Cell type-Specific Therapeutic Targeting of canonical WNT Pathway in Arrhythmogenic Cardiomyopathy

致心律失常性心肌病典型 WNT 通路的细胞类型特异性治疗靶向

• 批准号: 10594529
• 负责人: Ali J Marian
• 金额: $ 49.36万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-10 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594529
• 关键词: Apoptosis; Arrhythmia; Arrhythmogenic Right Ventricular Dysplasia; Biological; Biological Assay; Cardiac; Cardiac Myocytes; Cardiomyopathies; Caspase; Cell Death; Cells; Connexin 43; Data; Desmosomes; Development; Diagnosis; Disease; Echocardiography; Electrophysiology (science); Epicardium; Epithelium; Exposure to; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Transcription; Heart; Heart failure; Histologic; Human; In Situ Nick-End Labeling; Intercalated disc; Intervention; Knowledge; Malignant Neoplasms; Maps; Mechanics; Molecular; Monitor; Mus; Muscle Cells; Mutation; Myocardial; Myocardium; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Phenotype; Prognostic Marker; Proteins; Publishing; Regulatory Pathway; Reporter; Reporting; Risk; Role; Signal Transduction; Site; Sodium Channel; Sudden Death; Tamoxifen; Techniques; Therapeutic; Transcript; Uncertainty; Ventricular; Ventricular Arrhythmia; arrhythmogenic cardiomyopathy; beta catenin; causal variant; cell type; circulating biomarkers; desmoplakin; diagnostic biomarker; effective therapy; frizzled related protein-3; gain of function; heart dimension/size; heart function; inducible Cre; inhibitor; lipid biosynthesis; loss of function; mechanotransduction; mouse model; novel diagnostics; paracrine; postnatal; prognostic; single-cell RNA sequencing; therapeutic target; transcriptome sequencing

Arrhythmogenic cardiomyopathy (ACM) is a myocardial genetic disease whose cardinal manifestations are ventricular arrhythmias, sudden death, myocardial fibro-adiposis, cell death, and heart failure. ACM is caused primarily by mutations in genes encoding desmosome proteins, including desmoplakin (DSP). In the heart, desmosomes are mainly present in myocytes and are responsible for myocardial mechanical integrity. They are also hubs for mechanosensing and transduction, including the Hippo and canonical WNT (cWNT) pathways, which are involved in ACM. Desmosome proteins are also expressed in the epicardium, the site of initial manifestations of ACM phenotype. Specific role(s) of the epicardial cells in the pathogenesis of ACM is unknown. The cWNT pathway has emerged as an attractive therapeutic target in ACM. Experimental data, however, are conflicting, with a spectrum ranging from suppression to activation of the cWNT in ACM. The ambiguity along with potential fortuitous effects of systemic targeting of this major regulatory pathway have raised considerable trepidations, leading to an impasse on therapeutic targeting of the cWNT pathway in ACM. We provide data in human and mouse hearts with ACM suggesting that the ambiguity is in part due to determining the cWNT activity in cellularly heterogeneous myocardium. Accordingly, whereas the cWNT is activated in cardiac myocytes, the whole myocardium is exposed to increased levels of secreted cWNT inhibitors (cWNT-Is). The dysregulation is remarkable in the human hearts with ACM as transcript levels of 26 cWNT-Is are increased, including SFRP3 protein, a classic cWNT-I. To resolve the uncertainty, we will use an uncommitted approach of activating or suppressing the cWNT pathway specifically in cardiac myocytes and epicardial cells and determining the phenotypic effects, including effects on expression of secreted cWNT-Is. Inducible Cre-deleter mice will be used to delete Dsp in cardiac myocytes or epicardial cells post-natally while concomitantly suppressing or activating the cWNT using loss- and gain-of-function b-catenin mice. Cardiac function, arrhythmias, apoptosis, fibro-adipogenesis and gene expression, including secretome in myocytes and epicardial cells will be characterized. Dsp+/- epicardial cells will be conditionally tagged using the dual reporter (Rosa26mT/mG) mice and their differentiation to other cardiac cells will be analyzed by fate mapping. Tagged cells derived from Dsp+/- epicardial cells will be isolated and analyzed by single cell RNA-Seq to determine their transcriptional identity and identify secretome expressed by each subset of epicardial-derived cells. Likewise, effects of activation or suppression of the cWNT pathway on differentiation of the Dsp+/- epicardial cells to other cardiac cell types and the secretome will be determined using gain- and loss-of-function b-catenin mice. The findings will determine whether cell-type specific targeting of the cWNT is a beneficial therapeutic approach in ACM, will define the role of epicardial cells in ACM, and could lead to identification of secreted factors that might serve as diagnostic and prognostic biomarkers and therapeutic targets in ACM.

