UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP FOR RESEARCH OCTOBER 18 - 21, 2013

An International Scientific Symposium focused on the latest advances in Cardiovascular, Cancer and Neurobiology Research

Symposium and associated grants have been generously supported by: D. Dan & Betty Kahn Foundation | Meredith and David Kaplan A. Alfred Taubman Medical Research Institute and other generous supporters of the U-M – Israel Partnership for Research

TABLE OF CONTENTS

SYMPOSIUM AGENDA 1-5

ABSTRACTS (IN ORDER OF PRESENTATION) SUNDAY Ami Aronheim 8 Omer Berenfeld 9 Peleg Hasson & Santhi Ganesh 10 Marcelle Machluf & Jérôme Kalifa 11 Ofer Binah & Daniel Michele 12 Doron Aronson 13 Oren Schuldiner 15 Izhak Kehat 16 Shelly Tzlil 17 Orit Kollet 18 Zaid Abbasi 19 Yuval Shaked 20 Eran Elinav 21 Ronen Rubinshtein 22 Orly Reiner 23 Ofer Fainaru 24 MONDAY Adam Stein 26 Henry Paulson 27 Mordechai Choder 28 Michal Schwartz 29 José Jalife 30 Gad Rennert 31 Theodore Lawrence 32 Eran Hornstein 33 Ivan Maillard 34 Ariel Roguin 35 Zvulun Elazar 36 Michael Aviram 37 Eva Feldman 38 Zvi Livneh 39 David Pinsky 40 Amir Landesberg 41 Berta Strulovici 43

CONTACT INFORMATION University of Michigan 46 Technion Institute of Technology 48 Weizmann Institute of Science 51

EXTRA PAGES FOR NOTE TAKING 54-58 SYMPOSIUM AGENDA

SUNDAY, OCTOBER 20, 2013 SAMUEL AND JEAN FRANKEL CARDIOVASCULAR CENTER MARVIN AND BETTY DANTO AUDITORIUM

8:00-8:30 am CONTINENTAL BREAKFAST

8:30-8:40 am WELCOME & OPENING REMARKS David Pinsky (UM) MODERATORS: Donna Martin (UM) / Hector Valdivia (UM)

8:42-9:52 am PRESENTATIONS 8:42-8:54 Ami Aronheim (Technion): Adult cardiac expression of the activating transcription factor 3, ATF3, promotes ventricular hypertrophy Omer Berenfeld (UM): Murine Models of Two Novel Gain-of-Function Mutations in Kcnj2 Reproduce Distinct Clinical Arrhythmias 8:56-9:08 Peleg Hasson (Technion) & Santhi Ganesh (UM): TGF-beta and lysyl oxidase interactions in arterial development and remodeling 9:10-9:22 Marcelle Machluf (Technion) & Jérôme Kalifa (UM): Site Specific Ultrasound Enabled Cardiac Anti Fibrotic Gene Therapy 9:24-9:36 Ofer Binah (Technion) & Daniel Michele (UM): Induced pluripotent stem cell derived cardiac myocytes for testing therapeutic approaches for dystrophic cardiomyopathy 9:36-9:50 Doron Aronson (Technion): Patients with acute myocardial infarction and severe obstructive coronary atherosclerosis display distinct peripheral blood gene expression profiles

9:52-10:07 am BREAK MODERATORS: Daniel Michele (UM) / Ofer Binah (Technion)

10:07-11:28 am PRESENTATIONS 10:07-10:16 Oren Schuldiner (Weizmann): Mechanisms of neuronal remodeling 10:18-10:30 Izhak Kehat (Technion): Mechanisms of cardiac concentric and eccentric hypertrophy 10:32-10:44 Shelly Tzlil (Technion): Cardiac Cell Mechanosensing

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 1 10:46-10:58 Orit Kollet (Weizmann): Regulation of hematopoietic stem cells and the bone marrow microenvironment 11:00-11:12 Zaid Abbasi (Technion): Aorto-caval fistula in rat: A unique model of volume-overload congestive heart failure and cardiac hypertrophy 11:14-11:28 Yuval Shaked (Technion): Host response to cancer therapy may promote tumor re-growth and metastasis spread

11:28 am-12:08 pm WORKING LUNCH BREAK MODERATORS: Santhi Ganesh (UM) / Peleg Hasson (Technion)

12:08-1:02 pm PRESENTATIONS 12:08-12:20 Eran Elinav (Weizmann): Neighborly relations: host microbiota interactions in health and disease 12:22-12:34 Ronen Rubinshtein (Technion): Usefulness of Coronary Computed Tomography Angiography for Prediction of Late Cardiovascular Outcome Events in Patients with a Chest Pain Syndrome 12:36-12:48 Orly Reiner (Weizmann): Unconventional roles of the complement pathway in the developing brain 12:50-1:02 Ofer Fainaru (Technion): Angiogenesis in cancer and placentation

1:02-1:15 pm CLOSING REMARKS Boaz Golany (Technion)

2 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP MONDAY, OCTOBER 21, 2013 A. ALFRED TAUBMAN BIOMEDICAL SCIENCE RESEARCH BUILDING D. DAN AND BETTY KAHN AUDITORIUM

7:30-8:00 am CONTINENTAL BREAKFAST IN SEMINAR BREAKOUT ROOMS, LEVEL 1

8:00-8:20 am WELCOME & OPENING REMARKS David Pinsky (UM) / Michael Aviram (Technion) / Zvi Livneh (Weizmann) MODERATORS: Dean Brenner (UM) / Gad Rennert (Technion)

8:20-9:56 am PRESENTATIONS 8:20-8:32 Adam Stein (UM): The PTIP containing histone methyltransferase complex is necessary for the development of compensated cardiac hypertrophy. 8:34-8:46 Henry Paulson (UM): Toward understanding and treating polyglutamine neurodegeneration 8:48-9:00 Mordechai Choder (Technion): Gene Expression Is A Circular System 9:02-9:14 Michal Schwartz (Weizmann): A paradigm shift in the perception of brain-immune system dialogue in neurodegenerative diseases and implications for therapeutic vaccination for ALS and Alzheimer’s disease 9:16-9:28 José Jalife (UM): Dominant Frequency Increase Rate Predicts Transition from Paroxysmal to Long-Term Persistent Atrial Fibrillation 9:30-9:42 Gad Rennert (Technion): Gene-environment interactions APC I1307K as model of high prevalence, low penetrance genetic variant of clinical importance 9:44-9:56 Theodore Lawrence (UM): Combining Targeted Therapies with Chemoradiation in the Treatment of Pancreatic Cancer

9:56-10:10 am BREAK MODERATORS: Moshe Talpaz (UM) / Yehoash Raphael (UM)

10:10 am-12:00 pm PRESENTATIONS 10:10-10:22 Eran Hornstein (Weizmann): Causes and consequences of dysregulated microRNAs in ALS

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 3 10:24-10:36 Ivan Maillard (UM): Blockade of individual Notch ligands and receptors controls graft-versus-host disease 10:37-10:49 Ariel Roguin (Technion): Cardiac Imaging and Radiation 10:51-11:03 Zvulun Elazar (Weizmann): Ubiquitin like proteins and early stages of autophagosome formation 11:05-11:17 Michael Aviram (Technion): HDL - associated Paraoxonase1 (PON1) attenuates Atherosclerosis development: PON1 role in Cholesterol Efflux stimulation 11:19-11:31 Eva Feldman (UM): Dysregulation of miRNAs in sporadic amyotrophic lateral sclerosis 11:33-11:45 Zvi Livneh (Weizmann): DNA repair – from molecular mechanisms to cancer prevention 11:47-11:59 David Pinsky (UM): Protection Against Vascular Injury by Enzymatic Dissipation of Intravascular Nucleotides

12:00-1:00 pm LUNCH MODERATORS: Omer Berenfeld (UM) / Ami Aronheim (Technion)

1:00-1:25 pm REMARKS Dean Amir Landsberg (Technion) Dean Yehuda G. Assaraf (Technion) Dean Pini Bar Yoseph (Technion)

1:25-4:50 pm BREAKOUT SESSIONS A. ALFRED TAUBMAN BIOMEDICAL SCIENCE RESEARCH BUILDING LEVEL 1: SEMINAR ROOMS A, B, C The seminar room will be set-up for small group discussion with seating for 6 to 8 attendees at each table. Session hosts will provide a topic introduction which will be followed by small group discussion. A list of conversation starters will be provided at each table for use as needed. 1:25-1:30 INTRODUCTION FOR BREAKOUT SESSIONS: David Pinsky (UM) 1:30-3:05 BREAKOUT SESSION I Latest /Cutting-edge Advances in Personalized Medicine: From Gene Discovery to Designer Drugs Introductory Presentation & Small Group Discussions

BREAKOUT SESSION I HOSTS: David Sherman (UM) Hans W. Vahlteich Professor of Medicinal Chemistry Professor of Medicinal Chemistry

4 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP David Sherman (UM) (cont.) Professor of Microbiology & Immunology Professor of Chemistry Professor of Life Sciences James Shayman (UM) Professor of Pharmacology Professor of Internal Medicine Alan Saltiel (UM) Mary Sue Coleman Director, Life Sciences Institute John Jacob Abel Professor in the Life Sciences Professor of Internal Medicine Molecular and Integrative Physiology Berta Strulovici (Weizmann) Director, National Center for Personalized Medicine Vicki Ellingrod (UM) John Gideon Searle Professor Professor of Pharmacy Shaomeng Wang (UM) Warner-Lambert/Parke-Davis Professor of Medicine Professor of Internal Medicine Professor of Pharmacology Professor of Medicinal Chemistry

3:05-3:20 pm BREAK MODERATORS: Jérôme Kalifa (UM) / Marcelle Machluf (Technion)

3:20-4:50 pm BREAKOUT SESSIONS 3:20-4:50 BREAKOUT SESSION II Biomedical Device Entrepreneurship From Medical Ideas to the Medical Market Introductory Presentation & Small Group Discussions

BREAKOUT SESSION II HOSTS: Eliezer Shalev (Technion) Dean, Rappaport Faculty of Medicine, Technion Ron Ferber Co-founder and President of HoMedics

4:50-5:00 pm CLOSING REMARKS Alan Saltiel (UM)

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 5

SUNDAY, OCTOBER 20 , 2013

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 7 AMI ARONHEIM, PhD

Adult cardiac expression of the activating transcription factor 3, ATF3, promotes ventricular hypertrophy

