STEM CELLS AND DIABETIC CARDIOMYOPATHY PROJ 2 2009

干细胞和糖尿病心肌病项目 2 2009

• 批准号: 8168211
• 负责人: Jia-Qiang He
• 金额: $ 19.36万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168211
• 关键词: ABCB1 gene; Antigens; Apoptosis; Architecture; Biological Assay; Cardiac; Cell Death; Cell Proliferation; Cells; Computer Retrieval of Information on Scientific Projects Database; Diabetes Mellitus; Diabetic mouse; Echocardiography; Exhibits; Flow Cytometry; Freezing; Funding; Gene Chips; Gene Proteins; Grant; Growth; Heart; Heart Transplantation; Hyperglycemia; Immunohistochemistry; In Vitro; Infarction; Institution; Insulin-Dependent Diabetes Mellitus; Modeling; Mus; Myocardium; Non-Insulin-Dependent Diabetes Mellitus; Pattern; Protein Array; Proto-Oncogene Protein c-kit; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Source; Staining method; Stains; Stem cells; Structure; TP53 gene; Techniques; Testing; Tissues; Transplantation; United States National Institutes of Health; Western Blotting; cytotoxicity; diabetic; diabetic cardiomyopathy; immunocytochemistry; in vivo; non-diabetic; pluripotency; protein expression; research study; stemness

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. PROJECT #2: STEM CELLS AND DIABETIC CARDIOMYOPATHY The central hypothesis of this project is that diabetes produces negative effects on the growth reserve of cardiac stem cells (CSC) so that the enhanced cell's death cannot be counteracted by repopulating cells that preserve the architecture and function of the myocardium. To test this hypothesis, the murine models of type 1 and 2 diabetes (T1D, T2D) will be used in the following specific aims (SA): 1. SA1 is to examine whether CSC from normal mice will exhibit differences of stemness and pluripotency compared with CSC from diabetic mice. The whole heart of T1D and T2D mice will be used to isolate and identify CSC with c-kit positive, sca1 positive, MDR1 positive, also Lineage negative antigens and/or frozen tissue sections from those tissues will be stained for the markers. The total number of CSC, distribution in cardiac tissue, stem cell proliferation, and in vitro cardiac lineage differentiation will be compared between diabetic and non-diabetic groups. Flow cytometry, immunocytochemistry, immunohistochemistry, and echocardiography techniques will be used in these experiments. 2. SA2 is to investigate how diabetes and hyperglycemia impairs the stemness and cardiac lineage differentiation of CSC isolated from diabetic and non-diabetic hearts. Gene and protein expression patterns in c-kit, sca1, MDR1, AMPK, P53, PI3K/Akt, Nxk2.5, Glut1&4 of CSC from diabetic hearts will be compared with those from non-diabetic CSC. Cardiac lineage markers will be checked before and after in vitro differentiation. To mimic the in vivo diabetic condition, normal CSC of control hearts will also undergo in vitro hyperglycemia treatments. The underlying mechanism related with the genes and protein change under in vivo and in vitro conditions will be explored accordingly. Gene chip, protein array, Western blot, RT-PCR technique, and cell apoptosis/viability/cytotoxicity assays will be used in the analyses. 3. SA3 is to determine whether transplantation of CSC from normal mice will result in greater improvement in cardiac structure and function in infarcted diabetic mouse heart compared with CSC isolated from diabetic mouse heart. The normal mice and diabetic mice will be used to evaluate cardiac function and structural improvements with transplanted CSC. Echocardiography, histological, and pathological techniques will be used in the studies.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 项目＃2：干细胞和糖尿病心肌病 该项目的中心假设是糖尿病会对心脏干细胞（CSC）的生长储备产生负面影响，因此不能通过重新填充保留心肌体系结构和功能的细胞来抵消增强细胞的死亡。为了检验这一假设，将在以下特定目的（SA）中使用1型和2型糖尿病（T1D，T2D）的鼠模型： 1。SA1是要检查正常小鼠的CSC是否与糖尿病小鼠的CSC相比，是否会表现出干性和多能性的差异。 T1D和T2D小鼠的整个心脏将用于隔离和鉴定C-KIT阳性，SCA1阳性，MDR1阳性的CSC，也将对这些组织染色。将比较糖尿病和非糖尿病基团之间的CSC总数，心脏组织中的分布，干细胞增殖和体外心脏谱系分化。这些实验将使用流式细胞仪，免疫细胞化学，免疫组织化学和超声心动图技术。 2。SA2是为了研究糖尿病和高血糖如何损害与糖尿病和非糖尿病心脏分离的CSC的干性和心脏谱系分化。 C-KIT，SCA1，MDR1，AMPK，P53，PI3K/AKT，NXK2.5，CSC中CSC的基因和蛋白质表达模式将与非糖尿病心脏的CSC进行比较。 心脏谱系标记将在体外分化前后检查。为了模仿体内糖尿病状况，对照心脏的正常CSC也将接受体外高血糖治疗。将相应地探讨与体内和体外条件下基因和蛋白质变化有关的基本机制。分析将使用基因芯片，蛋白质阵列，蛋白质阵列，蛋白质阵列，RT-PCR技术以及细胞凋亡/生存力/细胞毒性测定。 3。SA3是为了确定与从糖尿病小鼠心脏分离的CSC相比，从正常小鼠接受CSC是否会导致心脏结构和功能的改善。正常小鼠和糖尿病小鼠将使用移植的CSC评估心脏功能和结构改善。超声心动图，组织学和病理技术将用于研究。