using clinically relevant animal models, human tissues made available at the time of cardiac transplantation and patient-based translational research. Studies typically involve multilevel physiological inquiries and employ isolated cardiac myocytes, muscle strips, isolated hearts and in vivo methodologies. Molecular analyses, including studies using high-throughput microarray techniques, complement these functional inquiries. Dr. Margulies is the Director of the Penn Heart Failure research program and has established extensive collaborations with Joe and Rob Gorman who direct the large animal cardiovascular surgery laboratory and Dr. Jonathan Epstein, Director of the Molecular Cardiology research program. Emile R. Mohler, MD Dr. Mohlers research examines the molecular pathogenesis of valvular heart disease and vascular proliferative syndromes. His laboratory has shown that inflammation may underlie common forms of degenerative valvular disease. They have studied excised human valves in tissue culture and documented that the same osteogenic program that is activated in atherosclerotic lesions is recapitulated in valvular heart disease. His laboratory is currently examining whether specific anti-inflammatory therapies may be efficacious in the treatment of aortic stenosis. Dr. Mohler is also interested in the function(s) of bone marrow-derived and/or circulating endothelial progenitor cells in vascular proliferative syndromes. In collaboration with Dr. Robert Wilensky, Mohler has generated a hypercholesterolemic, diabetic porcine model of type I diabetic vascular disease that recapitulates the progression from endothelial dysfunction to macrovascular disease observed in diabetic patients. Edward E. Morrisey, PhD The focus of the Morrisey laboratory is examining the transcriptional programs that control heart and lung development. Dr. Morrisey, while a postdoctoral fellow training with Dr. Parmacek, cloned and characterized the GATA-4/5/6 subfamily of transcription factors, each of which plays a unique role in the developing heart. Since establishing his laboratory, Morrisey has focused his research on defining the role of the Foxp1/2/4 subfamily of winged helix transcription factors play in the developing heart and lung. He has generated null mutations in ES cells and mice of each subfamily member and shown that each factor plays a unique and important role in the developing heart and/or lung. These studies led to the fundamentally important observation that heart chamber formation is pre-programmed in migrating cardiac progenitors and is not dependent upon heart tube fusion (Science 305:1619-1622, 2004). Warren S. Pear, MD The focus of the Pear laboratory is understanding the role of Notch signaling in cardiovascular and hematopoietic development and differentiation.. Notch proteins are a conserved family of receptors that regulate cell fate decisions in organisms ranging from Drosophila to humans. Both gain and loss of function studies have implicated Notch signaling in both cardiovascular and hematopoietic development. Using a variety of in vitro and in vivo approaches, the Pear laboratory has shown that Notch regulates hematopoietic stem cell homeostasis and T cell development. They have extended

their studies to cardiovascular development where they have found that Notch signaling plays key roles in vascular and cardiac development and function. In addition to their role in normal hematopoiesis, Notch proteins have also been implicated in human leukemia. Finally, they have developed a mouse model of Notch-related leukemia and are using this to study the signaling pathways that lead to oncogenic transformation. It is the ultimate goal of the Pear laboratory to translate our understanding of Notch signaling in development and disease into novel approaches that will lead to advances in diagnosing and treating cardiovascular and hematologic diseases. Ellen Pure, PhD Dr. Pure’s laboratory investigates the immunobiology of atherosclerosis. They have shown that the cell adhesion molecular CD44 is up-regulated, and its ligand, hyaluronan, accumulates in atherosclerotic lesions. They have also shown that CD44-/- x ApoE-/- mice have a marked decrease in atherosclerotic plaques compared to ApoE-deficient mice suggesting that CD44 is pro-atherogenic. They are currently investigating the molecular mechanisms regulating the atherogenicity of CD44. The Pure laboratory is also investigating the molecular mechanisms underlying the pathogenicity of oxidized LDL (OxLDL) in the vessel wall. They have determined that OxLDL does not promote macrophage production of pro-inflammatory cytokines including IL-12, TNF or IL-1. These data challenge the existing paradigm that OxLDL increases inflammation in the arterial wall. On-going research is investigating the molecular events that trigger the chronic inflammatory response that lead to atherosclerosis. Daniel J. Rader, MD Dr. Rader's laboratory is interested in inflammatory and genetic factors that regulate the metabolism of plasma lipoproteins and their interaction with the vessel wall in promoting and inhibiting atherogenesis. Both basic laboratory science and clinical research approaches are used to examine the role of lipids and other genetic modifiers in promoting atherosclerosis. The major projects can be divided into the following groups: 1) Inflammatory and genetic factors that regulate the in vivo metabolism of HDL and its plasma levels. 2) Molecular and cellular mechanisms by which HDL-associated proteins inhibit atherogenesis and induce regression of atherosclerotic lesions. Somatic gene transfer of HDL-associated proteins is used in mouse models of atherosclerosis in order to study their effects on atherogenesis in vivo. 3) Dietary and genetic regulation of hepatic lipoprotein production. Gene transfer and transgenic approaches are used to study the interaction between specific genes, such as the microsomal transfer protein, and dietary manipulation in the regulation of hepatic apoB production in mice. Lipoprotein kinetic studies are also performed in humans using endogenous labeling of apolipoproteins with stable isotopically labeled leucine. 4) Genetic factors associated with premature atherosclerotic disease and high or low levels of HDL cholesterol. Subjects with family history of premature coronary disease or with extremes of HDL cholesterol are recruited and phenotyped for cardiovascular risk factors and subclinical atherosclerosis. Candidate genes are investigated for their association with subclinical atherosclerosis. Muredach P. Reilly, MD Dr. Reilly's research interests include (1) mechanisms of atherosclerosis in metabolic syndrome,