Lilach Koren1, Izhak Kehat2, Yuval Shaked3 and Ami Aronheim1

1Department of Molecular Genetics 2Department of Physiology 3Department of Pharmacology

The Rappaport Family Institute for Research in the Medical Sciences, Technion-Israel Institute of Technology, 1 Efron St. Bat-Galim, Haifa 31096, Israel

Cardiac hypertrophy is an adaptive response to various mechanophysical and pathophysiological stresses. However, when chronic stress is sustained, the beneficial response turns into a maladaptive process that eventually leads to heart failure. Considerable efforts are being directed towards developing mice models and understanding the processes that lead to cardiac hypertrophy. ATF3 is an immediate early transcription factor whose expression is induced in various cardiac stress models but has been reported to have conflicting functional significance in cardiac hypertrophy. To address this issue, we generated a transgenic mouse line with tetracycline-regulated ATF3 cardiac expression. These mice allowed us to study the effect of ATF3 on development versus pathogenesis of cardiac dysfunction. Importantly, ATF3 expression in adult mice resulted in rapid ventricles hypertrophy, heart dysfunction, and fibrosis. When combined with a phenylephrine (PE) infusion pressure overload model, the ATF3 expressing mice displayed a severe outcome and heart dysfunction. In a complementary approach, ATF3 KO mice displayed a lower level of heart hypertrophy in the same pressure overload model. Following two weeks of PE infusion, we failed to detect ATF3 expression in wild type. Interestingly, the lack of ATF3 in the bone marrow was able to fully recapitulate the lower hypertrophy observed in whole body ATF3 KO mice. To study the role of ATF3 expression in cardiomyocytes in vitro, Lilach Koren from my lab visited Prof. Daniel Michele laboratory at Ann Arbor to adopt adult cardiomyocytes cell culture technique. Currently, we are able to study ATF3 role in cardiomyocytes cell culture in vitro in the absence of blood pressure and bone marrow cells using isolated cardiomyocytes derived from various ATF3 mice models. In summary, ATF3 expression promotes cardiac hypertrophy while ATF3 deletion protects the heart. ATF3 expression in bone marrow cells plays a role in cardiac hypertrophy.

8 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP OMER BERENFELD, PhD

Murine Models of Two Novel Gain-of-Function Mutations in Kcnj2 Reproduce Distinct Clinical Arrhythmias

Hassan Musa, Lena Carlton, Matthew Klos, Karen Vikstrom, Justus Anumonwo, Jose Jalife and Omer Berenfeld

Center for Arrhythmia Research, University of Michigan

Background: Two distinct families have been discovered expressing missense mutations in the KCNJ2 gene encoding Kir2.1, which is responsible for the major component of the strong inward rectifier potassium channel IK1. One family presented with inherited atrial fibrillation (AF, KCNJ2V93I), and the other with short QT syndrome (SQT3) and an elevated risk of ventricular fibrillation (KCNJ2D172N). Both mutations result in a gain-of-function in IK1. We set to investigate the mechanisms leading the two apparently similar gains of function mutations into two distinct arrhythmia phenotypes.

Methods and Results: Two mouse lines with the targeted knock-in mutations KCNJ2V93I and KCNJ2D172N were generated. Kcnj2V93I/WT animals exhibited several different rhythm disturbances including brief episodes of AF characterized by the lack of a p-wave, variable R-R intervals and chaotic atrial activity. Kcnj2V93I/WT animals also displayed ectopic activity and complete A-V dissociation. Kcnj2D172N/WT animals did not elicit comparable arrhythmogenic events but quantitative analysis of the ECG waveforms revealed a trend to a shortened mean QT interval relative to the WT littermates, consistent with a SQT3 phenotype. Control animals given isoproterenol demonstrated a transient rise in heart rate but otherwise unremarkable ECGs. Whole cell patch clamp analysis of V93I mutation demonstrated a gain of function of IK1 and unitary channel events that exhibited channel sub-states not seen in WT controls.

Conclusions: Two clinically relevant mutations in KCNJ2 resulting in similar gain of IK1 are associated with two distinct arrhythmia phenotypes. Two knock-in murine models expressing these mutations have recapitulated the clinical phenotypes with Kcnj2V93I/WT mice displaying both atrial and ventricular arrhythmias and Kcnj2D172N/WT mice presenting with only a shortened QT interval.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 9 PELEG HASSON, PhD SANTHI GANESH, MD

TGF-beta and lysyl oxidase interactions in arterial development and remodeling

Vascular development and maintenance of the arterial wall are complex processes. Signal-mediated crosstalk between cell types and the extracellular matrix (ECM) are required to form a scaffold of proteins, such as collagens and elastins, to surround the cells and provide both structural integrity and elasticity to the artery. A cardinal signal involved in vascular development and remodeling is the inflammatory molecule transforming growth factor beta (TGFβ). TGFβ has been implicated in a number of vascular diseases. It is crucial for ECM deposition and organization, and it interacts with lysyl oxidase (LOX), an enzyme which initiates collagen and elastin cross-linking in the ECM. The precise interactions between TGFβ and LOX are unknown. Using ex vivo angiogenesis assays, we show that Lox chemical inhibition can attenuate angiogenesis. Remarkably, this inhibition is overcome by activation of the TGF pathway, supporting a role for crosstalk between TGF signaling and LOX activity in the vasculature. Notably, our preliminary genetic studies performed on dermal fibroblasts derived from patients with fibromuscular dysplasia (FMD), a disease characterized by both arterial stenosis and aneurysms, further demonstrate abnormal TGF and altered LOX expression strengthening our hypothesis that the crosstalk between the two plays cardinal roles in vascular development and remodeling. We propose to carry out further genetic studies and functional assays that will determine the cellular mechanisms underlying the interactions between Lox and TGF and that will shed light on the vascular diseases they participate in.

10 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP MARCELLE MACHLUF, PhD JÉRÔME KALIFA, MD, PhD

Site Specific Ultrasound Enabled Cardiac Anti Fibrotic Gene Therapy

T. Haber1, I. Goldfracht1, Uma Avula2, J. Kalifa2 and M. Machluf1

1Biotechnology and Food Engineering, Technion – Israel Institute of Technology, Haifa, Israel 32000. 2Center for Arrhythmia Research, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109-2800.

Background: Fibrosis is the response of the heart and other organs to injuries. In the heart, excessive fibrosis is a pathophysiological factor leading to pump dysfunction and heart failure and is a substrate for atrial and ventricular arrhythmias.

TGF-β is a pro-fibrotic cytokine and represents a valuable target for therapies aiming at decreasing fibrotic progression.

Ultrasound (US) is a non-viral approach used to deliver genes into cells and tissue. Non-viral vectors are emerging as substitutes to the viral ones since they are considered safer, easier to prepare, lack immunogenic response, and do not have a limit in the size of gene introduced.

Our general objective is to develop and implement means of regional in-vivo delivery of the anti-fibrotic gene that encodes for Decorin, a natural antagonist of TGF-β.

Methods: TUS-transfection in-vitro was assayed on sheep atrial and ventricular fibroblast cells. pGFP with and without ultrasound contrast agents (USCA) was used to study transfection efficiency. The effect of TUS on the viability of the cells was measured using the AlamarBlue assay. The effect of TUS on cells’ morphology and the effect on cells’ surface markers were also studied.

The Decorin gene was inserted into pSELECT-GFPZeo plasmid. Sheep cardiac-fibroblast cells were TUS- transfected with the plasmid encodes for Decorin and the transfection levels were studied.

Results: TUS of 2W/cm2, 20% DC operated for 20 minutes led to the highest transfection rates, when using 10 μg/ml of cDNA. Adding USCA resulted in higher transfection efficiency. TUS did not affect significantly the viability of the cells and did not alter the surface markers of the cells.

Conclusions: Our study demonstrates that TUS can efficiently transfect cardiac fibroblast cells while maintaining their viability. These results suggest that TUS-transfected cardiac cells with plasmid encodes for Decorin (anti-fibrotic gene) might be alternate or adjuvant to existing procedures.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 11 OFER BINAH, PhD DANIEL MICHELE, PhD

Induced pluripotent stem cell derived cardiac myocytes for testing therapeutic approaches for dystrophic cardiomyopathy

1Department of Physiology and Biophysics, Technion-Israel Institute of Technology 2Department of Molecular and Integrative Physiology, University of Michigan

The long term goal of this project is to utilize induced Pluripotent Stem Cells (iPSC) to produce patient specific cardiomyocytes for studying mechanisms and testing therapies for the cardiomyopathies associated with inherited muscular dystrophies. Muscular dystrophies encompass a diverse group of diseases resulting in severe progressive striated muscle weakness and wasting, loss of ambulation, and early death. Genetic disruption of genes required for the function of the dystrophin glycoprotein complex (DGC) collectively represent the most common causes of muscular dystrophy in humans. Within this group nearly 100% of patients will be diagnosed with cardiomyopathy, and heart failure is the cause death in 20-40% of cases. With the support of this research program, we had a primary goal of characterizing for the first time the expression and function of the DGC during differentiation of healthy iPSC-derived cells into cardiac myocytes (iPSC-CM) as a starting point to develop this model system for patient-mutation specific studies. We show here that healthy volunteer-derived iPSCs induce gene expression of several key adult striated muscle specific components of the DGC during differentiation into cardiomyocytes, while repressing components of the DGC present in non-muscle tissues and early muscle development. Cardiomyocyte differentiation also quantitatively induces the function of the DGC as an extracellular matrix receptor on the cell surface to levels approaching the function of the DGC in adult cardiomyocytes. This data provides strong support for an expanded next phase of this project which will be focused on deriving muscular dystrophy patient’s iPSC, characterizing the cellular phenotypes of dystrophic iPSC-CM, and utilizing these cells for screening therapeutic compounds. As outcomes, we expect to define important patient specific cellular phenotypes with clinical relevancy that provide important mechanistic insights into the disease process and identify novel targets for developing therapies for cardiomyopathy in muscular dystrophy patients.

12 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP DORON ARONSON, MD, FESC

Patients with acute myocardial infarction and severe obstructive coronary atherosclerosis display distinct peripheral blood gene expression profiles

Background: The fact that patients with severe multivessel coronary artery disease (MV-CAD) remain stable for years without developing acute coronary events, while others develop myocardial infarction (MI) as the first manifestation of CAD despite mild coronary atherosclerosis remains poorly understood. We hypothesized that these two distinctive phenotypes may be associated with divergent gene expression profiles in peripheral blood. Methods: Whole genome microarray analysis (Illumina) was performed on peripheral-blood mononuclear cells (PBMCs) in 3 groups: 1) patients with angiographic MV- CAD (≥70% stenosis ≥ 2 vessels) but without prior MI (n=9); 2) patients with ST-elevation MI and angiographic evidence of 1-vessel disease with plaque rupture (n=14); 3) subjects with normal coronaries (NC) (n=11).

