Engineered Tissue for Biomechanical Phenotyping of Cardiomyopathy Patients

用于心肌病患者生物力学表型分析的工程组织

• 批准号: 8974854
• 负责人: STUART G CAMPBELL
• 金额: $ 20.81万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8974854
• 关键词: Affect; Age; Analysis of Variance; Anxiety; Authorship; Behavior; Biomechanics; Blood Tests; Cardiac; Cardiac Myocytes; Cardiology; Cardiomyopathies; Cell Line; Cells; Clinical; Collagen; Disease; Disease Progression; Early Diagnosis; Early identification; Engineering; Family; Family member; Family suidae; First Degree Relative; Gene Mutation; Genes; Genetic Markers; Genetic Screening; Genetic screening method; Goals; Health; Heart; Heart failure; Human; In Vitro; Individual; Inheritance Patterns; Inherited; Lasers; Life; Measurable; Measurement; Measures; Methods; Mind; Muscle Cells; Mutation; Myocardial; Myocardium; Myopathy; Nature; Neonatal; Parents; Patients; Pediatric Cardiomyopathy; Persons; Phenotype; Physicians; Pluripotent Stem Cells; Preventive care; Publishing; Rattus; Reporting; Reproducibility; Rest; Risk; Running; Scheme; Somatic Cell; Son; Source; Specimen; Staging; Stem Cell Factor; Stretching; Surrogate Markers; System; Technology; Test Result; Testing; Tissue Engineering; Tissues; Training; United States; Ventricular; Work; cell growth; design; disease phenotype; high risk; high throughput screening; improved; induced pluripotent stem cell; member; multidisciplinary; new technology; novel; novel strategies; scaffold; segregation; stem cell biology; success; sudden cardiac death

DESCRIPTION (provided by applicant): Cardiomyopathies are life threatening heart muscle disorders that often run in families. The disease takes many forms, all of which place patients at risk for pathological heart remodeling, sudden cardiac death, and heart failure. While no cure exists, physicians can reduce the impact of the disease if it is caught in its early stages. Early detection schemes currently rely on genetic testing, to see if an individual carries one of the ~1400 known cardiomyopathy mutations. If a new cardiomyopathy patient happens to carry a known mutation, their family members can be meaningfully screened. Those carrying the mutation will receive preventative care, while non-carriers can be safely excused from clinical follow-ups and relieved of undue anxiety. Unfortunately, only 40-60% of cardiomyopathy patients test positive for a verified mutation. The rest, comprising roughly half of all patient families, cannot benefit from genetic testing for early diagnosis unless further steps are taken. The options for such families are limited. Co-segregation analysis can be performed in families of sufficient size and may identify a novel gene mutation, but this result is not guaranteed. If th family is small or otherwise unsuited for co-segregation study, all first-degree relatives must be periodically tested for signs of emerging disease - even though there is a 50% chance that they do not carry the mutation. Hence, in cases where genetic testing results are indeterminate, a more robust, single-repetition method for early detection of cardiomyopathy inheritance is badly needed. Induced pluripotent stem cells (iPSC), derived from a person's own somatic cells by expression of pluripotent stem cell factors, may provide a solution. We envision an approach in which iPSC-derived cardiomyocytes (iPSC-CMs) are generated from a patient and formed into engineered heart tissue (EHT) for in vitro study. We hypothesize that abnormal behavior measured in EHTs will reflect direct effects of any mutations present. As such, these phenotypes could act as surrogate markers of inheritance. By testing EHTs of each family member, the pattern of inheritance could be established regardless of the subjects' ages, and without requiring a large family, as in co-segregation analysis. With this specific goal in mind, w have developed a novel strategy to generate EHTs that are ideal for biomechanical phenotyping. We produce these ribbon-like specimens by seeding cardiac cells into thin, laser-cut sections of decellularized porcine myocardium. Initial tests with neonatal rat ventricular myocytes have yielded spontaneously beating EHTs that produce measurable contraction forces and can survive months in culture. The goal of this project is to assess the true potential of this exciting new technology by generating EHTs with iPSC-CMs from control and diseased subjects. We can then test the ability of our system to reproducibly detect the phenotypic consequences of even clinically mild mutations. The successful completion of our aims will be an important step toward an alternative to genetic testing for the early identification of individuals who are at risk of developing cardiomyopathy.

描述（由申请人提供）：心肌病是一种危及生命的心肌疾病，通常在家族中遗传。这种疾病有多种形式，所有这些都使患者面临病理性心脏重塑、心源性猝死和心力衰竭的风险。虽然尚无治愈方法，但如果在疾病的早期阶段发现，医生可以减少疾病的影响。目前的早期检测方案依赖于基因检测，以确定个体是否携带约 1400 种已知心肌病突变中的一种。如果新的心肌病患者恰好携带已知突变，则可以对其家庭成员进行有意义的筛查。携带突变的人将接受预防性护理，而非携带者可以安全地免除临床随访并缓解过度的焦虑。不幸的是，只有 40-60% 的心肌病患者经验证的突变检测呈阳性。其余的人（约占所有患者家庭的一半）无法从早期诊断的基因检测中受益，除非采取进一步措施。这些家庭的选择是有限的。共分离分析可以在足够大的家族中进行，并且可以识别新的基因突变，但不能保证这个结果。如果家庭规模较小或不适合共同隔离研究，则所有一级亲属都必须定期接受新发疾病迹象的检测——即使他们有 50% 的可能性不携带突变。因此，在基因检测结果不确定的情况下，迫切需要一种更强大的单次重复方法来早期检测心肌病遗传。诱导多能干细胞（iPSC）是通过表达多能干细胞因子从人自身的体细胞中衍生出来的，它可能提供一种解决方案。我们设想了一种从患者体内产生 iPSC 衍生心肌细胞 (iPSC-CM) 并形成工程心脏组织 (EHT) 的方法，用于体外研究。我们假设 EHT 中测量到的异常行为将反映任何存在的突变的直接影响。因此，这些表型可以作为遗传的替代标记。通过测试每个家庭成员的 EHT，无论受试者的年龄如何，都可以建立遗传模式，并且不需要像共分离分析那样需要一个大家庭。考虑到这一具体目标，我们开发了一种新策略来生成非常适合生物力学表型分析的 EHT。我们通过将心肌细胞接种到脱细胞猪心肌的激光切割薄切片中来生产这些带状样本。对新生大鼠心室肌细胞的初步测试已经产生了自发跳动的 EHT，它产生可测量的收缩力，并且可以在培养物中存活数月。该项目的目标是通过使用来自对照和患病受试者的 iPSC-CM 生成 EHT，来评估这项令人兴奋的新技术的真正潜力。然后，我们可以测试我们的系统重复检测临床轻微突变的表型后果的能力。我们目标的成功实现将是迈向基因检测替代方案的重要一步，以早期识别有患心肌病风险的个体。