diabetes and obesity, (2) the role on innate immunity in promoting atherosclerotic risk in metabolic syndrome, (3) the functions of adipose tissue in regulation of HDL cholesterol and reverse cholesterol transport, (4) genetics of atherosclerosis and dyslipidemia in high risk populations including type 2 diabetes, subjects with family history of CAD, and in HIV infected populations (5) non-invasive imaging of atherosclerosis in humans. Dr Reilly employs a translational approach to his research including cell and animal based mechanistic studies, mechanistic patient-oriented, General Clinical Research Center studies as well as larger scale epidemiological studies in humans. Examples of his funded research projects include; a POR, GCRC based study of endotoxin as a model of inflammatory induced pro-atherosclerotic responses in humans with the metabolic syndrome; cross-sectional association study of novel genetic and biochemical markers of coronary atherosclerosis in asymptomatic type 2 diabetic patients; genetics of protease inhibitor induced dyslipidemia in HIV; effect of PKC modulation on mouse and human atherosclerosis; a clinical trial of HDL elevating therapy on carotid atherosclerotic plaque morphology at MRI. Joseph W. Sanger, PhD The focus of the Sanger laboratory is the molecular mechanisms underlying the assembly of myofibrils in living cardiac muscle cells. The assembly is visualized by either the microinjection of fluorescently labeled proteins or the transfection of cardiomyocytes with plasmids encoding cytoskeletal proteins linked to a fluorescent reporter molecule such as Green Fluorescent Protein or Ds Red protein. The appearance and position of the fluorescently labeled proteins are detected with low light level cameras. The data is analyzed with various image analysis programs. M. Celeste Simon, PhD The Simon laboratory focuses on understanding the transcriptional programs that control vascular and hematopoietic development. Dr. Simon leads the angiogenesis research program within the Abramson Family Cancer Research Institute. Using transgenic and gene knock-out technologies, Dr. Simon has shown that GATA-1 is required for differentiation of the erythroid cell lineage and ets-family members regulate specific steps in myeloid cell differentiation. Several on-going projects in Dr. Simons laboratory are focused on defining the transcription factors and signaling pathways that regulate vascular development and angiogenesis. They have demonstrated that mice harboring a null mutation in ARNT exhibit abnormal angiogenesis in response to glucose and oxygen deprivation. In addition, they are examining the signaling pathways that regulate HIF1a and HIF1b; transcription factors that coordinate response to tissue hypoxia including expression of the angiogenesis factor VEGF. These studies provide fundamental insights into vascular development. In addition, they are relevant to the understanding, and potentially therapy, of atherosclerosis and ischemic syndromes. Martin G. St. John Sutton, MBBS Dr. St. John Suttons research interests focus on evaluation of chamber architecture, function and the interaction of ventricular loading conditions on hypertrophy using non-invasive imaging techniques including echocardiography and cardiac MRI. This interest has stimulated the derivation and validation of equations and formulations employing ultrasound imaging to calculate circumferential stress, myocardial force development and load-independent measures of contractile performance. These non-invasive tools have been applied over the years in a number of models of pressure and volume overload in man before and following operative and pharmacologic interventions to

characterize the time and regression of ventricular hypertrophy and restoration of pump function. The magnitude and time course of left ventricular chamber remodeling has been related to the size of the reduction in loading conditions. He is currently studying mechanisms of hypertrophy and regression from hypertrophy using a unique model of myocardial infarction in sheep. Moreover, he is evaluating the long term effects of synchronous biventricular pacing on cardiac remodeling. Brian L. Strom, MD, MPH Dr. Strom's major research interest is pharmacoepidemiology; many of his research projects are relevant to cardiovascular medicine. He completed a large-scale case-control study on the effects of cholecystectomy on subsequent myocardial infarction. He completed a large-scale cohort study using data from Kaiser Permanente on the proper duration of oral anticoagulation after a pulmonary embolism, and a nested case-control study about the risk factors for bleeding from this anticoagulation. He collaborated on another large-scale case-control study of the relationship between oral contraceptive use and thromboembolism. He completed a large cohort study, using a large-scale Medicaid claims database, of the gastrointestinal toxicity of potassium chloride used in patients with hypertension or congestive heart failure. He has recently completed a large-scale population-based case-control study on risk factors for infective endocarditis. Dr. Strom is collaborating with Dr. Kimmel on a study of the incidence of and risk factors for protamine reactions, a series of case-control studies of nonsteroidal anti-inflammatory drugs and myocardial infarction, a large-scale population-based case-control study of nicotine patches and myocardial infarction, and a series of analyses of a nationwide registry of angioplasty, exploring predictors of complications from angioplasty. Craig B. Thompson, MD The Thompson laboratory studies a wide range of basic issues concerning cell proliferation, signal transduction, and apoptosis. The laboratory concentrates its studies on the lymphoid immune system focusing on basic aspects of development, cell division, and cell survival. Among the areas under investigation are: 1) Gene conversion as a mechanism to generate diversity within the immune system. 2) Recombination intermediates in the process of immunoglobulin rearrangement. 3) Studies of CD28 as a novel signaling pathway to regulate lymphocyte function. 4) Studies of CTLA-4 as a major negative regulator of lymphocyte cell proliferation. 5) Characterization of Bcl-x, to date the single most potent regulator of cell survival identified in vertebrates. 6) Studies of apoptotic control in the regulation of cancer invasion and metastasis. Major research programs ongoing in the Thompson laboratory include: 1) Characterizing the biochemical mechanisms by which Bcl-2 proteins promote cell survival. 2) Defining the role of CD28 and CTLA-4 signaling pathways in the regulation of lymphocyte homeostasis. 3) Determining how apoptosis resistance contributes to neoplastic transformation, particularly through the potentiation of genetic instability. Alexander Whitehead, PhD Hyperhomocysteinemia, a recognized risk factor for cardiovascular and other diseases, may be caused by suboptimal folate and B vitamin status and/or common functional mutations in enzymes of folate/Homocysteine(Hcy) metabolism. The Whitehead laboratory is characterizing the contribution of such mutations to atherothrombotic diseases by conducting a number of large scale genetic epidemiology studies in which the relationship between genetic, nutritional, biochemical and clinical

variables will be defined. In addition, in vitro and animal studies are performed to assess the impact of low folate and/or high Hcy on endothelial cell (EC) function, in particular, changes in the EC mRNA expression profile that might identify critical components in pathways that are involved in the pathogenesis of vascular diseases. Such components will be candidates for genetic analysis, and may be suitable targets for future novel therapeutic intervention. The Whitehead laboratory also investigates the cytokine-regulated expression of major acute phase proteins, specifically serum amyloid A (SAA) and C-reactive protein, each of which are predictors of myocardial infarction. In addition, the SAA promoter is being developed as a cytokine responsive controller of the expression of anti-inflammatory agents for use in gene therapy of inflammation associated diseases, including cardiovascular disease. Robert L. Wilensky, MD The research focus of the Laboratory for Experimental Interventional Cardiology, which is directed by Dr. Wilensky, centers on mechanisms of vascular injury and repair and approaches to modulating this response. Several studies in the past as well as ongoing studies have evaluated the response to balloon and stent injury in coronary and peripheral vasculature. Specifically the role of thrombus, coagulation proteins and inflammatory cells in the repair process have been addressed. The role of locally delivered pharmacologic and genetic agents in the modulation of the injury response mechanism has been evaluated and new approaches to increase intramural delivery and retention have been described. He has also evaluated the role of intracoronary stents as platforms for direct delivery of DNA and proteins into the arterial wall to effect the cell cycle and reduce cellular proliferation. The laboratory has branched out and is currently evaluating local myocardial delivery of mesenchymal stem cells to increase angiogenesis and/or improve contractile performance in large animal models of heart failure. In collaboration with Dr. Emile Mohler, Wilensky has developed a type I porcine model of diabetic, hypercholesterolemic vascular disease. Y. Joseph Woo, MD The focus of the Woo laboratory is the application of novel molecular- and cell-based therapies in the treatment of myocardial ischemia and cardiomyopathy. Three primary areas of interest include 1) the induction of angiogenesis to endogenously revascularize the failing heart, 2) myocardial reconstruction and regeneration through genetic modulation of cardiomyocytes and 3) the study of biochemical myocardial protection agents. Specific vasculogenesis projects include the overexpression or delivery of angiogenic agents such as hepatocyte growth factor and placental growth factor as well as the stimulation of and chemokine-directed targeting of endothelial progenitor cells to ischemic myocardium. Specific myocardial regeneration projects have entailed inducing localized, growth hormone-mediated somatotropism or cyclin-directed cell-cycle activation. Biochemical approaches to myocardial protection have included the study of a membrane permeable ethylated derivative of the glycolytic intermediary pyruvate and alternate high-energy phosphate compounds. Gary Webb, MD Dr. Webb is internationally recognized for his patient-oriented research and clinical expertise in the area of adult congenital heart disease (ACHD). He was recruited to Penn earlier this year from the University of Toronto where he directed the largest adult congenital heart disease program in North

