The Role of C-Kit Positive Cardiac Progenitors in Maternal Diabetes-Induced Heart Defects and the Therapeutic Values of These Cells

C-Kit 阳性心脏祖细胞在母亲糖尿病引起的心脏缺陷中的作用以及这些细胞的治疗价值

• 批准号: 9403962
• 负责人: Sunjay Kaushal
• 金额: $ 60.24万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9403962
• 关键词: Adult; Adverse effects; Affect; Antioxidants; Apoptosis; Autologous; Biological; Birth; Cardiac; Cell Cycle; Cell Proliferation; Cell Therapy; Cell physiology; Cells; Cellular Stress; Cellular biology; Congenital Abnormality; Congenital Heart Defects; Cyclin D1; DNA; DNA Methylation; DNA Modification Methylases; DNMT3a; Data; Deacetylase; Defect; Diabetes Mellitus; Diabetic mother; Embryonic Development; Embryonic Heart; Enzymes; Etiology; Fetal Therapies; Functional disorder; Glucose; Heart; Heart Abnormalities; Heart failure; Histone Acetylation; Hypermethylation; Impairment; In Vitro; Inositol; Mediating; Methylation; Modeling; Morphogenesis; Mothers; Myocardial; Myocardial Infarction; Myocardial dysfunction; Neonatal; Neural Crest Cell; Operative Surgical Procedures; Oxidative Stress; Patients; Phenylbutyrates; Pregnancy; Pregnancy in Diabetics; Proto-Oncogene Protein c-kit; Role; Sirtuins; Stem cells; Stress; Teratogens; Therapeutic; Transplantation; Up-Regulation; base; cardiogenesis; cyclin D3; design; diabetic; exosome; imprint; improved; in vivo; inhibitor/antagonist; maternal diabetes; mouse model; non-diabetic; offspring; overexpression; paracrine; phase 1 study; postnatal; progenitor; promoter; regenerative; repaired; structural heart disease; superoxide dismutase 1

Project Summary Congenital heart defects (CHDs) are the most common birth defects. Pregestational maternal diabetes is a noninherited factor associated with a five-fold increase in CHDs and cardiac dysfunction. The underlying mechanism of diabetes-induced CHDs and cardiac dysfunction is unknown but one mechanism may involve inhibition of cardiogenesis by high glucose levels. c-Kit+ cardiac progenitor cells (CPCs) are now being studied as a potential treatment option for adult heart failure patients for stimulating cardiac function. Our preliminary studies have determined that both diabetes and high glucose in vitro induce a spectrum of cellular dysfunction in c-kit+ CPCs, that is implicated in the etiology of diabetes-induced CHDs. Eliminating c- kit+ CPCs during cardiogenesis led to CHDs resembling those in diabetic pregnancy offspring. Equally important is to determine the adverse programming effect caused during maternal diabetic exposure on the postnatal derived c-kit+ CPCs which will be used in our upcoming autologous based c-kit+ CHD trial. Therefore, we hypothesize that high glucose in diabetes induces cellular dysfunction in c-kit+ CPCs, which contributes to cardiac septation defects and limits the remodeling effect of post- natal derived c-kit+ CPCs on damaged hearts. Reducing cellular stress or DNA methylation or histone acetylation in c-kit+ CPCs alleviates maternal diabetes-induced CHDs, and improves the therapeutic value of ex vivo expanded c-kit+ CPCs by restoring their paracrine function. Studies are designed specifically to reveal the diabetes or high glucose on c-kit+ CPC function. Aim 1 will determine whether cellular stress-induced c-kit+ CPCs dysfunction contributes to the teratogenicity of maternal diabetes. We hypothesize that diabetes triggers apoptosis and reduce cell proliferation of c-kit+ CPCs through cellular stress, which impairs cardiac septation and the function of critical cardiac septation regulators: second heart field progenitors and cardiac neural crest cells. Aim 2 will determine whether enhanced histone acetylation and DNA methylation in c-kit+ CPCs mediate the adverse effects of maternal diabetes on cardiogenesis and imprinting on these progenitors. We hypothesize that diabetes-reduced sirtuin deacetylase 2 (SIRT2) causes DNA hypermethylation leading to c-kit+ CPCs cellular dysfunction that critically involve in altered cardiac septation and adverse imprinting. Aim 3 will determine the therapeutic abilities of offspring derived c-kit+ CPCs and their exosomes from nondiabetic and diabetic mothers in a myocardial infarction model and embryonic hearts of diabetic pregnancy. We hypothesize that offspring derived c-kit+ CPCs from maternal diabetics have lower abilities in repairing CHDs and cardiac dysfunction due to miR-34a up-regulation, which alters secretome and exosome profiling compared with nondiabetic mothers, and retain high levels of cellular stress, histone acetylation and DNA methylation during CPC therapies.