Results: Analysis of differentially expressed genes (FDR<0.25, >1.3 Fold-change) demonstrated gene expression changes occurring predominantly within the MV-CAD vs. NC and MV-CAD vs. MI comparisons. These included genes encoding proteins previously found to be associated with atherosclerosis such as COX-2, EGR¬-1 and JUNB; pro-inflammatory cytokines (IL-1β, oncostatin M, visfatin), superoxide dismutase 2, and toll-like receptors (TLR4 and TLR6). Other differentially expressed genes included genes involve in the regulation of immune and inflammatory responses ZFP36( [also known as TTP or tristetraprolin], interleukin-1 receptor antagonist [IL1RN], BCL3, AIF1, S100A11, IL-15, NFIL3, TNFAIP6 and FPR2 [Formyl peptide receptor 2]). Several cell cycle and apoptosis-related genes were also differentially regulated including tumor necrosis factor (ligand) superfamily member 10 [TNFSF10, also known as TNF-related apoptosis inducing ligand, TRAIL], TNF receptor superfamily, member 10B (TNFRSF10B, also known as KILLER/DR5 or TRAILR2), and tumor necrosis factor receptor superfamily member 8 (TNFRSF8).

Pathway analysis identified a graded increase in enrichment in the inflammation, cell death and free radical-related pathways: the lowest degree of enrichment was observed when MI patients (mild atherosclerosis) were compared with NC. Higher enrichment was observed when MV CAD (severe atherosclerosis) was compared with MI patients (mild atherosclerosis), and the greatest enrichment when MV CAD patients were compared with NC, suggesting that several inflammatory pathways may be linked to the severity of atherosclerosis. Network analysis was used to identify interactions between the genes that were differentially regulated between the study groups. This revealed a network containing multiple inflammation and apoptosis-related genes, withIL-1β being the hub molecule in the highest scored pathway. In addition, gene expression interactome identified of the differentially expressed genes identified nuclear factor (NF)-κB as an upstream modulator of inflammatory transcripts upregulated in PBMCs of patients with MV-CAD.

Continued on page 14

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 13 Conclusion: Gene expression in PBMCs revealed systemic inflammation with stronger activation of inflammatory and apoptosis-related pathways in MV-CAD as compared with MI, reflecting the extent of CAD. Robust systemic inflammation is not inconsistent with long-term stability of severe obstructive CAD, suggesting that some inflammatory pathways may also play a role in plaque stabilization.

14 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP OREN SCHULDINER, PhD

Endocytosis promotes axon pruning by down-regulating inhibitory signals

Noa Issman-Zecharia and Oren Schuldiner

Remodeling of neuronal networks during development is widely conserved and is essential for mature connectivity in vertebrates and invertebrates. One mechanism of remodeling - axon pruning - involves the elimination of specific neuronal connections during development. Understanding the molecular mechanisms regulating axon pruning should provide a broad insight into the mechanisms of axon fragmentation during development, disease and after injury. The developmental remodeling of the Drosophila mushroom body (MB) γ neurons is a unique model to investigate these mechanisms due to its stereotypy and wide spectrum of genetic tools available. From a mosaic forward genetic screen, we found that UVRAG, a tumor suppressor gene, is cell autonomously required for the pruning of MB γ neurons. UVRAG can function as a subunit in the Phosphoinositide 3-kinase class III (PI3K-CIII) complex, previously shown to promote autophagy and/or endocytosis. Loss of function experiments determined that the entire PI3K-CIII complex is required for pruning by promoting endocytosis. Thus, we suggest that PI(3)P on endosomes recruits a PI(3)P binding protein(s) that in turn promotes pruning. Indeed, we found that Hrs, which contains an FYVE domain that binds to PI(3)P, is important for normal pruning progression. Hrs is a subunit of the ESCRT-0 complex, promoting the maturation of early endosomes into multi-vesicular bodies, suggesting that its role is to attenuate a signaling molecule that normally inhibits pruning. Indeed, we found that patched, the hedgehog receptor, is at least partially the mediator of this process. Its expression is upregulated in UVRAG mutant clones, its force expression results in an exacerbated phenotype and, most importantly, its mutation partially suppresses the pruning defects of PI3K-cIII mutants. Taken together, we suggest that during normal development patched signaling inhibits pruning and is downregulated by the endosomal-lysosomal pathway.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 15 IZHAK KEHAT, MD, PhD

Mechanisms of cardiac concentric and eccentric hypertrophy

Congestive heart failure is a worldwide epidemic. Heart failure develops mainly after myocardial infarction, chronic arterial hypertension, or diseases of the valves. Associated with all these different disease entities is a profound alteration of the heart shape and function, which is triggered by mechanical overload. Stimuli that lead to pressure overload like chronic arterial hypertension primarily induce concentric myocardial growth in which the heart-cells grow in width because new contractile units (sarcomeres) in the cells are added in parallel. In contrast, volume overload induces eccentric cardiac growth in which the heart cells grow in length because new contractile units in the cells are added in series. Sarcomere addition most likely restores the ability of the heart to generate forces in proper magnitude and rate. However, this remodeling usually have destructive outcome. We identified the protein kinase ERK as a very specific inducer of a cardiac concentric growth pattern. We used mice lacking all ERK1/2 protein in the heart (Erk1–/–;Erk2fl/fl-Cre) and mice expressing activated MEK1 in the heart to induce ERK1/2 signaling, as well as mechanistic experiments in cultured myocytes to assess cellular growth characteristics associated with this signaling pathway. While loss of all ERK1/2 from the heart did not block the cardiac hypertrophic response per se, meaning that the heart still increased in weight with both aging and pathologic stress stimulation, it did dramatically alter how the heart grew. For example, adult myocytes from hearts of Erk1–/–;Erk2fl/fl-Cre mice showed preferential eccentric growth (lengthening) while myocytes from MEK1 transgenic hearts showed concentric growth (width increase). Isolated adult myocytes acutely inhibited for ERK1/2 signaling by adenoviral gene transfer showed spontaneous lengthening while infection with an activated MEK1 adenovirus promoted constitutive ERK1/2 signaling and increased myocyte thickness. Thus, ERK signaling represents a nodal point in the concentric/eccentric growth decision.

16 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP SHELLY TZLIL, PhD

Cardiac Cell Mechanosensing

Interactions between cells and their surrounding matrix play defining roles in biological processes. It is clear by now that substrate mechanical properties strongly influence cell behavior. Furthermore, recent lines of evidence indicate that cells can respond to mechanical deformations generated by neighboring cells. The basis for this phenomenon and the role of mechanical communication between cells is unknown. In my talk, I will describe the progress made in our lab focusing on the role of cell mechanosensing in cardiac cell synchronized beating and in cardiac cell response to infarct scar tissue formation.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 17 ORIT KOLLET, PhD

Regulation of hematopoietic stem cells and the bone marrow microenvironment

Lapidot’s group focuses on regulation of hematopoietic stem cell (HSC) migration and development by the dynamic bone marrow (BM) microenvironment. HSCs replenish the blood and immune system with new cells throughout life. They dynamically change their features and location, shifting from quiescent and stationary cells anchored via adhesion interactions in their supporting BM niches, into cycling and motile cells which egress to the circulation. These dynamic changes are driven by stress signals, activation of proteolytic enzymes, cleavage of adhesion interactions, and generation of reactive oxygen species (ROS). ROS levels are dynamic. ROShigh are short term repopulating HSC and ROSlow are long term repopulating HSC. Low levels of ROS are maintained by the cytokines SCF, FGF-2 and PGE2 produced by Cox2 expressing monocytes/macrophages. High ROS accumulation, which is observed following injury, bleeding and infection; and also after clinical G-CSF-mediated stem cell mobilization, induce enhanced stem cell migration and differentiation, involving AKT, p38, HGF and S1P signaling. The chemokine CXCL12 (SDF-1) and its major receptor CXCR4 play a pivotal role in regulation of HSC function and this axis is prominent in our studies. HSC functionally express CXCR4 and are interacting with BM stromal cells expressing high levels of CXCL12 for their survival and maintenance. HSC follow CXCL12 signals to home into the BM or leave it and egress or mobilize to the circulation, which are controlled by signals coming from the nervous system. Current projects include deciphering the role of the coagulation system in regulation of HSC migration and development via thrombin/PAR-1 and EPCR shedding, or in maintaining stem cell adhesion to BM stromal cells via APC/EPCR and PAR-1. We also investigate the roles of the dynamic blood-BM endothelial barrier permeability and FGF-2 signaling in promoting hematopoietic, mesenchymal and leukemic stem cell proliferation and expansion, versus migration and differentiation.

18 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP ZAID ABBASI, MS, DSC

Aorto-caval fistula in rat: A unique model of volume- overload congestive heart failure and cardiac hypertrophy

Congestive heart failure (CHF) is the major cause of morbidity and mortality in the Western world, thus posing a major health and economic burden. Despite the continuous progress in our understanding of the pathogenesis of CHF and its management, the mortality remains high. Therefore, the development of reliable experimental models of CHF and cardiac hypertrophy are essential to better understand the progression of the disease and elaborate new therapy. One of the oldest models is the aorto-caval fistula (ACF), which was first described in dogs almost a century ago. More than 2 decades ago, our group adopted this experimental model in small animals including rats and mice. Although considered to be a model of high cardiac output heart failure, its long term manifestations are similar to those seen in patients with low cardiac output CHF. These include increased activity of vasoconstrictor/ anti-natriuretic neurohormonal systems such as: renin-angiotensin-aldosterone, sympathetic nervous, endothelin and arginine vasopressin on one hand, and activation of compensatory vasodilating/ natriuretic systems including atrial and brain natriuretic peptides and nitric oxide, on the other. Likewise, rats with ACF-induced CHF exhibit renal and cardiac manifestations similar to those reported in clinical CHF, including Na+ retention and cardiac hypertrophy. Previous data from our laboratory and other groups suggest that cardiac hypertrophy and urinary sodium excretion in this situation is largely determined by the balance these two opposing hormonal systems. In decompensated CHF, enhanced activities of the vasoconstrictive/Na+-retaining systems overwhelm the effects of vasodilatory/natriuretic systems, leading to a net reduction in Na+ and water excretion and severe cardiac hypertrophy. Based on that, this experimental model of CHF serves as a simple, cheap, and reproducible platform to investigate the pathogenesis of CHF and most importantly to examine the efficacy of new therapeutic approaches for management of clinical CHF.

