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Programs of Excellence in Nanotechnology

Topic focus: Third Annual Intra-PEN Meeting

IN THIS ISSUE

4 Spotlight 6 Research Collaborations 7 Upcoming

Events 12 3rd Annual Intra-PEN

Meeting: UCB, UCSB,WUSTL

14 2nd Annual Inter-PEN Meeting

PEN members in attendance at the 1st Annual Inter-PEN Conference, on the campus of Washington University in Saint Louis, October 6-8, 2006.

Second Annual

Inter-PEN Meeting

N a t i o n a l H e a r t , L u n g & B l o o d I n s t i t u t e

The Second Annual Inter-PEN Meeting will be held on October 19-21, 2007,

at the University of California at Santa Barbara (UCSB). Please see the back cover of this newslet-ter for an overview of the meeting agenda.

We have selected the Best Western South Coast Inn - Santa Barbara-Goleta, CA as the preferred hotel for the meeting. The guest room rate is $135.00 single/double + 10% tax, for a total of $148.50. A shuttle service to and from the University of California, Santa Barbara will be arranged by UCSB.

For questions or information re-garding the Inter-PEN Meeting,

please contact either Eileen A. Cler at Washington Uni-versity in Saint Louis at “ea-cler@wustl.edu” or James Lee at the Burnham Institute at“ j a m e s l e e @ b u r n h a m . o rg ” . You may also visit the In-ter-PEN website at www.nhl-bi-pen.net for more details.

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EMORY UNIVERSITY, GEORGIA TECHN a n o t e c h n o l o g y : D e t e c t i o n & A n a l y s i s o f P l a q u e F o r m a t i o n

Use of Quantum Dots to Evaluate Atherosclerotic DiseasePresence and Activity

By W. Robert Taylor, Don Giddens, Shuming Nie and Gang Bao

(gang.bao@bme.gateach.edu)

One of the fundamental and unresolved problems in cardiovascular biology is the in-

vivo detection of atherosclerotic disease and the evaluation of atherosclerotic disease activity. Current technology limits clinicians to diagnostic techniques that either image or functionally asses the signifi cance of large obstructive vascular lesions. However, current imaging modalities do not allow for the possibility of imaging atherosclerotic disease at its earliest stages nor do available techniques allow for routine assessment of atherosclerotic lesions susceptible to rupture and/or thrombosis. This is of particular clinical signifi cance given that myocardial infarctions and other consequences of atherosclerotic disease are just as likely to occur from small non-obstructive lesions as from larger obstructive lesions. Thus, the existing paradigm of diagnosing and treating obstructive coronary

artery disease based on the degree of luminal obstruction is fundamentally fl awed. The PEN at Emory University and Georgia Tech is aimed at developing newer methodologies that are capable of identifying earlier atherosclerotic lesions as well as atherosclerotic lesions that are active or unstable.

As an initial step in developing these methodologies, we have adapted a quantum dot-based approach coupled with two-photon excitation laser scanning microscopy to

localize expression of pro-atherogenic proteins on the intimal surface of the arterial wall [1]. Our studies have demonstrated that this novel fl uorescence en face technique has considerable advantages (no autofl uorescence, sensitive

Figure 1: Multiplexing with one excitation wavelength. An Intercostal branch of an apoE-/- mouse sectioned at the level of the media. ECs were targeted with a biotinyl-ated anti-VCAM-1 Ab and Qdot-585nm streptavidin con-jugates (pseudocolored green for contrast). Macrophages were stained with a monoclonal rat anti-mouse Ab. (MAC-3) and Qdot-655nm goat anti-rat IgG conjugate (red). Nuclear counterstaining was done with Hoechst (blue). The elongat-ed nuclei correspond to smooth muscle cells. The emission spectra of the fluorophores are on the left. Only one wave-length (750nm) was used to excite all three fluorophores.

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quantifi cation and 3D reconstruction) over traditional methods, especially for examining early infl ammatory changes of the endothelium at sites exposed to different fl ow patterns. As can be seen in Figure 1, we are able to visualize expression of the adhesion molecule VCAM-1 and adherent macrophages at the origin of an intercostal artery in a mouse with early atherosclerosis.

