CARDIAC MITOHORMESIS PROTECTS AGAINST DIABETIC CARDIOMYOPATHY THROUGH MITOPHAGY: EVALI ADMINISTRATIVE SUPPLEMENT

项目总结 本申请是根据NOT-HL-19-724号文件提交的。电子烟的使用一直很迅速 在美国，年轻人的人数正在增加，但长期使用电子烟对健康的影响尚不清楚。这个 电子烟的毒性最近因最近大量的急性肺损伤病例而凸显出来。 与电子烟使用或蒸发有关的疾病，共导致2 807例电子烟感染住院或死亡。是否 这些案件与含有非法化合物的电子液体或广泛使用的尼古丁的蒸发有关- 含有电子液体的情况仍有待确定。重要的是，EVALI患者的特征 首次出院后再次住院或死亡显示存在一种或多种并发症，包括心脏 疾病、呼吸道疾病和糖尿病，这表明电子烟使用者的某些队列正在增加 发生EVALI的风险。在亲本R00 HL130416的这一补充中，我们建议利用患者来源的人类 诱导多能干细胞(IPSC)平台在确定EVALI发病机制中的作用 关于电子烟或其成分的风险，以及易感因素的存在可能如何使结果恶化。我们 假设与健康细胞相比，患病的IPSC来源的细胞将表现出更高的细胞毒性 暴露在电子烟及其成分中。目标1将以IPSCs的区别为基础 不同的危险因素(健康、单独糖尿病、糖尿病合并心肌病)转化为内皮细胞和肺泡 上皮细胞。当这些细胞暴露在电子烟或维生素E醋酸酯及其有毒衍生酮中时， 将进行细胞健康的表型表征，以确定这些成分中的哪一种诱导 毒性最大的。在目标2中，我们将研究电子烟诱导的内皮功能障碍的机制和 关注p62/NRF2/Keap1抗氧化应激途径，因为我们观察到只有健康细胞才有 暴露后持续的p62水平。我们还将对分离出的外切体进行蛋白质组学分析 暴露时健康和患病的IPSC-ECs，以确定潜在预测的新生物标志物 EVALI的发展。总而言之，这项行政补编提案将创造新的机会 建立以人为基础的IPSC储存库，代表可用于评估的不同风险因素 蒸发造成的健康后果。生物化学方法与蛋白质组学的结合也将 允许对某些患者对EVALI易感性增加背后的机制有更深的了解 并提供可预测EVALI发展的生物标志物。

PROJECT SUMMARY This application is being submitted in response to NOT-HL-19-724. The use of e-cigarettes has been rapidly increasing among young adults in the US but the health effects of prolonged e-cigarette use are unknown. The toxicity of e-cigarette was recently highlighted by the large number of recent cases of acute lung injury associated with e-cigarette use or vaping (EVALI), resulting in a total of 2,807 hospitalized EVALI cases or deaths. Whether these cases are linked to the vaping of illicit compounds-containing e-liquids or the widely used nicotine- containing e-liquids remain to be determined. Importantly, characterization of EVALI patients who had to be rehospitalized or died after initial discharge revealed the presence of one or more comorbities including cardiac disease, respiratory diseases and diabetes which suggests certain cohort of e-cigarette users are at increased risk of EVALI. In this Supplement to the Parent R00 HL130416, we propose to leverage patient-derived human induced pluripotent stem cell (iPSC) platform towards determing the pathogenesis of EVALI as a consequence of e-cigarette or its constituents and how the presence of susceptibility factors may worsen the outcome. We hypothesize diseased iPSC-derived cells will exhibit increased cytotoxicity compared to healthy cells when exposed to e-cigarette and its constituents. Aim 1 will be based on the differentiation of iPSCs representing different risk factors (healthy, diabetes alone, diabetes with cardiomyopathy) into endothelial cells and alveolar epithelial cells. Upon exposure of these cells to e-cigarette or vitamin E acetate and its toxic derivative ketene, phenotypic characterization of cellular health will be performed to determine which of these constituents induces the most toxicity. In Aim 2, we will investigate the mechanism of e-cigarette-induced endothelial dysfunction and focus on the p62/NRF2/KEAP1 anti-oxidative stress pathway since we observed that only healthy cells had sustained levels of p62 upon exposure. We will also perform proteomic profiling of exosomes isolated from healthy and diseased iPSC-ECs upon exposure in order to identify novel biomarkers that can potentially predict the development of EVALI. In summary, this Administrative Supplement proposal will create novel opportunities to establish a human-based iPSC repository representing different risk factors that can be used to evaluate health outcomes from vaping. The combination of biochemical methodologies coupled with proteomics will also allow a deeper mechanistic understanding behind the increased susceptibility of certain patients against EVALI and provide biomarkers that may be predictive of EVALI development.