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.

项目摘要 先天性心脏缺陷（CHD）是最常见的先天缺陷。孕产妇糖尿病是 与CHD和心脏功能障碍增加五倍相关的非秘密因素。基础 糖尿病引起的CHD和心脏功能障碍的机制尚不清楚，但一种机制可能 涉及高葡萄糖水平抑制心脏病。 C-KIT+心脏祖细胞（CPC）现在为 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葡萄糖在体外诱导了一系列 C-KIT+ CPC中的细胞功能障碍与糖尿病诱导的CHD的病因有关。消除c- 心脏病发生过程中的试剂盒+ CPC导致类似于糖尿病妊娠后代的CHD。同样 重要的是要确定母体糖尿病暴露期间对 产后衍生的C-KIT+ CPC将用于我们即将进行的自体C-KIT+ CHD试验。 因此，我们假设糖尿病中的高葡萄糖会诱导C-kit+的细胞功能障碍 CPC有助于心脏分离缺陷，并限制了后的重塑作用 纳塔尔派生的c-kit+ CPC在受损的心脏上。减少细胞应激或DNA甲基化或 C-KIT+ CPC中的组蛋白乙酰化减轻了母体糖尿病诱导的CHD，并改善 体内的治疗值通过恢复其旁分泌功能扩展了C-KIT+ CPC。研究 专门设计用于在C-KIT+ CPC功能上揭示糖尿病或高葡萄糖。目标1意志 确定细胞应力诱导的C-KIT+ CPCS功能障碍是否有助于致畸性 母体糖尿病。我们假设糖尿病会触发凋亡并减少C-KIT+的细胞增殖 CPC通过细胞应激，会损害心脏分离和关键心脏分离的功能 调节剂：第二心脏野外祖细胞和心脏神经rest细胞。 AIM 2将确定是否 C-KIT+ CPC中增强的组蛋白乙酰化和DNA甲基化介导 母体糖尿病是心脏病发生的，并在这些祖细胞上印记。我们假设这一点 糖尿病还原的Sirtuin脱乙酰基酶2（SIRT2）引起DNA高甲基化，导致C-KIT+ CPCS 严重涉及改变心脏分离和不良印记的细胞功能障碍。目标3意志 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母亲和糖尿病的胚胎心脏 怀孕。我们假设母体糖尿病患者的后代派生的C-kit+ CPC具有较低的能力 在修复由于miR-34a上调引起的CHD和心脏功能障碍时，会改变分泌和 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疗法期间的乙酰化和DNA甲基化。