项目概要 先天性心脏病（CHD）是最常见的出生缺陷。妊娠期母亲糖尿病是 一种与 CHD 和心功能障碍增加五倍相关的非遗传因素。底层的 糖尿病诱发冠心病和心功能不全的机制尚不清楚，但可能有一种机制 涉及高葡萄糖水平抑制心脏发生。 c-Kit+ 心脏祖细胞 (CPC) 现已 正在研究作为成人心力衰竭患者刺激心脏功能的潜在治疗选择。 我们的初步研究已经确定，糖尿病和高血糖在体外都会诱发一系列的 c-kit+ CPC 中的细胞功能障碍，与糖尿病引起的 CHD 的病因有关。消除 c- 心脏发生过程中的 kit+ CPC 导致的 CHD 类似于糖尿病妊娠后代的 CHD。同样 重要的是确定孕产妇暴露于糖尿病期间对 产后衍生的 c-kit+ CPC 将用于我们即将进行的基于自体的 c-kit+ CHD 试验。 因此，我们假设糖尿病中的高血糖会导致 c-kit+ 细胞功能障碍 CPCs，导致心脏间隔缺陷并限制术后重塑效果 受损心脏上的出生衍生 c-kit+ CPC。减少细胞应激或 DNA 甲基化或 c-kit+ CPC 中的组蛋白乙酰化可减轻母体糖尿病诱发的 CHD，并改善 离体扩增的 c-kit+ CPC 通过恢复其旁分泌功能具有治疗价值。研究 专门设计用于揭示糖尿病或高血糖对 c-kit+ CPC 功能的影响。目标1将 确定细胞应激诱导的 c-kit+ CPC 功能障碍是否会导致致畸性 母亲患有糖尿病。我们假设糖尿病会引发细胞凋亡并减少 c-kit+ 的细胞增殖 CPC 通过细胞应激，损害心脏间隔和关键心脏间隔的功能 调节因子：第二心脏区祖细胞和心脏神经嵴细胞。目标 2 将确定是否 c-kit+ CPC 中组蛋白乙酰化和 DNA 甲基化增强介导了以下不良反应 母亲糖尿病对心脏发生的影响以及对这些祖细胞的影响。我们假设 糖尿病减少的沉默调节蛋白脱乙酰酶 2 (SIRT2) 导致 DNA 高甲基化，导致 c-kit+ CPC 细胞功能障碍严重涉及心脏间隔改变和不良印记。目标3将 确定后代衍生的 c-kit+ CPC 及其外泌体的治疗能力 心肌梗死模型中的非糖尿病和糖尿病母亲以及糖尿病的胚胎心脏 怀孕。我们假设来自母亲糖尿病患者的后代 c-kit+ CPC 的能力较低 修复由于 miR-34a 上调导致的冠心病和心脏功能障碍，从而改变分泌组和 与非糖尿病母亲相比，外泌体分析，并保留高水平的细胞应激、组蛋白 CPC 治疗期间的乙酰化和 DNA 甲基化。