Figure 2: Spatial and Temporal Distribution of WSS and VCAM-1 expression in the Mouse Aorta.

Left Panel: Depiction of the computed, time-varying WSS vectors at 3 points corresponding to the sites of VCAM-1 expression in the right panel. The color coding indicates the mean WSS distribution.. The vector diagrams indicate the time-varying changes in the direction and magnitude of WSS through-out the cardiac cycle. Zero degrees indicates the direction of shear near peak systole; the magnitude scales of vectors are in dynes/cm2. There is a greater variation in the direction of the instantaneous WSS vectors in areas (a) and (b), as can be seen in the polar plots of WSS. Note the relatively low mean WSS zone at the inner curvature of the aortic arch (area b), has instantaneous magnitudes of WSS of approximately 150 dynes/cm2, while values exceeding 600 dynes/cm2 were found in the ascending aorta (region c). The expression of VCAM-1 was greatest at the sites of low or oscillatory shear stress.

In additional studies we have used this technique to examine the fundamental relationships that exists between hemodynamics and vascular infl ammation.[2] Using this technique we have mapped infl ammatory gene expression in the intimal surface of the aorta on to a computational fl uid mechanics model of blood fl ow that was developed using a combination of high resolution MRI, micro CT and ultrasound. As can be seen in fi gure 2, these studies demonstrated that areas of low or oscillatory shear stress were associated with enhanced expression of infl ammatory proteins. These approaches will allow us the opportunity to more fully explore the relationship between

hemodynamics and atherosclerosis in order to provide better insight into the relevant molecular mechanisms of atherogenesis and atherosclerotic plaque stability.

“This is of particular clinical signifi cance given that myocardial infarctions and other consequences of atherosclerotic disease are just as likely to occur from small non-obstructive lesions as from larger obstructive lesions. Thus, the existing paradigm of diagnosing and treating obstructive coronary artery disease based on the degree of luminal obstruction is fundamentally fl awed.”

References1. Ferrara, D.E., Weiss, D., Carnell, P.H., Vito, R.P., Vega, D., Gao, X., Nie, S., and Taylor, W.R. (2006), “Quantitative 3D fl uorescence technique for the analysis of en face preparations of arterial walls using quantum dot nanocrystals and two-photon excitation laser scanning microscopy”. Am J Physiol Regul Integr Comp Physiol., 290(1), R114-23.

2. Suo, J., Ferrara, D.E., Sorescu, D., Guldberg, R.E., Taylor, W.R., and Giddens, D.P. (2007), “Hemodynamic shear stresses in mouse aortas: implications for atherogenesis”. Arterioscler Thromb Vasc Biol., 27(2), 346-51.

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P E N S P O T L I G H T O N :U C S B G r a d u a t e S t u d e n t , J e a n - L u c F r a i k i n

Jean-Luc Fraikin started his under-graduate career at McGill University majoring in mechanical engineering, and switched

to physics after a couple of years before graduating with a joint honors degree in math and physics. That may sound like a meandering educational path but when viewed through the prism of nanomedicine with all of its integrated fi elds, it was perhaps an ideal training regimen.

Fraikin works in Andrew Cleland’s lab at the University of California Santa Barbara as part of the “Nanotherapy for Vulnerable Plaque” grant under the direction of Principal Investigator Jeffrey W. Smith. Now a third-year graduate student, Fraikin is helping develop nanoscale electronic sensors that detect conformational changes in tagged protein switches.

“This campus is great for this type of project,” says Fraikin. “There is a collaborative attitude here that I don’t think is present at other schools. Our work requires physics, biology and chemistry, and it is very easy to work with these other departments at UCSB. Everyone is helpful and ready to lend a hand.”

Dr. Cleland’s group is using radio frequency refl ectometry to probe the sensors, a measurement technique that works very well in a bio-setting. “We want to be able to detect a change immediately and allow for changes to be implemented in real time,” Fraikin explains. “Radiofrequency measurements are ideally suited for this purpose.”

Although the sensors have been working to some extent, the challenge of late has been to increase sensitivity and specifi city. “We are trying to incorporate nature’s molecular specifi city into our technology, and to do so we need to improve our chemistry. That part of the project is headed up by Mike Requa, Ph.D., in the lab.”