Keywords: Animals models; Heart failure; Cardiac hypertrophy; Aortocaval fistula; Neurohormonal systems; Kidney function; Rat.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 19 YUVAL SHAKED, PhD

Host response to cancer therapy may promote tumor re-growth and metastasis spread

A major obstacle in clinical oncology is that tumors eventually acquire resistance to their therapy. The mechanisms by which resistance arises are usually associated with changes that occur in the tumor cells in response to the treatment. However, recent studies demonstrated that following almost any kind of treatment the host may also react to the therapy, and induce pro-tumorigenic and/or pro-metastatic effects which can lead to tumor re-growth, angiogenesis, metastases, and drug resistance. These host response effects can minimize or even negate the initial benefit of the anti-tumor activity achieved by the conventional therapy. Several pro-tumorigenic and/or pre-metastatic host factors have already been identified in the plasma of mice and patients following therapy. For example, metalloproteinase 9 (MMP9) is upregulated specifically in bone marrow derived cells which colonize treated tumors hence contributing to tumor cell dissemination from the primary tumor leading to metastasis spread; several pro-inflammatory key cytokines are overexpressed in the plasma of mice treated with chemotherapy and further promote tumor cell invasiveness by inducing macrophage skewing from M1 to M2 pro- metastatic phenotype; a variety of cell types located at distant sites react to the therapy by homing to the treated tumor site and promoting systemic angiogenesis. My laboratory focuses on identifying host pro- tumorigenic and pro-metastatic effects found following conventional cancer therapies and designing new treatment strategies to block them in order to improve currently used cancer treatments.

20 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP ERAN ELINAV, MD, PhD

Neighborly relations: host microbiota interactions in health and disease

The distal intestine contains tens of trillions of microbes, a community that is dominated by members of the domain Bacteria but also includes members of Archaea, Eukarya, and viruses. The vast repertoire of this microbiome functions in ways that benefit the host. The mucosal immune system co-evolves with the microbiota beginning at birth, acquiring the capacity to tolerate components of the community while maintaining the capacity to respond to invading pathogens. In the small intestine, Paneth cells sense bacterial products through receptors for microbe-associated molecular patterns (MAMPs), resulting in regulated production of bactericidal molecules, and leading to relative paucity of the microbiome in this region. The large intestinal regulatory counterpart remains largely unknown. It has been recently highlighted that dysregulation of the delicate balance between the microbiota community and the mucosal immune system have been linked to pathologies ranging from chronic inflammation, obesity and the metabolic syndrome. We have identified various possible mechanisms for the reciprocal regulation between the mucosal innate immune system and the intestinal microbial ecosystem, including inflammasomes and the newly recognized nod-like receptor (NLRs) family member NLRP6, which has been implicated in the pathogenesis of intestinal auto-inflammation, the metabolic syndrome and inflammation-associated colorectal cancer. Deciphering the mechanisms by which inflammasomes and NLRs regulate intestinal homeostasis and host-microbiota interactions has profound implications on our understanding of multi-factorial disease.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 21 RONEN RUBINSHTEIN, MD, FACC, FESC

Usefulness of Coronary Computed Tomography Angiography for Prediction of Late Cardiovascular Outcome Events in Patients with a Chest Pain Syndrome

Risk stratification plays a major role in the diagnosis and approach to management of patients with a chest pain syndrome which is possibly ischemic in origin. A normal or “low risk” non-invasive test by exercise treadmill testing, myocardial perfusion scintigraphy or stress echocardiography predicts a low cardiac event rate during follow-up. Contrast-enhanced multi-channel coronary computed tomographic angiography (CCTA) allows rapid scanning of the heart and great vessels, requires minimal patient co- operation and has high diagnostic accuracy for diagnosing anatomic coronary stenoses and coronary atheromatous plaques.

We have examined the predictive value of CCTA for risk stratification regarding later (2-8 years) major adverse cardiovascular events (MACE, cardiac death or myocardial infarction) in several patient populations undergoing CCTA as part of the investigation of a chest pain syndrome.

Among patients without previously known coronary artery disease (CAD), MACE occurred more frequently (>10%) (early 30 day CCTA-driven events excluded) in patients with ≥1 coronary luminal narrowing(s) of ≥70%, less frequently in patients with non obstructive atheromas and in less than 1% of patients with no coronary luminal narrowing. MACE was not increased in patients with the frequent anatomic variant of myocardial bridging (but without obstructive CAD). In patients with a history of coronary artery bypass surgery, MACE rate was increased (>7% / year) if the patient had > 2 obstructed grafts and especially if an arterial graft was occluded.

In conclusion, CCTA is useful to predict late clinical outcome events in a wide range of patient populations undergoing evaluation of a chest pain syndrome.

22 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP ORLY REINER, PhD

Molecular Mechanisms Regulating Neuronal Migration

The main interest of our lab is the process of neuronal migration that occurs during embryonic brain development. In the developing brain neurons are born in one position and have to migrate to their final destination by active cell migration. This is a very dynamic process that is regulated via the concerted action of multiple gene products. In humans this process occurs over the period of several months.

Aberrant neuronal migration may result in devastating consequences, such as severe brain malformation, mental retardation, epileptic seizures and early death. We have concentrated on one severe form of brain malformation, known as lissencephaly, which means “smooth brain”. Abnormal neuronal migration has been also associated with Schizophrenia and autism. Interestingly, the same gene products that are involved in regulation of neuronal migration participate in additional fundamental cell processes such as neuronal transport. Therefore, our studies have implications on neurodegenerative diseases as well.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 23 OFER FAINARU, MD, PhD

Angiogenesis in cancer and placentation

CD11b+Gr1+ cells are a heterogeneous population of hematopoietic cells that consist of immature myeloid cells (IMCs). In healthy individuals, IMCs that are generated in the bone marrow differentiate into mature granulocytes, macrophages or dendritic cells (DCs). In contrast, under pathological conditions, such as cancer, a block in differentiation of IMCs results in the expansion of this population. IMCs have been shown to actively promote tumor growth by modulating the cytokine environment and by promoting angiogenesis. Accumulating evidence suggest that the proliferative, invasive, and immune tolerance mechanisms that malignant tumors use to establish a nutrient supply and evade the immune response, originate from those used by the developing placenta. We have shown that ~30% of bone marrow derived cells infiltrating the mouse and human placenta are IMCs. This population showed striking similarity to that observed in tumors, in terms of their relative abundance, morphology and location adjacent to blood capillaries. Importantly, Matrigel plug assays and human endothelial tube formation assays indicated that IMCs derived from both placenta and tumor tissues are proangiogenic in vivo. We thus hypothesized that IMCs that populate mouse tumors and placentas share gene expression profiles to account for their similar phenotype and function. We analyzed gene expression profiles of placenta- and tumor derived CD11b+Gr1+IMCs using gene microarrays and qPCR. Our findings indicate that CD11b+Gr1+IMCs that infiltrate mouse placentas share approximately 500 highly expressed genes with those that infiltrate malignant tumors. This gene set is enriched with proangiogenic and inflammatory genes. Smaller, however unique gene expression sets for tumor IMCs and placenta IMCs were also detected. We thus suggest that similar molecular signatures of IMC genes derived from tumors and placentas indicate that these cells share similar origins and functions. Unique gene-sets that are differentially expressed in tumor vs. placenta derived IMCs, may be required for specific interactions between IMCs and the hosting tissue.

24 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP MONDAY, OCTOBER 21, 2013 ADAM STEIN, MD

The PTIP containing histone methyltransferase complex is necessary for the development of compensated cardiac hypertrophy

PTIP is a key component of a histone methyltransferase (HMT) complex that regulates H3K4me3 marks at the 5’ region of actively expressed genes. We hypothesized that the PTIP-HMT complex is necessary for the development of adaptive cardiac hypertrophy. Using a tamoxifen inducible cardiac specific Cre murine model and a floxed allele of PTIP, we generated inducible PTIP null mice (PTIP- ) and control mice (PTIP+). In order to test the importance of the PTIP-HMT in the development of cardiac hypertrophy, PTIP+ and PTIP- mice were subjected to 2wk of transverse aortic constriction (TAC) or sham surgery (Sham). PTIP- TAC and PTIP+ TAC hearts demonstrated similar amounts of cardiac hypertrophy as compared to PTIP+ Sham and PTIP- Sham hearts. H&E staining of ventricular cross sections revealed that PTIP-TAC hearts are dilated when compared to PTIP+TAC hearts. Echocardiography revealed that PTIP-TAC mice develop a significant increase in LVEDD and a significant decrease in ejection fraction when compared with PTIP- Sham and PTIP+TAC mice. Picrosirius red staining revealed that PTIP-TAC hearts have more cardiac fibrosis than PTIP+TAC hearts. In order to define the mechanism responsible for the maladaptive response in PTIP- hearts, ChIP- seq was performed to identify PTIP target genes. ChIP-seq and gene expression studies suggest that PTIP regulates the expression of HIF1A, MEF2A and ATP2A2. These genes are known to be important for the development of adaptive cardiac hypertrophy. Studies are being performed to confirm our ChIP-seq data and define the mechanism whereby PTIP- mice develop maladaptive heart failure.

