Thomas Jefferson University
Sidney Kimmel Medical College
Department of Medicine

Research Activity

The Cardeza Center pursues a broad range of basic research that can be divided into three focus areas: basic research, clinical/translational and core laboratories.

Paul Bray, MD

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Dr. Bray’s laboratory studies the role of platelets in cardiovascular disease and disorders of bleeding and excessive blood clotting.  He identifies and characterizes novel variations in platelet genes and in platelet gene expression that affect both clinical and platelet phenotypes. His laboratory utilizes genomic and transcriptomic approaches, and molecular and cell biology approaches to address these important areas of research. He is a clinician-scientist who has been continuously funded by the National Institutes of Health from 1986 to the present. He is a member of the Genomics and Cancer Biology graduate program. Current areas of investigation include:

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  • Studies on demographic effects on platelet function and thrombosis

  • Studies on microRNA function in human megakaryocytes

  • Pharmacogenetic studies:

    • Testing for interactions between genetic variations and medications for clinical outcomes of myocardial infarction, stroke or bleeding

    • Testing for interactions between genetic variations and medications for platelet functional responses

Visit Plateletomics.com for more information.

Protocols

Publications

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

MicroRNA expression differences in human hematopoietic cell lineages enable regulated transgene expression

Human platelet microRNA-mRNA networks associated with age and gender revealed by integrated plateletomics

The human platelet: Strong transcriptome correlations among individuals associate weakly with the platelet proteome

PDK1 selectively phosphorylates Thr(308) on Akt and contributes to human platelet functional responses

Racial differences in human platelet PAR4 reactivity reflect expression of PCTP and miR-376c

Dengue platelets meet sir arthur conan doyle

Platelet Genomics

Erosive Arthritis and Hepatic Granuloma Formation Induced by Peptidoglycan Polysaccharide in Rats Is Aggravated by Prasugrel Treatment

MicroRNAs in platelet production and activation

The complex transcriptional landscape of the anucleate human platelet

Small RNAs as potential platelet therapeutics

Fractalkine is expressed in early and advanced atherosclerotic lesions and supports monocyte recruitment via CX3CR1

Human genome-wide association and mouse knockout approaches identify platelet supervillin as an inhibitor of thrombus formation under shear stress

Human genome-wide association and mouse knockout approaches identify platelet supervillin as an inhibitor of thrombus formation under shear stress

Platelet biology and response to antiplatelet therapy in women: Implications for the development and use of antiplatelet pharmacotherapies for cardiovascular disease

Identification of a specific intronic PEAR1 gene variant associated with greater platelet aggregability and protein expression

Transfection of human platelets with short interfering RNA

MicroRNAs in platelet production and activation

Platelet microRNA-mRNA coexpression profiles correlate with platelet reactivity

Members

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Paul Bray, MD
Professor

edestein

Leonard Edelstein, PhD
Research Assistant Professor

kong

Xiango Kong, MD
Research Associate

lindsay

Corey Lindsay, PhD
Research Assistant

bhatlekar

Seema Bhatlekar, PhD
Postdoctoral Fellow

whitley

Michael Whitley
Graduate Student

ma

Lin Ma
Research Technician

Past Members

trueuel-montoya

Raul Trueuel-Montoya

Leonard Edelstein, PhD

Dr. Edelstein’s laboratory studies gene expression in megakaryocytes and platelets and their role in cardiovascular disease and thrombopoiesis. Current research includes:

  • Activated platelets release microparticles (MPs) which contain RNA and protein. These MPs have been shown to deliver miRNAs to endothelial cells in culture and alter the level of RNAs in these cells. We are studying this effect in a flow chamber in which the endothelial cells are cultured under shear stress and then analyzing the effect on genes regulated by inflammation or shear stress, two processes important to atherosclerotic plaque formation.

  • Genome wide association studies have identified ~50 loci in the human genome associated with heart attacks. However, neither the genetic variant responsible for the increased risk nor the cell type in which it functions has been identified. We are developing methods to identify which locus-linked genes are functional in platelets and the effect the common variants have on their expression and function.

  • The DLK1-DIO3 locus in an imprinted region of the genome containing 54 maternally-expressed miRNAs. Among hematopoietic cells, these miRNAs are platelet specific. We have generated data that expression of these miRNAs increases during megakaryopoiesis and that early overexpression inhibits megakaryopoiesis. We are investigating the role of these miRNAs in megakaryocyte development.

  • Arterial hypertension is associated with thrombotic events due to platelet activation. The exact mechanism by which hypertensive platelets become more sensitive to agonist stimulation is unknown. We have collected platelet miRNA from healthy and pre-hypertensive subjects and found association between miRNA level and blood pressure.  We will use this data to identify differentially expressed genes responsible for the increased function in platelets.

