Multiomics and Functional Characterization Establish Druggable Targets for PVC-Driven Idiopathic VF

项目摘要 在美国，心脏性猝死(SCD)每年夺走30万人的生命，尽管有激进的尝试 表型-基因型相关性，约50%的原发性电性SCD患者不符合诊断标准 用于任何SCD综合征，并被标记为特发性室颤(IVF)。这种“包罗万象”的诊断 排除包括一组表型不明的个体，表现为心律失常，即 机械上没有被探索过。此外，如果没有明确的基因型-表型特征，临床实践 试管受精指南建议对异质性疾病进行同种治疗。认识到基因连锁 关于试管受精的研究已经失败，我们建议进行范式转换，以解决这些具有挑战性的知识差距。我们 建议将综合电生理和多组学输出的计算模型集成到 明确与浦肯野触发的室性早搏相关的体外受精亚型的机制基础。 体外受精)。与波尔多集团深度表型努力合作，后者最初描述了这些新出现的 体外受精、PVC-IVF患者的亚型被招募来创造诱导多能干细胞(IPSCs) 来自威斯康星州和波尔多。我们的IPSC实验系统具有明显的优势，包括集成PVC- 体外受精iPS-心肌细胞(iPS-CMS)与混合(心室肌细胞和浦肯野)细胞群 反映特定区域表型的计算心肌细胞模型。我们团队为iPSCs实验设计的 结合促进电气和功能成熟度的平台创新，包括整合iPS- 心脏成纤维细胞(iPS-CFs)及其在矽肺成人中的计算模型分析 肌细胞和组织。在我们的试验数据中，我们展示了使用IPSCs区分试管受精和 实验证据，并将这些数据集成到计算建模方法中，以获得机理 对细胞心律失常扰动的洞察。我们的首要目标是检查机械基础 并确定特定的互补和协同治疗靶点。在目标1中，我们将整合 我们先进的模型系统的实验功能读数，旨在概括自然心脏环境 结合微米和纳米尺度的线索和“自下而上”的计算模型来解开特定的 导致观察到的功能PVC-IVF读数的细胞扰动。目标2的重点是将 广泛、无偏见的数据驱动数据集，来自PVC-IVF患者特定的iPS-CMS的多组特征 计算系统药理学框架，用于定义协同致心律失常途径和 抗心律失常综合疗法，我们预测这将比单一疗法更安全和有效。 最后，计算跨细胞翻译器将预测成人心脏的反应。随着我们目标的完成， 我们将定义细胞心律失常的特征；解开下游的转录组，蛋白质表达 更改和翻译后修改；并将实验读数与计算建模相结合 创建可操作的数据并为PVC-IVF心律失常预防寻找可用药的靶点。

PROJECT ABSTRACT Sudden cardiac death (SCD) claims >300,000 lives yearly in the US and despite aggressive attempts at phenotype-genotype correlations, ~50% of patients with primary electrical SCD do not meet diagnostic criteria for any SCD syndrome and are labeled Idiopathic Ventricular Fibrillation (IVF). This “catch-all” diagnosis of exclusion encompasses a cohort of individuals that are undefined phenotypically with arrhythmia that is mechanistically unexplored. Moreover, without defined genotype-phenotype characterization, clinical practice guidelines for IVF suggest homogenous treatment for a heterogenous disorder. Recognizing that genetic linkage studies have failed for IVF, we propose a paradigm shift to address these challenging gaps in knowledge. We propose to integrate computational modeling of comprehensive electrophysiologic and multiomic outputs to identify the mechanistic underpinnings of an emerging IVF subphenotype related to Purkinje-triggered VF (PVC- IVF). In collaboration with the Bordeaux group Deep Phenotype efforts, who originally described these emerging subphenotypes of IVF, PVC-IVF patients have been recruited to create induced pluripotent stem cells (iPSCs) from UW and Bordeaux. Our iPSC experimental system has distinct advantages including integration of PVC- IVF iPS-cardiomyocytes (iPS-CMs) with a mixed (ventricular myocyte and Purkinje) cell population with computational myocyte models reflecting region-specific phenotypes. Our group’s design for iPSCs experiments combine innovation of platforms that promote electrical and functional maturity, including incorporation of iPS- cardiac fibroblasts (iPS-CFs), and analysis by computational modeling of iPS-CMs and in silico adult human myocytes and tissue. In our pilot data we demonstrate the effectiveness of using iPSCs to differentiate IVF with experimental evidence and integrate this data into computational modeling approaches to gain mechanistic insight into cellular arrhythmic perturbations. Our overarching goal is to examine the mechanistic underpinning of PVC-IVF and identify specific complementary and synergistic therapeutic targets. In Aim 1 we will integrate experimental functional readouts from our advanced model system designed to recapitulate native cardiac milieu with a combination of micro and nanoscale cues with “bottom-up” computational modeling to unravel the specific cellular perturbations that cause the observed functional PVC-IVF readout. The focus of Aim 2 is to incorporate a broad, unbiased data-driven dataset from multiomic characterization of PVC-IVF patient-specific iPS-CMs into a computational systems pharmacology framework to define synergistic arrhythmogenic pathways and antiarrhythmic polytherapy, which we predict to be safer and more effective than monotherapy approaches. Finally, computational cross-cell translators will predict responses in the adult heart. With completion of our aims, we will define the cellular arrhythmic signature; unravel the down-stream transcriptome, protein expression changes and post-translational modifications; and integrate experimental readouts with computational modeling to create actionable data and find druggable targets for PVC-IVF arrhythmia prevention.