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

基本信息

批准号：
10651896
负责人：
Daysha Ferrer-Torres
金额：
$ 9万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-22 至 2024-08-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10651896
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 Cytoprotection DNA Damage Data Disease Drug Metabolic Detoxication Drug Screening Enzymes Epithelium Esophageal Adenocarcinoma Esophageal Diseases Esophageal Tissue Esophageal injury Esophagitis Esophagus European FDA approved Fellowship GSTT2 gene Gastric Acid Gastroesophageal reflux disease Gene Expression Profiling Genes Genetic 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 Postdoctoral Fellow Predisposition Prevalence Proliferating Reactive Oxygen Species Reporting Research Resources Role Stomach Techniques Testing Tissue Sample Tissues Training Universities Work biobank biological adaptation to stress cell injury cohort data acquisition design drug discovery experimental study fitness gain of function genetic manipulation genome editing 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 derived tissues stem cell model stem cells tool transcriptomic profiling transcriptomics two-dimensional vector

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 Cytoprotection DNA Damage Data Disease Drug Metabolic Detoxication Drug Screening Enzymes Epithelium Esophageal Adenocarcinoma Esophageal Diseases Esophageal Tissue Esophageal injury Esophagitis Esophagus European FDA approved Fellowship GSTT2 gene Gastric Acid Gastroesophageal reflux disease Gene Expression Profiling Genes Genetic 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 Postdoctoral Fellow Predisposition Prevalence Proliferating Reactive Oxygen Species Reporting Research Resources Role Stomach Techniques Testing Tissue Sample Tissues Training Universities Work biobank biological adaptation to stress cell injury cohort data acquisition design drug discovery experimental study fitness gain of function genetic manipulation genome editing 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 derived tissues stem cell model stem cells tool transcriptomic profiling transcriptomics two-dimensional vector

展开

项目摘要

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.

项目摘要/摘要非洲裔美国人（AA）和欧洲美国人（EA）的胃道胃反射率类似疾病（GERD）。尽管如此，与EA相比，AA显示出较低的食道损伤事件，化生和食管腺癌。人口遗传学和分子研究已经实施这些差异的特定基因；但是，缺乏多样化的人类食道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化的干细胞/器官生物库和一个基于图像的高素质筛选分析以询问胆汁酸损伤反应。结果表明AA食管细胞反应与EA衍生细胞的明显不同，反映组织分析和临床发现。此外，我们以前已经报道过，关键酶，谷胱甘肽转移酶theta-2（GSTT2）负责灭活活性氧，从而减少了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，我们显示了GSTT2低水平的关键关联损伤较高，与原发性人体组织对损伤的反应一致。但是，GSTT2在这种反应和机制/药物维持上皮稳态和对食管细胞的适应性尚待阐明。假设：来自非裔美国人的食管组织对胃的反应不同酸/胆汁损伤由于较高的排毒酶GSTT2表达而引起的，并且可以稳定的化合物 GSTT2将保护细胞免受损伤。在本提案中进行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，以验证分子特征和损伤的差异 EA和AA细胞在单细胞水平上的响应（AIM 1），GSTT2的基因操纵对确定免受伤害保护的直接调解（AIM 2）和高通量公正损伤反应的表征与药物筛查相结合以确定会抑制的化合物胆汁/酸损伤（AIM 3）。 Ferrer-Torres博士在K99.R00计划中的主要研究目标是发展高吞吐技术将使她能够研究祖先多样化的人群及其对伤害的反应。因此，已经计划在干细胞遗传修饰和高含量的情况下训练K99阶段基于表型的药物发现。在此问题阶段，Ferrer-Torres博士将与Jason Spence博士合作以及乔纳森·塞克斯顿（Jonathan Sexton）博士的联合学。指导的K99计划是为Ferrer-Torres博士设计的在这些领域获得专业知识。此外，Jules Lin博士和Marcia Cruz-Correa博士将担任咨询博士后委员会成员和临床沉浸式顾问。这将使用密歇根大学提供的卓越资源。这将冲动Ferrer-Torres博士的目标和帮助她建立她的独立研究计划，重点是食管疾病中的种族分布她的R00阶段。