26 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP HENRY PAULSON, MD, PhD

Toward understanding and treating polyglutamine neurodegeneration

Henry L. Paulson, MD, PhD Lucile Groff Professor of Neurology Director, Michigan Alzheimer’s disease Center Co-Director, Protein Folding Diseases Initiative

My lab explores why the brain degenerates in disorders caused by abnormal protein misfolding, with a major focus on the polyglutamine diseases including Spinocerebellar Ataxia type 3 (SCA3). We investigate why polyglutamine disease proteins are toxic to neurons and explore how the neuron’s protein quality control machinery counteracts this toxicity. To address these questions, we employ techniques ranging from recombinant protein analysis to engineered mouse models, with the long- term goal of understanding disease processes so that we can develop therapies for these currently untreatable disorders. I will briefly describe three recent advances in our lab. We created a novel knock- in mouse model for SCA3 that recapitulates important disease features and should shed light on disease pathogenesis. In translational studies, we developed cell-based assays with which we are identifying compounds that lower steady state levels of the SCA3 disease protein ATAXN3 or its oligomerization, with plans soon to begin testing promising compound in mouse models. Finally, we recently completed an in vivo trial of viral-mediated RNA interference as a potentially powerful, gene-selective therapeutic strategy for SCA3, underscoring the potential for nucleotide-based therapies in this, and other, dominantly inherited neurodegenerative diseases.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 27 MORDECHAI CHODER, PhD

Gene Expression is a Circular System

Choder, M.1, Haimovich, G.1, Bregman, A1., Harel-Sharvit, L.1, Eldad, N.1, Barkai, O.1, Duek, L.1, Goler-Baron, V.1, Seletrinnik, M.1, Lotan, R.1, Avraham-Kelbert, M.1, Suissa, L.1, Guterman, A.1, Garber, M.2, Causse, S.3, Darzacq, X.3, Millán-Zambrano, G.4, Chávez, S.4, Medina, D.5, and Pérez-Ortín J.E.5

1Department of Molecular Microbiology, Rappaport Faculty of Medicine, Technion – Israel Institute of Technology, Haifa, Israel. 2University of Massachusetts Medical School, 55 Lake Avenue North, Worcester MA 01655 USA. 3Functional Imaging of Transcription, Ecole Normale Supérieure, CNRS, IBENS, 46 rue d’Ulm 75230 Paris cedex 05, France. 4Departamento de Bioquímica y Biología Molecular, Facultad de Biológicas and ERI Biotecmed, Universitat de València, Dr Moliner 50, E-46100 Burjassot, Valencia, Spain. 5Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Avda. Reina Mercedes 6, E-41012 Sevilla, Spain.

Gene expression is traditionally viewed as a linear process divided into distinct stages (e.g., transcription, translation). We have shown that this view is oversimplified. First, RNA polymerase II controls mRNA translation and decay, via a mediator – Rpb4/7 (1-2). Second, many transcripts are “tagged” with factors co-transcriptionally, one of these tags is Rpb4/7, which later regulate the mRNA localization, translatability and decay. We name this tagging “mRNA imprinting”. Remarkably, promoters, DNA elements known to control only transcription, regulate also “mRNA imprinting”, thus affecting the mRNA fate in the cytoplasm (3). Third, we found that the major mRNA ‘decaysome”, known to degrade mRNAs in the cytoplasm, also functions as a transcription activator by physically associating with chromatin (4). Significantly, the capacity of the decaysome to function in the synthesis of a certain mRNA in the nucleus dependends on its ability to complete degrading this mRNA in the cytoplasm. Our findings demonstrate that gene expression is a circular process in which the hitherto first and last stages are interconnected. Finally, we propose that Rpb4/7 represents a novel class of factors, “mRNA coordinators”, which integrate all stages of the gene expression process into a system (2). Hence, the many decisions made during the expression of a given gene seem to be the result of coordination between all “distinct” stages.

1. Goler-Baron, V., et al. (2008). Genes & Dev. 22, 2022-2027. 2. Harel-Sharvit, et al. (2010) Cell 143, 552-563. 3. Bregman, et al (2011). Cell 147, 1473-83. 4. Haimovich et al. (2013). Cell 153, 1000-11.

See http://choder.net.technion.ac.il for references and further information

28 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP MICHAL SCHWARTZ, PhD

A paradigm shift in the perception of brain-immune system dialogue in neurodegenerative diseases and implications for therapeutic vaccination for ALS and Alzheimer’s disease

Neurodegenerative diseases are associated with neuronal loss occurring within the central nervous system (CNS), a compartment behind walls. Thus, it was assumed that these tissues are unable to benefit from the assistance provided by the immune system. Moreover, signs of inflammation accompanying brain pathology were believed to indicate immune cell infiltration from the circulation that should be mitigated. However, we have shown that both circulating blood macrophages and T lymphocytes are needed for recovery, and that these two cell types orchestrate neuroprotection and cell renewal. In healthy animals, we demonstrated that T cells recognizing CNS antigens reside in the CNS borders and are needed for the maintenance of brain plasticity, including cognitive ability, neurogenesis, and coping with stress as well as for facilitating recruitment of monocytes under injurious conditions through special portal gate, rather than due to a breached blood-brain-barrier. We found that such T cells, including effector CD4+ T cells and regulatory T cells (Treg), are part of a dynamic network that provides life-long brain maintenance and repair. These and other studies have led us to propose that neurodegenerative diseases may remain dormant long before their onset as long as the circulating immune cells can contain the pathology-induced deviation; disease onset indicates that either the deviation overrides the ability of the immune cells to counterbalance the rise of the risk factors, or that the immune system becomes exhausted and deteriorates concomitantly with the disease process. Recently, we identified the unique routes through which immune cells travel to infiltrate to the traumatized CNS; we showed that immune cell infiltration does not necessitate breakdown of barriers, but rather activation of a physiological route of entry. The cells enter through the choroid plexus epithelium, which we identified as a site that controls the fate of the infiltrating cells to enable a risk free reparative effect. We further showed in the healthy CNS, that the same site, the choroid plexus epithelium, hosts adaptive immune cells (T cells), which remotely control CNS plasticity; the activity of these cells can change from supportive to detrimental with aging. Lately, we found that in animal model of both ALS and Alzheimer’s disease, trafficking through the CP is suppressed relative to that of healthy wild type mice. To activate this “gate”, we vaccinated the mice with an immunogenic peptide derived fthat cross-react with CNS antigens. The vaccination elicited an immune response that induced the expression of trafficking molecules by the CP, enabling infiltration of leukocytes to the CSF, and initiated an active programmed immune response that culminated with recruitment of inflammation-resolving cells to the diseased parenchyma and contributed to disease arrest and even reverse. Taken together, our recent advances reveal a new therapeutic target for neurodegenerative diseases.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 29 JOSÉ JALIFE, MD

Dominant Frequency Increase Rate Predicts Transition from Paroxysmal to Long-Term Persistent Atrial Fibrillation

Raphael P. Martins, MD, Kuljeet Kaur, PhD, Elliot Hwang, BSc, Rafael J. Ramirez, PhD, B. Cicero Willis, MD, David Filgueiras-Rama, MD, Steven R. Ennis, PhD, Yoshio Takemoto, MD, PhD, Daniela Ponce- Balbuena, PhD, Manuel Zarzoso, PhD, Ryan O’Connell, PhD, Hassan Musa, PhD, Guadalupe Guerrero- Serna, PhD, Uma Mahesh R. Avula, MD, Michael F. Swartz, PhD, Sandesh Bhushal, BSc, Makarand Deo, PhD, Sandeep V. Pandit, PhD, Omer Berenfeld, PhD, José Jalife, MD

Center for Arrhythmia Research, Department of Internal Medicine/Cardiovascular Medicine, University of Michigan

Little is known about the mechanisms underlying the transition from paroxysmal to persistent atrial fibrillation (AF). In an ovine model of long-standing persistent AF (LS-PAF) we tested the hypothesis that the rate of electrical and/or structural remodeling, assessed by dominant frequency (DF) changes, determines the time at which AF becomes persistent.

Methods and Results: Self-sustained AF was induced by atrial tachypacing. Seven sheep were sacrificed after 11.5±2.3 days in self-sustained AF without reversal to sinus rhythm (SR); 7 sheep were sacrificed after 341.3±16.7 days of LS-PAF. Seven sham-operated animals were in SR for 1 year. DF was monitored continuously in each group. RT-PCR, western blotting, patch-clamping and histological analyses were used to determine changes in functional ion channel expression and structural remodeling. Atrial dilatation, mitral valve regurgitation, myocyte hypertrophy, and atrial fibrosis occurred progressively and became statistically significant after the transition to persistent AF, with no evidence for left ventricular dysfunction. DF increased progressively during the paroxysmal-to-persistent AF transition and stabilized when AF became persistent. Importantly, the rate of DF increase (dDF/dt) correlated strongly with the time to persistent AF. Significant action potential duration (APD) abbreviation, secondary to functional

ion channel protein expression changes (CaV1.2, NaV1.5 and KV4.2 decrease; Kir2.3 increase), was already present at the transition and persisted for one-year follow up.

Conclusions: In the sheep model of LS-PAF, the rate of DF acceleration predicts the time at which AF stabilizes and becomes persistent, reflecting changes in APD and densities of sodium, L-type calcium and inward rectifier currents.

30 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP GAD RENNERT, MD, PhD

Gene-environment interactions APC I1307K as model of high prevalence, low penetrance genetic variant of clinical importance

Gad Rennert, M.D., Ph.D. Clalit National Cancer Control Center (NICCC), Carmel Medical Center and Technion Faculty of Medicine

Background: Our genes interact with our exposures to influence the probability of developing cancer (initiation, promotion and progression). Identifying penetrant prevalent gene variants and understanding their interaction with the environment can lead to the development of personalized remedies for prevention, detection (screening) and treatment and influence the clinical outcome. TheAPC I1307K variant is common in Ashkenazi Jews leading to doubling of the risk of colorectal cancer (CRC). Data regarding risk factors, clinical behavior and prognosis, and means of their modification, can help form a paradigm for handling cancer affected and unaffected people with this and other SNPs.

Materials and Methods: The Molecular Epidemiology of colorectal cancer (MECC) population-based case-control study in Northern Israel is one of several such studies (breast, kung, ovary, endometrium, pancreas, liver with a total of 25,000 participants) conducted by NICCC with the aim of developing large cohorts with data on genetic variation and environmental/behavioral exposures and clinical outcomes. Data on diet, smoking, physical activity, use of medications, medical history, and family history of cancer are self-reported and validated. Blood and tissue samples are collected. All participants are followed up. Molecular investigations take the form of searching for germline and somatic point mutations employing a variety of techniques such as TaqMan, Sanger sequencing, MySeq NGS, GWAS and whole exome sequencing.