America. Since joining the Penn faculty, he has established a joint Penn-CHOP adult congenital heart disease program that includes research into the genetics and molecular biology of adult congenital heart disease. The Penn-CHOP ACHD database will include clinical profiles of all patients referred with ACHD and in consenting patients will include DNA samples for genetic analyses. Trainees interested in translational/POR in adult congenital heart disease will have the opportunity to perform 1-2 years of research in the Molecular Cardiology laboratories focusing on cardiovascular development (Epstein, Kahn, Morrisey, Parmacek) that they may apply to the large group of patients referred to the Penn-CHOP ACHD program. Faculty Interactions and Collaborations One of the unique features of the current cardiovascular research program at the University of Pennsylvania is the interdepartmental and interdivisional nature of the collaborative efforts in this area. Such collaborations will be further facilitated by the fact that many of the faculty preceptors will occupy geographically contiguous space within the Penn Cardiovascular Institute. Large-scale faculty collaborations have been established at the University of Pennsylvania in the following programmatic areas: vascular biology/atherosclerosis (Parmacek, Davies, FitzGerald, Rader, Wilensky, Pure, Simon, Whitehead, Gooch, Brass, Kahn, High, Koretzky), cardiovascular development/genetics (Parmacek, Epstein, Birnbaum, Kahn, Koretzky, Morrisey, Pear, Simon, Webb) heart failure/muscle function (Sweeney, Margulies, Cappola, Dries, Birnbaum, Callans, Goldman, Lazar, Marchlinski, Sanger, St. John Sutton, Thompson) and health services research (Kimmel, Strom, Cappola, Marchlinski, Callans). A number of multi-investigator NIH-sponsored grants link many of the faculty together including: 1) SCCOR in Congenital Heart Disease (P50 HL61014) 2) SCOR in Thrombosis Regulation and Dysregulation of GPCR signaling (P50 HL54500) 3) SCOR in Oxidative Modifications and Atherogenesis (P50 HL62250) 4) Program Project on Vascular Signal Transduction and Atherosclerosis (PO1 HL62250) 5) Program Project in Muscle and Motility (PO1 HL15835) 6) Program Project on Vascular Heterogeneity (PO1 HL75380) 7) Program Project on Subcellular Mechanisms of Platelet Activation (PO1 HL40387) 8) Program Project on Integration and Control of Molecular Motors (PO1 AR051174) 9) Program Project on Gene Therapy for Hemophilia (PO1 HL64190) 10) Program Project on Wnt Signaling and Lung Vascular Development (PO1 HL075215) 11) Program Project on Airway Smooth Muscle, Inflammation and Asthma (PO1 HL67663) 12) Program Project on Cellular and Molecular Biology of Lipoprotein Metabolism (PO1 HL22633)

Proposed Training Overview The design of this program is based on its twin goals: First, to prepare MD/PhD graduate students and physician fellows for research careers in academic cardiology, and second, to attract and train in promising PhD graduate students and postdoctoral fellows in cardiovascular biology and medicine. The specific rationale underlying the design of this program is that training in molecular and cell biology will serve as a foundation for careers in basic, translational and patient-oriented cardiovascular investigation. Particular emphasis was placed on identifying trainers with outstanding research programs that are relevant to cardiovascular biology and medicine. Support is requested for six postdoctoral fellows (MD, MD/PhD, PhD) and two graduate students (MD/PhD or PhD) per year who will normally devote two to three years to laboratory or translational research. Size of program: For the past five years, the training program has supported six postdoctoral fellows per year, 75% of which had MD or MD/PhD degrees (Table VIII). As discussed above, to attract talented MDs, MD/PhDs or PhDs into careers in academic Cardiology, we believe that it is important to provide research training early in their careers. We therefore, are requesting that two additional pre-doctoral training slots be phased into the program. With the expansion of the MD/PhD program and the Clinical Investigator Pathway in the Internal Medicine residency program, the University of Pennsylvania Department of Medicine has attracted increasing numbers of residency applicants with an interest in cardiovascular research. Many of these residents have made a clear commitment to careers in basic and translational cardiovascular research. Support by this training grant provides a means to attract them into this training program. Graduate students will be supported for a maximum of 3 years after completion of course requirements in the MD/PhD program and/or their basic science department. Postdoctoral fellows will be supported for 2 years, except in rare cases where basic science background is weak, requiring substantial coursework. Cardiology fellows (and other postdoctoral fellows) will not be supported during clinical training. It is expected that support will be generated through the individual advisers research program either at the end of the first year, or at the latest, at the end of the second year of training grant support. Postdoctoral fellowships by PhDs pursuing training in cardiovascular biology or medicine will be supported for a maximum of 2 years. Distribution of positions: a mix of physician and non-physician trainees. As recommended by the NIH guidelines, we have found that bringing physician and non-physician trainees together helps to create the attitudes that foster collaborative research in the future (and leads to better training of both MDs and PhDs). This is accomplished in several ways. First, by