As for his future, Fraikin doesn’t know if he will end up in academia or industry. “I think there will be great opportunities in this fi eld because it is new and fi lled with a lot of interesting problems. I just don’t know in what capacity I’ll be involved.”

By James Lee Project Manager Nanotherapy for Vulnerable Plaque

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P E N S P O T L I G H T O N :W U S T L S e n i o r I n v e s t i g a t o r , P a m e l a K . W o o d a r d

Principal Investigator, Pamela K. Woodard, M.D., an associate professor at Mallinckrodt Institute, will be leading a cardiac imaging staff to work in

conjuction with Benico Barzilai, M.D., a professor in the Department of Medicine’s Division of Cardiovascular Disease. “The partnership is one of such a handful of such interdisciplinary efforts in the country.”

Woodard was featured in the Mallinckrodt Institute of Radiology, Focal Spot Magazine, Spring 2007, in an article titled “A Window Into the Heart: A Joint Venture for Radiology and Cardiology” by Anne Kessen Lowell. Some highlights from the article are outlined here.

* * * * *

“Recent advances in cardiovascular imaging technology promise quicker diagnosis of hidden health threats in some patients. The partnership combines the top-level expertise of radiologists and cardiologists with the cutting-edge technology of computed (CT) and magnetic resonance imaging (MRI) will provide a greater understanding of coronary artery disease.”

“The advent of the 64-slice CT scanner in 2003 opened a wealth of new applications in cardiac imaging.” ... “The speed and resolution of these newest CT scanners has altered the diagnostic landscape. The 64-slice CT scanner takes 64 images (in scanner nomenclature, these are known as slices) in a single revolution around the tube in which the patient lays supine for the exam. The duration that patients must hold their breath during the exam (known as the “breath-hold”) takes only a few seconds. The resulting images are richly detailed, showing the intricacies of the cardiovascular system, including the exterior of cornonary arteries and vessels. In contrast, invasive techniques can only reveal what is going on in the interior of the structures.”

“Cardiac CT angiography is evolving into a powerful tool in the battle against one of the deadliest threats to heart health-coronary artery disease. Sudden cardiac deaths, primarily from heart attack and stroke, number about one-half million in the United States”.

For more information about Pamela K. Woodard’s research, see her article on page 10. For a full copy of this article, visit the MIR website at www.mir. wustl.edu.

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R E S E A R C H C O L L A B O R A T I O N

N a n o t e c h n o l o g y f o r V u l n e r a b l e T h e r a p yBURNHAM, SCRIPPS RESEARCH, UCSB

U C - S B , U C - B a n d W U - S T LI n t e g r a t e d N a n o s y s t e m s f o r D i a g n o s i s a n d T h e r a p y

Targeting Platelet Cells for Prevention of Thrombosis

By Ashley Mynar, Craig J. Hawker, Jeffrey W. Smith, Zaverio M. Ruggeri

jsmith@burnham.org, klwooley@artsci.wustl.edu

Multi-discipline, cross-cutting research is at the heart of the NHLBI PEN program and provides the

most effi cient mechanism for the discovery and translation of nanotechnology research to clinical materials. It also provides an unprecedented opportunity to expose students to interdisciplinary research, not only through strong intra-PEN research collaborations but also in direct collaboration with other PENs. This sharing of materials, techniques and concepts across the PENs is a major goal and illustrates the versatility and wide promise of these nanomaterials.

To illustrate these efforts, a collaboration has recently been formed between researchers at the University of California at Santa Barbara (UCSB)/Washington University (WU) and the Burnham Institute for Medical Research. The objective of this multi-disciplinary project is to develop nanodevices that halt the aggregation of blood platelets, the cells which adhere to ruptured plaque and precipitate heart attack. Initial work will examine nanoparticles synthesized by the Hawker group at UCSB that will be used as drug delivery vehicles to

target the αIIbβ3 integrin which is expressed exclusively on platelets.1

This work is based on recent results in collaboration with the Wooley, Welch and Anderson groups at Washington University, from which it has been determined that well-defi ned nanoparticles synthesized by the Hawker group and conjugated with RGD-based peptide sequences exhibited strong binding to the αvβ3 integrin, over-expressed in cardiovascular injury. In addition, these materials have been shown to have controllable blood circulation times that can be accurately varied from minutes to days.

