Integration Free IPS Cells-Derived Progenitors for Cardiac Regeneration

用于心脏再生的免整合 IPS 细胞衍生祖细胞

• 批准号: 8839043
• 负责人: Muhammad Ashraf
• 金额: $ 73.37万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8839043
• 关键词: Animals; Apoptosis; Autologous; Behavior; Biology; Blood Vessels; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Line; Cell Lineage; Cell Therapy; Cell surface; Cells; Characteristics; Chromatin; Cicatrix; DNA; Data; Development; Engraftment; Enzymes; Epigenetic Process; Experimental Animal Model; Family suidae; Fibroblasts; Fibrosis; Generations; Genetic; Genome; Goals; Health; Hearing problem; Heart; Heart Diseases; Human; Hypoxia; Infarction; Ischemic Preconditioning; Methods; Modeling; Molecular Biology; Mus; Muscle Cells; Myoblasts; Myocardial; Myocardial Infarction; Myocardium; Natural regeneration; Nature; Patients; Physiology; Pluripotent Stem Cells; Population; Process; Proteins; Recombinants; Research; Risk; Signal Transduction; Skeletal Muscle; Skeletal Myoblasts; Source; Staging; Stem cells; Techniques; Testing; Therapeutic; Tissues; Transferase; Transplantation; Ultrasonography; Up-Regulation; Vascular Endothelial Growth Factors; Viral; Viral Vector; attenuation; base; cardiac regeneration; design; experience; functional outcomes; heart function; improved; in vivo; induced pluripotent stem cell; inhibitor/antagonist; innovation; interdisciplinary approach; pluripotency; pre-clinical; preconditioning; progenitor; public health relevance; regenerative therapy; repaired; small molecule; stem; stem cell therapy; transcription factor; tumor; vector-induced

DESCRIPTION (provided by applicant): The generation of induced pluripotent stem (iPS) cells is an innovative approach for generating autologous pluripotent stem cell lines for individualized cell therapy. Our research will use human skeletal muscle derived myoblasts rather than terminally differentiated fibroblasts for non-viral generation of iPS and their differentiation int cardiac progenitor cells. The hypothesis is that skeletal myoblasts (SMs) are superior candidates for induction to pluripotent state with fewer factors either alone or in combination with treatment with small molecules. Thus iPS derived cardiac progenitors may be readily generated with the use of cardiogenic small molecules, purified to generate off shelf universal cardiac cells. The direct generation of progenitors from iPS cells with specific small molecule may be a major current paradigm shift in stem cell therapy. We further propose that use of iPS derived cardiovascular progenitors will allow successful regeneration of infarcted myocardium without the risk of tumorgenecity. The hypotheses will be tested in the following specific Aims. Specific Aim-1will generate iPS cells from human SMs using small molecules; Specific Aim-2 will focus on developing strategies to direct iPS cells to cardiac and vascular progenitors; Specific Aim-3 will exploit the power of Ischemic preconditioning signaling in regulating survival and engraftment of iPS -progenitors in the ischemic tissue for effective regeneration; Specific Aim 4 will test that transplantation of iPS - progenitors and preconditioned progenitors effectivel regenerates infarcted myocardium and reverses fibrosis in murine and pre-clinical porcine heart models. The end points of the in vivo studies will be reversal of fibrosis through myoangiogenic differentiation of the engrafted progenitor cells, functional integration of developing cardiac myocytes into the host heart, attenuation of infarct size and the functional benefits in terms of improved global heart function. These studies will involve multidisciplinary approach which will employ state of the art molecular biology, histochemical and immunohistochemical techniques and well integrative physiology involving well established experimental animal model, and transthoracic ultrasonography for animal heart function. These studies are expected to facilitate robust cardiac differentiation and cardiomyocyte purification resulting in generation of unlimited number of cardiac progenitor cells from SM-iPS for restoring damaged myocardium without the risk of tumor formation.

描述(申请人提供)：诱导多能干细胞(IPS)的产生是一种用于个体化细胞治疗的产生自体多能干细胞系的创新方法。我们的研究将使用人骨骼肌源性成肌细胞而不是终末分化的成纤维细胞进行iPS的非病毒生成和向心脏前体细胞的分化。假设骨骼肌母细胞(SMS)是诱导多能状态的更好的候选细胞，无论是单独还是结合小分子治疗，诱导的因素更少。因此，iPS来源的心脏祖细胞可以很容易地通过使用生心小分子来生成，并进行纯化以生成现成的通用心肌细胞。从具有特定小分子的iPS细胞直接产生祖细胞可能是当前干细胞治疗中的一个重大范式转变。我们进一步提出，使用iPS来源的心血管祖细胞将允许成功地再生梗死心肌，而不会有肿瘤发生的风险。这些假设将在以下具体目标中得到检验。特异性目标-1将利用小分子从人的短信中产生iPS细胞；特异性目标-2将专注于开发策略，将iPS细胞定向至心脏和血管前体细胞；特异性目标-3将利用缺血预适应信号调节iPS-前体细胞在缺血组织中的存活和植入，以实现有效的再生；特异性目标4将在小鼠和临床前猪心脏模型中测试iPS-前体细胞和预适应前体细胞移植是否有效地再生梗死心肌并逆转纤维化。体内研究的终点将是通过移植的前体细胞的肌肉血管生成分化逆转纤维化、发育中的心肌细胞与宿主心脏的功能整合、缩小梗死面积以及改善整体心脏功能方面的功能益处。这些研究将涉及多学科方法，将使用最先进的分子生物学、组织化学和免疫组织化学技术，以及良好的综合生理学，包括已建立的实验动物模型，以及动物心脏功能的经胸超声检查。这些研究有望促进强大的心脏分化和心肌细胞纯化，从而从SM-iPS中产生无限数量的心脏前体细胞，用于修复受损的心肌，而不会有肿瘤形成的风险。