accepting as trainees on this grant a mixture of MDs, MD/PhDs and PhDs. Of the available training slots on this grant (2000-2004), 13 were filled by MDs, 6 were filled by MD/PhDs and 6 were filled by PhD postdoctoral fellows (Table VIII). Second, by encouraging physician trainees to work in laboratories where there are graduate students and PhD post-docs, and, third, by requiring all trainers to participate in laboratory meetings, seminars and journal clubs where they will come in contact with other investigators with varied backgrounds and experiences. In the next funding period we will seek to obtain the same ratio of MD and MD/PhD to PhD ratio of approximately 4:1 (19 or 24 positions). Research Mentoring Helping our postdoctoral trainees achieve their goal of becoming independent investigators is very important to us. Although the details are tailored to the individual, in general this is accomplished through a combination of 1) assistance with mentor selection, 2) defining the role and responsibilities of the research mentor, 3) constructive feedback, 4) career advising and 5) skills training. Each of the trainees has two mentors: their research mentor in whose lab they are working and a member of the Oversight Committee with whom they are expected to meet several times per year. The proportion of former trainees who have successfully established an academic career speaks to the high quality of our trainees, but also suggests that our efforts have succeeded. We plan to do even better. Research mentor selection Trainees select mentors and projects based upon their research interests and the interests of their mentors laboratory. The 36 trainers on the grant provide a broad range of choices and represent the basic science departments and research institutes on campus, as well as the clinical departments of Medicine, Pediatrics and Surgery. Physicians in the clinical programs (M.D. and M.D.-Ph.D.) learn about potential mentors through informal contacts, by attending the weekly research seminars held in both of the clinical hematology-oncology divisions and through meetings with the chief of their division and other senior members of the faculty including the PI of this grant. Many of the clinical fellows have had extensive prior research experience and have a pretty good idea of the area in which they would like to resume their research training. Others require more guidance. Ph.D. trainees are usually recruited directly into laboratory groups, either in response to advertisements placed by a faculty member and the Office of Postdoctoral Programs or through prior exposure to the faculty members publications. In general, we encourage all trainees to work in successful laboratories in which there is a mix of M.D. and Ph.D. trainees guided by a mentor or mentors who have a track record of scientific accomplishment and active participation in the guidance of pre- and postdoctoral trainees. We steer trainees (particularly physician trainees) away from laboratories in which the faculty member is not an active participant in daily events. 2) The role and responsibilities of the research mentor. The research mentor is to be more than a temporary employer and provider of a lab bench. In general, the faculty research mentor serves as a research supervisor, teacher, career counselor and role model. As the trainee begins an intensive research experience in the preceptors laboratory, the preceptor helps the trainee select appropriate courses, research seminars and scientific meetings. Over time,

the preceptor may also recommend that the trainee spend some time in another specialized laboratory on campus. Typically this includes the laboratories of other investigators (at Penn or elsewhere) with whom the research mentor has either an ongoing collaborative relationship or sufficient previous contact to insure that the trainee will receive the necessary attention. Research mentor and trainees jointly plan the specifics of the trainees individual experience. Trainee progress and accomplishments are evaluated by the research mentor on a continual basis, as they are for other members of the mentors laboratory. The research mentor and the Oversight Committee mentor guide the trainee informally and formally and together they make adjustments in research activities or curricula as required. Constructive Feedback The progress of each trainee is monitored by the assigned mentor from the Oversight Committee. The intent is that the Oversight Committee should help to guide the trainees progress and, when necessary, intervene. Trainees meet with the committee as a whole on at least an annual basis and summarize their progress in a written report and an oral presentation. The written report includes a description of research results, copies of manuscripts that have been submitted or published. If research progress is problematic or other concerns are expressed by the trainee or preceptor during the formal review or at any other time during the training period, then the Oversight Committee may recommend a change in research direction, research mentor or course work. The trainee may request a special session with the Principal Investigator or with the Committee as a whole in order to resolve problems regarding their training. Feedback works both ways and we have always made a point of finding out from the postdoctoral trainees what their experience has been with their research mentor and used the information to help trainers improve their mentoring skills. Career advising and resources Career advising is an important part of helping our postdoctoral trainees take the next step in their careers. For some this step will be to apply for junior faculty positions. For others, it may be to apply for a second term of postdoctoral training within the five year limit on postdoctoral training time at Penn. Career advising is done jointly by the research mentor and by the members of the Oversight Committee. When and where appropriate, all trainees are encouraged to submit NIH K award proposals timed to help them with the transition from postdoc to faculty member. Senior members of the training faculty exploit their contacts at other institutions in behalf of the trainees. The Office of Biomedical Postdoctoral Programs also organizes a Career Workshop Series that addresses basic job-search skills, careers in industry, academia and alternative careers. Every year a Biomedical Career Fair is co-organized with Career Services and BGS to provide postdocs the opportunity to network with prospective employees. Skills training Success in the laboratory is seen as being only one part of building a successful career in academic research. Additional essential skills include manuscript writing, seminar presentations, grant

preparation, manuscript reviewing, grant reviewing and laboratory management. Postdoctoral trainees in this program are expected to have had considerable exposure to these skills if they completed an MD-PhD or PhD program, but less so if they only attended medical school. As already described, the Office of Biomedical Postdoctoral Programs offers a set of workshops that address many of these issues. In addition, research mentors are expected to provide trainees with experience in each of these areas. This would include (within the accepted bounds of confidentiality), opportunities to help review manuscripts and grants. It also includes applying for individual postdoctoral fellowships and NIH K awards for career transition. Selection of Mentors Trainers are selected by the Program Directors and Oversight Committee. Selection of faculty as trainers is for a three-year period of time with periodic review to make certain that they continue to meet the criteria for being a trainer. Another function of the periodic review is to make sure that the areas of research of trainers continue to match the overall direction of the training program and the research interests and needs of the students. As new faculty with vital research programs in the appropriate areas of science are recruited to the University, they will be considered for appointment to the training grant. Incumbent trainers whose focus of research is no longer germane to the goals of the training program or whose research programs have lost their vitality will not be re-appointed. These decisions will be made by the Director of the training grant with the advice of the Oversight Committee. Consequently, the composition of the program faculty will be dynamic, and their percent effort will fluctuate: some faculty will train more students than others; some will teach more than others; some will lecture simultaneously to students from a variety of programs and disciplines; committee members have administrative duties, etc. Of the faculty listed as trainers (Table II), all but the newest faculty members are participating or have recently participated in research training and/or teaching (Table III). Basic and Translational/POR Curriculum Basic Research Curriculum: Each of the trainees will meet with the Program Director before the beginning of their training period to design a training program that is tailored to the needs of that trainee. MD/PhD and PhD graduate students will fulfill the basic science requirements of their specific training program. Postdoctoral MD, MD/PhD and PhD trainees will be expected to possess or obtain working knowledge in Biochemistry, Cell Biology and Molecular Biology. The following three courses or their equivalents will fulfill this initial requirement: 1. Introduction to Biochemistry (Biochem 500) 2. Cell Biology (Cell 600) 3. Molecular Genetics (MolB/Biol 421) The Director, Co-Director and Oversight Committee of the proposed training program would take an

