Physiologic stress in advanced tissue culture models of cardiomyopathy

项目摘要/摘要 多种形式的肌营养不良以心脏受累为特征，可能危及生命， 包括杜氏肌营养不良症(DMD)和LMNA相关性肌营养不良症。这些疾病是 分别由DMD和LMNA突变引起。心力衰竭是发病率和死亡率的主要原因 这些疾病的死亡率。骨骼肌和心肌中的生理性机械应力是导致 这些形式的肌肉营养不良的疾病。此外，心律失常，不成比例地向左 心功能不全，在LMNA相关疾病中是突出的。新的治疗策略，如外显子 跳过、基因治疗和CRISPR-Cas9基因编辑正在开发中或已被批准用于这些 疾病。患者特异性诱导的多能干细胞可以被创造并分化为心肌细胞 (IPSC-CMS)，提供了研究患者特定突变和测试碟子中的治疗方法的机会。 然而，IPSC-CMS受到组织不成熟、非生理性应激源和非生理性输出的限制。 工程心脏组织是通过将IPSC-CMS种植在由柔性支架支撑的水凝胶基质中而形成的 帖子。通过这些组织结构，可以实现更高级的细胞成熟，并且 可以很容易地确定收缩速度和幅度等输出。本K08奖项的具体目标1 目的探讨生理性机械应力对IPSC-CMS的影响。具体目标2将改善 应用患者来源评估新型工程化心脏组织结构的致心律失常潜能 携带DMD和LMNA突变的IPSC-CMS。将对这两种疾病的新疗法进行评估。 提供的初步数据表明DMD IPSC-CMS对机械的敏感性增加 一种新的治疗方法对这种表型的压力和缓解。一种工程心脏组织原型是 这将允许对这些组织进行直接电生理讯问。我们预料到了这一点 原型，这将允许更均匀的起搏，将是有用的，特别是在表征新的治疗方法 对LMNA IPSC-CMS的影响。这款K08将支持圆周派多米尼克·富伦坎普的职业发展计划 医学博士，他接受了生物医学工程的研究生培训，是内科科学家、心脏病学家。 富伦坎普博士的长期目标是成为一名独立的研究员，成为一名内科科学家 心血管生物学和工程学的接口，并使用这个接口来测试新的治疗方法。在……里面 除了提出的研究之外，富伦坎普博士的目标是利用这一奖项进一步发展他在 在伊丽莎白·麦克纳利医学博士和阿尔弗雷德·乔治医学博士的指导下的心血管生物学 西北大学的经验丰富和协作的导师。他的发展计划将得到 与他的研究兴趣直接相关的内部和外部课程和临床活动。

PROJECT SUMMARY/ABSTRACT Multiple forms of muscular dystrophy are characterized by cardiac involvement which can be life-threatening, including Duchenne muscular dystrophy (DMD) and LMNA-related muscular dystrophy. These diseases are caused by mutations in DMD and LMNA, respectively. Heart failure is a leading cause of morbidity and mortality in these diseases. Physiologic mechanical stress in skeletal and cardiac muscle is a main driver of disease in these forms of muscular dystrophy. In addition, cardiac arrhythmias, disproportionate to left ventricular dysfunction, are prominent in LMNA-related disease. New therapeutic strategies, such as exon skipping, gene therapy, and CRISPR-Cas9 gene editing, are in development or have been approved for these diseases. Patient-specific induced pluripotent stem cells can be created and differentiated into cardiomyocytes (iPSC-CMs), offering the opportunity to study patient-specific mutations and test therapeutics in the dish. However, iPSC-CMs are limited by tissue immaturity, non-physiologic stressors, and non-physiologic outputs. Engineered heart tissues are created by seeding iPSC-CMs in a hydrogel matrix that is supported by flexible posts. With these tissue constructs, a more advanced degree of cellular maturation can be achieved, and outputs such as contraction velocity and amplitude can be readily determined. Specific Aim 1 of this K08 award aims assess the effect of physiologic mechanical stress on iPSC-CMs. Specific Aim 2 will improve the assessment of arrhythmogenic potential in novel engineered heart tissue constructs using patient-derived iPSC-CMs harboring DMD and LMNA mutations. Novel therapeutics for both disease entities will be assessed. Preliminary data is presented that demonstrates increased susceptibility of DMD iPSC-CMs to mechanical stress and mitigation of this phenotype by a novel therapeutic. An engineered heart tissue prototype is presented that will allow for the direct electrophysiologic interrogation of these tissues. We expect this prototype, which will allow for more uniform pacing, will be useful, especially in characterizing novel therapeutic effects on LMNA iPSC-CMs. This K08 will support the career development plan for the PI, Dominic Fullenkamp MD PhD, who received his graduate training in biomedical engineering and is physician-scientist cardiologist. Dr. Fullenkamp's long-term objective is to become an independent investigator as a physician scientist at the interface of cardiovascular biology and engineering, and use this interface to test novel therapeutics. In addition to the research proposed, Dr. Fullenkamp aims to use this award to further develop his training in cardiovascular biology under the mentorship of Elizabeth McNally MD PhD and Alfred George MD, two experienced and collaborative mentors, at Northwestern University. His development plan will be supported by internal and external coursework and clinical activity that directly correlates with his research interests.