Mechanisms enhancing functional coupling between native and embryonic stem cell d

增强天然干细胞和胚胎干细胞之间功能耦合的机制

• 批准号: 8686485
• 负责人: Karen Maass
• 金额: $ 16.76万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-26 至 2015-09-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8686485
• 关键词: Adherens Junction; Adult; Affect; Animals; Autologous; Biological; Breeding; Cardiac; Cardiac Myocytes; Cell Communication; Cell Size; Cell Transplantation; Cell Transplants; Cells; Coculture Techniques; Collaborations; Connexin 43; Connexins; Coupling; Data; Desmosomes; Engraftment; Failure; Freezing; Funding; Funding Opportunities; Future; Gap Junctions; Genes; Goals; Grant; Heart; Heart Diseases; Hurricane; Intercalated disc; Knockout Mice; Label; Lead; Mechanics; Medicine; Molecular; Mus; Mutant Strains Mice; Myocardial Infarction; Myocardium; Natural regeneration; Neonatal; Pathway interactions; Pilot Projects; Pluripotent Stem Cells; Proteins; Rattus; Regulation; Replacement Therapy; Research; Research Personnel; Sampling; Source; Stem cells; Testing; Therapeutic; Therapeutic Uses; Tissue Sample; Tissues; Transplantation; Treatment Efficacy; base; embryonic stem cell; falls; gap junction channel; heart function; high throughput screening; improved; in vivo; intercellular connection; interest; medical schools; mutant; public health relevance; research study; restoration; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): The goal of this application is to obtain funding through the "Restoration of New Investigator Pilot Projects Adversely Affected by Hurricane Sandy" opportunity. In October 2012, I had collected strong preliminary data and was preparing an R01 application for the June 2013 deadline. My research was seriously impacted by the after-effects of Super-storm Sandy. My losses include a colony of connexin43 (gap junction) mutant mice, generated as recipients for cell-transplantation studies, a breeding colony of rats, as well as precious frozen cell lysates and tissue samples awaiting molecular and histological analyses. Experiments proposed in this application will restore pilot project data towards a competitive grant submission, anticipated for summer/fall 2014. My long-term research interest is improving the therapeutic use of pluripotent stem cells (PSC) for heart diseases. PSC allow gene manipulation and can be differentiated into functional heart muscle cells in the dish. Already, PSC are being used for personalized medicine approaches, and PSC will be a suitable source for autologous cell replacement therapy in the future. To be seriously contemplated for therapeutic applications, however, several challenges remain, including long-term survival and functional engraftment of transplanted cells. Engraftment of cells is enhanced by intercellular connections, or cell-cell contacts, formed by specialized proteins. Cell contacts provide structural tissue support (mechanical junctions: adherens junctions; desmosomes) and allow fast impulse propagation and the synchronous contraction of the heart muscle (electrical junctions: gap junction channels). Indeed, transplantation of cardiomyocytes derived from PSC lacking the gap junction protein connexin43, lead to increased arrhythmogenesis in mice. Our preliminary data demonstrate that stem cell derived cardiomyocytes form very few functional gap junction contacts with native cardiomyocytes. The overarching hypothesis of this proposal is that an increase in gap junction channel formation and function will improve the therapeutic efficacy of stem cell derived cardiomyocytes. I will employ three different approaches to test this hypothesis. Based on preliminary data, I will investigate how the formin protein Daam1 enhances gap junction formation. Further, I will analyze the influence of forced gap junction channel formation for cell engraftment using PSC expressing a mutant gap junction channel, K258stop. Additionally I will use a high throughput screen to identify small molecules regulating gap junction expression in stem cell derived cardiomyocytes. While experiments proposed in this study will elucidate ways to enhance the formation of gap junctions in stem cell derived cardiomyocytes, the mechanisms identified might very well also be applicable and therapeutically relevant to the treatment of cardiac diseases related to changes in gap junction formation.

描述（由申请人提供）：本申请的目标是通过“恢复受飓风桑迪影响的新调查员试点项目”的机会获得资金。2012年10月，我收集了大量的初步数据，并准备在2013年6月的截止日期前提交R 01申请。我的研究受到超级风暴桑迪的严重影响。我的损失包括一群连接蛋白43（间隙连接）突变小鼠，作为细胞移植研究的受体，一群繁殖的大鼠，以及珍贵的冷冻细胞裂解物和等待分子和组织学分析的组织样本。本申请中提出的实验将恢复试点项目数据，以提交竞争性赠款，预计将于2014年夏季/秋季提交。我的长期研究兴趣是改善多能干细胞（PSC）用于心脏病的治疗用途。PSC允许基因操作，并且可以在培养皿中分化成功能性心肌细胞。PSC已经被用于个性化医疗方法，PSC将成为未来自体细胞替代疗法的合适来源。然而，要认真考虑治疗应用，仍然存在一些挑战，包括移植细胞的长期存活和功能性植入。细胞的植入通过由专门蛋白质形成的细胞间连接或细胞-细胞接触来增强。细胞接触提供结构组织支持（机械连接：粘附连接;桥粒），并允许快速脉冲传播和心肌同步收缩（电连接：间隙连接通道）。事实上，移植来源于缺乏差距连接蛋白连接蛋白43的PSC的心肌细胞，导致小鼠中心肌细胞生成增加。我们的初步数据表明，干细胞衍生的心肌细胞与天然心肌细胞形成非常少的功能性间隙连接接触。该提议的首要假设是间隙连接通道形成和功能的增加将改善干细胞衍生的心肌细胞的治疗功效。我将采用三种不同的方法来测试这一点 假说.基于初步的数据，我将调查如何的蛋白质Daam 1增强间隙连接的形成。此外，我将使用表达突变间隙连接通道K258 stop的PSC分析强制间隙连接通道形成对细胞植入的影响。此外，我将使用高通量筛选来鉴定调节干细胞衍生的心肌细胞中间隙连接表达的小分子。虽然本研究中提出的实验将阐明增强干细胞衍生的心肌细胞中间隙连接形成的方法，但所确定的机制也可能非常适用于与间隙连接形成变化相关的心脏疾病的治疗。