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Principal Investigator: Alec Stepanian
Organization: TUFTS UNIVERSITY BOSTON
Fiscal Year: 2024
Award: $44,724
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY/ABSTRACT
The development of first generation Abl-tyrosine kinase inhibitor (Abl-TKI) imatinib (Im) to treat chronic
myelogenous leukemia (CML) in 2001 has changed this previously lethal cancer into a chronic illness. Despite
initial success, Im intolerance and resistance necessitates development of newer generation Abl-TKIs.
Ponatinib (Po), a newer Abl-TKI that is lifesaving for T315I mutation CML, is associated with a four-fold
increased risk of acute arterial occlusive events, including life-threatening heart attack or stroke, compared to
Im, which has nullified any cancer survival benefit. With a rapidly growing prevalence of over 100,000 survivors
and 10,000 new cases annually in the US alone, there exists an urgent need to determine the mechanism of
Abl-TKI toxicity to improve CML outcomes. Previous published studies assessing direct effects of Po on
platelets and clotting are inconclusive. Because CML patients present with high prevalence of cardiovascular
risk factors and underlying atherosclerotic disease, we introduce the novel hypothesis that Po acts to instead
inflame pre-existing atherosclerotic plaques and prime them for rupture and subsequent thrombosis.
Endothelial cells (ECs) lining the vasculature are anti-inflammatory but become proinflammatory in the setting
of injury, expressing adhesion molecules that induce leukocyte trafficking and subsequent plaque
inflammation. As these patients also exhibit elevated serum proinflammatory cytokine TNFa despite remission,
this proposal also tests the hypothesis that Po sensitizes ECs to TNFa signaling via increased expression of
TNF receptor (TNFR) on cell surface, resulting in increased adhesion molecule expression on human ECs in
vitro, increased leukocyte trafficking into vessels in vivo, and thus more inflamed atherosclerotic plaques in
vivo. Preliminary data confirms that Po: 1) induces expression of adhesion molecules in ECs in vitro and that
this increase is prevented by a TNFR inhibitor, 2) increases leukocyte trafficking by intravital microscopy in
vivo, and 3) increases plaque leukocyte content, a marker of plaque inflammation, measured by flow cytometry
in atherogenic mice (ApoE-KO). This proposal tests this hypothesis for Im, Po, and asciminib which is recently
approved but vascular safety is unknown. Aim 1 explores the mechanism by which Po induces EC adhesion
molecule expression and the role of EC TNFR signaling in vitro using genetic and pharmacologic approaches
to block TNFR. Aim 2 examines the impact of Po on leukocyte trafficking and plaque inflammation in vivo via
intravital microscopy in EC-TNFR1-KO mice and on plaque inflammation in ApoE-KO mice by aortic arch flow
cytometry. Successful completion of these aims will test a recently approved Abl-TKI for toxicity and provide
novel insight into Po’s mechanism of vascular toxicity, which can lead to support for an EC protective strategy
that will prevent arterial thrombosis and improve CML outcomes. The detailed training plan will prepare the PI
for a career as an independent academic physician-scientist through rigorous training in molecular and
vascular biology and guidance from a translational and transdisciplinary advisory team.
Terms: <(TNF)-α><ABL Tyrosine Kinase><Acute><Adhesion Molecule><Adhesions><Advisory Committees><Affect><Age><American><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Aorta><Apo-E><ApoE><ApoE protein><Apolipoprotein E><Apoplexy><Arterial Fatty Streak><Arterial Fatty Streaks><Atheroma><Atheromatous><Atheromatous degeneration><Atheromatous plaque><Atherosclerosis><Atherosclerotic Cardiovascular Disease><BB2><BCR-ABL Kinase><Bcr-Abl tyrosine kinase><Biology><Blood Plasma><Blood Platelets><Blood Serum><Blood Vessels><Blood leukocyte><Brain Vascular Accident><CD 120a Antigen><CD120a Antigens><CD54><CD62P Antigens><Cachectin><Cachectin Receptors><Cancer Treatment><Cancers><Cardiac artery><Cardiac infarction><Cardiovascular><Cardiovascular Body System><Cardiovascular Organ System><Cardiovascular system><Cell Adhesion Molecule Gene><Cell Adhesion Molecules><Cell Communication and Signaling><Cell Line><Cell Signaling><Cell surface><CellLine><Cerebral Stroke><Cerebrovascular Apoplexy><Cerebrovascular Stroke><Cessation of life><Chromosomal dislocation><Chromosomal translocation><Chronic><Chronic Disease><Chronic Granulocytic Leukemia><Chronic Illness><Chronic Myelocytic Leukemia><Chronic Myelogenous