Cardiac Functional and Structural Implications of Lamin A/C Mutations

Lamin A/C 突变对心脏功能和结构的影响

基本信息

批准号：
9266679
负责人：
Anna Grosberg
金额：
$ 42.87万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-04 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9266679
关键词：
Affect Age of Onset Animal Model Architecture Arrhythmia Biological Assay Biomedical Engineering Cardiac Cardiac Myocytes Cardiomyopathies Cardiovascular Diseases Cardiovascular Models Cell Nucleus Cell physiology Cells Cellular Structures Clinical Complex Cytoskeleton DNA DNA Sequence Alteration Data Defect Dermal Development Diagnosis Diagnostic Disease Disease model Doctor of Philosophy Etiology Family Fibroblasts Frequencies Gene Expression Gene Mutation Genes Genetic Genomics Genotype Goals Heart Heart Abnormalities Heart Diseases Heart failure In Vitro Individual Inherited Knowledge Laboratories Lamin Type A Lead Length Mechanics Medical Methods Morbidity - disease rate Morphology Muscle Cells Muscle Weakness Mutation Nuclear Nuclear Envelope Nuclear Lamina Nuclear Matrix Nuclear Protein Organ Pathogenicity Pathway interactions Patients Phenotype Physiological Play Research Risk Role Severities Signal Transduction Stress Structure Structure-Activity Relationship Sudden Death Supporting Cell Symptoms Techniques Testing Therapeutic Tissue Engineering Tissues Translational Research Variant cell motility exome sequencing fluorescence imaging genomic variation heart cell heart function improved induced pluripotent stem cell innovation insight practical application programs public health relevance response screening self assembly sudden cardiac death trait transcriptome transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Cardiomyopathies and arrhythmia are conditions with high morbidity and limited therapies. Although a vast number of genes have been discovered to contribute to the etiology of these diseases, translational research- the practical application of genetic knowledge to improve screening, diagnosis, and treatment for affected individuals and their families-has been limited. One major obstacle is the lack of functional studies to understand the relationship between genotype and emergent phenotype at multiple physiological scales (cells to tissues) and to identify factors that cause clinical variability between and within families. The proposed study focuses on three affected families each with different mutation in the Lamin A/C (LMNA) gene. LMNA encodes the main protein of the nuclear lamina, the structural matrix of the nuclear envelope that interacts with both the cell nucleus and cytoskeleton. In this proposal, we will test the hypothesis that LMNA mutations are associated with defects in the structure, organization, and function at multiple length-scales. Between families, the severity of the defects is associated with the type of LMNA mutation; and within families, severity is modified by additional genetic factors. Our long term goals are to develop in vitro disease models directly from patients to understand how proper cell structure, tissue organization, and contractile function are affected by the mutation and genetic modifiers. In Specific Aim 1, we will use exome sequencing and in vitro tissue engineering techniques to evaluate genomic variation and defects in cell and nuclear morphology, intracellular architecture, motility, and tissue self-assembly and architecture in fibroblasts from LMNA patients and controls. In Specific Aim 2, we will derive induced pluripotent stem cells (iPS) from fibroblasts and analyze cell structure and tissue organization of iPS-derived cardiomyocytes from LMNA patients and controls. In Specific Aim 3, we will use RNA sequencing to test for altered gene expression and the "Heart-on-a-Chip" to characterize contractility function (frequency, systolic, diastolic, and twitch stresses) of iPS-derived cardiomyocytes from LMNA patients and controls. These results combined with information of genetic and phenotypic variances will imply pathways and functionalities we will further study. Understanding of the complex relationship between genotype and emergent phenotype and identifying modifying factors will provide insight into the mechanism of heart disease and may assist in the development of new preventative, diagnostic, and therapeutic strategies.

描述（由申请人提供）：心肌病和心律失常是发病率高且治疗方法有限的疾病，尽管已发现大量基因与这些疾病的病因学有关，但转化研究 - 遗传知识的实际应用以改善筛查，对受影响个人及其家人的诊断和治疗一直受到限制，一个主要障碍是缺乏功能研究来了解多个生理尺度（细胞到组织）的基因型和出现的表型之间的关系并识别。导致家庭之间和家庭内部临床变异的因素。拟议的研究重点关注三个受影响的家庭，每个家庭的核纤层蛋白 A/C (LMNA) 基因编码核膜的主要蛋白质，即核膜的结构基质。在本提案中，我们将测试 LMNA 突变与多个长度尺度的结构、组织和功能缺陷相关的假设。与类型LMNA 突变；在家庭中，严重程度会受到其他遗传因素的影响，我们的长期目标是直接从患者身上开发出体外疾病模型，以了解突变和遗传修饰因素如何影响适当的细胞结构、组织组织和收缩功能。具体目标 1，我们将使用外显子组测序和体外组织工程技术来评估 LMNA 患者和对照的成纤维细胞的基因组变异和细胞和核形态、细胞内结构、运动性以及组织自组装和结构的缺陷。在具体目标 2 中，我们将从成纤维细胞中衍生出诱导多能干细胞 (iPS)，并分析来自 LMNA 患者和对照的 iPS 衍生心肌细胞的细胞结构和组织组织。在具体目标 3 中，我们将使用 RNA 测序来测试改变的基因。表达和“芯片上的心脏”来表征来自 LMNA 的 iPS 衍生心肌细胞的收缩功能（频率、收缩压、舒张压和抽搐应力）这些与遗传和表型差异信息相结合的结果将意味着我们将进一步研究基因型和出现的表型之间的复杂关系并识别修饰因素，这将有助于深入了解心脏病的机制。开发新的预防、诊断和治疗策略。