Dysregulated Polyamine Metabolism in H. pylori-associated Gastric Inflammation and Disease Progression

幽门螺杆菌相关胃炎症和疾病进展中多胺代谢失调

• 批准号: 10572035
• 负责人: Keith T. Wilson
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10572035
• 关键词: 3-Dimensional; Animal Model; Antibiotics; Area; Attenuated; Back; Bacteria; Biological; Biological Markers; Biological Models; Biopsy; CRISPR/Cas technology; Cancer Etiology; Carcinogens; Cell physiology; Cessation of life; Chemoprevention; Chronic; Chronic Gastritis; Clinical; DNA Damage; Development; Disease; Disease Progression; Dysplasia; Effectiveness; Enzymes; Epithelial; Epithelial Cells; Etiology; Exhibits; Exposure to; FVB/N Mouse; Functional disorder; Gastric mucosa; Gastrins; Gastritis; Gene Mutation; Generations; Genomic Instability; Genomics; Gerbils; Grant; Health; Helicobacter Infections; Helicobacter pylori; Helicobacter pylori induced carcinogenesis; Human; Induced Mutation; Infection; Infectious Agent; Inflammation; Insulin; Lead; Lesion; Link; Malignant Neoplasms; Mediating; Metabolism; Microbe; Military Personnel; Modeling; Molecular; Mucositis; Mus; Mutation; Neoplastic Cell Transformation; Organoids; Ornithine Decarboxylase; Pathogenicity; Pathway interactions; Patients; Peptic Ulcer; Phenotype; Plants; Polyamines; Population; Prevalence; Process; Proteins; Proteomics; Putrescine; Reporter; Research; Risk; Risk Assessment; Role; Spermidine; Spermine; Stomach; Stomach Diseases; System; Testing; Tissues; Transgenes; Translations; Validation; Veterans; Woman; base; beta catenin; cancer risk; carcinogenesis; carcinogenicity; chemokine; deoxyhypusine synthase; disorder prevention; exome sequencing; experience; gastric carcinogenesis; gastric organoids; high risk; in vivo; inhibitor; innovation; insight; interest; malignant stomach neoplasm; mutant mouse model; novel; pathogen; polyamine oxidase; premalignant; prevent; programs; recruit; response; service member; tissue injury; tool

Helicobacter pylori infection of the stomach and the resulting clinical consequences of chronic gastritis, peptic ulcer disease, and disease progression to gastric cancer remains a major health concern for Veterans. This pathogen infects half of the world’s population, and gastric cancer is the third leading cause of cancer deaths worldwide. H. pylori prevalence in Veterans is high; deployment-related infection is a problem, especially acquisition of strains associated with higher risk for carcinogenesis. The three polyamines, putrescine (Put), spermidine (Spd), and spermine (Spm), are ubiquitous molecules with many biological effects. Put is synthesized by the rate-limiting enzyme, ornithine decarboxylase (ODC), and is converted sequentially to Spd and Spm, which is back-converted to Spd by spermine oxidase (SMOX). Our Lab has had a long interest in the role of polyamines in GI inflammation and carcinogenesis, and we have the following exciting findings pertinent to this renewal grant: 1) Smox–/– mice infected with H. pylori exhibit decreased gastric inflammation, DNA damage, and chemokine expression, associated with depletion of Spd; 2) These phenotypes are recapitulated in H. pylori- infected 2D organoids from Smox–/– mice; 3) A new link between the H. pylori-induced pro-carcinogenic activation of b-catenin and SMOX, including findings that Smox–/– mice and gerbils treated with a SMOX inhibitor have loss of b-catenin activation in vivo; 4) Use of human gastric organoid cultures to demonstrate the effectiveness of a novel and potent SMOX inhibitor in blocking H. pylori-induced b-catenin activation, associated with Spd depletion; 5) Implication of a unique form of protein translation, hypusination, via the action of deoxyhypusine synthase (DHPS), a process which has been linked to carcinogenesis and appears to be attenuated by Smox deletion; 6) Demonstration of somatic genomic abnormalities by whole exome sequencing (WES) in gastritis and dysplasia tissues of INS-GAS mice, a model of H. pylori-induced carcinogenesis. We will now make effective use of important molecular tools, including valuable mutant mouse models, a key SMOX inhibitor, and advanced use of 3D and 2D gastric organoids from mice and VA patients. We hypothesize that dysregulated polyamine metabolism, due to SMOX and associated generation of Spd and hypusination, provides a molecular pathway leading to risk for gastric disease progression to carcinogenesis. Our specific aims are: 1) To directly determine the role of SMOX and Spd in gastric carcinogenesis. We will test: A) The effect of SMOX in cancer-prone INS-GAS mice, analyzing H. pylori-induced carcinogenesis, DNA damage, b-catenin activation, and GEC function in FVB/N INS-GAS Smox–/– mice +/- Spd; B) A novel, potent, second-generation SMOX inhibitor, SLH150-54, in INS-GAS mice and gerbils; C) The effect of SMOX/Spd on the formation of somatic genomic abnormalities using whole exome sequencing. 2) To determine if epithelial DHPS mediates deleterious effects of SMOX/Spd in gastric carcinogenesis. We will analyze: A) The role of SMOX in hypusination during H. pylori infection in C57BL/6 and FVB/N INS-GAS Smox–/– vs. WT mice +/- H. pylori +/- Spd; B) The effect of hypusination in gastric epithelial cells (GECs) on inflammation and carcinogenesis, using C57BL/6 and FVB/N INS-GAS mice with specific deletion of Dhps in GECs; C) the effect of DHPS/hypusination on somatic genomic instability. 3) To utilize gastric organoid reporter systems in tissues from VA patients to establish human biomarkers for carcinogenesis. H. pylori-induced DNA damage, b-catenin activation, and pathways identified in Aims 1 and 2 will be studied, related to: A) Induction and role of SMOX, using CRISPR/CAS9-mediated deletion of SMOX and SMOX inhibitors; and B) Induction and role of DHPS, using CRISPR/CAS9-mediated deletion of DHPS and the DHPS inhibitor, GC7. This research program will provide crucial new insights into H. pylori- induced inflammation and disease progression along the pathway to gastric carcinogenesis. The translation of findings from valuable animal models into robust human organoid systems is expected to facilitate the validation of new strategies for cancer risk assessment, chemoprevention, and treatment for VA patients.