active role in helping trainees select an appropriate mentor after exploring the interests of the trainee. Following didactic course work, the major element of each training program will consist of a research project carried out in the laboratory of one or more of the training preceptors. During their time in the laboratory, clinical requirements for cardiology fellows will be restricted to a maximum of one-half day per week of outpatient clinic. Over the two to three years of laboratory research training program, the trainee will be expected to develop an independent research question that will be pursued with less supervision from the preceptor. A list of projects of recent trainees is shown in Table IX. Emphasis will be placed upon gaining a prior understanding of the literature relevant to the trainee's interest, defining an important scientific problem, and designing a logical and feasible experimental approach to this problem. The molecular and cellular methods needed to address the problem will be learned both in the preceptor's laboratory and those of other faculty members in the training program. As the trainees interests become focused on a specific area, they have the opportunity to take advanced seminar courses in their area of interest. Trainees will be expected to organize and present their data both at individual laboratory meetings and at the Molecular Cardiology monthly research seminar series. Trainees will also be encouraged to prepare both abstracts and manuscripts for submission to peer reviewed journals and for presentation at national meetings. An illustrative example of the basic science curriculum for a Cardiology fellow interested in muscle biology is shown below: First Year: 1. Laboratory Research 2. Participate in Bioethics course and workshop 3. Participate in Muscle Development Seminar Course 4. Participate in Cell Motility Journal Club 5. Attend weekly Cardiovascular Medicine Grand Rounds & P.M.I. Seminars 6. Attend national meeting germane to research interests Second Year: 1. Laboratory Research 2. Participate in Muscle Physiology and Disease Seminar Course 3. Attend Cardiovascular Journal Club 4. Attend weekly Cardiovascular Grand Rounds & P.M.I. Seminars 5. Present research findings at P.M.I. retreat 6. Present research findings at national meeting Translational/Patient-Oriented Research (POR) Curriculum:

The Translational/POR training program is a two-year program during which time MD or MD/PhD postdoctoral trainees will devote at least 80% of their time to research training. This includes didactic coursework and completion of a research project under the mentorship of a trainer. Trainees performing translational research are required to fulfill the basic science course requirement or their equivalents described above and in addition to attend the Translational Medicine Core Curriculum sponsored through the ITMAT. This Core Curriculum in patient-oriented research (POR) consists of four modules: 1) MTR 601: Fundamentals of Patient Oriented Research 2) MTR 602 Scientific and Ethical Conduct 3) MTR 520: Introductory Biostatistics 4) MTR 603: Disease Measurement Trainees may elect to enroll in the Masters of Science in Translational Research (MTR) degree program. The curriculum requires the satisfactory completion of 12 Course Units including the four Core Courses. Trainees must also complete four elective courses in basic science departments and four laboratory units (MTR 999). In addition, all MTR students are required to engage in a clinical research project of their design under the supervision of a primary mentor that upon completion will serve as their Masters thesis. Muredach Reilly, a former trainee and now faculty member, completed the MTR program (Table IX). Alternatively, trainees focusing their research in the direction of clinical epidemiology and/or clinical trials must complete the CCEB-sponsored Clinical Research Certificate program. This program provides didactic course work. The curriculum for this program requires pre-specified courses in: 1. EP 510 Introductory Epidemiology 2. EP 520 Introductory Biostatistics 3. EP 530 Informatics for Clinical Epidemiology 4. EP 531 Informatics for Clinical Epidemiology I 5. EP 540 Health Social Science Research Methods The first year of the translational/POR track provides the formal course work and the formulation and initiation of the research project. The second year provides for the completion of the research project. Trainees will be chosen to participate in the Translational/POR program approximately 6 to 9 months before beginning the formal program. The Director will review the different clinical research programs and preceptors with the trainee. For those fellows selected from the Cardiovascular Fellowship Program of the University of Pennsylvania, they will have participated in rotations through each of the clinical laboratories during their clinical training, and thus will be familiar with possible research projects and with the preceptors.

For trainees selected from Cardiovascular Fellowship Programs outside the University of Pennsylvania, the trainees will have selected a general area of research (i.e. Heart Failure/Transplantation) before enrolling in the program, and will select a specific preceptor in consultation with the Director, Co-Director and Advisory Committee. This is an illustrative example of the training program for a Cardiology fellow who seeks advanced training in patient-oriented research with particular interest in clinical epidemiology and/or clinical trials: First-year curriculum The POR trainee will have a Clinical Advisory Committee consisting of the Director of the training program, the Director of the Cardiology Fellowship Training Program (Dr. St. John Sutton), the preceptor chosen from the training faculty, and a faculty member of the Center for Clinical Epidemiology and Biostatistics or ITMAT. This committee will participate with the trainee in the selection of didactic course work, and will approve the choice of research projects. Trainees will enroll in two courses selected from the following courses offered by the Department of Clinical Epidemiology and Biostatistics: EP 510 Introductory Epidemiology EP 520 Introductory Biostatistics EP 530 Informatics for Clinical Epidemiology EP 531 Informatics for Clinical Epidemiology II EP 540 Health Social Science Research Methods These courses are designed to provide the basic epidemiology training necessary to initiate a research project, and to effectively collaborate with clinical epidemiologists and biostatisticians. It is anticipated that the fellow will develop a research project concurrent with participating in the formal course work. A preceptor will be selected at the beginning of the research year, and will work with the fellow to develop the research project. The preceptor will assist in the design of the project, and in supervising data acquisition and interpretation. It is anticipated that 50% of the training time will be devoted to course work, and 50% to designing and carrying out the research project. During this year of training, the fellow will participate in clinical activities for no greater that 20% effort. The clinical activities may involve the initial acquisition of advanced training in the chosen clinical sub-subspecialty (e.g. Cardiovascular Imaging, Heart Failure/Cardiac Transplantation, Cardiac Catheterization, Electrophysiology), and participation in a longitudinal general cardiovascular clinic. The participation in the limited clinical activities will allow the fellow to maintain clinical skills and to help to identify clinical questions to pursue during the research portion of the training.

Second year curriculum During the second year of the training program, the trainee will complete the specified clinical research project, and formulate plans for subsequent research. During the second year of the advanced clinical research program, the fellow will be support by either a research fellowship from a national organization or the training grant. The fellow will spend at least 80% of his/her time directly involved in research. Over the course of the two-year program, the fellow will have acquired the skills to initiate a career in clinical research in a subspecialty area of cardiovascular medicine. It is anticipated that during the first 2 to 3 years following this training program, the trainee will have the clinical skills necessary to join the faculty of an academic cardiovascular program, and during this time will gain further training in clinical research through senior faculty mentors. Research Conferences: Trainees will be expected to attend a variety of research conferences offered by the Division of Cardiovascular Medicine, the CCEB, the ITMAT and the Penn Cardiovascular Institute. A partial list of these conferences and seminar series include: 1) Cardiology Grand Rounds (see Appendix for list of speakers) 2) Cardiology/Pharmacology/Thrombosis Seminar Series (see Appendix for list of speakers) 3) Institute for Medicine and Engineering Seminar Series (see Appendix for list of speakers) 4) Molecular Cardiology Speaker Series (monthly) 5) Department of Physiology Weekly Seminar 6) Center for Experimental Therapeutics/Pharmacology Weekly Seminar 7) Pennsylvania Muscle Institute Weekly Seminar 8) Diabetes Center Weekly Seminar 9) Gene Therapy Weekly Seminar All trainees must attend the weekly Cardiovascular Medicine/Molecular Cardiology journal club at which trainees learn to critically review relevant literature. Trainees are paired with specific mentors for these presentations. As described in the Appendix, these research conferences include presentations by University of Pennsylvania faculty as well as internationally recognized guest speakers working in basic, translational and patient-oriented cardiovascular research. In addition to these general conferences, there are several more specialized conferences that will be attended by subsets of the trainees in each of the programs. Finally, the Penn Cardiovascular Institute sponsors an annual retreat focused on one particularly important area in cardiovascular research. In addition to these group activities, trainees will be expected to present their research at regularly scheduled lab meetings supervised by their preceptor.

