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的中心目的是利用一个原始的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。惠普通常会引发炎症 导致胃肠化生(GIM)的过程，这是一种可能演变为浸润性癌的情况。 然而，并不是所有的GIM患者都会患上胃癌。该项目提出了一个实验平台，以 调查高危GIM的特点。它的假设是，高危的GIM病变具有上皮干细胞 具有易于肿瘤进展的独特性质，特别是在Hp感染的背景下。项目2 有三个目的来研究这一假设： (1)高危牙周炎上皮祖细胞的特征。 (2)从高危GIM中模拟基因组改变。 (3)阐明幽门螺杆菌与高危GIM的胃有机物质之间的相互作用。 对于目标1，阿米耶娃博士的团队将开发一个干细胞丰富的顶端取出的胃类器官的图书馆，这些器官代表 胃肿瘤横跨胃内不同部位的光谱。其特点是 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鉴定。在……里面 目标2，通过CRISPR，有机物将被选定的基因改变(在项目1中确定)所干扰 基因编辑。随后基因表达的变化可能提供对高危与非高危疾病生物学的洞察 低风险的牙膏纸巾。最后，在目标3中，研究人员将感染来自目标1的胃尖外器质。 已知基因型别和表型的人源性幽门螺杆菌菌株。单细胞基因组学将使研究人员能够 区分不同时间点、感染状态和 炎症，并使对细菌基因的研究成为可能，而细菌基因对定殖化和寄主扰动至关重要。 虽然项目2将在基础研究实验室进行，但其目标和结果与 提高对幽门螺杆菌在人类胃中的行为以及启动组织的分子变化的全面了解 用于致癌。