Michael Holinstat, PhD

The Holinstat lab focuses on understanding complex platelet signaling mechanisms that regulate hemostasis and thrombosis. The work focuses on three primary areas of platelet research spanning from a basic science and drug discovery program to clinical and translational projects, including a clinical trial focused on platelet function in type 2 diabetes mellitus. This work is fully funded by three National Institutes of Health grants (NIH R01 grants), support from the American Heart Association (AHA), the Parenteral Drug Association Foundation (PDAF) and the Cardeza Foundation for Hematologic Research at Thomas Jefferson University.

One project in the lab focuses on identifying the inherent racial differences in platelet function between blacks and whites. This clinical project investigates some of the underlying genetic differences in blacks and whites that predispose some people to a higher risk for thrombosis and stroke while sparing others. Through this effort, we will be able to determine for the first time how to differentially treat patients based on their genetic/racial background. This area of research, often termed “individualized medicine,” has the potential to shift our mode of treatment from disease to patient by understanding that not all treatments will equally benefit every patient who presents with a thrombotic event, and more importantly prevention of thrombotic events may differ based on genetic or racial background. Early findings in this study have already shown that blacks express a higher level of the thrombin receptor PAR4 and have uncovered a novel protein in the human platelet (absent in the mouse platelet), phosphatidylcholine transfer protein (PCTP), which appears to play an important role in platelet activation and is highly expressed in platelets from blacks compared to whites.

A second area of focus in the lab is identifying how platelet activation is regulated by the enzyme 12-lipoxygenase (12-LOX). Similar to COX-1, 12-LOX oxidizes free fatty acids in order to form bioactive metabolites (called eicosanoids). Our lab has recently shown that the 12-LOX eicosanoids derived from arachidonic acid in the platelet are pro-thrombotic and we, along with our collaborators across the country, have developed the first selective inhibitor against human 12-LOX activity. This new class of inhibitors prevents agonist-mediated platelet activation and clot formation in human platelets. Subsequently, we also identified that 12-LOX oxidizes Omega-6 fatty acids to produce a novel eicosanoid, 12-HETrE, which is a potent inhibitor of platelet function. We are currently trying to identify the underlying mechanisms by which 12-HETrE protects against platelet activation, clot formation and stroke.

A third focus of the lab takes advantage of our basic scientific observations that altering the fatty acid content on the platelet may allow for formation of 12-LOX metabolites which would protect type 2 diabetics from suffering a thrombotic event. This translational project investigates in animal and patients the potential for omega-3 and omega-6 fatty acids and their 12-LOX eicosanoids to prevent platelet activation and unwanted thrombosis. This is an important area in diabetes research as 65% of deaths in type 2 diabetes mellitus are due to thrombosis and stroke.

Publications

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

Platelet 12-LOX is essential for FcγRIIa-mediated platelet activation

Racial differences in resistance to P2Y12receptor antagonists in type 2 diabetic subjects

MicroRNA expression differences in human hematopoietic cell lineages enable regulated transgene expression

Human platelet microRNA-mRNA networks associated with age and gender revealed by integrated plateletomics

Synthesis and structure-activity relationship studies of 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide derivatives as potent and selective inhibitors of 12-lipoxygenase

Mechanism of race-dependent platelet activation through the protease-activated receptor-4 and Gq signaling axis

The emerging role of oxylipins in thrombosis and diabetes

Racial differences in human platelet PAR4 reactivity reflect expression of PCTP and miR-376c

12-lipoxygenase activity plays an important role in PAR4 and GPVI-mediated platelet reactivity

MicroRNAs in platelet production and activation

Dichotomous effects of exposure to bivalirudin in patients undergoing percutaneous coronary intervention on protease-activated receptor-mediated platelet activation

Investigations of human platelet-type 12-lipoxygenase: Role of lipoxygenase products in platelet activation

Protein kinase C regulation of 12-lipoxygenase-mediated human platelet activation

Rap1-Rac1 circuits potentiate platelet activation

We can do it together: PAR1/PAR2 heterodimer signaling in VSMCs

Protein kinase Cα phosphorylates the TRPC1 channel and regulates store-operated Ca 2+ entry in endothelial cells (Journal of Biological Chemistry (2004) 279, (20941-20949))

Discovery of potent and selective inhibitors of human platelet-type 12-lipoxygenase

12-lipoxygenase: A Potential Target for Novel Anti-Platelet Therapeutics

Protease-activated receptor signaling in platelets activates cytosolic phospholipase a2α differently for cyclooxygenase-1 and 12-lipoxygenase catalysis

