Project 2: Ex Vivo Modeling and Analysis of Gastric Precancerous Lesions

项目2：胃癌前病变的离体建模与分析

• 批准号: 10715763
• 负责人: MANUEL R AMIEVA
• 金额: $ 32.26万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715763
• 关键词: 3-Dimensional; Address; Anatomy; Antibiotics; Apical; Atrophic; Bacterial Genes; Basic Science; Behavior; Biological; Biology; Biopsy; Cancer Etiology; Carcinoma; Categories; Cell Communication; Cell Polarity; Cell model; Cells; Cellular biology; Cessation of life; Characteristics; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Confocal Microscopy; Correa cascade; DNA Sequence Alteration; Data; Development; Disease Progression; Dysplasia; Endoscopic Biopsy; Epithelial Cells; Epithelium; Experimental Models; Exposure to; Gastric Glands; Gastric Tissue; Gastritis; Gene Expression; Gene Expression Profile; Genes; Genomics; Genotype; Gland; Growth; Helicobacter Infections; Helicobacter pylori; Human; Immune response; In Situ; In Vitro; Infection; Inflammation; Inflammatory; Intestines; Laboratory Research; Lesion; Libraries; Location; Malignant Neoplasms; Measures; Methods; Microscopy; Modeling; Molecular; Monitor; Mutation; Organoids; Pathogenesis; Patients; Peptide Initiation Factors; Persons; Phenotype; Precancerous Conditions; Process; Property; Quantitative Microscopy; Recrudescences; Research; Research Personnel; Resolution; Resources; Risk; Risk Factors; Risk Marker; Role; Sampling; Site; Stomach; Stomach Carcinoma; Surface; System; Techniques; Testing; Time; Tissue Model; Tissues; Tumor Stem Cells; Virulence Factors; Visualization; carcinogenesis; chronic infection; epithelial stem cell; experimental study; fitness; gastric intestinal metaplasia; gastric organoids; genetic selection; high risk; human tissue; improved; insight; malignant stomach neoplasm; mortality; neoplastic; next generation sequencing; novel; premalignant; programs; response; risk stratification; single-cell RNA sequencing; stem; stem cell niche; stem cell population; stem cells; tissue culture; tumor progression

ABSTRACT – PROJECT 2 The central purpose of Project 2 is to utilize a primary human tissue model to investigate the features of gastric stem cells and how the cells interact with Helicobacter pylori (Hp). Hp commonly triggers an inflammatory process that leads to gastric intestinal metaplasia (GIM), a condition that can evolve into invasive carcinoma. However, not all people with GIM develop gastric cancer. This project proposes an experimental platform to investigate the features of high-risk GIM. Its hypothesis is that high-risk GIM lesions possess epithelial stem cells with unique properties predisposed to neoplastic progression, especially in the setting of Hp infection. Project 2 has three aims to investigate this hypothesis: (1) Characterizing the epithelial progenitor cells of high-risk GIM. (2) Modeling genomic alterations from high-risk GIM. (3) Elucidating interactions between Hp and gastric organoids from high-risk GIM. For Aim 1, Dr. Amieva’s group will develop a library of stem-cell rich, apical-out gastric organoids representing the gastric neoplastic spectrum across various topographical locations in the stomach. The characteristics of these stem cell organoids will be identifying using next generation sequencing and advanced microscopy. In Aim 2, the organoids will be perturbed with select genetic alterations (identified in Project 1) through CRISPR gene editing. Subsequent changes to gene expression may provide insight into the biology of high-risk versus low-risk GIM tissues. Lastly in Aim 3, the investigators will infect apical-out gastric organoids (from Aim 1) with human-derived Hp strains of known genotypes and phenotypes. Single-cell genomics will allow investigators to distinguish the unique transcriptional profiles of organoids at different time points, states of infection and inflammation, and enable studies into bacterial genes that are crucial for colonization and host perturbations. While Project 2 will be conducted in a basic research laboratory, its aims and results are directly related to improving overall understanding of Hp behavior in the human stomach, and molecular changes that prime tissue for carcinogenesis.

摘要 – 项目 2 项目2的中心目的是利用原代人体组织模型来研究胃的特征 干细胞以及细胞如何与幽门螺杆菌 (Hp) 相互作用通常会引发炎症。 导致胃肠化生 (GIM) 的过程，这种情况可以演变成浸润性癌。 然而，并非所有患有 GIM 的人都会患上胃癌。该项目提出了一个实验平台来治疗胃癌。 研究高危GIM的特征，其假设是高危GIM病变具有上皮干细胞。 具有易于肿瘤进展的独特特性，尤其是在 Hp 感染的情况下（项目 2）。 研究这一假设有三个目的： (1) 表征高危 GIM 的上皮祖细胞。 (2) 对高风险 GIM 的基因组改变进行建模。 (3) 阐明 Hp 与高危 GIM 胃类器官之间的相互作用。 对于目标 1，Amieva 博士的团队将开发一个富含干细胞的顶端出胃类器官库，代表 跨越胃不同部位的胃肿瘤谱。 这些干细胞类器官将使用下一代测序和先进的显微镜进行识别。 目标 2，类器官将通过 CRISPR 受到选择性基因改变（在项目 1 中确定）的干扰 基因编辑的后续变化可能会提供对高风险与高风险的生物学的深入了解。 最后，在目标 3 中，研究人员将用目标 1 感染顶端外的胃类器官。 已知基因型和表型的人源 Hp 菌株将使研究人员能够进行单细胞基因组学研究。 区分类器官在不同时间点、感染状态的独特转录谱 炎症，并能够研究对定植和宿主扰动至关重要的细菌基因。 虽然项目 2 将在基础研究实验室进行，但其目标和结果与 提高对人胃中 Hp 行为以及启动组织的分子变化的整体了解 对于致癌作用。