Results and conclusions: APC I1307K was detected in 514 of 9,472 participants in the MECC study; with a prevalence of 6% in healthy Ashkenazi Jews, and 12% of Ashkenazi cases, a risk magnitude of 1.9. Risk was found strongly negatively associated with consumption of vegetables, physical activity and use of common medications such as aspirin, NSAIDS, statins, and found to develop more in the right colon and have more synchronous tumors/polyps. These have major implication for tailoring specific prevention, detection and clinical handling protocols for these carriers.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 31 THEODORE LAWRENCE, MD, PhD

Combining Targeted Therapies with Chemoradiation in the Treatment of Pancreatic Cancer

Despite intensive research efforts, the median survival of patients with locally advanced pancreatic cancer (LAPC) remains approximately 1 year. Patients die of failure to control both the original tumor (1/3 of patients) as well as the development of metastases (2/3 of patients). We have taken 2 approaches toward improving outcome. The first involves conducting preclinical studies using both cultured cells and patient-derived xenografts to determine if inhibitors of the G2 checkpoint can increase the effectiveness of radiation therapy and gemcitabine, one of the most widely used drugs in the treatment of pancreatic cancer. Normal cells react to DNA damage by arresting to repair DNA damage in both G1 (prior to DNA synthesis) and G2 (prior to mitosis), whereas pancreatic cancer cells lack the G1 checkpoint. Therefore, treatment with radiation and gemcitabine plus a G2 checkpoint inhibitor should cause G1 arrest and DNA damage repair in normal cells, but progression into mitosis with damaged DNA, and subsequently, cell death, in pancreatic cancer cells. Our preclinical studies confirm that G2 checkpoint inhibitors selectively potentiate the anti-cancer effects of radiation (to improve local control) and gemcitabine (to improve systemic control). The second approach is aimed at improving the efficacy of radiation therapy by intensifying treatment. We have shown that radiation doses can be safely increased by using sophisticated radiation therapy techniques that limit the dose to the sensitive surrounding normal tissue and that this increased dose improves local control. Our newest study, which has just received approval from our Cancer Center Protocol Review Committee, will combine both the use of a G2 checkpoint inhibitor (Wee1 kinase inhibitor MK1775) and intensified radiation therapy, with the goal of improving local control, systemic control, and survival for patients with LAPC.

Supported by NCI grants R01 CA138723, P30 CA130810, P30 CA46592, and a Taubman Scholar Award

32 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP ERAN HORNSTEIN, MD, PhD

Causes and consequences of dysregulated microRNAs in ALS

Dysregulation of RNA–related processes emerge as novel and fundamental molecular component in the pathogenesis of ALS and in other neuromuscular diseases. We explored the mechanism and consequences of dysregulation in microRNA in several forms of ALS (TDP-43, FUS), by employing ALS patient specimen, molecular biology approaches and mouse genetics. Our data reveal that microRNA expression is globally downregulated in human ALS motoneurons and that impairment of Dicer– complex activity is a primary reason for loss of microRNA expression in ALS. Accordingly, loss of Dicer1 and microRNAs activity in mice resulted in neurodegeneration of spinal motoneurons and in denervation–dependent muscle atrophy. We further uncover the role of stress signaling in controlling Dicer and microRNA maturation by characterizing molecular pathways for control of microRNAs in stressed neurons. These data reveal that stress pathways regulate Dicer activity and suggest that a microRNA-based common molecular mechanism may be involved in several forms of ALS. The study is providing a framework for understanding how regulatory RNAs may be involved in the pathogenesis of neurodegeneration.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 33 IVAN MAILLARD, MD, PhD

Blockade of individual Notch ligands and receptors controls graft-versus-host disease

Ivy Tran, Ashley Sandy, Jooho Chung, Christen Ebens, Alexis Carulli, Ann Friedman, Ute Koch, Freddy Radtke, Pavan Reddy, Philip C. King, Linda Samuelson, Minhong Yan, Christian Siebel, Ivan Maillard

Life Sciences Institute; Departments of Medicine, Pediatrics, Physiology, Microbiology and Immunology, University of Michigan, Ann Arbor, MI; EPFL, Lausanne, Switzerland; Genentech, South San Francisco, CA

Graft-versus-host disease (GVHD) is a life-threatening complication of allogeneic bone marrow transplantation (allo-BMT) mediated by donor-derived T cells. Using a genetic approach, we discovered a major function for Notch signaling in alloreactive T cells mediating acute GVHD. Allo-BMT recipients of Notch-deprived T cells had decreased GVHD incidence and severity, leading to markedly improved survival. Notch inhibition blocked the production of inflammatory cytokines and tissue damage induced by alloreactive T cells, while preserving T cell expansion, cytotoxic functions and graft-versus-tumor effects. Thus, Notch inhibition led to beneficial immunomodulation after allo-BMT, without causing global immunosuppression. To define safe and effective approaches to target Notch signaling in allo-BMT recipients, we identified Notch1/2 as the key Notch receptors and Delta-like1/4 (Dll1/4) as the key Notch ligands in GVHD, with dominant roles for Notch1 and Dll4. Systemic Dll1/4 blockade with humanized monoclonal antibodies inhibited Notch signaling in alloreactive T cells and efficiently prevented GVHD. Transient Dll1/4 inhibition was sufficient to confer long-term protection. Unlike gamma-secretase inhibitors, anti-Dll1/4 antibodies were well tolerated after allo-BMT. Our observations identify Dll1/4 Notch ligands as an attractive new therapeutic target after allo-BMT. We are currently investigating the molecular mechanisms of Notch action in alloreactive T cells, the cellular source of Notch ligands and the role of Notch signaling in other T cell-mediated immune disorders, including organ rejection and autoimmunity. In addition, we are exploring preclinical strategies of Notch inhibition to develop Notch- based therapeutics for human patients.

34 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP ARIEL ROGUIN, MD, PhD

Cardiac Imaging and Radiation

The Rambam Interventional Cardiology Division is one of the leading centers in Israel. We perform more than 3500 procedures each year which include coronary interventions, TAVI procedures, carotid artery stenting, Mitral valvuloplasties, Endomyocardial biopsies and hemodynamic studies. We have a busy 24/7 Primary PCI service for acute MI patients. We also perform congenital heart disease interventions [PFA and ASD closure] and electrophsiological studies and device implantation.

We were involved in MRI safety in patients with implantable pacemakers and ICDs, and published extensively in this area. We are the national referral center for MRI of device patients and several of the protocols used today worldwide were developed here.

We are involved in numerous clinical and basic research activities. The basic research areas carried out in collaboration with the Technion Faculty of Medicine are in the areas of angiogenesis, collateral formation and repair of damaged vascular vessels. We studied genetic markers for restenosis as VEGF, HIF and Haptoglobin.

Our research in the field of PCI were intended to answer some of the key questions in this field. We conducted several of the pilot studies on nitinol self-expanding coronary stents and balloon expandable stents. We published on small vessel stenting, staged Vs non-staged angioplasty, intracoronary radiation, the interaction between stent design and results, and recently on robotic assistance in PCI and safety of live case demonstrations.

We were one of the pioneers in multislice CT imaging and published on its accuracy and ease of use as well as the importance of a Cardiologist in the process. Furthermore we studied the use of SPECT/CT and its use adjunctive to IVUS or as a worldwide novel application, as a co-registration in the cath lab.

The topic of radiation is a major concern. Occupational radiation exposure, and the orthopedic complica¬tions from wearing the heavy leaded aprons necessary to limit such exposure risk, has become a major concern among physicians per¬forming interventional procedures. We conducted a large trial on reducing radiation exposure in our working area - the cath lab.

We recently reported on brain cancer among interventional cardiologists. As of October 2013, our series includes 34 physicians with brain and neck malignancies: 26 interventional cardiologists, two electro¬physiologists and six interventional radiologists. All worked for prolonged periods (latency period 12-32 years, mean 23.5±5.9 years) in active interventional practice with exposure to ionizing radiation in the catheterization laboratory. Tumors included 17 (52%) cases of gliobalstoma multifome (GBM), two (7%) astrocytomas and five (16%) meningiomas. The malignancy was left-sided [the area with more radiation] in 88% cases.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 35 ZVULUN ELAZAR, PhD

Autophagy in health and disease

Department of Biological Chemistry, The Weizmann Institute of Science

Autophagy is an evolutionarily-conserved catabolic process initiated by the engulfment of cytosolic components in a crescent-shaped structure, called phagophore that expands and fuses to form a closed double-membrane vesicle, the autophagosome. Autophagosomes are subsequently targeted to the lysosome/vacuole with which they fuse to degrade their content. The formation of autophagosome is carried out by a set of autophagy-related (Atg) proteins, highly conserved form yeast to mammals. The Atg8s are ubiquitin-like (Ubl) proteins that play an essential role in autophagosome biogenesis. This family of proteins comprises a single member in yeast and several mammalian homologues grouped into three subfamilies: LC3, GABARAP and GATE-16. The Atg8s are synthesized as cytosolic precursors but can undergo a series of post-translational modifications leading to their tight association with autophagosomal structures upon autophagy induction. Autophagy dysfunction has been implicated in a group of progressive neurodegenerative diseases, and has been reported to play a major role in the pathogenesis of these disorders. We have recently reported a recessive mutation in TECPR2, an autophagy-implicated WD repeat-containing protein, in five individuals with a novel form of monogenic hereditary spastic paraparesis (HSP). We found that diseased skin fibroblasts had a decreased accumulation of the autophagy-initiation protein MAP1LC3B (LC3), and an attenuated delivery of both LC3 and the cargo-recruiting protein SQSTM1 (p62) to lysosomal degradation. The discovered TECPR2 mutation reveals for the first time a role for aberrant autophagy in a major class of Mendelian neurodegenerative diseases, and suggests mechanisms by which impaired autophagy may impinge on a broader scope of neurodegeneration.

36 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP MICHAEL AVIRAM, DSC

HDL - associated Paraoxonase1 (PON1) attenuates Atherosclerosis development: PON1 role in Cholesterol Efflux stimulation

Michael Aviram, The Lipid Research Laboratory, Technion Rappaport Faculty of Medicine and Rambam Medical Center, Haifa, Israel

Macrophage foam cells, the hallmark of early atherogenesis, are filled with oxidized lipids which stimulate cholesterol influx and biosynthesis rate, and inhibit HDL – mediated cholesterol efflux from the cells. HDL – associated Paraoxonase 1 (PON1) possesses hydrolyzing activities against atherogenic oxidized lipids, and these activities are related to PON1 histidine (115 &134) diad active site. We have sown that serum PON1 activities are inversely related to atherosclerosis development as studied in macrophage cell lines , as well as in PON1 – manipulated (knockout and over - expressed) mice. PON1 increased HDL binding to macrophages and consequently, stimulated cholesterol efflux from these cells. PON1 stimulation of HDL – mediated cholesterol efflux was found to involve the ABCA1, but not the SRB1 transporter on macrophage. Upon injection of PON1 to mice, monocyte – to – macrophage differentiation was inhibited and macrophage foam cell formation was substantially attenuated, in parallel to anti atherogenic changes that were observed in cholesterol fluxes, and mainly - cholesterol efflux stimulation, by up to 65%.

Next, we questioned PON1 regulation and since PON1 is inactivated by oxidative stress we used a most potent antioxidant – the pomegranate - derived polyphenol hydrolyzable tannin – punicalagin, and showed ( in vitro and in vivo) that PON1 activity and stability (PON1 - HDL association), significantly increased by punicalagin.

