The role of TGFβs and cFAPs in Cardiac Pathology from RNA Toxicity

TGFβ 和 cFAP 在 RNA 毒性心脏病理学中的作用

• 批准号: 10717904
• 负责人: Mani Subramaniam Mahadevan
• 金额: $ 80.7万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717904
• 关键词: Ablation; Adult; Affect; Alleles; Antibodies; Antisense Oligonucleotides; Arrhythmia; Attention; Behavior; Biological Assay; Biological Markers; Breeding; Cardiac; Cardiac Myocytes; Cause of Death; Child; Clinical; Clinical Pathology; Clinical Research; Collaborations; Data; Defect; Degenerative Disorder; Development; Dominant Genetic Conditions; Dose; Doxycycline; Electrocardiogram; Fatty Acids; Fibrosis; Future; Gene Deletion; Gene Expression; Genes; Genetic Diseases; Goals; Grant; Heart; Heart Abnormalities; Heart Diseases; Histology; Individual; Infiltration; Ligands; Lipid Inclusion; LoxP-flanked allele; Maintenance; Mediating; Molecular; Morbidity - disease rate; Mus; Muscle; Muscular Dystrophies; Myotonic Dystrophy; Myotonic dystrophy type 1; Nature; Nuclear; Organ; Outcome; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Pharmacologic Substance; Phenotype; Platelet-Derived Growth Factor alpha Receptor; Pre-Clinical Model; Production; Proliferating; Protein Isoforms; RNA; Reporting; Role; Signal Transduction; Stains; Sudden Death; Symptoms; System; Tamoxifen; Testing; Therapeutic; Therapeutic Effect; Therapeutic Studies; Time; Tissues; Toxic effect; Transforming Growth Factor Beta 2; Transforming Growth Factor beta; Transgenes; autosome; behavioral study; cardiac magnetic resonance imaging; coronary fibrosis; functional outcomes; inducible Cre; insight; interest; interstitial; mortality; mouse model; mutant; myotonic dystrophy protein kinase; next generation; novel; progenitor; receptor; response; stem cells; targeted treatment; therapeutic target; tool; treatment response

Project Summary: Myotonic dystrophy (DM1), the most common form of muscular dystrophy in adults and children, is an autosomal dominant genetic disorder caused by an expanded CTG repeat in the DM protein kinase (DMPK) gene that leads to nuclear retention of the mutant RNA and subsequent RNA toxicity. The heart is one of the primary organs affected in DM1. Cardiac conduction problems are present in up to 75% of adult DM1 cases, and sudden death due to cardiac arrhythmias is one of the most common causes of death in DM1. Unfortunately, the pathogenesis of cardiac manifestations in DM1 is not well understood. Clinical focus for cardiac disease in DM1 has been on arrhythmias and conduction abnormalities. Of note, the pathology of cardiac defects in DM1 has been historically associated with interstitial fibrosis, and fatty infiltration and fibrosis of cardiac conduction tissues. We reported the first inducible mouse model of RNA toxicity and cardiac conduction defects and demonstrated the potential for reversibility of DM1 phenotypes by silencing toxic RNA production. Recently using the DM200 mouse model, we showed for the first time, the potential for antisense oligonucleotides (ASOs) to treat cardiac disease in DM1. We also found evidence for fibrotic changes associated with RNA toxicity in the heart. In the past decade, the advent of new cardiac MRI (CMR) studies has led to evidence of and an increased interest in understanding cardiac fibrosis in DM1 and its role in the clinical pathology of DM1. We propose to use the DM200 mouse model as a tool for developing and investigating these ideas and concepts in a pre-clinical model and to try to understand the cellular and molecular drivers of fibro-adipogenic changes in the heart. We will do this through four independent but complementary aims. First, based on preliminary evidence of increased TGFβs expression in the heart, we will determine the role of TGFβs from cardiomyocytes, in RNA toxicity in the heart. Second, we will study the behavior of cardiac fibro-adipogenic progenitors (cFAP) cells and the role of TGFβ signaling in cFAPs in pathologic responses to RNA toxicity. Third, we will study the therapeutic response to therapies targeting TGFβs (isoform specific antibodies against TGFβ2 and TGFβ3). Fourth, we will evaluate next generation ASOs that target the toxic RNA. With both these classes of therapy, we will assess their therapeutic effect on fibroadipogenic changes, cFAPs, TGFβ expression and cardiac outcomes, by using ECGs and CMR and functional testing. The ultimate goal of this proposal is to elucidate critical players in the primary pathologies associated with RNA toxicity in the heart, and to identify pathways and therapies that may mitigate morbidity and mortality associated with cardiac disease in DM1.

项目概要： 强直性肌营养不良 (DM1) 是成人和儿童最常见的肌营养不良症，是一种常染色体遗传性肌营养不良症 由 DM 蛋白激酶 (DMPK) 基因中 CTG 重复扩增引起的显性遗传疾病，该基因导致 突变体 RNA 的核保留和随后的 RNA 毒性。心脏是主要器官之一 DM1 受到影响。高达 75% 的成人 DM1 病例存在心脏传导问题，并导致猝死 因心律失常是DM1最常见的死亡原因之一。不幸的是，发病机制 DM1 的心脏表现尚不清楚。 DM1 型心脏病的临床重点一直是 心律失常和传导异常。值得注意的是，DM1 心脏缺陷的病理学在历史上一直被认为是 与间质纤维化、心脏传导组织的脂肪浸润和纤维化有关。我们报道了 第一个诱导 RNA 毒性和心脏传导缺陷的小鼠模型，并证明了其潜力 通过沉默有毒 RNA 的产生来实现 DM1 表型的可逆性。最近使用DM200鼠标型号， 我们首次展示了反义寡核苷酸 (ASO) 治疗 DM1 型心脏病的潜力。 我们还发现了与心脏 RNA 毒性相关的纤维化变化的证据。在过去的十年里， 新的心脏 MRI (CMR) 研究的出现带来了证据并增加了人们对理解的兴趣 DM1 中的心脏纤维化及其在 DM1 临床病理学中的作用。我们建议使用 DM200 鼠标型号 作为在临床前模型中开发和研究这些想法和概念的工具，并尝试 了解心脏纤维脂肪生成变化的细胞和分子驱动因素。我们将通过以下方式做到这一点 四个独立但互补的目标。首先，基于 TGFβ 表达增加的初步证据 在心脏中，我们将确定心肌细胞中的 TGFβ 在心脏 RNA 毒性中的作用。第二，我们 将研究心脏纤维脂肪祖细胞 (cFAP) 的行为以及 TGFβ 信号在 cFAPs 对 RNA 毒性的病理反应。第三，我们将研究对疗法的治疗反应 靶向 TGFβ（针对 TGFβ2 和 TGFβ3 的异构体特异性抗体）。四、接下来我们来评价 生成针对有毒 RNA 的 ASO。对于这两种疗法，我们将评估它们的治疗效果 通过使用心电图和 CMR 对纤维脂肪生成变化、cFAP、TGFβ 表达和心脏结果的影响 和功能测试。该提案的最终目标是阐明主要病理学中的关键参与者 与心脏中的 RNA 毒性相关，并确定可以减轻发病率和 DM1 中与心脏病相关的死亡率。