Building upon these ground-breaking results, the same nanoparticle scaffolds are being modifi ed to display the KQAGDV sequence,

Figure 1. Formation of a platelet thrombus on the surface of an atherosclerotic plaque can lead to vessel occlusion and heart attack. Inhibition of platelet aggregation will halt the growth of the thrombus and prevent heart attack.

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Second Annual Inter-PEN Meeting

October 19-21, 2007Santa Barbara, California

Executive Committee Meetings

2007 - September 7 December 7

U P C O M I N G E V E N T S F O R 2 0 0 7 / 2 0 0 8

which binds the platelet’s αIIbβ3 integrin. These new nanoparticles are currently being examined in the Smith (Burnham Institute) and Ruggeri (Scripps) laboratories. Initial studies will be conducted to assess the ability of these nanoparticles to incorporate into platelet aggregates using in vitro fl ow systems that mimic vessel physiology. Once these analyses have been completed the nanoparticles will be loaded with anti-platelet drugs and tested for the ability to halt and reverse platelet aggregation in vitro and in vivo.

A critical aspect of this Inter-PEN collaboration is to share expertise and material platforms across the PEN programs as well as to develop the next generation of scientists who will push the frontier of nanomedicine. For example, the students synthesizing the functionalized nanomaterials in the Hawker group at UCSB are being sent to both Washinton University and the Burnham Research Institute to learn the cell biology and imaging techniques essential to

References1. Ruggeri, Z.M., “Platelets in athero-thrombosis”, Nat Med., 2002, Nov; 8(11): 1227-34.

Figure 2. Schematic design of the versatility of these nanoparticles.

the translation of these materials to the clinic. This interactive approach allows the students to have a more complete understanding of the overall goal of their project while learning new techniques and ensuring a highly innovative and valuable learning experience.

2008 - March 7 June 13 September 5 December 5

R E S E A R C H C O L L A B O R A T O R S

Ashley MynarUniversity of California

Santa Barbara

Craig J. HawkerUniversity of California

Santa Barbara

Jeffrey W. SmithThe Burnham InstituteUniversity of California

Santa Barbara

* Not pictured, Zaverio M. Ruggeri, UCSB

8 Volume 1, Issue 3 Aug 2007

M G H , H A R VA R D , M I T , B W HT r a n s l a t i o n a l P r o g r a m o f E x c e l l e n c e i n N a n o t e c h n o l o g y

Dual Channel Optical Tomographic Imaging of Leukocyte Recruitment

and Protease Activity in the Healing Myocardial Infarct1

By Jason McCarthy,Ralph Weissleder

(weissleder@helix.mgh.harvard.edu)

The effi ciency of infarct healing after myocardial infarction (MI) correlates with prognosis. In cases in which

healing is ineffi cient, infarct expansion and heart failure may occur, leading to high mortality. This healing process is driven by sequential infl ammatory responses and recruitment of leukocytes to the damaged myocardium. These neutrophils and macrophages clear away cellular debris, a process which is characterized by the secretion of a number of proteinases (MMPs) and proteases (cathepsins). Until recently, evidence of these processes was based mainly upon in vitro and ex vivo observations. Thus, a group led by Drs. Matthias Nahrendorf and David Sosnovik utilized f l u o r e s c e n c e - m e d i a t e d molecular tomography (FMT) to systematically explore the healing process in an in vivo mouse model of myocardial infarction.1 More specifi cally, the investigators used dual channel imaging to examine the recruitment of neutrophils and macrophages to the infarct and cathepsin B protease activity. The presence of phagocytes in the lesion can be imaged using CLIO–AF750, a magnetofl uorescent iron oxide nanoparticle bearing AlexaFluor 750, that has previously demonstrated avid uptake by phagocytes.2 Protease activity was monitored by ProSense

