Physiologic stress in advanced tissue culture models of cardiomyopathy

心肌病高级组织培养模型中的生理应激

基本信息

批准号：
10592151
负责人：
Dominic Edward Fullenkamp
金额：
$ 16.41万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-15 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10592151
关键词：
3-Dimensional Affect Annexin A6 Arrhythmia Award Biological Markers Biology Biomedical Engineering CRISPR/Cas technology Cardiac Cardiac Myocytes Cardiology Cardiomyopathies Cardiovascular system Cell Line Cell Nucleus Cell membrane Cells Clinical Code Collaborations Complex Data Defect Development Development Plans Dilated Cardiomyopathy Disease Doctor of Philosophy Duchenne muscular dystrophy Dystrophin Effectiveness Electrophysiology (science)Engineering Ensure Extracellular Matrix Genes Genetic Genetic Diseases Genetic Medicine Heart Heart failure Human Hydrogels Injury Intermediate Filaments Lamin Type A Lead Left Ventricular Dysfunction Life Link Measurement Measures Mechanical Stress Mechanics Membrane Mentors Mentorship Methods Modeling Morbidity - disease rate Muscular Dystrophies Mutation Myocardium Nuclear Envelope Nuclear Lamina Output Pathologic Patients Phenotype Physicians Physiological Play Predisposition Proteins Recombinants Research Research Personnel Role Sarcomeres Scientist Signal Transduction Signaling Protein Skeletal Muscle Stress Structural Protein System Technology Testing Therapeutic Effect Tissue constructs Tissues Training Troponin Universities Work bench to bedside cardiac tissue engineering career development clinical translation clinically relevant comparison control disease-causing mutation drug testing electrical measurement exon skipping experience flexibility flexible electronics gene therapy improved individualized medicine induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes inhibitor interest male monolayer mortality mutant novel novel therapeutic intervention novel therapeutics protein complex prototype response stressor therapeutic evaluation tissue culture transmission process two-dimensional

项目摘要

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.

项目概要/摘要多种形式的肌营养不良症的特点是心脏受累，可能危及生命，包括杜氏肌营养不良症 (DMD) 和 LMNA 相关肌营养不良症。这些疾病是分别由 DMD 和 LMNA 突变引起。心力衰竭是发病的主要原因这些疾病的死亡率。骨骼和心肌的生理机械应力是主要驱动因素这些形式的肌营养不良症。另外，心律失常，向左不成比例心室功能障碍在 LMNA 相关疾病中尤为突出。新的治疗策略，例如外显子跳跃、基因治疗和 CRISPR-Cas9 基因编辑正在开发或已被批准用于这些领域疾病。可以创建患者特异性诱导多能干细胞并分化为心肌细胞（iPSC-CM），提供了研究患者特异性突变并在培养皿中测试疗法的机会。然而，iPSC-CM 受到组织不成熟、非生理应激源和非生理输出的限制。工程心脏组织是通过将 iPSC-CM 接种到由柔性材料支撑的水凝胶基质中来创建的帖子。通过这些组织构建体，可以实现更高级的细胞成熟度，并且可以轻松确定收缩速度和幅度等输出。 K08 奖项的具体目标 1 目的评估生理机械应力对 iPSC-CM 的影响。具体目标 2 将改善使用源自患者的新型工程心脏组织结构评估致心律失常的可能性含有 DMD 和 LMNA 突变的 iPSC-CM。将评估这两种疾病的新疗法。初步数据表明 DMD iPSC-CM 对机械损伤的敏感性增加通过一种新的治疗方法来缓解和缓解这种表型。工程心脏组织原型是提出将允许对这些组织进行直接电生理检查。我们期待这个原型将允许更统一的节奏，将是有用的，特别是在表征新的治疗方法方面对 LMNA iPSC-CM 的影响。该 K08 将支持 PI Dominic Fullenkamp 的职业发展计划医学博士，博士，接受了生物医学工程研究生培训，是医师科学家心脏病专家。 Fullenkamp 博士的长期目标是成为一名独立研究者，成为一名医学科学家心血管生物学和工程学的接口，并使用该接口来测试新的疗法。在除了拟议的研究之外，富伦坎普博士还希望利用该奖项进一步发展他在以下领域的培训：心血管生物学在伊丽莎白·麦克纳利医学博士和阿尔弗雷德·乔治医学博士（两位）的指导下西北大学经验丰富且协作的导师。他的发展计划将得到以下方面的支持与他的研究兴趣直接相关的内部和外部课程以及临床活动。