胃幽门螺杆菌感染及其引起的慢性胃炎、消化性胃炎的临床后果 溃疡病和胃癌进展仍然是退伍军人的主要健康问题。这 病原体感染世界一半人口，胃癌是癌症死亡的第三大原因 全世界。退伍军人中幽门螺杆菌的患病率很高；与部署相关的感染是一个问题，尤其是 获得与较高致癌风险相关的菌株。三种多胺，腐胺（Put）， 亚精胺 (Spd) 和精胺 (Spm) 是普遍存在的分子，具有许多生物效应。 put 是合成的 被限速酶鸟氨酸脱羧酶 (ODC) 催化，并依次转化为 Spd 和 Spm， 它被精胺氧化酶 (SMOX) 反向转化为 Spd。我们的实验室长期以来对以下角色感兴趣： 多胺在胃肠道炎症和致癌作用中的作用，我们有以下与此相关的令人兴奋的发现 更新资助：1) 感染幽门螺杆菌的 Smox–/– 小鼠表现出胃部炎症、DNA 损伤和 趋化因子表达，与 Spd 的消耗相关； 2）这些表型在幽门螺杆菌中得到了概括- 来自 Smox–/– 小鼠的感染 2D 类器官； 3）幽门螺杆菌诱导的致癌激活之间的新联系 b-连环蛋白和 SMOX 的含量，包括用 SMOX 抑制剂治疗的 Smox–/– 小鼠和沙鼠的发现 体内β-连环蛋白激活； 4) 使用人胃类器官培养物来证明 新型有效的 SMOX 抑制剂，可阻断幽门螺杆菌诱导的与 Spd 相关的 β-catenin 激活 消耗; 5) 通过脱氧马尿素的作用实现蛋白质翻译的独特形式——马尿素化 合成酶 (DHPS)，一种与致癌有关的过程，并且似乎会被 Smox 减弱 删除； 6) 通过全外显子组测序 (WES) 证明胃炎和胃炎的体细胞基因组异常 INS-GAS 小鼠的发育不良组织，幽门螺杆菌诱导的致癌模型。我们现在将有效 使用重要的分子工具，包括有价值的突变小鼠模型、关键的 SMOX 抑制剂和先进的 使用来自小鼠和 VA 患者的 3D 和 2D 胃类器官。我们假设多胺失调 由于 SMOX 以及相关的 Spd 和 hypusination 的产生，新陈代谢提供了一种分子 导致胃病进展为致癌风险的途径。我们的具体目标是： 1) SMOX和Spd在胃癌发生中的作用直接决定。我们将测试： A) SMOX 在 易患癌症的 INS-GAS 小鼠，分析幽门螺杆菌诱导的致癌作用、DNA 损伤、β-连环蛋白激活， FVB/N INS-GAS Smox–/– 小鼠 +/- Spd 中的 GEC 功能； B) 一种新颖、有效的第二代 SMOX INS-GAS 小鼠和沙鼠中的抑制剂 SLH150-54； C) SMOX/Spd 对体细胞形成的影响 使用全外显子组测序的基因组异常。 2) 确定上皮DHPS是否介导有害物质 SMOX/Spd 在胃癌发生中的作用。我们将分析：A) SMOX 在 H. C57BL/6 和 FVB/N INS-GAS Smox–/– 与 WT 小鼠的幽门螺杆菌感染 +/- 幽门螺杆菌 +/- Spd； B）效果 使用 C57BL/6 和 FVB/N 观察胃上皮细胞 (GEC) 中的催眠作用对炎症和癌变的影响 GEC 中 Dhps 特异性缺失的 INS-GAS 小鼠； C) DHPS/hypusination 对体细胞基因组的影响 不稳定。 3) 利用 VA 患者组织中的胃类器官报告系统来建立人类 致癌的生物标志物。幽门螺杆菌诱导的 DNA 损伤、β-连环蛋白激活以及在 将研究目标 1 和 2，涉及：A) 使用 CRISPR/CAS9 介导的删除来诱导 SMOX 及其作用 SMOX 和 SMOX 抑制剂； B) DHPS 的诱导和作用，使用 CRISPR/CAS9 介导的删除 DHPS 和 DHPS 抑制剂 GC7。该研究计划将为幽门螺杆菌提供重要的新见解 沿着胃癌发生途径诱导炎症和疾病进展。的翻译 从有价值的动物模型到强大的人类类器官系统的研究结果有望促进验证 VA 患者的癌症风险评估、化学预防和治疗的新策略。