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）：心肌病是危及生命的心脏肌肉疾病，通常在家庭中运行。这种疾病有多种形式，所有这些形式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。虽然没有治愈方法，但如果在早期阶段被发现，医生可以减少疾病的影响。早期检测方案目前依赖于基因检测，以确定个体是否携带约1400种已知的心肌病突变之一。如果一个新的心肌病患者碰巧携带一个已知的突变，他们的家庭成员可以进行有意义的筛查。那些携带突变的人将得到预防性护理，而非携带者可以安全地免于临床随访，并减轻过度的焦虑。不幸的是，只有40-60%的心肌病患者对经验证的突变检测呈阳性。其余约占所有患者家庭的一半，除非采取进一步措施，否则无法从早期诊断的基因检测中获益。这些家庭的选择有限。共分离分析可以在足够大的家族中进行，并且可以鉴定新的基因突变，但不能保证该结果。如果家庭规模较小或不适合进行共分离研究，则必须定期对所有一级亲属进行检测，以确定是否有新出现的疾病迹象-即使他们有50%的机会不携带突变。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），来自一个人自己的体细胞通过表达多能干细胞因子，可能提供一个解决方案。我们设想了一种方法，其中iPSC衍生的心肌细胞（iPSC-CM）从患者中产生并形成工程心脏组织（EHT）用于体外研究。我们假设在EHT中测量的异常行为将反映存在的任何突变的直接影响。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，而不需要像共分离分析那样需要一个大家庭。考虑到这一特定目标，我们开发了一种新的策略来产生理想的生物力学表型的EHT。我们通过将心肌细胞接种到薄的、激光切割的去细胞猪心肌切片中来制备这些带状标本。用新生大鼠心室肌细胞进行的初步试验已经产生了自发跳动的EHT，它能产生可测量的收缩力，并能在培养物中存活数月。该项目的目标是通过从对照和患病受试者中产生iPSC-CM来评估这种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结果的能力。我们的目标的成功完成将是一个重要的一步，以替代基因检测的早期识别的个人谁是在发展心肌病的风险。