Members

holinstat

Michael Holinstat, PhD
Associate Professor

tourdot

Benjamin Tourdot, PhD
Postdoctoral Fellow

ren

Jin Ren, PhD

niisuke

Katrin Niisuke, PhD

yeung

Jennifer Yeung, MS

conaway

Stanley Conaway

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Johnny Yu

vesci

Joanne Vesci

Steven McKenzie, MD PhD

Dr. McKenzie is a Hematology physician-scientist with clinical expertise in adult and pediatric non-malignant hematologic disorders and a scientific expertise in translational research. He works in the Cardeza Hemophilia and Thrombosis Center and also in the Hereditary Anemias Center. He is a member of the Sidney Kimmel Cancer Center (SKCC), in the Molecular Biology and Genetics Program. He is a member of two Graduate Programs:

  • Genetics, Genomics and Cancer Biology

  • Immunology and Microbial Pathogenesis

He is a physician mentor for Thomas Jefferson University MD/PhD program.  

Dr. McKenzie directs two major laboratory research projects. The first project has a focus on immune-mediated thrombocytopenia and thrombosis syndromes (see McKenzie and Sachais, Current Opinion in Hematology, September 2014). This work led to the first and only mouse model of heparin-induced thrombocytopenia and thrombosis (HIT). The current work explores Novel Therapeutics in HIT, in an NIH-supported Program Project grant with Drs. Poncz and Rauova at Children’s Hospital of Philadelphia, Drs. Cines, Sachais and Cuker at University of Pennsylvania, and Dr. Arepally at Duke. In another subproject, Dr. McKenzie is co-funded on an NIH R01 with Dr. Bergmeier at UNC Chapel Hill and Dr. Woulfe at University of Delaware to explore platelet signaling mechanisms.

The second major McKenzie lab project focuses on the Genomics and Molecular Genetics of inter-individual variation in human platelet activation via FcgammaRIIa. This molecule has dual functions, as a receptor for IgG immune complexes and as a transmembrane adapter in integrin “outside-in” signaling. In collaboration with Dr. Paul Bray, who led the PRAX1 study, our team has identified differentially expressed mRNAs and miRNAs as well as genomic variants that regulate platelet reactivity. Team members of our longstanding Platelet Interest Group are Drs. Bray, McKenzie, Holinstat, Edelstein and Naik at Thomas Jefferson University, Drs. Rigoutsos and Londin at Thomas Jefferson University Computational Medicine Center, Dr. Fortina of SKCC Genomics, Drs. Shaw and Simon at Baylor, and Dr. Kunapuli at Temple. Dr. McKenzie is also co-funded, with Dr. Holinstat as PI, on work that involves 12-LOX and diabetes vascular biology.  Our foci in the McKenzie lab moving forward are novel molecular genetic pathways for determination of receptor levels, protein tyrosine phosphatase activity and oligo-ubiquitylation in platelet FcgammaRIIa  functions.

Publications

Platelet 12-LOX is essential for FcγRIIa-mediated platelet activation

MicroRNA expression differences in human hematopoietic cell lineages enable regulated transgene expression

Human platelet microRNA-mRNA networks associated with age and gender revealed by integrated plateletomics

The human platelet: Strong transcriptome correlations among individuals associate weakly with the platelet proteome

Identification of a developmental gene expression signature, including hox genes, for the normal human colonic crypt stem cell niche: Overexpression of the signature parallels stem cell overpopulation during colon tumorigenesis

CalDAG-GEFI deficiency protects mice from FcγRIIa-mediated thrombotic thrombocytopenia induced by CD40L and β2GPI immune complexes

Advances in the pathophysiology and treatment of heparin-induced thrombocytopenia

Tyrosine phosphorylation on spleen tyrosine kinase (Syk) is differentially regulated in human and murine platelets by protein kinase C isoforms

MicroRNAs in platelet production and activation

Cooperative integrin/ITAM signaling in platelets enhances thrombus formation in vitro and in vivo

The complex transcriptional landscape of the anucleate human platelet

Monocytes in HIT: An evolving story

CalDAG-GEFI deficiency protects mice in a novel model of FcγRIIA-mediated thrombosis and thrombocytopenia

Safety and pharmacokinetics of subcutaneously administered recombinant activated factor VII (rFVIIa)

PRT-060318, a novel Syk inhibitor, prevents heparin-induced thrombocytopenia and thrombosis in a transgenic mouse model

Amelioration of murine immune thrombocytopenia by CD44 antibodies: A potential therapy for ITP?