We thus conclude that PON1 is a potent HDL – associated antioxidant, and anti atherosclerotic hydrolase due to its ability to breakdown specific oxidized lipids in oxidized lipoproteins, in arterial macrophages, and in the atherosclerotic lesion. Up regulation of PON1 by hypo cholesterolemic and anti - oxidants means, could be the target for atherosclerosis development attenuation.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 37 EVA FELDMAN, MD, PhD, FAAN, FANA

Dysregulation of miRNAs in sporadic amyotrophic lateral sclerosis

Claudia Figueroa-Romero1, Junguk Hur1, Diane E. Bender1, Michael D. Cataldo1, Guillermo Flores1, Andrea L. Smith1, Raymond Yung2, Brian C. Callaghan1, and Eva L. Feldman1

Department of Neurology1 and Department of Internal Medicine2, University of Michigan, Ann Arbor, MI, USA

Amyotrophic lateral sclerosis (ALS) is a progressive, lethal, late-onset neurodegenerative disorder. The majority (90-95%) of cases are sporadic (sALS) with unknown causes. Previous work on postmortem spinal cord tissue identified several differentially expressed genes (DEGs) related to sALS; however, the molecular mechanisms responsible for these variations are mostly unknown. Recent evidence over the past decade associating increased sALS risk with long-term adverse environmental exposures suggests that epigenetic mechanisms, such as methylation and microRNAs (miRNAs), may play a role in neurodegeneration. miRNAs are small, non-coding RNAs that negatively regulate the expression of multiple genes; therefore, altered expression of only a few miRNAs can impact many genes and promote aberrant effects on multiple biological pathways and lead to disease. Our goal is to address the role of miRNAs in the pathogenesis of sALS. We hypothesize that differential expression of miRNAs in human sALS spinal cord promotes dysregulation of key genes and biological pathways leading to sALS. Systems biology analyses combining TaqMan OpenArray and Affymetrix GeneChip Human Genome U133 Plus 2.0 on postmortem human spinal cord tissue identified 90 differentially expressed miRNAs and 1,182 DEGs. Since miRNAs negatively regulate gene expression, we identified inverse correlations between differentially expressed miRNAs and mRNAs. Potential biologically relevant miRNA/DEG target pairs were obtained by compiling and assessing multiple miRNA prediction target databases. Gene and miRNA expression was confirmed by qPCR. Our analysis identified aberrant down-regulation of miR- 577, members of the let-7 family of miRNAs, miR-133b, miR-148-5p, and miR-140b-3p as potential regulators of genes previously implicated in neuronal homeostasis and/or the pathogenesis of ALS. Experimental validation of miRNA-DEG pairs is underway. We contend that changes in miRNAs and their corresponding targets may, in part, represent pathogenic epigenetic mechanisms leading to sALS.

Funding support: NIEHS P30 Pilot Grant U035448; Katherine Rayner Fund; A. Alfred Taubman Medical Research Institute

38 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP ZVI LIVNEH, PhD

DNA repair – from molecular mechanisms to cancer prevention

Zvi Livneh, Omer Ziv, Isadora Cohen, Ilan Vered, Umakanta Swain, Nataly Mirlas-Neisberg, Dana Davidovsky, Yael Leitner-Dagan and Tamar Paz-Elizur

Dept. of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel

DNA repair is a major mechanism for eliminating DNA damage, preventing mutation, and ensuring genomic stability. Under condition where DNA damage cannot be accurately repaired, e.g., when present on a single-stranded DNA during replication, cells utilize DNA damage tolerance mechanisms. Two such mechanisms are known: translesion DNA synthesis (TLS; also termed error-prone repair) and homology-dependent repair (HDR; also termed postreplication repair or template switch repair). We are studying the molecular mechanism of TLS in mammalian cells using assay systems based on site-specific DNA damages in plasmid substrates, or in the chromosomal context. We have recently demonstrated that both HDR and TLS operate in human cells (PNAS USA 2013, 110:E1462-9), that TLS occurs primarily at the G2 phase of the cell cycle (Nucleic Acids Res. 2012, 40:170-80), and the existence of a TLS pathway which is independent of PCNA-monoubiquitination (PLoS Genetics 2011 Sep;7(9):e1002262). We are currently investigating the effect of nuclear architecture on TLS, and a series of novel TLS genes that we have identified using an siRNA-based screen.

An additional direction in our lab is harnessing molecular mechanisms of DNA repair for cancer risk assessment, prevention and early detection. We focus on functional assays of DNA repair enzymes that act on oxidative DNA damage. We have identified three DNA repair risk factors for lung cancer: Low activity of OGG1 (J Natl Cancer Inst. 2003, 95:1312-9), high activity of MPG (J Natl Cancer Inst. 2012, 104:1765-9) and low activity of APE1 (submitted) (in collaboration with Gad Rennert, Technion, and Laurence Freedman, Gertner Institute). An integrated DNA repair score, composed of the three risk factors, was strongly associated with lung cancer risk (submitted). Ongoing experiments are directed towards expanding the panel of risk factors and examining various types of cancer, with the goal of applying DNA repair-based cancer prevention to public health.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 39 DAVID J. PINSKY, MD

Protection Against Vascular Injury by Enzymatic Dissipation of Intravascular Nucleotides

David J. Pinsky, Nadia R. Sutton, Yogen Kanthi, Scott H. Visovatti, Dana Petrovic-Djergovic, C. Anuli Anyanwu, Amy Baek, Sascha Goonewardena, Matthew C. Hyman, and Hui Liao

University of Michigan Health Systems, Ann Arbor, Michigan

Inflammation and coagulation are triggered in settings of vascular injury, ranging from blood flow interruption to atherosclerosis. To maintain the essential homeostatic intravascular milieu, specific enzymes have evolved on the surface of vascular cells to dissipate nucleotide tri-, di-, and mono- phosphates. These typically intracellular purinergic nucleotides are released by activated endothelium and dead or dying parenchymal cells into the intravascular environment, where they can act in paracrine fashion on nearby cells to signal danger. Chief among the enzymes which tonically degrade intravascular nucleotides are CD39 and CD73, which decorate the surface of vascular endothelium as well as leukocytes. These ectoenzymes, so-called because their catalytically-active sites face the cell exterior and hence, are active at the endovascular lumen, sequentially phosphohydrolyze ATP, ADP, and AMP to form the nucleoside adenosine. We have shown that these ectoenzymes can be detected on microparticles liberated into the bloodstream of patients with pulmonary vascular disease, where they may signal danger to cells in the vicinity of an injured vessel. Our laboratory has focused on the role of CD39 on the leukocyte surface as an endogenous regulator of leukocyte trafficking and platelet- driven thrombosis, in the setting of both arterial and venous disease. CD39 serves as an important endogenous protective mechanism against the development of atherosclerosis in a hypercholesterolemic environment, with expression levels regulated by laminar shear. CD73, which cleaves AMP to adenosine, is also important in maintaining immunologic equipoise following vascular injury. The absence of either ectoenzyme results in an increase in ischemic injury in the setting of middle cerebral arterial occlusion. Reconstitution of genetically altered mice which lack either CD39 or CD73 protects them against ischemic cerebral injury. This presentation will review work of our laboratory focused on mechanisms by which these ectoenzymes act naturally to protect the vessel wall from inflammatory and coagulation events, with the hope of harnessing their properties to protect against arterial, venous, and pulmonary vascular diseases.

40 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP AMIR LANDESBERG, MD, DSC

Modulation of the actomyosin Cross-Bridge Kinetics by Sarcomere Velocity; A New View on Muscle Dynamics, Clinical Implications and Technological Applications

Explanations for the basic skeletal and cardiac muscle characteristics, such as the force–velocity relation, the Fenn effect and the high contractile efficiency, are still vague. These properties are dominated by the actomyosin cross-bridge (XB) dynamics. Numerous models have been suggested to describe the XB dynamics and its dependence on the mechanical loading conditions. These studies suggest that XB dynamics is plausibly dependent on one of three variables: displacement, load or velocity. Theories of displacement-dependent XB kinetics stem from Huxley’s classic theory of muscle contraction. Load- dependent motor kinetics was suggested by recent studies of single molecular motors. However, there is a gap between our detailed knowledge of isolated single-molecule assays and the sarcomere dynamics and energetics. A third hypothesis suggests that the XB kinetics are simple functions of filament sliding velocity.

We have scrutinized the three putative theories mentioned (displacement, velocity and the load dependent kinetics) by testing the effects of both shortening and lengthening on the generated force and stiffness, in isolated trabeculae from rat right ventricles. Sarcomere length was measured by laser diffraction. Changes in the number of strong XBs (NXB) were evaluated by measuring the dynamic stiffness. Ramp stretches and releases at different velocities and onset times were imposed over isometric sarcomere contractions. Interestingly, stretches yielded parallel increases in force and stiffness at all stretch velocities. This observation is incongruent with the classical displacement dependent kinetics. The force per XB during stretch was constant, independent of the velocity, and equal to the isometric force per XB. The latter observation is incongruent with a load dependent kinetics. The difference in stiffness between ramp perturbations and isometric regime, normalized by the instantaneous isometric stiffness, (the normalized stiffness difference) was a linear function of velocity and ramp duration; Moreover, the normalized stiffness difference was independent of ramp onset time; Normalized stiffness development rate was linearly dependent on the lengthening velocity, with a slope of 6.73±0.98 µm-1. During shortening, the normalized stiffness decline rate depended linearly on the shortening velocity, with an almost identical slope of -6.70±1.43 µm-1. The revealed symmetrical dependence of the normalized stiffness development rate on the filament sliding velocity, for both stretches and releases, is conveniently explained only by the velocity-dependent hypothesis, a mechanism that also explains the muscle high contractile efficiency. By exclusion, the velocity dependent XB kinetics remains the plausible explanation for the velocity-dependent, and onset time-independent rates of change in normalized stiffness during ramp stretches and releases. This body of evidence strongly supports the theory that XB turnover rate from strong to weak conformation (weakening rate) is a linear function of filament sliding velocity. Stretch increases the force and stiffness (and decreases energy consumption) by decreasing the XB weakening rate, whereas shortening symmetrically increases the XB weakening rate, at the same velocity dependent manner.

