Clonal Reconstruction and Targeting of the Correa Sequence

Correa 序列的克隆重建和靶向

• 批准号: 10470091
• 负责人: FRANK D. MCKEON
• 金额: $ 55.08万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-19 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10470091
• 关键词: Address; Adenocarcinoma; Automobile Driving; B-Lymphocytes; Barrett Esophagus; Barrett' s Adenocarcinoma; Biochemical; Cancer Patient; Cancerous; Carcinoma; Cell Differentiation process; Cell model; Cells; Clone Cells; Coculture Techniques; Diagnosis; Disease; Distal; Dose; Dysplasia; Endoscopic Biopsy; Esophageal Adenocarcinoma; Esophagus; Event; Evolution; Gastric Adenocarcinoma; Gastrointestinal tract structure; Genetic; Goals; Growth; Human; Immunodeficient Mouse; In Vitro; Incidence; Intervention; Intestinal Metaplasia; Laboratories; Lead; Lesion; Libraries; Life Expectancy; Malignant - descriptor; Malignant Neoplasms; Minor; Modeling; Molecular Genetics; Multipotent Stem Cells; Mus; Mutation; Pathology; Patient Care; Patients; Pharmaceutical Preparations; Phylogenetic Analysis; Population; Premalignant Cell; Process; Resolution; Sampling; Technology; Testing; Therapeutic; Transplantation; Tumorigenicity; Undifferentiated; Validation; Work; Xenograft Model; Xenograft procedure; base; cell type; drug discovery; exome sequencing; genetic analysis; high throughput screening; improved; in vivo; in vivo Model; inhibitor; innovation; insight; lead candidate; new technology; nonsynonymous mutation; novel; premalignant; prevent; promoter; reconstruction; regenerative growth; regenerative tissue; response; screening; small molecule; stem cell population; stem cells; tumor; tumor xenograft

PROJECT SUMMARY/ABSTRACT It is now well established that epithelial cancers arise in a multi-decade process from precancerous lesions. Esophageal adenocarcinoma, a cancer whose incidence has risen 5-fold since 1950, initiates with precancerous lesions known as “Barrett's esophagus”, progresses to dysplasia, and finally emerges as malignant esophageal adenocarcinoma along a path that parallels the Correa Sequence for gastric adenocarcinoma. As the average life expectancy of patients diagnosed with esophageal adenocarcinoma is approximately one-year, considerable efforts are underway to define its Correa sequence, and especially its pre-malignant stages, as potential targets for interdiction ahead of the onset of malignant disease. Toward this end, we have applied novel stem cell cloning technologies originally developed in our laboratories for normal human gastrointestinal tract stem cells to reconstructing the Correa sequence in patients with early esophageal adenocarcinoma. Significantly, each of these patient-matched endoscopic biopsies of Barrett's, dysplasia, and esophageal adenocarcinoma yields discrete populations of stem cells that respectively yield intestinal metaplasia, dysplasia (but not tumors), and aggressive adenocarcinoma following transplantation to immunodeficient mice. From a detailed molecular genetics analysis of nearly 100 independent clones from across the Barrett's, dysplasia, and adenocarcinoma lesions of one patient, we have been able to reconstruct, at unprecedented resolution, both the direct phylogenetic sequence that led to this tumor as well as identify “dead-ends” at both the Barrett's and dysplasia stages that did not contribute to the final tumor. Moreover, each of the cloned stem cells of Barrett's, dysplasia, and adenocarcinoma lesions represent permanent lines that have enabled powerful approaches to drug discovery that has culminated in leads that selectively target the entire Correa sequence while sparing normal esophageal stem cells. In three specific aims, we will 1.) clonally reconstruct the Correa sequence from 10 patients with early esophageal adenocarcinoma; 2.) establish high-throughput screens involving co-cultures of normal esophageal and Correa sequence stem cells for lead discovery; and 3.) develop in vivo xenograft models of patient-matched normal esophageal and Correa sequence stem cells for validating lead combinations targeting these lesions. Based on extensive preliminary studies, we anticipate the analysis of patient-matched stem cells of these progressive lesions will provide fundamental insights into the evolution of esophageal adenocarcinoma and as well as epithelial cancers in general. From the standpoint of filling important gaps in patient care, the drug discovery enabled by these sets of lesional stem cells offers promising and novel interventions to prevent to onset of esophageal adenocarcinoma as well as ones to address disease that has already taken hold.

项目摘要/摘要 现在已经很好地确定，上皮癌是在癌前病变的多年过程中出现的。 食管腺癌是一种癌症，其事件自1950年以来的事件增长了5倍 被称为“巴雷特的食道”，发展为发育不良，最终成为恶性食管腺癌 沿着与胃腺癌的Correa序列相似的路径。作为患者的平均预期寿命 被诊断为食管腺癌的诊断为一年，正在努力定义其 Correa序列，尤其是其恶性前阶段，作为拦截的潜在目标。 恶性疾病。为此，我们应用了最初在我们的新型干细胞克隆技术中开发的新型干细胞克隆技术 正常人胃肠道干细胞的实验室，以重建早期患者的裂殖序列 食管腺癌。值得注意的是，这些患者匹配的内窥镜检查中的每一个，发育不良和 食管腺癌产生分别产生肠道化生的干细胞的离散群体 移植到免疫缺陷小鼠后，发育不良（但不是肿瘤）和攻击性腺癌。来自 详细的分子遗传学分析来自巴雷特的近100个独立克隆，发育不良和 一名患者的腺癌病变，我们能够以前所未有的分辨率重建 导致该肿瘤的系统发育序列，并在巴雷特和发育不良阶段识别“死末端” 没有贡献最终肿瘤。此外，巴雷特的每个克隆干细胞，发育不良和腺癌瘤 病变代表永久性线，使得有效的药物发现方法达到领先地位 这在放大正常食管干细胞的同时有选择地靶向整个裂殖序列。在三个具体目标中，我们 将1.）克隆重建10名早期食管腺癌患者的Corre序列； 2.）建立 高通量筛选涉及正常食管和沟序干细胞共培养以发现铅发现； 和3.）开发患者匹配的正常食管和折叠序列干细胞的体内特征模型 验证针对这些病变的铅组合。基于广泛的初步研究，我们预计 这些进行性病变的患者匹配的干细胞将提供对食管进化的基本见解 腺癌以及上皮癌。从填补患者护理中重要空白的角度来看， 这些病变的干细胞启用的药物发现提供了希望和新颖的干预措施，以防止发作 食管腺癌以及解决已经存在的疾病的腺瘤。