Dominant expression of the inhibitory FcγRIIB prevents antigen presentation by murine plasmacytoid dendritic cells

The clot thickens (or not)

Thrombopoietin following transfusion of platelets in preterm neonates

The yin and yang of Fcγ receptors in ITP

Members

mckenzie

Steven E. McKenzie, MD PhD

abraham

Shaji Abraham, PhD

zhou

Yuhang Zhou

Past Members

andre

Pierette Andre

Ulhas P. Naik, PhD

The Naik laboratory is focused on developing therapeutic strategies to interrupt the progress of cardiovascular disease and cancer. In this regard, the team has identified several novel gene products that play key regulatory roles in the progression of these diseases. Using cell and molecular biological approaches, the team has characterized the potential role of calcium- and integrin-binding protein family and junctional adhesion molecule family members in physiological and pathological settings. Cutting edge technologies, such as the yeast two-hybrid system, siRNA, transgenic mouse models, systems biology, nanotechnology and fluid dynamics modeling are routinely employed in the laboratory.

Some specific projects involve:

  • Positive and negative regulatory mechanisms of platelet activation during thrombosis

  • Mechanism of new blood vessel formation (angiogenesis) during ischemia and cancer

  • Mechanism of breast and prostate cancer cell metastasis

  • Use of nanotechnology to deliver drugs to specifically targeted sites

Current Projects

  • 2P20 RR015588-10

  • 1R01 HL113118-02

  • 1R01 HL119374-01

  • AHA Grant-in-Aid

  • Fraunhofer-UD Research Grant

Publications

Bacteria exploit platelets

Junctional adhesion molecule-A suppresses platelet integrin αIIbβ3 signaling by recruiting Csk to the integrin-c-Src complex

Junctional Adhesion Molecule-A Regulates Vascular Endothelial Growth Factor Receptor-2 Signaling-Dependent Mouse Corneal Wound Healing

Fucoidan is a novel platelet agonist for the C-type lectin-like receptor 2 (CLEC-2)

Effects of JAM-A deficiency or blocking antibodies on neutrophil migration and lung injury in a murine model of ALI

A spatial model for integrin clustering as a result of feedback between integrin activation and integrin binding

CASK interacts with PMCA4b and JAM-A on the mouse sperm flagellum to regulate Ca2+ homeostasis and motility

Pericyte-endothelial cell interaction: A survival mechanism for the tumor vasculature

JAM-A protects from thrombosis by suppressing integrin α IIbβ 3-dependent outside-in signaling in platelets

Calcium- and integrin-binding protein 1 regulates megakaryocyte ploidy, adhesion, and migration

Contra-regulation of calcium- and integrin-binding protein 1-induced cell migration on fibronectin by PAK1 and MAP kinase signaling

Efficient implementation of the proper outlet flow conditions in blood flow simulations through asymmetric arterial bifurcations

Probabilistic modeling and analysis of the effects of extra-cellular matrix density on the sizes, shapes, and locations of integrin clusters in adherent cells

An impedance model for blood flow in the human arterial system. Part I: Model development and MATLAB implementation

Application of 1D blood flow models of the human arterial network to differential pressure predictions

Calcium-dependent inhibition of polo-like kinase 3 activity by CIB1 in breast cancer cells

Calcium- and integrin-binding protein 1 regulates endomitosis and its interaction with polo-like kinase 3 is enhanced in endomitotic dami cells

Calcium- and integrin-binding protein 1 regulates microtubule organization and centrosome segregation through polo like kinase 3 during cell cycle progression

Integrin αIIbβ3: A novel effector of Gα13

PTEN: Not just a tumor suppressor

Members

Ulhas P. Naik, PhD

Brendan Bachman, M.S.

Randall Derstine, MS

Vilas Desai, PhD

Kalyan Golla, PhD

Meghna U. Naik, MSc

Pravin Patel, BS

Cardeza Seminars

All Seminars in Cardeza Library (Curtis Building Room 903) at 12:00 pm unless otherwise noted.

2015

January 28 – Athan Kuliopulos (Tufts University)

February 11 – Rajanikanth Vadigepalli (Thomas Jefferson University)

February 25 – Khadija Rafiq (Thomas Jefferson University)

March 11 – Robert Flaumenhaft (Harvard University)

March 25 – Jean Pierre Issa (Temple University)

April 8 – Shawn Jobe (Milwaukee Blood Research Institute)

April 22 – Jerry Nadler (EVMS, Chair of Medicine)

May 13 – Ian Blair (University of Pennsylvania)

May 27 – TBA

2014

September 3 – Caleb Kallen (Thomas Jefferson University)

September 17 – Shey-Shing Sheu (Thomas Jefferson University)

October 1 – Deepak Deshpande (Thomas Jefferson University)

October 15 – Domenico Pratico (Temple University)

October 29 – John Semple (University of Toronto; St. Michael’s Hospital)

November 12 – John Weisel (University of Pennsylvania)