The Role of PRDM16 in Cardiac Development and Cardiomyopathy

PRDM16 在心脏发育和心肌病中的作用

基本信息

批准号：
10408821
负责人：
Sihem Boudina
金额：
$ 68.64万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10408821
关键词：
1p36 Adult Affect Age Alleles Arrhythmia Binding Brown Fat Calcium Cardiac Cardiac Myocytes Cardiac development Cardiomyopathies Cell Death Cell Respiration Cells Cessation of life ChIP-seq Child Chromosome Deletion Clinical Clustered Regularly Interspaced Short Palindromic Repeats Code Data Deletion Mutation Deterioration Development Diagnosis Dilated Cardiomyopathy Disease Etiology Fatty Acids Fibrosis Future Gene Deletion Gene Expression Genes Genetic Screening Genetic Transcription Heart Heart Hypertrophy Heart failure Hematopoiesis Human Hypertrophic Cardiomyopathy Individual Knock-in Knock-in Mouse Knockout Mice Lead Left ventricular non-compaction Mediating Metabolic Mitochondria Molecular Morbidity - disease rate Morphology Mus Mutation Myocardial dysfunction Non-compaction cardiomyopathy Nonsense Mutation Nucleotides Oxidative Phosphorylation Pathologic Pathology Pathway interactions Patients Perinatal mortality demographics Phenotype Play Proteins RNA Splicing Regulation Research Role Syndrome Tamoxifen Testing Translational Research Transplantation Variant Work Zinc Fingers arrhythmogenic cardiomyopathy cardiogenesis de novo mutation early onset genomic locus heart function induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes innovation insight mortality mouse model multidisciplinary mutant novel pediatric patients proband programs promoter stem cell function sudden cardiac death therapy development transcription factor transcriptome sequencing treatment strategy young adult

项目摘要

Cardiomyopathies are the major cause of morbidity and mortality in children, with death or transplant occurring in nearly half the patients within the first 2 years of diagnosis. Cardiomyopathies are genetically heterogeneous and mutations in genes encoding sarcomeric, cytoskeletal, mitochondrial, and calcium-handling proteins have been associated with the disease. We identified two pediatric patients with de novo non-sense and splice variant mutations in the transcription factor PR domain 16 (PRDM16) presenting with early onset left ventricular non-compaction (LVNC) that progressed to dilated cardiomyopathy (DCM) in one patient. Furthermore, cardiomyocytes-specific loss of Prdm16 in mice causes LVNC and perinatal lethality while heterozygous mice develop cardiac hypertrophy. Similarly, heterozygous CRISPR knock-in of the non-sense mutation also developed cardiac hypertrophy. To overcome the perinatal lethality in these mice and to characterize the role of PRDM16 in adult cardiac cells, we also generated mice with cardiomyocytes-specific tamoxifen inducible deletion of this gene and show that, similar to the germline heterozygous mice and the Prdm16 knock-in mice, these mice also developed cardiac hypertrophy that progressed into cardiac dysfunction with age. Our preliminary characterization of human induced pluripotent stem cell-derived cardiomyocytes from one proband and cardiomyocytes-specific Prdm16 KO mice reveal a dual role for this transcription factor in gene transcription. Lack of Prdm16 repressed metabolic and oxidative phosphorylation genes whereas it increased fibrotic gene expression. The forgoing scientific premise leads us to hypothesize that PRDM16 plays transcriptional roles pertinent to heart development, metabolic programing and cardiac remodeling and its loss results in LVNC/DCM phenotypes in humans and mice. Our specific aims are: (1) to elucidate the mechanisms underlying cardiomyopathy phenotypes consequent to PRDM16 mutations/deletion in humans and mice and (2) to identify transcriptional mechanisms by which PRDM16 regulates different gene programs in the heart. We have assembled a multidisciplinary team to conduct this highly translational research. This study is novel and highly significant as it shows that PRDM16 is an orchestrator of gene expression in the heart and that its loss in humans and in mice causes LVNC/DCM. Identifying PRDM16 targets and their involvement in cardiomyopathies is key towards a better understanding of the etiology of LVNC/DCM and for the development of therapies for these heterogeneous diseases.

心肌病是儿童发病和死亡的主要原因，近一半的患者在诊断后的头两年内发生死亡或移植。心肌病具有遗传异质性，编码肌节、细胞骨架、线粒体和钙处理蛋白的基因突变与该疾病有关。我们发现两名儿科患者的转录因子 PR 结构域 16 (PRDM16) 中存在从头无义和剪接变异突变，表现为早发性左心室致密化不全 (LVNC)，其中一名患者进展为扩张型心肌病 (DCM)。此外，小鼠心肌细胞特异性缺失 Prdm16 会导致 LVNC 和围产期死亡，而杂合子小鼠则出现心脏肥大。同样，无义突变的杂合 CRISPR 敲入也导致了心脏肥大。为了克服这些小鼠的围产期致死率并表征 PRDM16 在成年心肌细胞中的作用，我们还生成了心肌细胞特异性他莫昔芬诱导删除该基因的小鼠，并表明，与种系杂合子小鼠和 Prdm16 敲入相似小鼠中，这些小鼠还出现了心脏肥大，并随着年龄的增长而发展为心脏功能障碍。我们对来自一名先证者的人诱导多能干细胞来源的心肌细胞和心肌细胞特异性 Prdm16 KO 小鼠的初步表征揭示了该转录因子在基因转录中的双重作用。 Prdm16 的缺乏会抑制代谢和氧化磷酸化基因，同时会增加纤维化基因的表达。上述科学前提使我们假设 PRDM16 发挥与心脏发育、代谢程序和心脏重塑相关的转录作用，其缺失导致人类和小鼠的 LVNC/DCM 表型。我们的具体目标是：(1) 阐明人类和小鼠中 PRDM16 突变/缺失导致的心肌病表型的机制；(2) 确定 PRDM16 调节心脏中不同基因程序的转录机制。我们组建了一个多学科团队来进行这项高度转化的研究。这项研究是新颖且非常重要的，因为它表明 PRDM16 是心脏中基因表达的协调器，并且它在人类和小鼠中的缺失会导致 LVNC/DCM。确定 PRDM16 靶点及其与心肌病的关系是更好地了解 LVNC/DCM 病因和开发这些异质性疾病疗法的关键。