680, an activatable fl uorescent polymeric substrate bearing VivoTag 680, a Cy 5.5 analogue. Both probes can be simultaneously injected into the same mouse, as the fl uorescence signals can be spectrally resolved. In this study, cohorts of mice received MI by left coronary artery ligation, and were injected with the probes 24 hours prior to imaging. The researchers found that MI causes substantial recruitment of phagocytic cells to the damaged myocardium, which is readily imaged using FMT. In fact, the time course of phagocytic infi ltration could be followed using the CLIO-AF750 probe, which demonstrated maximum uptake in the lesion 6 days post infarct (Figure 1E). Four days post-MI, ProSense activation also reaches its maximum, with focal signal localized to the cardiac region of the FMT images (Figure 1A-D, F). These results were further confi rmed by histological analysis

Figure 1. Prosense-680 is specifi cally activated in the infarct. A, FMT of mouse with myocardial infarction after injection of Prosense-680. Panels A through C are windowed identically. B, FMT of mouse without myocardial infarction (sham surgery involves thoracotomy only), 24 hour after injection of Prosense-680. C, FMT of mouse with myocardial infarction after injection of saline. D, Fluorescence refl ectance image of excised heart 4 days after MI. The fusion of fl uorescence image with the white light image shows Prosense activation mainly in the thin infarct scar (top). Almost no signal is observed in the remote myocardium in the septum and right ventricle (bottom). E, Time course of CLIO-uptake after coronary ligation as assessed by FMT. F, Time course of cathepsin activity as assessed by activation of the protease sensor Prosense- 680 by FMT.

WASHINGTON UNIVERSITY UNIVERSITY OF CALIFORNIA - SANTA BARBARAUNIVERSITY OF CALIFORNIA - BERKELEY

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of the heart. Activated ProSense 680, as imaged by fl uorescence microscopy was colocalized with immunoreactive cathepsin B, whereas CLIO-AF750 was found in neutrophils and macrophages. This was further confi rmed by fl ow cytometry of digested infarcts (Figure 2). In this experiment, cells were labeled with a cocktail of fl uorescently labeled antibodies in order to ascertain the specifi c cell types. Once gated on the appropriate cells, the associated fl uorescence from both CLIO-AF750 and ProSense 680 were determined. The results showed that monocytes/macrophages were the major contributor to the fl uorescence signal in both channels, while neutrophils also contributed, albeit much less, to the observed fl uorescence. All other cell types demonstrated no cell-associated fl uorescence. This study has established that in vivo imaging can be used to monitor the myocardial response to ischemia. Following the invasion of neutrophils to the lesion, monocytes are recruited to the area of the insult, and differentiate to macrophages, which are responsible for degrading the existing extracellular matrix and make room for the newly forming collagen scaffold.1 This infi ltration can be monitored by FMT, and may be used in the future to proactively assess the healing of myocardial infarctions.

References1. Nahrendorf, M.; Sosnovik, D. E.; Waterman, P.; Swirski, F. K.; Pande, A. N.; Aikawa, E.; Figueiredo, J. L.; Pittet, M. J.; Weissleder, R. “Dual channel optical tomographic imaging of leukocyte recruitment and protease activity in the healing myocardial infarct.”, Circ. Res. 2007, 100(8), 1218-1225.2. Pande, A. N.; Kohler, R. H.; Aikawa, E.; Weissleder, R.; Jaffer, F. A. “Detection of macrophage activity in atherosclerosis in vivo using multichannel, high-resolution laser scanning fl uorescence microscopy.”, J Biomed Opt 2006, 11(2), 021009.

The Four PENs and

their Principal Investigators

Nanotechnology: Detection & Analysis of Plaque

FormationEmory University

Georgia Institute of Technology

P.I. - Gang Bao, Ph.D. Director , Emory-GT, Program of Excellence in

Nanotechnology

CoE Distinguished Professor

The Wallace H. Coulter Department of

Biomedical Engineering

Georgia Institute of Technology

Emory University

Nanotherapy for Vulnerable PlaqueThe Burnham Institute

University of California - Santa Barbara

The Scripps Institute

P.I. - Jeff rey W. Smith, Ph.D.