Trainee Candidates Source of predoctoral trainees Graduate Students (MD/PhD or PhD) will be recruited from the MD/PhD program at the University of Pennsylvania or from the pool of graduate students in the comprehensive Biomedical Graduate Studies (BGS) program at the University of Pennsylvania. The qualifications of pre-doctoral candidates in the BGS and the MD/PhD program are shown in Tables VIa and VIb, respectively. There are currently 162 MD/PhD students enrolled at the University of Pennsylvania many of whom are interested in pursuing careers in academic cardiology. More than 92% of our MD/PhD program graduates hold positions at academic medical centers or research institutions. Pre-doctoral trainees will be recruited through the pool of MD/PhD and PhD graduate students through three mechanisms: i) a formal presentation and brochure describing the program at the time of MD/PhD and graduate student recruitment, ii) an annual Saturday Brunch presentation to all MD/PhD and BGS graduate students who have not as yet selected mentors and thesis projects, and iii) direct solicitation of qualified trainees and projects from the trainers on this application. Source of postdoctoral trainees Postdoctoral trainees will be recruited from one of four groups: 1) MD (or MD/PhD) fellows who have completed the 2-year core clinical curriculum of the Cardiovascular Medicine Training Program 2) MD (or MD/PhD) fellows who have completed core cardiovascular clinical training in another Cardiovascular training program 3) MD (MD/PhD) fellows who have completed clinical training in another subspecialty relevant to cardiovascular biology and medicine (i.e. CT Surgery, Vascular Surgery, Vascular Medicine, Diabetes/Metabolism) 4) PhD postdoctoral fellows pursuing academic careers in cardiovascular research The list of applicants for the postdoctoral training positions in FY04 is shown in Table VIIa. This list is representative of most years where approximately 50% of applicants are derived from the Cardiology fellowship training program and 50% from other divisions, departments or from outside the University

of Pennsylvania. Of note, Cardiology fellows at Penn represent a very select group. Each year, approximately 400-500 applications are received for 6-7 available positions (see Table V). As part of this process Drs. Parmacek and St. John Sutton have made a special effort to identify, recruit, and support fellows interested in careers in basic and translational cardiovascular research. Scholarship is judged by performance during medical school and during internal medicine training. Academic commitment is judged by undergraduate and medical school research activities and letters of support from previous research advisors, and by a personal interview by the Director of the Cardiovascular Division, Dr. Parmacek. Applicants with obvious research potential are matched with leading cardiovascular researchers at Penn during the interview process. Appointment Process: Trainees will be selected for support on this training grant by the Oversight Committee after reviewing nominations obtained from all listed trainers. Selection will be based on the relevance of the trainee's interests to the goals of this program, excellence in past performance, and high potential for research. Support on this grant will generally be limited to the first two years of training except when MD postdoctoral fellows require remedial course work. After being selected by the Oversight Committee for support on the training grant, each trainee will meet the Co-Directors of the training grant in order to be advised as to the requirements of the training program. For MD fellows interested in basic research but with inadequate basic science background, this will involve setting a program of coursework that fulfills the requirements of the training grant. Before the trainee is allowed to go onto the training grant, this coursework plan must be reviewed and approved by the Oversight Committee. In the case of all trainees, the semiannual evaluation procedure will be explained, along with the requirements that must be met in order to have continued support on the training grant. Trainee Qualifications Upon Completion of Training: For postdoctoral fellows, with their clinical training completed and research training acquired through this program, trainees will not only be board eligible in their specialty (MD or MD/PhD), but will also be qualified to pursue the next step in career development, attaining independent funding of their research and appointment as junior faculty members. All postdoctoral fellows will be advised to apply for the appropriate career development award during their final year of training. Graduate students will either complete their medical training (MD/PhD) or apply for post-doctoral research fellowships (PhD). Recruitment of Individuals from Under Represented Racial/Ethnic Groups One of the major goals of Biomedical Graduate Studies at the University of Pennsylvania is the education of scientists who will assume leading roles in medical research as under represented minorities. This is a critical mission in light of the exceptionally poor representation of minorities in biomedical research and the anticipated need to replenish the faculties of medical centers in the next decade. Both the American Heart Association and American College of Cardiology have identified training of underrepresented minorities to be cardiologists and CT surgeons as a national priority.

The disproportionate under representation of certain minorities (African-American, Hispanic/Latino, and Native American) in research (and in Cardiology and CT Surgery) is of grave national concern. The causes for this dilemma are deeply rooted in the cultural biases, socioeconomic inequities, and educational anachronisms of American society. Aggressive strategies are necessary if we are to reverse this situation. The University of Pennsylvania School of Medicine and Biomedical Graduate Studies have been at the forefront of efforts to increase the recruitment and training of under represented minorities in medicine and biomedical research. Table VIc describes the Biomedical Graduate Studies (BGS) predoctoral underrepresented minority applicant pool broken out by graduate group over the past five years. Table VId describes the number of underrepresented minority (URM) applicants and the percentage of URM applicants accepted and enrolled in the University of Pennsylvania School of Medicine MD/PhD degree program. Table VIIc describes the number of applicants, positions offered and enrollees for the University of Pennsylvania MD program from 2000-2004. Table VIId lists the URM minorities enrolled in the University of Pennsylvania Cardiology fellowship training program as well as other URM applicants for postdoctoral positions on this training grant. Our minority recruitment and retention effort contains five elements: 1) Education, 2) Recruitment, 3) Financial Assistance, 4) Individualized Program Instruction, and 5) Peer and Mentor Support. 1. Education: A major objective of the minority program is to diminish the impact of reduced educational opportunities and environmental disadvantages on minority high school students in Philadelphia. This program extends our existing local outreach efforts addressing the fundamental cause of the under representation of minorities in biomedical research careers. The accepted premise is that a long-term solution must improve the educational opportunities and exposure of minority students from the earliest stages onward. To meet this need, we have developed a comprehensive, multi-component program that increases minority interest and competence in biomedical science through hands-on laboratory experiences, supplemented by basic science instruction and career counseling. The program is now in its 17th year. Participants include college undergraduates and medical students. Our ultimate goal, through all of these programs, is to increase the pool of interested and academically prepared minority students for careers in academic medicine and biomedical research. 2. Recruitment: Aggressive strategies are necessary if we are to expand the pool of minority applicants and interest them in pursuing medical and/or research training. Biomedical Graduate Studies staff participate in national symposia sponsored by MARC (Minority Access to Research Careers) and SACNAS (Society for the Advancement of Chicanos and Native Americans in Science). Staff also attend annual minority undergraduate career conferences at Morehouse College and in Philadelphia. Minority doctoral and medical students in the program make recruitment trips to these conferences, to local Philadelphia public high schools, and to historically black colleges and universities to meet with students and encourage them to consider careers in biomedical research. In addition, Biomedical Graduate Studies communicates annually by mail with approximately 125 MARC and MBRS program directors; they receive an updated catalog, newsletters, and program brochures,

