external advisory board

The Joseph Stokes, Jr., Research Institute

The NHLBI Specialized Center of Clinically Oriented Research (SCCOR) in Pediatric Heart Disease at the Children's Hospital

of Philadelphia: P50-HL74731

Program Title: Genetic Mechanisms in Pediatric Heart Disease

http://stokes.chop.edu/programs/sccor/ Program Director: Robert J. Levy, M.D.

http://stokes.chop.edu/programs/sccor/






External Advisory BoardElazer R. Edelman, MD, PhD, FACCDirector, Harvard-MIT Biomedical Engineering CenterThomas D. and Virginia W. Cabot ProfessorHealth Sciences and TechnologyMassachusetts Institute of Technology & Harvard Medical School, Attending Cardiologist, Brigham and Women’s Hospital

David H. Ledbetter, Ph.D., Robert W. Woodruff Professor of Human GeneticsDirector, Division of Medical Genetics Emory University School of Medicine

Jurg Ott, Ph.D.Professor and HeadLaboratory of Statistical GeneticsRockefeller University


The CHOP SCCOR—Programmatic Hypothesis

“The CHOP SCCOR is a direct outgrowth of productive researchat our Institution over the past decade that was based on the hypothesis thatcongenital heart abnormalities are caused by gene defects”

“Basic discoveries concerning gene abnormalities and related patterns ofgene expression can be applied to a unifying approach for both understandingthe complex basis for cardiac dysmorphogenesis as well as providing therapeuticinsights for translational directions.”


THE CHOP SCCOR GENETIC MECHANISMS IN PEDIATRIC HEART DISEASE

PRO JECT TITLE PROJECT LEADER Academic Unit1 Biocompatible Heterograft Biomaterials Robert Levy, M.D. Pediatric Cardiology2 Genetic Analysis of Human Outflow Tract Malformations Deborah Driscoll, M.D. Genetics3 Genotype and Clinical Outcome in Conotruncal Defects Elizabeth Goldmuntz, M.D. Pediatric Cardiology4 Molecular Analysis of Human Subtelomeric Rearrangements Ian Krantz, M.D. Genetics5 Chromosomal Rearrangements and Cardiac Candidate Genes Beverly Emanuel, Ph.D. Genetics

CORE TITLE PROJECT LEADERA Administrative Robert Levy, M.D. Pediatric CardiologyB Clinical Elizabeth Goldmuntz, M.D. Pediatric CardiologyC Cell Culture, DNA and Microarray Beverly Emanuel, M.D. GeneticsD Cardiac Morphology, Gene Expression, and Histology Kenneth Ryan, Ph.D. Pediatric CardiologyE Bioinformatics and Data Analysis Peter White, Ph.D. Bioinformatics


Project 1:Biocompatible Heterograft Biomaterials Investigations concerning novel surgical therapies for congenital

cardiac malformations.Project Leader: Robert J. Levy, M.D., Professor of Pediatrics

and Pharmacology, University of Pennsylvania School of MedicineSpecific Aims1. Triglycidyl Amine (TGA)—A new crosslinking reagent for preparing

heart valve bioprostheses: Chemical and biological mechanisms

2. TGA-Matricellular interactions: Cellular and molecular biology studies related to anticalcification mechanisms.

3. Mechanisms responsible for TGA-mediated inhibition of heart valve calcification: Biomechanics, biocompatibility, & changes in gene

expression patterns.

Subcontract PI: Joseph Gorman, M.D., Asst.Prof. Surg. Univ.Penn Sch.Med.

Subcontract PI: Michael Sacks, Ph.D., Prof. of Bioengineering, Univ.Pitt.


Carpentier-Edwards

Hancock[Medtronic]

Ionescu-Shiley

Porcine aorticvalve

Porcine aorticvalve

Bovine pericardium

Bioprosthetic Heart Valves


Calcified Bioprosthesis


Triglycidyl Amine (TGA)

O

CH2Cl

HOCl

N

Cl

OHHO

Cl

NH3

i-PrOH-water

O

NO

O

aq. NaOH, toluene-THF

-HCl

TGA

Epichlorohydrin

Project 1

•Reacts irreversibly with lysine, methionine, cystine, histidine

•Results in biomechanical properties superior to glutaraldehyde

•Biocompatibility—supports cellular growth of all cardiovascular cell types


Project 2:Genetic Analysis of Human Outflow Tract Malformations

Project Leader: Deborah Driscoll, M.D., Professor and Chair,Department of Obstetrics and Gynecology, University of

Pennsylvania School of Medicine

Specific Aims1. Developing a map of single nucleotide polymorphisms (SNP’s) in

selected genes implicated in cardiac development.

