Restoration of myocardial reparative function of diabetic progenitor cells by epigenetic modulation

通过表观遗传调节恢复糖尿病祖细胞的心肌修复功能

• 批准号: 9903831
• 负责人: Raj Kishore
• 金额: $ 56.03万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-10 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9903831
• 关键词: American; Animals; Attenuated; Azacitidine; Blood Vessels; Bone Marrow; CD34 gene; Cardiac; Cardiovascular Diseases; Cell Therapy; Cell division; Cell physiology; Cells; Clinical; Clinical Trials; Coupled; DNA Modification Methylases; DNMT3a; Diabetes Mellitus; Diabetic mouse; Endothelium; Enzymes; Epigenetic Process; Functional disorder; G9a histone methyltransferase; Gene Expression; Genes; Goals; HDAC1 gene; Heart Diseases; Heart failure; Hematopoietic stem cells; Histone Acetylation; Histone Deacetylase; Histones; Human; Hyperglycemia; Impairment; In Vitro; Inherited; Lysine; Mediator of activation protein; Memory; Methylation; Methyltransferase; MicroRNAs; Modeling; Modification; Molecular; Morbidity - disease rate; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Natural regeneration; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Patients; Pattern; Perfusion; Pharmaceutical Preparations; Phenotype; Physiological; Process; Property; Public Health; Publishing; Research; Role; Sequential Treatment; Testing; Therapeutic; Tissues; Valproic Acid; Vascular Diseases; Vascular Endothelium; adult stem cell; angiogenesis; base; critical limb Ischemia; daughter cell; db/db mouse; diabetic; diabetic patient; differential expression; disability; endothelial stem cell; exosome; experimental study; functional disability; gene repression; genome wide methylation; heart disease risk; histone modification; improved; in vivo; inhibitor/antagonist; ischemic injury; limb ischemia; mortality; mouse model; neovascularization; novel strategies; novel therapeutic intervention; pre-clinical research; progenitor; programs; promoter; regenerative; repaired; restoration; small molecule; small molecule inhibitor; stem cells; targeted agent; tissue repair

In the absence of effective endogenous repair mechanisms after ischemic injury, cell-based therapies have emerged as a potential novel therapeutic approach in ischemic tissue repair. After the initial characterization of putative bone marrow-derived endothelial progenitor cells (EPC) and their potential to promote cardiac and critical limb ischemia neovascularization and to attenuate ischemic injury, more than a decade of intense preclinical research, led to the BM progenitors/EPC-based clinical trials. However, despite early enthusiasm, cell based therapies yielded modest clinical results. Modest clinical outcomes of cell-based therapies may reflect the cellular dysfunction that is known to ensue in EPC/progenitor cells obtained from diabetic animals as well as patients. Compelling evidence indicates that EPCs dysfunction represents a mechanism for impaired vascular repair and angiogenesis in diabetes, subsequently leading to vascular dysfunction. Therefore, understanding the molecular basis of diabetes-induced EPC dysfunction and potentially reversing EPC dysfunction may represent a strategy to enhance cell-based therapeutics for myocardial/ischemic limb repair in diabetic patients. Increasing evidence also indicates that the mechanism underlying hyperglycemic memory and EPC dysfunction may involve epigenetic mechanisms involving enhanced epigenetic repressive marks on vascular genes leading to their epigenetic silencing. Moreover, since hyperglycemic memory is inherited through cell division, and altered epigenetic patterns in diabetic EPCs can be transmitted to daughter cells. Understanding the epigenetic basis of EPC dysfunction in diabetics and epigenetic reprogramming of diabetic EPCs, is therefore, of paramount importance for cell based therapies in diabetic patients. Therefore, our central hypothesis is that epigenetic repressive marks in diabetic EPCs render them dysfunctional and epigenetic modifying agents targeting those repressive marks can reprogram diabetic EPCs to a more functional and reparative phenotype. This project aims to study, in detail, phenotypic, epigenetic and molecular characterization of reprogrammed diabetic EPCs from diabetic mice as well as human CD34+ hematopoietic stem cells from diabetic patients and their exosome derivatives and test the ischemic myocardial repair capacity of these reprogrammed diabetic EPCs in physiologically relevant model of MI and hind limb ischemia. This overall aim will be achieved by conducting experiments organized under the following three specific aims: 1) To evaluate phenotypic stability and reparative potential of epigenetically reprogrammed diabetic EPCs; 2) To determine the specific epigenetic modifications in reprogrammed EPCs and establish a role of HDAC1 and G9a methytransferase in reprogramming process and 3) To establish epigenetic reprogramming rescues functional and reparative deficits in human CD34+ stem cells from patients with Type 2 diabetes.

在缺血性损伤后没有有效的内源性修复机制的情况下，基于细胞的疗法已经出现 作为缺血组织修复中潜在的新型治疗方法。在推定骨的最初表征之后 骨髓来源的内皮祖细胞（EPC）及其促进心脏和关键肢体缺血的潜力 新血管形成和减弱缺血性损伤，十多年的激烈临床前研究导致了 BM祖细胞/基于EPC的临床试验。然而，尽管早期热情，基于细胞的疗法却谦虚 临床结果。基于细胞的疗法的适度临床结果可能反映出已知的细胞功能障碍 随后在从糖尿病动物和患者获得的EPC/祖细胞中。令人信服的证据表明 EPCS功能障碍代表了糖尿病中血管修复和血管生成受损的机制，随后 导致血管功能障碍。因此，了解糖尿病诱导的EPC功能障碍的分子基础 EPC功能障碍可能逆转EPC功能障碍可能代表了增强基于细胞的治疗剂的策略 糖尿病患者的心肌/缺血性肢体修复。越来越多的证据也表明机制 潜在的高血糖记忆和EPC功能障碍可能涉及涉及增强的表观遗传机制 血管基因上的表观遗传抑制标记导致其表观遗传沉默。此外，由于高血糖 记忆是通过细胞分裂遗传的，并且可以将糖尿病EPC中的表观遗传模式改变为 子细胞。了解EPC功能障碍在糖尿病患者和表观遗传重编程中的表观遗传基础 因此，糖尿病EPC对糖尿病患者的基于细胞的疗法至关重要。因此，我们的 中心假设是糖尿病性EPC中的表观遗传抑制作用使它们功能失调和表观遗传学 修改靶向这些抑制标记的剂可以将糖尿病EPC重新编程为功能更强和更高的 表型。该项目旨在详细研究表型，表观遗传和分子表征 来自糖尿病小鼠以及糖尿病的人类CD34+造血干细胞的重编程糖尿病EPC 患者及其外泌体衍生物，并测试这些重新编程的缺血性心肌修复能力 MI和后肢缺血的生理相关模型中的糖尿病EPC。这个总体目标将由 进行以下三个特定目的组织的实验：1）评估表型稳定性和 表观遗传重编程的糖尿病EPC的修复潜力； 2）确定特定的表观遗传学 对重编程的EPC进行修改，并在重编程中建立HDAC1和G9A甲基转移酶的作用 过程和3）建立表观遗传重编程挽救人CD34+的功能和修复缺陷 来自2型糖尿病患者的干细胞。