and they are asked to encourage their students to consider Penn for graduate or professional training. In September 1993, the Medical School administration developed a summer program designed to strengthen Penn's ties with minority undergraduates at four of the leading predominantly and historically black colleges and universities: Howard University, Xavier University of Louisiana, Lincoln University, and Spelman College. In 1994, faculty from these institutions visited Penn for a period of time, determined by their schedules and the nature of their research project, and brought along with them a science student from their home institution. These ties also led to the submission of a proposal to the Fogarty Foundation to support Minority International Research Training (MIRT). The Medical School was the recipient of the largest MIRT grant offered. This program provides brief overseas research experiences for all minority PhD and combined degree students. In 2003, the Division of Cardiovascular Medicine formed a research and educational collaboration with Cheney University, historically an African-American college, focused on disparities in the management of hypertension in underrepresented minorities. As part of this Commonwealth sponsored program Cheney University undergraduate students come to Penn for didactic class work in clinical epidemiology and participate in this ongoing research study under the direction of Stephen Kimmel, MD, MSCE, a trainer on this grant. 3. Financial Assistance: Financial assistance is provided in terms of sponsored research through appropriate categorical grants and through institutional resources. Undergraduates may be supported for 10-week summer research projects through an NIH sponsored Minority Undergraduate Training Grant or for foreign travel through the MIRT grant described above. Graduate students are supported through predoctoral training grants, individual NRSA awards, or through the University of Pennsylvania Fontaine Minority Fellowship. As a result of this cooperative effort, all minority graduate and combined degree students training in biomedical research currently receive full tuition and stipend support for their entire program. In addition, medical students are supported for short- term research via Short-Term Research Experience grants from NIH. Research of longer duration is provided by private or institutional fellowships. 4. Individualized Program Opportunities: There are numerous individualized program opportunities for medical or graduate students and fellows. All research programs for minority students, regardless of category, are coordinated through the Office of Biomedical Graduate Studies. The Office maintains a dedicated, full time Minority Affairs Officer who works closely with the Director of Minority Affairs at the Medical School. The special opportunities of individual training programs are best described in the specific research handbooks and brochures. In general, students are encouraged to develop their own programs with the guidance of faculty and staff. The existence of a central administration for all student/fellow research enables students to select laboratories throughout the medical school. 5. Peer and Mentor Support: Peer and mentor support plays a crucial role in nurturing young students. Students are individually assigned student and faculty mentors. Mentors play an important role in fostering the development of

individual programs, provide important role models for development assessment of career goals, and establish an important link for the recognition of special needs. In this latter regard, the program has a well-established academic tutor service that is available without charge to all students who identify a need for help. Finally, in coordination with the Center for Excellence, the program organizes numerous social activities that bring program participants, medical and graduate, and faculty together to discuss concerns. Achievements: Table VIc and VId describe the number of minority applicants, the number of minority applicants offered positions, and the number of graduate students successfully recruited to the BGS and MD/PhD programs, respectively over the last five years. Over this period, the number of URM students accepted has increased from 7.0% to 12.0% and the capture rate (number accepted/enrolled) has increased from 7.0% to 15%. Table VIIc shows the number of underrepresented minority applicants that were accepted and matriculated in the M.D. program at the University of Pennsylvania School of Medicine. In 2004, 20.6% of the incoming class medical school class represented underrepresented minorities. Table VIIc lists the URM enrolled in the Penn Cardiology fellowship training program and URM applicants sponsored by this training grant. Over the past five years 6 of 33 (18.2%) fellows entering Cardiology fellowship training at Penn were underrepresented minorities. Three underrepresented minority postdoctoral fellows (two MDs and one PhD) have been sponsored by the training grant. As described in Table IX, Joseph Poku, MD, an African-American, completed his cardiac electrophysiology subspecialty training and then performed a two-year research fellowship with Dr. Francis Marchlinski focused in patient-oriented research in the area of cardiac electrophysiology. He published several manuscripts during his training and presented his research at national meetings (Table X). Merielle Moise, MD, an African-American, is training to become a CT Surgeon, she spent one year training in the laboratory of Dr. Joseph Woo investigating stem cell therapies in animal models of vascular disease. She received an NRSA and is currently completing her research and clinical training in the Department of Surgery at Penn. Fiona Mack, PhD, an African-American, is a postdoctoral research fellow in the laboratory of Dr. Jonathan Epstein, Director of the Molecular Cardiology program, this year. She recently obtained her PhD in the laboratory of Dr. Celeste Simon, an HHMI investigator and trainer on this grant. Proposed Plans: A significant change in this application, is our request to train graduate students (versus only postdoctoral fellows) in cardiovascular sciences. For the reasons described in the Background Section, we believe that it is especially important to introduce talented underrepresented minority MD/PhDs and PhDs to cardiovascular biology early in their career, prior to obtaining formal training in clinical cardiology. As described above, the University of Pennsylvania Combined (MD/PhD) and BGS degree granting programs actively recruit under-represented minority students. Using the recruiting

