Utilizing a human stem cell model of the esophagus to understand racial disparities during injury repair

项目总结/摘要 非裔美国人（AA）和欧洲裔美国人（EA）的胃食管反流患病率相似 疾病（GERD）。尽管如此，与EA相比，AA显示食管损伤的发生率较低， 化生和食管腺癌。群体遗传学和分子研究表明 人类组织中这些差异的特定基因;然而，缺乏种族多样性的人类食管 模型阻碍了对机制和潜在治疗方案的进一步研究。我们开发了一个 人类食管的祖先多样性干细胞/类器官生物库和一个高含量的基于图像的 筛选测定以询问胆汁酸损伤应答。结果表明，AA食管细胞响应 与EA衍生的细胞显著不同，反映了组织分析和临床发现。而且我们 我以前曾报道过一种关键酶，谷胱甘肽转移酶θ-2（GSTT 2），负责 失活活性氧，从而减少DNA损伤，并在AA中高度表达。 食道利用祖先多样性干细胞模型，我们显示GSTT 2低水平的关键关联 损伤程度更高，与人体组织对损伤的反应一致。然而，GSTT 2在 这种反应和机制/药物维持上皮稳态和适应食管细胞 仍有待阐明。假设：来自非裔美国人的食管组织对胃肠道反应不同 由于解毒酶GSTT 2的更高表达导致的酸/胆汁损伤，以及可以稳定 GSTT 2可以保护细胞免受损伤。本提案中要调查的三个具体目标将涉及 原代组织和干细胞衍生的体外培养物，以验证损伤的分子特征和差异 EA和AA细胞之间的反应，在单细胞水平（目的1），与GSTT 2的遗传操作， 确定保护免受伤害的直接介导（目标2），以及高通量无偏 损伤反应的表征与药物筛选相结合，以确定将抑制 胆汁/酸损伤（目的3）。Ferrer-Torres博士在K99.R00项目中的主要研究目标是开发高 通量技术，这将使她能够研究祖先不同的人群和他们对伤害的反应。 因此，K99阶段已计划在干细胞遗传修饰和高含量的培训 基于表型的药物发现在这个指导阶段，Ferrer-Torres博士将与Jason Spence博士合作 乔纳森·塞克斯顿博士的共同导师指导K99计划是为Ferrer-Torres博士的 掌握这些领域的专业知识。此外，Jules Lin博士和Marcia Cruz-Correa博士将担任顾问 博士后委员会成员和临床沉浸顾问。这将利用 密歇根大学提供的特殊资源。这将推动费雷尔-托雷斯博士的目标， 帮助她建立她的独立研究计划，重点是食管疾病的种族差异， R 00阶段

PROJECT SUMMARY/ABSTRACT African Americans (AA) and European Americans (EA) have a similar prevalence of gastro-esophageal reflux disease (GERD). Nonetheless, when compared to EA, AA show a lower incidence of esophagus damage, metaplasia, and esophageal adenocarcinoma. Population genetics and molecular studies have implicated specific genes for these differences in human tissue; however, a lack of racially diverse human esophagus models hinders further investigation into the mechanisms and potential treatment options. We developed an ancestrally diverse stem cell/organoid biobank of human esophagus and a high-content, image-based screening assay to interrogate bile-acid injury response. Results showed that AA esophageal cells responded significantly differently than EA-derived cells, mirroring tissue profiling and clinical findings. Furthermore, we have previously reported that a key enzyme, glutathione-transferase theta-2 (GSTT2), is responsible for inactivating reactive oxygen species, thus reducing DNA damage, and is highly expressed in the AA esophagus. Utilizing the ancestrally diverse stem cell model, we show key associations of GSTT2 low levels with higher injury, consistent with primary human tissue response to injury. However, a direct role of GSTT2 in this response and mechanism/drugs to maintain epithelial homeostasis and fitness to esophageal cells remains to be elucidated. Hypothesis: Esophageal tissue from African Americans respond differently to gastric acid/bile injury due to higher expression of detoxifying enzyme GSTT2, and compounds that can stabilize GSTT2 will protect cells against injury. The three specific aims to be investigated in this proposal will involve primary tissue and stem cell-derived in vitro cultures to validate the molecular profiles and differences in injury response between EA and AA cells at the single-cell level (Aim 1), with genetic manipulation of GSTT2 to determine direct mediation of protection against injury (Aim 2), and a high-throughput unbiased characterization of injury response coupled with a drug screen to determine compounds that will inhibit bile/acids injury (Aim 3). Dr. Ferrer-Torres’ primary research goals in the K99.R00 program is to develop high throughput techniques that will allow her to study ancestrally diverse populations and their response to injury. Therefore, the K99 phase has been planned to train in stem cell genetic modifications and high-content phenotypic-based drug discovery. For this mentored phase, Dr. Ferrer-Torres will work with Dr. Jason Spence and co-mentor by Dr. Jonathan Sexton. The mentored K99 program has been designed for Dr. Ferrer-Torres’ gain expertise in these areas. In addition, Dr. Jules Lin and Dr. Marcia Cruz-Correa will serve as advisory postdoctoral committee members and advisors for clinical immersions. This will be carried out utilizing the exceptional resources available at the University of Michigan. This will impulse Dr. Ferrer-Torres’ goals and help her establish her independent research program focusing on racial disparities in esophageal diseases in her R00 phase.