致心律失常性心肌病（ACM）是一种心肌遗传性疾病，其主要表现 包括室性心律失常、猝死、心肌纤维脂肪变性、细胞死亡和心力衰竭。 ACM 引起 主要是由编码桥粒蛋白（包括桥粒斑蛋白（DSP））的基因突变引起的。在心里， 桥粒主要存在于肌细胞中，负责心肌的机械完整性。他们是 也是机械传感和转导的中心，包括 Hippo 和规范 WNT (cWNT) 通路， 涉及ACM。桥粒蛋白也在心外膜中表达，即初始的部位 ACM表型的表现。心外膜细胞在 ACM 发病机制中的具体作用尚不清楚。 cWNT 通路已成为 ACM 中有吸引力的治疗靶点。实验数据， 然而，这些都是相互矛盾的，ACM 中 cWNT 的范围从抑制到激活。这 这一主要监管途径的系统性目标的模糊性以及潜在的偶然影响已经引起了人们的关注。 相当大的恐惧，导致 ACM 中 cWNT 通路的治疗靶向陷入僵局。 我们通过 ACM 提供了人类和小鼠心脏的数据，表明这种模糊性部分是由于 确定细胞异质心肌中的 cWNT 活性。因此，而 cWNT 是 在心肌细胞中被激活，整个心肌暴露于分泌的 cWNT 抑制剂水平升高的环境中 （cWNT-Is）。在 ACM 人类心脏中，26 个 cWNT-Is 的转录水平的失调非常显着 增加，包括 SFRP3 蛋白，一种经典的 cWNT-I。为了解决不确定性，我们将使用 未明确的激活或抑制 cWNT 通路的方法，特别是在心肌细胞和 心外膜细胞并确定表型效应，包括对分泌的 cWNT-Is 表达的影响。 诱导型 Cre-deleter 小鼠将用于在出生后删除心肌细胞或心外膜细胞中的 Dsp 同时使用功能丧失和获得功能的 b-连环蛋白小鼠抑制或激活 cWNT。心脏 功能、心律失常、细胞凋亡、纤维脂肪生成和基因表达，包括肌细胞中的分泌组和 将表征心外膜细胞。 Dsp+/- 心外膜细胞将使用双报告器进行有条件标记 (Rosa26mT/mG) 小鼠及其与其他心脏细胞的分化将通过命运图谱进行分析。标记细胞 源自 Dsp+/- 心外膜细胞的细胞将被分离并通过单细胞 RNA-Seq 进行分析，以确定其 转录身份并识别由心外膜衍生细胞的每个子集表达的分泌组。同样地， 激活或抑制 cWNT 通路对 Dsp+/- 心外膜细胞分化为其他细胞的影响 将使用获得和丧失功能的β-连环蛋白小鼠来确定心肌细胞类型和分泌组。 这些发现将确定 cWNT 的细胞类型特异性靶向是否是一种有益的治疗方法 ACM 中的方法将定义心外膜细胞在 ACM 中的作用，并可能导致识别分泌的 可能作为 ACM 诊断和预后生物标志物以及治疗靶点的因素。

项目成果

Evaluation of Mavacamten in Symptomatic Patients With Nonobstructive Hypertrophic Cardiomyopathy.

Mavacamten 在有症状的非梗阻性肥厚型心肌病患者中的评价。

• 发表时间: 2020-06-02
• 影响因子: 24
• 作者: Ho, Carolyn Y;Mealiffe, Matthew E;Bach, Richard G;Bhattacharya, Mondira;Choudhury, Lubna;Edelberg, Jay M;Hegde, Sheila M;Jacoby, Daniel;Lakdawala, Neal K;Lester, Steven J;Ma, Yanfei;Marian, Ali J;Nagueh, Sherif F;Owens, Anjali;Rader, Florian
• 通讯作者: Rader, Florian

DNA double-stranded breaks, a hallmark of aging, defined at the nucleotide resolution, are increased and associated with transcription in the cardiac myocytes in LMNA-cardiomyopathy.

DNA 双链断裂是衰老的标志，以核苷酸分辨率定义，在 LMNA 心肌病中，DNA 双链断裂增加并与心肌细胞的转录相关。

• DOI: 10.1093/cvr/cvae063
• 发表时间: 2024-04-05
• 期刊: Cardiovascular research
• 影响因子: 10.8
• 作者: Benjamin Cathcart;Sirisha M Cheedipudi;Leila Rouhi;Zhongming Zhao;Priyatansh Gurha;A. J. Marian
• 通讯作者: A. J. Marian

A tribute to james thornton willerson, M.D.

向医学博士詹姆斯·桑顿·威尔森致敬

• 发表时间: 2020-10-18
• 影响因子: 10.8
• 作者: Marian; Aj
• 通讯作者: Aj