2. Evaluating SNP’s for potential functional alterations.

3. Determining the genetic contribution of selected genes to the development of outflow tract malformations using family-based linkage disequilibrium testing.

4. Identifying modifiers of the cardiac phenotype in patients with 22q11 deletions


VEGF Related Directions Animal models demonstrate VEGF capable of

influencing pharyngeal arch patterning SNPs with reduced VEGF expression are associated with

cardiovascular defects in 22q11 deletion syndrome Identified an “at risk” haplotype for cardiovascular defects

among individuals with 22q11 deletion Suggests risk of CHD in the fetus with 22q11 deletion

increases when VEGF levels fall below critical threshold needed for proper development of pharyngeal arch arteries, severity may be due to degree of vascular impairment


Project 3:Genotype and Clinical Outcome in Conotruncal Defects

Project Leader: Elizabeth Goldmuntz, M.D., Associate Professor of Pediatrics, University of Pennsylvania School of Medicine

Specific Aims1. Investigations of the contribution of NKX2.5 and related genes to the

etiology of controtruncal defects using mutation analyses and family-based association studies.

2. Investigating whether subsets of patients with transposition of the great arteries or double outlet right ventricle share a common genetic etiology with the heterotaxy syndrome: Studies of CFC1 mutations & other genes (NODAL, ZIC3,LEFTY1, ACVRIIB)

3. Studies of the relationship between genetic etiology and clinical variability/outcome in subjects with conotruncal defects


Impact of Genotype on Clinical Status

Cross sectional study Subjects with TOF, Truncus or IAA Ages 8-18 yo Clinical Assessment

Exercise study Echocardiogram Cardiac MRI Child health questionnaire

Project 3: Aim 3


Project 4:Molecular Analysis of Human Subtelomeric Rearrangements

Project Leader: Ian Krantz, M.D., Assistant Professor of Pediatrics and Genetics, University of Pennsylvania School of Medicine

Specific Aims1. Identify individuals with subtelomeric chromosomal deletions and

congenital heart defects.

2. Develop a diagnostic assay that targets the critical region and sizes subsequent rearrangements of each of the telomeres.

3. Defining the critical regions and identifying candidate disease related genes for specific clinical phenotypes by mapping the extent and composition of the associated rearrangements.


Project 5:Chromosomal Rearrangements (CR) and Cardiac Candidate Genes

Project Leader: Beverly S. Emanuel, Ph.D., Professor and Chair of

Genetics (at CHOP), University of Pennsylvania School of Medicine

Specific Aims

1. Identify and characterize CR’s in patients with congenital heart disease by high-resolution cytogenetics and molecular cytogenetic analysis

2. Develop PCR-based mapping strategies using the human genomic sequence to identify the translocation BP’s

3. Characterize the genomic DNA from normal chromosomes at the chromosomal breakpoints

in order to identify mechanisms of rearrangement

4. Identify the candidate genes disrupted or deleted at the translocation BP’s as candidates for early cardiac morphogenesis.

5. Determine whether mutations in the candidate genes are associated with the specific cardiac defect in other patients in the SCCOR Clinical Core.


The Clinical Core (a continuing resource from the SCOR’s)

Director: Elizabeth Goldmuntz, M.D.

Objectives:1. Molecular analyses of the genetic etiology of conotruncal defects 2. Molecular analyses of bioprostheses3. Impact of genotype on cardiac anatomy and clinical outcome

Services1. Ascertain Subjects2. Acquisition of clinical data3. Acquisition of relevant samples4. Review of pertinent medical records5. Coordination of clinical studies


The Cell Culture, DNA, and Microarray Core (a continuing resource from the SCOR’s)