Professor

Director, Center on Proteolytic Pathways

Director, Program of Excellence in

Nanotechnology

Burnham Institute for Medical Research

Translational Program of Excellence in

NanotechnologyHarvard University

Massachusetts General Hospital

Massachusettes Institute of Technology

Brigham and Women’s Hospital

P.I. - Ralph Weissleder, M.D., Ph.D. Director, Center for Molecular Imaging Research

Professor of Radiology, Harvard Medical School

Massachusetts General Hospital

Integrated Nanosystems for Diagnosis and TherapyWashington University in Saint Louis

University of California - Santa Barbara

University of California - Berkeley

P.I. - Karen L. Wooley, Ph.D.

James S. McDonnell Distinguished University

Professor in Arts & Sciences

Professor, School of Arts & Sciences,

Department of Chemistry

Professor, School of Medicine,

Department of Radiology

Faculty member in the

Center for Materials Innovation

10 Volume 1, Issue 3 Aug 2007

U C S B , U C B a n d W U S T LI n t e g r a t e d N a n o s y s t e m s f o r D i a g n o s i s a n d T h e r a p y

64Cu-C-ANF NatriureticPeptide (NP) Fragment as a PET Tracer for Atherosclerotic Plaque

Molecular Imaging

by Raffaella Rossin, Dana R. Abendschein,

Jie Zheng, Geoffrey E. Woodard, Kyle McCommis, P. McQuade,

Michael J. Welch, Pamela K. Woodardwoodardp@mir.wustl.edu

Atherosclerosis and plaque rupture is one of the principal causes of sudden death in the western world. While there are

numerous methods of detecting the presence of atherosclerotic plaque, including x-ray and CT angiography, no method accurately predicts whether a plaque is at “at risk.”

The natriuretic peptide receptor C (NPR-C) is present diffusely on vascular endo-thelium. It has been shown to be upregulated in injured endothelium and atherosclerotic plaque, its presence decreasing with plaque stability (1-3). The C-type natriuretic peptide (CNP) is a paracrine hormone present in high concentrations in vascular endothelial cells. CNP has a direct effect on immune cell recruitment in vivo, is a potent inhibitor of vascular smooth muscle cell migration and proliferation. It also is vasoactive, causing vascular dilatation.

C-ANF (4-18) is a synthetic ring deleted analog of CNP used in studies to compete with other

natriuretic peptides that bind to NPR-C. It binds selectively to NPR-C without eliciting bioactivity from the NPR-C receptor.

We have investigated the use of a 64Cu-labeled C-ANF as a possible tool for non-invasive plaque imaging with PET (4). C-ANF (atrial natriuretic factor), a C-type NP analog, was functionalized with DOTA and labeled with 64Cu in 0.1 M citrate buffer (pH 5.5). After DTPA challenge and Sep-Pak C-18 purifi cation, the radiochemical purity of 64Cu-DOTA-ANF was >95%, as measured by radio-HPLC. The right femoral artery of male New Zealand white rabbits fed a cholesterol-rich diet was double-injured by air desiccation and balloon overstretching.

C-ANF (4-18) is a synthetic ring deleted analog used in studies to compete with other natriuretic peptides that bind to NPR-C. It is able to bind selectively to NPR-C without eliciting bioactivity from the NPR-C receptor.

Rabbits injected with 64Cu-DOTA-C-ANF (2-4 mCi, ca. 1 mCi/nmol) at each time point (n = 3)

0-60 min dynamic scan in microPET Focus 220 scanner

MRI is used to confirm the presence of the plaque be-fore microPET (Siemens 3T Allegra Head-only clinical system with a home-made volumetric coil)

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The presence of atherosclerotic lesion was confi rmed by 3T MRI 1 hr after administration of a plaque-targeted contrast agent (Gadofl urine M, Schering AG). A rabbit was injected with 64Cu-DOTA-ANF (2.8 mCi, ca. 7.5 μg peptide) and a 20 min dynamic scan was acquired on the microPET Focus-220. Fiducial markers attached to the animal bed and fi lled with a 64Cu aqueous solution were used to correlate the MRI and microPET images. Histopathologic imaging and immunohistochemistry (IHC) of

Injured artery Neointimal thickening

Images obtained on a laser confocal microscope (20X) show C-type NP receptor presence on the surface of the neointima

Primary stain: rabbit anti-human polyclonal antibody designed against the first 100 AA of the NPR-C

Secondary stain: FITC conjugated to goat anti rabbit IgG, affinity purified and blocked with 2% goat serum

the ex-vivo atherosclerotic artery were performed (Fig 1). IHC demonstrated the presence of NP receptor type-C (Fig 2) .