techniques described above, we will make a concerted effort to identify talented minority applicants early in the recruiting process. In addition, Drs. Lawrence Brass and Gary Koretzky, trainers on this grant who oversee the MD/PhD training program, and Dr. Jonathan Epstein, coordinator of the Clinician-Investigator Pathway (CIP), will identify potential minority trainees to the Advisory Committee. We believe that if we are able to recruit under-represented minority applicants as MD/PhD or PhD graduate students, they are extremely likely to pursue successful careers in cardiovascular science and medicine. The recruitment of minorities into the Cardiovascular Fellowship Training Program has been deemed a high priority by the director of this program (Dr. Michael S. Parmacek). Minority applicants are identified as follows in several ways. First, the application form for clinical fellowship asks the applicant to specify whether he/she is a member of a minority group. All minority applicants from academic internal medicine training programs who meet the qualifications for consideration for admission to the clinical cardiology program are personally contacted by Dr. Parmacek. These applicants interview schedule is specifically arranged to address the interests of the candidate and an informal meeting is arranged with other minority applicants in the Cardiovascular Medicine Fellowship Training program. Another step taken to identify minority applicants is to contact members of the Association of Black Cardiologists to indicate the specific interests in this training program in attracting African-American applicants. Finally, all advertisements for the training program contain a statement encouraging minority applicants. Plan for Instruction in the Responsible Conduct of Research All of the postdoctoral trainees supported by this grant are required to attend the medical schools training program in the Ethical Conduct of Biomedical Research. This program has three major educational components: The Symposium in Bioethics; the program literature and reference source guide; and small group discussion workshops. Participation in all three phases is required. As described in the training program above, trainees performing translational and patient-oriented research will also be required to participate in either the ITMAT- or CCEB-sponsored Bioethics Courses. The training program is introduced during the trainees first year through the Symposium in Bioethics. The Symposium is designed to provide all participants with an introduction to bioethics and, more specifically, ethics in biomedical research. The Symposium is divided into four one-hour sessions presented by the Center for Bioethics that is directed by Dr. Arthur Caplan. The topics presented include: 1) Ethics & Research: The Big Picture, 2) Ethical Issues in Human Subjects Research, 3) A Graduate Students Bill of Rights, 4) The Art of Authorship, 5) Where the Rubber Hits the Road: Research Collaboration, 6) Technology Transfer and You, and

7) Conflicts of Interest. Each year the presentations are videotaped, and copies of the program are held on reserve and maybe viewed in the Biomedical Library. The program literature consists of two primary documents: ON BEING A SCIENTIST: Responsible Conduct in Research (National Academy Press, Washington, DC) ETHICAL CONDUCT IN BIOMEDICAL RESEARCH: A handbook for Biomedical Graduated Students and Research Fellows (Biomedical Graduate Studies (BGS), University of Pennsylvania). The organization and intent of these two documents are similar. The BGS handbook includes topics that are not included in ON BEING A SCIENTIST but are judged to be important to the training of graduate students and research fellows at the University of Pennsylvania. These topics include discussions of peer review, mentoring, and research on human and animal subjects. The BGS handbook also includes a practical guide to assist students and fellows in the guidelines and procedures regarding alleged misconduct at the University of Pennsylvania and to identify the appropriate source for contact when questions arise. Copies of these handbooks, and more detailed reference materials, are disseminated by the BGS Office. Small group discussions on the major topics of ethical conduct in biomedical research, as outlined in the documents, are discussed using the case study approach. Small group workshops of 12-15 trainees are organized with two faculty preceptors each. The workshops meet for two hours. Successful case based learning offers considerable workshop flexibility. One of its benefits is that student and faculty become engaged in a process of discovery together. Research integrity includes not only learning rules, but also exploration of the preceptors own attitudes and prejudices and recognition of conflicting ethical principles. This method also provides the opportunity to directly illustrate the avoidance of misconduct through good laboratory practice. The participation of active investigators is essential in this exercise. Indeed, participation in bioethics training is required of all BGS faculty. Their involvement lends credibility to the process and may influence their own research practices. E. Progress Report Accomplishments of the training program (years 05-09). This training program was established in 1996, but was revised significantly in 2000 when Dr. Parmacek became the P.I. The list of all fellows enrolled in this training program including their current positions and grants obtained is shown in Table IX. While it is too soon to determine whether this training program (i.e. especially the program put in place in FY00) has been successful, as measured by the % of trainees who establish successful careers as independent investigators with research interests in cardiovascular science or medicine, obtainable early measures of success including:

i) applicant academic profile, ii) record of publication during and after training (Table X), and iii) % of applicants staying in academics (Table IX); suggest strongly that this program will ultimately be judged successful. We believe that the number of first authored publications in outstanding journals reflects the overall quality of training and research experiences offered through this program. Trainees have been lead authors of publications published in Cell, JCI, Molecular and Cellular Biology, Development, JBC, PNAS, JAMA, JACC and Circulation (Table X). Since inception of the revised training program in FY00, fifteen of nineteen (79%) trainees have obtained faculty positions and/or are in the process of completing their fellowship training. This is significantly above the % of Cardiology fellows nationally that stay in academic positions. In addition, graduates of this program and trainees have obtained NIH RO-1, K23, K08 and NSRA awards. We are very encouraged by this upward trend in the % of trainees pursuing academic careers over the past five years and the % of trainees obtaining NIH support. Importantly, graduates from this training program have obtained fulltime faculty positions at leading academic institutions including: University of Pennsylvania (4), UCLA, Columbia, University of Washington, University of Florida and the University of West Virginia. Assignment of awarded positions In the current project period, this grant provided training for 6 postdoctoral trainees per year. Table VIII documents for each year of the current project period, the programs actual assignment of awarded trainee positions including: (1) the total number of positions awarded in each year, (2) the number of postdoctoral trainees appointed, their degrees, and their length of appointment. As detailed in Table VIII, 23/24 awarded training grant positions have been filled though one student left the program this year upon obtaining a NRSA. This includes successful recruitment of 6 MD/PhDs, 13 MDs and six PhDs. This also includes successful recruitment of three URM research fellows. Trainees outcomes During the five-year training period from 7/1/00 to 6/30/05, a total of 19 trainees were supported by this grant. Follow-up information on all of them is included in Table IX. Table IX lists all past and present trainees supported by this training grant including: (1) name, (2) the year of entry into the training program, (3) prior institution, and degree at entry, (4) source of support during each year of training, (5) research mentor, (6) research topic, and

(7) current position and institutional affiliations (for those completing training). Of those trainees who have completed their training during the history of this award (FY96-04) eight of sixteen have obtained fulltime faculty positions at other Medical Schools and two additional trainees have joined bio-pharmaceutical companies. Trainee publications, current research projects Table IX also includes a brief summary of the research conducted by each trainee supported and Table X is a list of research publications emanating from this work. 23 of 26 trainees supported by this grant have published one or more manuscripts. All trainees in the past five years have published one or more manuscript and many have one prestigious awards. Human Subjects All of the projects involving human subjects in which trainees might participate are subject to annual review and approval by the institutional human studies committee. In addition, the committee issues an overall approval of the training grant itself. This was last done on 2/2/04 and approval extends through 2/28/05. Vertebrate Animals All of the projects involving animal studies in which trainees might participate are subject to annual review and approval by the institutional animal studies committee. In addition, the committee issues an overall approval (or disapproval) of the training grant itself. This was last done on 8/13/04 and approval extends through 8/13/05.