Director: Beverly Emanuel, Ph.D.Objectives:1. Provide cell culture, DNA isolation, cytogenetic and DNA analysis support for all of the projects2. Provide microarray resources3. Training and consultation services to all Projects andCoresServices:1. Establishing lympholastoid cell lines from patientswith congenital heart defects.2. Isolation of DNA from established cell lines, peripheral lymphocytes3. Perform FISH to screen for 22q11.2 deletions4. Regionally localize newly identified human cDNAs by FISH5. Provide genotyping services for the SCCOR


Cardiac Morphology, Gene Expression and Histology Core

Director: Kenneth Ryan, Ph.D., Assistant Professor of Pediatrics, University of Pennsylvania School of MedicineObjectives:1. Analysis of gene (mRNA and protein) expression2. Histological support3. Breed mice & xenopus for harvesting embryos for whole-mount in situ hybridizations and sectioning re. cardiac gene expression patterns

Services1. Generate and bank frozen staged mouse & xenopus embryo RNA samples. 2. Generate as blocks and slides embedded mouse and xenopus embryos.3. Share expertise in and perform in situ hybridizations using antisense RNA probes. 4. Perform immunohistochemistry and related histology


Bioinformatics and Data Analysis Core

Director: Peter White, Ph.D., Assistant Prof. Ped.,Univ.Penn.School of Medicine, Director, CHOP’s Bioinformatics CoreCo-Director: Charles Scott, Ph.D.

Objectives:1. State of the art bioinformatics & biostatistics resources2. Experimental design support3. Data base development and data management support4. The integration of bioinformatics and biostatistics data on SCCOR subjectsServices1. Provide assistance in study design, database design, and data storage2. To provide infrastructure, hardware, software, technical support, for analyzing results of molecular biology experiments3. Statistical analysis and data interpretation


NHLBI PEDIATRIC HEART DISEASE SCCOR’s: 2005 Programmatic Meeting

At the NHLBI, December 13, 2005Children’s Hospital, BostonPI: Jane Newburger, M.D.

Children’s Hospital of CincinnatiPI: Woody Benson, M.D., Ph.D.

Children’s Hospital of PittsburghSteve Webber, M.D., Ph.D.


Jane W. Newburger, M.D., M.P.H.Children’s Hospital, Boston

Harvard Medical School

From Molecular Mechanisms to

Improved Outcomes in TOF


Children’s Hospital, Boston:ProjectsProject 1: Neurologic and developmental

outcome in TOF (Newburger)

Project 2: Randomized trial of pulmonary valve replacement in TOF (Geva)

Project 3: Human mutations that cause TOF (Seidman)

Project 4: Mitochondria in hypertrophied RV and surgical ischemia (McGowan)

Project 5: Functional analysis of cardiac transcription factor NKX2.5 (Izumo/Jay)

Project 6: Cardiac regeneration in zebrafish (Keating)


CoresCore A: Research support and statistics (Newburger)

Core B: Microarray core (Schinke)

Core C: Children’s Hospital-Harvard TOF registry

(Breitbart)

Core D: Skills development core: The pathology of CHD (Collins/Jurazek)


Children’s Hospital of Cincinnati: SCCOR in Pediatric Heart Disease: PI Woody Benson

“Molecular mechanisms of valve development and disease”PI: D. Woodrow Benson, MD, PhD

Project 1 – Benson—Genetic Studies of Valvular Heart DiseaseProject 2 – Gelb—To Identify PTPN11 Defects Associated with Noonan’s syndrome and other forms of congenital heart disease.Project 3 – Yutzey--Regulation of valvuloseptal development by DSCR1Project 4 – Robbins-- Mechanisms of Cardiac pathogenesis in Noonan Syndrome, effects of SHP-2Gln79Arg, a common PTPN11 mutation associated with Noonan’s syndrome.


Children’s Hospital of Pittsburgh: Children’s Hospital of Pittsburgh: Optimizing Outcome after Pediatric Heart Transplantation

SCCOR ProgramS.A. Webber, PI Consultants

CORESPROJECTS

Project 1Thymic ToleranceS. A. Webber, PI

Project 2Transplant EBV Disease

D. T. Rowe, PI

Project 3Genetic Contributions to

Transplant OutcomesA. Zeevi, PI

AdministrationS. A. Webber, PI

Immunological Monitoring D. Metes, PI

Biostatistics and Data ManagementS. Kelsey, PI

Clinical Skills TrainingB. B. Keller, PI

external advisory board

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