MicroPET images clearly showed tracer accumulation at the injury site, with an average SUV of 2.1 during the acquisition scan (3-fold higher than surrounding muscle) (Fig 3). In the same time frame, the uninjured control artery was not observable. This preliminary imaging study suggests that 64Cu-DOTA-ANF is

a promising candidate tracer for PET imaging of atherosclerotic plaques.

References1. Woodard G.E., Rosado J.A., Brown J., ”Expression and control of C-type natriuretic peptide in rat vascular smooth muscle cells”, Am J Physiol Regul Integr Comp Physiol., 2002, Jan; 282 (1): R156-65.2. Casco, V.H., Veinot, J.P., Kuroski de Bold, M.L., Masters, R.G., Stevenson,

M.M., de Bold, A.J., “Natriuretic peptide system gene expression in human coronary arteries”, J Histochem Cytochem., 2002, Jun; 50(6):799-809. 3. Brown J, Chen Q. “Regional expression of natriuretic peptide receptors during the formation of arterial neointima in the rabbit”, Circ Res., 1995, Nov; 77(5):906-18.4. Rossin R., Abendschein D., Zheng J., Sharp T., Woodard G.E., McQuade P., Welch M.J., Woodard, P.K. “64Cu-Labeled natriuretic peptide: a promising tracer for atherosclerotic plaque molecular imaging with PET”, J Nucl Med , 2007, 48 (Suppl. 2), 103P).

Control arteryFigure 2 - Plaque Immunohistochemistry

Internal elastic lamina (autofluoresence)

Figure 3 - Imaging of Rabbit #1621(6 weeks after 1st injury)

This preliminary imaging study suggests that 64Cu-DOTA-ANF is a promising candidate tracer for PET imaging of athero-sclerotic plaques.

coronaltransaxial

MRI: rabbit injected with Gadofluorine M (Schering, ) 0.5 μmol/kg bw) and imaged 1 h p.i.MicroPET: rabbit injected with 64Cu-DOTA-C-ANF (3.8 mCi, ca. 1 mCi/nmol), 0-60 min p.i. dynamic scan

12 Volume 1, Issue 3 Aug 2007

UC - Santa Barbara, UC - Berkeley, Wash U. - STL

Jul y 6 -7 , 2007

So many new faces!

In t eg r a t ed Nanosy s t ems fo r D iagnos i s and Therapy

Ke Zhang (far left, Wooley lab), and John Taylor (far right) talk to three new Ph.D. candidates in the Taylor group: (from left to right), Ziyan Zhang, Ronald Chang and Zhenghui Wang at the Friday evening Welcome Reception.

Walter Akers (Achilefu group-WUSM) and Adah Almutairi (Fréchet group-UCB) discuss their collaboration which involves developing small nanoprobes for near infrared imaging. The nano-probes are activated upon in vivo interactions with diseased tissue.

Jasmine Hunt (Hawker group), Aviv Hagooly and Yongjian Liu (Welch group) and Ashley Fiamengo.(Anderson group).

Hawker group - (left to right) Ryosuke Sakai, Ashley Mynar and Jos Paulusse. Ashley has been working with Jeff Smith’s PEN at the Burnham Institute and wrote the article under “Research Collaborations”, with input from Jeff Smith’s group.

A w a r d R e c o g n i t i o n

Going the Extra Mile - A handful of small “door prizes” were given out to one or two members in each of the four PEN subgroups, ALI, AVI, FUND & SKILLS for their excellent work on the PEN.

AdahAlmutairi

JessicaCohen

KenyaPowell

EricPressly

MonicaShokeen

Derekvan der Poll

Raffaella Rossin filled in for Michael Welch at the Senior In-vestigator Meeting, in addition to giving three of the talks.