Leukemia><Chronic Myeloid Leukemia><Clinical><Clotting><Coagulation><Coagulation Process><Coronary artery><DNA Molecular Biology><Data><Death><Development><Disease remission><Drugs><Endothelial Cells><Event><Exhibits><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Future><GMP-140><Generations><Genetic><Genetic Alteration><Genetic Change><Genetic Translocation><Genetic defect><Goals><Heart Vascular><Heart artery><High Prevalence><Histology><Human><ICAM><ICAM-1 Gene><ICAM1><ICAM1 gene><Imatinib><Immunoblotting><Immunofluorescence><Immunofluorescence Immunologic><Immunoglobulin Enhancer-Binding Protein><In Vitro><In vivo analysis><Inflammation><Inflammatory><Injury><Intracellular Communication and Signaling><KO mice><Knock-out Mice><Knockout Mice><LECAM-3><Leukocyte Trafficking><Leukocyte Trafficking Induction><Leukocytes><Leukocytes Reticuloendothelial System><Life><Macrophage-Derived TNF><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Marrow leukocyte><Marrow platelet><Measures><Mediating><Medication><Mice><Mice Mammals><Modeling><Modern Man><Molecular><Molecular Biology><Monocyte-Derived TNF><Murine><Mus><Mutation><Myocardial Infarct><Myocardial Infarction><NF-kB><NF-kappa B><NF-kappaB><NFKB><Nuclear><Nuclear Factor kappa B><Nuclear Transcription Factor NF-kB><Null Mouse><Outcome><P-Selectin><P150><PTK Inhibitors><Pathway interactions><Patients><Peripheral><Pharmaceutical Preparations><Physicians><Plasma><Plasma Serum><Platelet alpha-Granule Membrane Protein><Platelets><Prevalence><Protein Tyrosine Kinase Inhibitors><Proto-Oncogene Proteins c-abl><Proto-Oncogene Tyrosine -Protein Kinase ABL><Publishing><Quantitative RTPCR><Quantitative Reverse Transcriptase PCR><Receptor Inhibition><Receptor Signaling><Remission><Resistance><Reticuloendothelial System, Serum, Plasma><Risk><Role><Rupture><Safety><Scientist><Selectins><Serum><Short interfering RNA><Signal Transduction><Signal Transduction Systems><Signaling><Small Interfering RNA><Strains Cell Lines><Stroke><Survivors><TK Inhibitors><TNF><TNF A><TNF Alpha><TNF Receptor Family Protein><TNF Receptor Superfamily><TNF Receptor p55><TNF Receptors><TNF gene><TNF-sR55><TNF-α><TNF-α receptor><TNFA><TNFAR><TNFR><TNFR p60><TNFR, 55-kD><TNFR, 60-kD><TNFR-I><TNFR1><TNFR55><TNFR60><TNFRSF1A><TNFRSF1A Receptor><TNFRSF1A gene><TNFalpha receptor><TNFα><TNFα receptor><Task Forces><Testing><Thrombocytes><Thrombosis><Time><Toxic effect><Toxicities><Training><Transcription Factor NF-kB><Tumor Necrosis Factor><Tumor Necrosis Factor Receptor><Tumor Necrosis Factor Receptor 1><Tumor Necrosis Factor Receptor 55><Tumor Necrosis Factor Receptor Family><Tumor Necrosis Factor Receptor Superfamily><Tumor Necrosis Factor-alpha><Tyrosine Kinase Inhibitor><Umbilical vein><Western Blotting><Western Immunoblotting><White Blood Cells><White Cell><abl Proto-Oncogene Products><abl Proto-Oncogene Proteins><advisory team><ages><anti-cancer therapy><aortic arch><atheromatosis><atherosclerosis plaque><atherosclerotic disease><atherosclerotic lesions><atherosclerotic plaque><atherosclerotic vascular disease><biological signal transduction><brain attack><c-ABL Protein><cancer survival><cancer therapy><cancer-directed therapy><cardiac infarct><cardioprotectant><cardioprotection><cardioprotective><cardiovascular risk><cardiovascular risk factor><career><cell adhesion protein><cerebral vascular accident><cerebrovascular accident><chromosome dislocation><chromosome translocation><chronic disorder><circulatory system><coronary attack><coronary infarct><coronary infarction><cultured cell line><cytokine><developmental><drug/agent><flow cytophotometry><genetic approach><genetic strategy><genome mutation><heart attack><heart infarct><heart infarction><improved><in vitro testing><in vivo><in vivo evaluation><in vivo testing><inhibitor><injuries><insight><intercellular cell adhesion molecule><intra-vital microscopy><intravital microscopy><kappa B Enhancer Binding Protein><knock-down><knockdown><leukemia><leukemia treatment><leukemic therapy><malignancy><mouse model><murine model><neoplasm/cancer><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><nuclear factor kappa beta><pathway><pharmacologic><prevent><preventing><promoter><promotor><protein blotting><qRTPCR><resistant><safety assessment><siRNA><side effect><social role><stroke event><stroked><strokes><success><survivorship><thrombotic><thrombotic disease><thrombotic disorder><tumor necrosis factor alpha receptor><tumor necrosis factor receptor 1A><tumor necrosis factor α receptor><tyrosine kinase ABL1><vascular><vascular inflammation><vulnerable plaque><white blood cell><white blood corpuscle>