Defective endothelial progenitor cells (EPCs) may contribute to the progression of coronary artery disease. Defective EPCs and a reduced responsiveness of aged/diseased tissues may also be reasons for the limited success of clinical trials of autologous CD34+ bone marow cels for the treatment of acute myocardial infarction and peripheral artery disease (PAD). In the Preliminary Results section of the proposal we present the first micro!array and micro!RNA (miR) profiles of human CD34+/Lin! EPCs, comparing healthy volunteers with coronary artery disease (CAD) and age!matched non!CAD patients. The results reveal down!regulation of multiple angiogenic, growth and survival factors including VEGF, integrin!1V, Akt, and eNOS in the CAD group, and >3!fold increased expression of miR!92a, a global anti!angiogenesis miR that targets integrin!1V and Akt, miR!16, that targets VEGF and cell cycle genes CCDN!1 and 2, and miR!21 that targets cell migration factors and stem cell self renewal and has also been associated with neointimal formation. Here we will test the hypothesis that ischemia can be effectively and safely resolved on multiple disease backgrounds by tissue engineering with hypoxia!regulated AAV9 vectors expressing pro!angiogenic growth factors, combined with EPC therapy after engineering the EPCs with selective micro!RNAs (premirs) and micro!RNA inhibitors (antagomirs). We have created and tested a series of unique AAV vectors that express pro!angiogenic genes under the direction of hypoxia!regulated, conditionally silenced promoters. When expressing VEGF, these vectors support directional vessel growth, arteriogenesis and stable reperfusion of ischemic hind limbs, and are protective against AMI in a mouse model. The vectors are highly regulated due to the presence of silencer elements that switch off gene expression under physiological oxygen tension. We propose to profile miR expression in different CD34+ and CD133+ cell populations from CAD patients with and without type 2 diabetes and investigate the role of serum factors in regulating miRs expression, and whether normal regulation and function can be recovered pharmacologically. We will determine whether EPC functions can be recovered by molecular genetic engineering of EPCs, and test engineered EPC therapy in multiple models of ischemia on backgrounds of age and atherosclerosis.
In Aims 1 and 2 we will profile miRs, their targets and functions in EPCs from CAD patients 1 T2D and investigate pathways to regain function.
In Aim 3 we will characterize the cellular and molecular events that promote revascularization in response to optimal gene and engineered cell therapy in each model of ischemia and each age/disease background.

Public Health Relevance

Studies are proposed to determine why bone marrow cells from patients with coronary artery disease and peripheral artery disease have defective functions and do not provide efficient therapy when they are collected from a patient's bone marrow, purified and delivered back to the same patient for stem cell therapy. Experiments are proposed to correct the defects in these cells and optimize a cellular bioengineering protocol that will improve stem cell therapy for patients with peripheral and coronary artery disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL072924-07
Application #
8461966
Study Section
Myocardial Ischemia and Metabolism Study Section (MIM)
Program Officer
Reid, Diane M
Project Start
2003-04-01
Project End
2015-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
7
Fiscal Year
2013
Total Cost
$438,424
Indirect Cost
$151,872
Name
University of Miami School of Medicine
Department
Pharmacology
Type
Schools of Medicine
DUNS #
052780918
City
Coral Gables
State
FL
Country
United States
Zip Code
33146
Ma, QunChao; Xia, Xiangyang; Tao, Quanwei et al. (2016) Profound Actions of an Agonist of Growth Hormone-Releasing Hormone on Angiogenic Therapy by Mesenchymal Stem Cells. Arterioscler Thromb Vasc Biol 36:663-72
Hu, Xinyang; Chen, Panpan; Wu, Yan et al. (2016) MiR-211/STAT5A Signaling Modulates Migration of Mesenchymal Stem Cells to Improve its Therapeutic Efficacy. Stem Cells 34:1846-58
Graham, Regina M; Thompson, John W; Webster, Keith A (2015) BNIP3 promotes calcium and calpain-dependent cell death. Life Sci 142:26-35
Hu, Xinyang; Zhang, Ling; Jin, Jing et al. (2015) Heparanase released from mesenchymal stem cells activates integrin beta1/HIF-2alpha/Flk-1 signaling and promotes endothelial cell migration and angiogenesis. Stem Cells 33:1850-62
Thompson, John W; Wei, Jianqin; Appau, Kweku et al. (2015) Bnip3 Binds and Activates p300: Possible Role in Cardiac Transcription and Myocyte Morphology. PLoS One 10:e0136847
Adi, Nikhil; Adi, Jennipher; Cesar, Liliana et al. (2015) Role of Micro RNA-205 in Promoting Visceral Adiposity of NZ10 Mice with Polygenic Susceptibility for Type 2 Diabetes. J Diabetes Metab 6:
Hu, Xinyang; Wu, Rongrong; Shehadeh, Lina A et al. (2014) Severe hypoxia exerts parallel and cell-specific regulation of gene expression and alternative splicing in human mesenchymal stem cells. BMC Genomics 15:303
Hu, Xinyang; Wu, Rongrong; Jiang, Zhi et al. (2014) Leptin signaling is required for augmented therapeutic properties of mesenchymal stem cells conferred by hypoxia preconditioning. Stem Cells 32:2702-13
Graham, Regina M; Thompson, John W; Webster, Keith A (2014) Inhibition of the vacuolar ATPase induces Bnip3-dependent death of cancer cells and a reduction in tumor burden and metastasis. Oncotarget 5:1162-73
Omran, Ahmed; Elimam, Dalia; Webster, Keith A et al. (2013) MicroRNAs: a new piece in the paediatric cardiovascular disease puzzle. Cardiol Young 23:642-55

Showing the most recent 10 out of 32 publications