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TheHome Team

Our Sen ior Inve s t i ga tor s *

Program Official Denis Buxton

Principal InvestigatorKaren L. Wooley

Dana R.Abendschein

Samuel I.Achilefu**

Joseph J. Ackerman

Carolyn J. Anderson

Jean M.Fréchet

Craig J. Hawker

Jason S.Lewis

Daniel P. Schuster

John-StephenTaylor

Robert J. Gropler**

Geoffrey E. Woodard, Ph.D.,is a fellow and our External Consultant from the National Institute of Allergy and Infec-tious Diseases of the National Institutes of Health. He gave a presentation on “Measurement and Significance of Natriuretic Peptides in Cardiovascular Disease”.

Guest Attendees Albert J. Sinusas, M.D.,of Yale University, School of Medicine, Professor of Medicine and Diagnostic Radiology. He is one of our four External Advisory Board members. Al shared his feedback both throughout the day as he asked questions, and again at the end of the day before the Wrap Up Session. We appreciated his input and time commit-ment to the PEN.

Jie Zheng**

Pamela K.Woodard

** Images provided by Focal Point Magazine and WUMS

Michael J. Welch**

14 Volume 1, Issue 3 Aug 2007

NHLBI Inter-PEN QuarterlyNHLBI Inter-PEN Quarterlywww.nhlbi-pen.net

Program Official

Denis Buxton

National Heart, Lung, and Blood Instituteemail: buxtond@nhlbi.nih.govhttp://www.nhlbi.nih.gov

Principal Investigators

Gang Bao

Emory UniversityGeorgia Institute of TechnologyEmail: gang.bao@bme.gatech.eduhttp://pen.bme.gatech.edu/

Jeffrey W. Smith

The Burnham InstituteThe Scripps Research InstituteUniversity of California, Santa BarbaraEmail: jsmith@burnham.orghttp://www.pennvp.org

Ralph Weissleder

Harvard UniversityMassachusetts General HospitalMassachusettes Institute of TechnologyBrigham and Women’s HospitalEmail: rweissleder@partners.orghttp://cmir.mgh.harvard.edu/nano/nano-tpen.

php?menuID_=223

Karen L. Wooley

Washington University in Saint LouisEmail: klwooley@wustl.eduwww.nhlbi-pen.info

Production Designer, Editor, Photographer

Eileen A. Cler

Web Designer and DeveloperProgram Coordinator for NHLBI-PEN GrantWashington University in Saint LouisEmail: eacler@wustl.edu

Principal Investigator, Jefffrey W. Smith, of The Burnham Institute for Medi-

cal Research, University of Califor-nia in Santa Barbara, will host the upcoming Second Annual Inter-PEN Meeting.

2ND ANNUAL INTER-PEN MEETINGS a n t a B a r b a r a , C a l i f o r n i a O c t o b e r 1 9 - 2 1 , 2 0 0 7

Location – The meeting will be held on the University of California - Santa Barbara campus in the Engineering Sciences Building, Room 1001. For a UCSB campus map, go to http://www.aw.id.ucsb.edu/maps/).

Accommodations – We have reserved a block of rooms at the Best Western South Coast Inn in Goleta, the small coastal community that surrounds the campus. Please mention the PEN conference when you make a reservation. The hotel is about three miles from the campus. UCSB will provide a shuttle service between the hotel and the UCSB campus.

Meeting Agenda Friday 6:00 p.m. Dinner - Elephant Bar on Firestone Road (less than a mile from the airport & the hotel). Informal gathering. This is an opportunity to meet others attending the conference in a low-key atmosphere. Saturday 8:30 a.m. Breakfast 9:00 a.m. Opening Remarks Presentations12:30 p.m. Lunch 1:30 p.m. Presentations (cont.) 5:30 p.m. Presentations conclude 6:00 p.m. Dinner (same location) 8:00 p.m. Poster session

Sunday 9:00 a.m. Executive Committee Meeting (Principal Investigators & Denis Buxton)11:00 a.m. Meeting concludes

By James Lee The Burnham Institutejameslee@burnham.org

Airport - The Santa Barbara Airport is adjacent to UCSB, and just a few miles from the hotel.