Continued on page 42

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 41 The velocity-dependent kinetics provides analytical expressions for the Hill’s force–velocity relationship, muscle efficiency, economy and power. The suggested theory elucidates the relationship between oxygen utilization and mechanical energy output observed in both isolated fibers and whole hearts and enlightens the riddle of the high and load-independent contractile efficiency of the cardiac muscle. This theory can explain the function and energetics of an inhomogenous myocardium with myocardial infarction. This new insight can be used for optimal planning of cardiac resynchronization therapy and for the development of ventricular assist device that works in cadence with the failing heart.

42 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP BERTA STRULOVICI, PhD

Israel National Center for Personalized Medicine (INCPM): An Integrated -Omics approach to basic and clinical research

Berta Strulovici, Ph.D. Director, INCPM

INCPM aims to build an infrastructure of state-of-the-art tools and technologies in Genomics, Computational biology, Proteomics and Drug Discovery (Chemical Genomics) and to provide access to academic and clinical researchers to further basic and medical research in Israel. Our vision is that the INCPM activities will create interdisciplinary research opportunities such as that between basic, pharmaceutical and clinical research, will promote many collaborations between research labs at Israeli campuses, and eventually will contribute to the further advancement of scientific technology in Israel through collaborations with universities, other research institutes and hospitals. The Center is being built as a scalable organization, located in temporary facilities within The Weizmann Institute, looking forward to moving into its dedicated 11, 345 square feet space by Jan. 2015. The uniqueness of INCPM is in the integration of highly enabling technological platforms to advance biomedical research in Israel. We aim, after 3 years of operation, to provide academic and clinical researchers data that will uncover extensive, dynamic changes in diverse molecular components and biological pathways across healthy and diseased conditions, reveal medical risks and enable a potentially more precise, more effective, more experimental and personalized approach to medical treatment. Specific examples will be provided describing collaborations between academic and clinical researchers in Israel with our young Center.

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 43

CONTACT INFORMATION UNIVERSITY OF MICHIGAN CONTACT INFORMATION

UNIVERSITY OF MICHIGAN, CARDIOVASCULAR CENTER 1500 EAST MEDICAL CENTER DRIVE, ANN ARBOR, MI 48109

NAME AND TITLE EMAIL ADDRESS Omer Berenfeld, PhD [email protected] Associate Professor of Internal Medicine Associate Professor of Biomedical Engineering Dean Brenner, MD [email protected] Kutsche Memorial Chair of Internal Medicine Professor of Internal Medicine Professor of Pharmacology Vicki Ellingrod, Pharm D, BCPP [email protected] John Gideon Searle Professor Professor of Pharmacy Professor of Psychiatry Eva L. Feldman, MD, PhD, FAAN, FANA [email protected] Russell N. DeJong Professor of Neurology Santhi Ganesh, MD [email protected] Assistant Professor of Internal Medicine Assistant Professor of Human Genetics José Jalife, MD [email protected] Cyrus and Jane Farrehi Professor of Cardiovascular Research Professor of Internal Medicine Professor of Molecular and Integrative Physiology Co-Director, Center for Arrhythmia Research Jérôme Kalifa, MD, PhD [email protected] Assistant Professor of Internal Medicine Theodore Lawrence, MD [email protected] Isadore Lampe Collegiate Professor of Radiation Oncology Ivan Maillard, MD, PhD [email protected] Associate Professor of Life Sciences Assistant Professor of Internal Medicine Assistant Professor of Cell and Developmental Biology

(CONTINUED ON PAGE 47)

46 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP U OF M CONTACT INFORMATION (CONT.) NAME AND TITLE EMAIL ADDRESS Donna Martin, MD, PhD [email protected] Donna B. Sullivan MD Research Professor of Pediatrics and Communicable Diseases Associate Professor of Pediatrics and Communicable Diseases Associate Professor of Human Genetics Daniel Michele, PhD [email protected] Associate Professor of Molecular and Integrative Physiology Associate Professor of Internal Medicine Henry Paulson, MD, PhD [email protected] Lucile Groff Chair of Neurology for Alzheimers Disease and Related Disorders Professor of Neurology David J. Pinsky, MD [email protected] J. Griswold Ruth MD & Margery Hopkins Ruth Professor of Internal Medicine Professor of Molecular and Integrative Physiology Senior Taubman Scholar Chief, Cardiovascular Medicine Director, Samuel and Jean Frankel Cardiovascular Center Yehoash Raphael, MS, PhD [email protected] R. Jamison and Betty J. Williams Professor of Otolaryngology-Head and Neck Surgery Alan Saltiel, PhD [email protected] Mary Sue Coleman Director, Life Sciences Institute John Jacob Abel Professor in the Life Sciences Professor of Internal Medicine Molecular and Integrative Physiology James Shayman, MD [email protected] Professor of Pharmacology Professor of Internal Medicine David Sherman, PhD [email protected] Hans W. Vahlteich Professor of Medicinal Chemistry Professor of Medicinal Chemistry Professor of Microbiology & Immunology Professor of Chemistry Professor of Life Sciences

(CONTINUED ON PAGE 48)

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 47 U OF M CONTACT INFORMATION (CONT.) NAME AND TITLE EMAIL ADDRESS Adam Stein, MD [email protected] Assistant Professor of Internal Medicine Moshe Talpaz, MD [email protected] Alexander J. Trotman Professor of Leukemia Research Professor of Internal Medicine Hector Valdivia, MD, PhD [email protected] Frank Norman Wilson Professor of Cardiovascular Medicine Professor of Internal Medicine Professor of Molecular and Integrative Physiology Co-Director, Center for Arrhythmia Research Shaomeng Wang, PhD [email protected] Warner-Lambert/Parke-Davis Professor of Medicine Professor of Internal Medicine Professor of Pharmacology Professor of Medicinal Chemistry

TECHNION INSTITUTE OF TECHNOLOGY CONTACT INFORMATION

TECHNION-ISRAEL INSTITUTE OF TECHNOLOGY THE RUTH AND BRUCE RAPPAPORT FACULTY OF MEDICINE 1 EFRON STREET, BAT-GALIM, P.O. BOX 9649 HAIFA 31096 ISRAEL

NAME AND TITLE EMAIL ADDRESS Michael Aviram, DSc [email protected] Professor of Biochemistry Head, The Lipid Research Laboratory Tanenbaum Chair in Preventive Medicine Technion Rappaport Faculty of Medicine and Rambam Medical Center Zaid Abassi, DSc [email protected] Department of Physiology, Technion Rappaport Faculty of Medicine and Rambam Medical Center

(CONTINUED ON PAGE 49)

48 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP TECHNION CONTACT INFORMATION (CONT.) NAME AND TITLE EMAIL ADDRESS Ami Aronheim, PhD [email protected] Associate Professor Department of Molecular Genetics Technion Rappaport Faculty of Medicine Doron Aronson, MD, FESC [email protected] Associate Professor of Medicine (Cardiology) [email protected] Technion Rappaport Faculty of Medicine Director, Intensive Care Unit, Division of Cardiology Rambam Health Care Campus Yehuda Assaraf, PhD [email protected] Dean, Technion Faculty of Biology Pini Bar Yoseph, DSc [email protected] Dean, Technion Faculty of Mechanical Engineering Ofer Binah, PhD [email protected] Professor of Physiology Head, The Cardiac Research Laboratory Henry Taub Chair in Life Sciences Department of Physiology Technion Rappaport Faculty of Medicine Mordechai Choder, PhD [email protected] Technion Rappaport Faculty of Medicine Ofer Fainaru, MD, PhD [email protected] Technion Rappaport Faculty of Medicine Hillel Yaffe Medical Center in Hadera Boaz Golany, DSc [email protected] Technion Vice President for External Relations and Resources Development Peleg Hasson, PhD [email protected] Technion Rappaport Faculty of Medicine Izhak Kehat, MD, PhD [email protected] Assistant Professor, Department of Physiology Technion Rappaport Faculty of Medicine Amir Landbserg, MD, DSc [email protected] Dean, Technion Faculty of Biomedical Engineering

(CONTINUED ON PAGE 50)

2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 49 TECHNION CONTACT INFORMATION (CONT.) NAME AND TITLE EMAIL ADDRESS Marcelle Machluf, PhD [email protected] Associate Professor and Vice-Dean, Technion Faculty of Biotechnology and Food Engineering Gad Rennert, MD, PhD [email protected] Director, National Cancer Control Center and National Personalized Medicine Program Professor and Chairman, Department of Community Medicine and Epidemology, Carmel Medical Center and Technion Rappaport Faculty of Medicine Ariel Roguin, MD, PhD [email protected] Head, Interventional Cardiology [email protected] Rambam Medical Center, Technion Rappaport Faculty of Medicine Ronen Rubinshtein, MD, FACC, FESC [email protected] Assistant Professor Department of Cardiovascular Medicine Lady Davis Carmel Medical Center Technion Rappaport Faculty of Medicine Yuval Shaked, PhD [email protected] Technion Rappaport Faculty of Medicine Eliezer Shalev, MD [email protected] Dean, Technion Rappaport Faculty of Medicine Shelly Tzlil, PhD [email protected] Assistant Professor, Technion Faculty of Mechanical Engineering

50 | 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP WEIZMANN INSTITUTE OF SCIENCE CONTACT INFORMATION

WEIZMANN INSTITUTE OF SCIENCE 234 HERZL STREET, REHOVOT 7610001 ISRAEL

NAME AND TITLE EMAIL ADDRESS Israel Bar-Joseph, PhD [email protected] The Jane and Otto Morningstar Professorial Chair in Physics Vice President, Resource Development and Public Affairs Weizmann Institute of Science Zvulun Elazar, PhD [email protected] Department of Biological Chemistry Weizmann Institute of Science Eran Elinav, MD, PhD [email protected] Senior Scientist, Department of Immunology Weizmann Institute of Science Eran Hornstein, MD, PhD [email protected] Department of Molecular Genetics Weizmann Institute of Science Orit Kollet, PhD [email protected] Department of Immunology Weizmann Institute of Science Zvi Livneh, PhD [email protected] Dean, Faculty of Biochemistry Department of Biological Chemistry Weizmann Institute of Science Orly Reiner, PhD [email protected] Incumbent of the Berstein-Mason Professional Chair of Neurochemistry Department of Molecular Genetics Oren Schuldiner, PhD [email protected] Dept of Molecular Cell Biology Weizmann Institute of Science Michal Schwartz, PhD [email protected] Department of Neurobiology Weizmann Institute of Science Berta Strulovici, PhD [email protected] Director, National Center for Personalized Medicine Weizmann Institue of Science 2013 UNIVERSITY OF MICHIGAN – ISRAEL PARTNERSHIP | 51

NOTES NOTES

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