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

利用人类食道干细胞模型来了解损伤修复过程中的种族差异

• 批准号: 10514960
• 负责人: Daysha Ferrer-Torres
• 金额: $ 9.13万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-22 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10514960
• 关键词: 3-Dimensional; Acids; Address; African American population; American; Area; Artificial Intelligence; Barrett Esophagus; Bile Acids; Bile fluid; Biological Assay; Biological Models; CRISPR/Cas technology; Cell Death; Cell Line; Cells; Clinical; Committee Members; Coupled; DNA Damage; Data; Dimensions; Disease; Drug Compounding; Drug Screening; Enzymes; Epithelial; Esophageal Adenocarcinoma; Esophageal Diseases; Esophageal Tissue; Esophageal injury; Esophagus; European; FDA approved; Fellowship; GSTT2 gene; Gastric Acid; Gastroesophageal reflux disease; Gene Expression Profiling; Genes; Genetic; Genome; Goals; Health; Homeostasis; Human; Image; Image Analysis; Immersion; In Vitro; Incidence; Individual; Injury; Investigation; Lead; Libraries; Machine Learning; Mediating; Mediation; Mentors; Metaplasia; Methods; Michigan; Modeling; Modification; Molecular; Molecular Profiling; Molecular Target; Natural Products; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Play; Population Genetics; Population Heterogeneity; Prevalence; Reactive Oxygen Species; Reporting; Research; Resources; Role; Stomach; Techniques; Testing; Tissue Sample; Tissues; Training; Universities; Work; acid stress; base; biobank; cell injury; cohort; data acquisition; design; drug discovery; experimental study; fitness; genetic manipulation; human stem cells; human tissue; imaging modality; in vivo; injury and repair; loss of function; prevent; programs; racial disparity; racial diversity; repaired; response; response to injury; screening; severe injury; single-cell RNA sequencing; stem cell model; stem cells; tissue stem cells; tool; transcriptomics; two-dimensional; vector

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.

项目概要/摘要 非裔美国人 (AA) 和欧洲裔美国人 (EA) 胃食管反流的患病率相似 疾病（胃食管反流病）。尽管如此，与 EA 相比，AA 的食道损伤发生率较低， 化生和食管腺癌。群体遗传学和分子研究表明 人体组织中这些差异的特定基因；然而，人类食道缺乏种族多样性 模型阻碍了对其机制和潜在治疗方案的进一步研究。我们开发了一个 人类食道的祖先多样化干细胞/类器官生物库和高内容、基于图像的 询问胆汁酸损伤反应的筛选试验。结果显示 AA 食管细胞有反应 与 EA 衍生细胞显着不同，反映组织分析和临床结果。此外，我们 之前报道过一种关键酶，谷胱甘肽转移酶 theta-2 (GSTT2)，负责 灭活活性氧，从而减少 DNA 损伤，并在 AA 中高度表达 食管。利用祖先多样化的干细胞模型，我们展示了 GSTT2 低水平的关键关联 具有较高的损伤，与人体组织对损伤的主要反应一致。然而，GSTT2 的直接作用 这种反应和维持上皮稳态和食管细胞适应性的机制/药物 仍有待阐明。假设：非裔美国人的食管组织对胃的反应不同 由于解毒酶 GSTT2 表达较高而导致的酸/胆汁损伤，以及可以稳定的化合物 GSTT2 将保护细胞免受损伤。该提案要研究的三个具体目标将涉及 原代组织和干细胞来源的体外培养物，以验证分子特征和损伤差异 EA 和 AA 细胞在单细胞水平上的反应（目标 1），通过 GSTT2 的基因操作 确定针对伤害的保护的直接调解（目标 2），以及高通量无偏见 损伤反应的表征与药物筛选相结合，以确定能够抑制的化合物 胆汁/酸损伤（目标 3）。 Ferrer-Torres 博士在 K99.R00 计划中的主要研究目标是开发高 吞吐量技术将使她能够研究祖先不同的人群及其对伤害的反应。 因此，K99阶段已规划用于干细胞基因修饰和高含量的训练 基于表型的药物发现。在这一指导阶段，Ferrer-Torres 博士将与 Jason Spence 博士合作 乔纳森·塞克斯顿博士担任联合导师。受指导的 K99 计划是为 Ferrer-Torres 博士设计的 获得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 密歇根大学提供卓越的资源。这将推动 Ferrer-Torres 博士的目标 帮助她建立独立的研究计划，重点关注食管疾病的种族差异 她的R00阶段。