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

项目摘要 本申请是根据NOT-HL-19-724提交的。电子烟的使用已经迅速 在美国，电子烟在年轻人中的使用率正在上升，但长期使用电子烟对健康的影响尚不清楚。的 电子烟的毒性最近被大量的急性肺损伤病例所强调， 使用电子烟或电子烟（EVALI），共导致2，807例EVALI住院病例或死亡。是否 这些案件与吸食含有非法化合物的电子液体或广泛使用的尼古丁有关， 是否含有电子液体仍有待确定。重要的是，必须接受EVALI治疗的患者的特征 初次出院后再次住院或死亡，显示存在一种或多种合并症，包括心脏病 疾病，呼吸系统疾病和糖尿病，这表明某些电子烟使用者群体正在增加 风险评估。在母药R 00 HL 130416的补充中，我们建议利用患者源性人源性 诱导多能干细胞（iPSC）平台确定EVALI的发病机制， 电子烟或其成分的影响，以及易感因素的存在如何使结果恶化。我们 假设患病iPSC衍生细胞与健康细胞相比将表现出增加的细胞毒性， 接触电子烟及其成分。目标1将基于iPSC的分化， 不同的风险因素（健康、单纯糖尿病、糖尿病伴心肌病）进入内皮细胞和肺泡 上皮细胞当这些细胞暴露于电子烟或维生素E乙酸酯及其有毒衍生物乙烯酮时， 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 最大的毒性。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， 我们关注p62/NRF 2/KEAP 1抗氧化应激途径，因为我们观察到只有健康细胞才有 p62在暴露后的持续水平。我们还将对分离的外来体进行蛋白质组学分析， 健康和患病的iPSC-EC，以鉴定可以潜在地预测 EVALI的发展。总之，这项行政补充提案将创造新的机会 建立一个基于人类的iPSC储存库，代表不同的风险因素， vaping的健康结果。生物化学方法与蛋白质组学的结合也将 允许更深入地了解某些患者对EVALI敏感性增加背后的机制 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.