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: 10620757
• 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 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10620757
• 关键词: 3-Dimensional; Animal Model; Animals; 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; Classification; Clinical; DNA Damage; Development; Disease; Disease Progression; Dysplasia; Effectiveness; Enzymes; Epithelial Cells; Epithelium; 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; active duty; 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.

胃的幽门螺杆菌感染以及慢性胃炎的临床后果 溃疡疾病和胃癌的疾病进展仍然是退伍军人的主要健康问题。这 病原体感染了世界一半的人口，胃癌是癌症死亡的第三主要原因 全世界。退伍军人的幽门螺杆菌患病率很高。与部署相关的感染是一个问题，尤其是 获取与更高的癌变风险相关的菌株。三个多胺，perrescine（put）， 精子（SPD）和精子（SPM）是无处不在的分子，具有许多生物学作用。放置合成 通过限速酶，鸟氨酸脱羧酶（ODC），然后依次转换为SPD和SPM， 通过精子氧化酶（SMOX）将其反向转化为SPD。我们的实验室对 胃肠道炎症和癌变中的多胺，我们有以下令人兴奋的发现 更新授予：1）幽门螺杆菌感染的Smox - / - 小鼠胃感染降低，DNA损伤和 趋化因子表达，与SPD的消耗有关； 2）这些表型在幽门螺杆菌中概括 从Smox - / - 小鼠感染的2D类器官； 3）幽门螺杆菌诱导的促癌激活之间的新联系 B-catenin和smox的大量，包括Smox –/ - 小鼠和用SMOX抑制剂处理的沙鼠的发现 B-catenin激活体内的激活； 4）使用人类胃器官培养物来证明A的有效性 与SPD相关的幽门螺杆菌诱导的H.幽门杆菌诱导的B-catenin激活方面的新颖和潜在的SMOX抑制剂 消耗; 5）通过脱氧蛋白的作用，蛋白质翻译的独特形式的含义 合酶（DHP），该过程与癌变有关，似乎被SMOX衰减 删除； 6）在胃炎和 Ins-Gas小鼠的发育不良组织，幽门螺杆菌诱导的癌变模型。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SMOX抑制剂和高级 使用来自小鼠和VA患者的3D和2D胃癌。我们假设多胺失调 由于SMOX和相关的SPD和非偶然产生，代谢提供了分子 途径导致胃病的风险发展为癌变。我们的具体目的是：1） 直接确定SMOX和SPD在胃癌发生中的作用。我们将测试：a）Smox在 易受癌症的INS-GAS小鼠，分析了幽门螺杆菌诱导的癌变，DNA损伤，B-钙蛋白激活， FVB/N INS-GAS SMOX - / - 小鼠+/- SPD中的GEC功能； b）一种新颖的，潜力的第二代SMOX 抑制剂，SLH150-54，在Ins-Gas小鼠和沙鼠中； c）SMOX/SPD对躯体形成的影响 使用整个外显子组测序的基因组异常。 2）确定上皮DHP是否介导有害 SMOX/SPD对胃癌发生的影响。我们将分析：a）SMOX在H期间的非偶然性中的作用。 幽门螺杆菌感染C57BL/6和FVB/N INS-GAS SMOX - / - 与WT小鼠+/- H. Pylori +/- SPD; b）效果 使用C57BL/6和FVB/N INS-GAS小鼠在GEC中具有特异性DHP的特异性缺失； c）DHP/非胰岛对体细胞基因组的影响 不稳定。 3）在VA患者的组织中利用胃器官的记者系统来建立人类 癌变的生物标志物。幽门螺杆菌诱导的DNA损伤，B-catenin激活和在 AIM 1和2将研究： Smox和Smox抑制剂； b）使用CRISPR/CAS9介导的删除DHP的诱导和作用 DHP和DHP抑制剂GC7。该研究计划将为幽门螺杆菌提供至关重要的新见解 沿胃癌发生的途径引起的炎症和疾病进展。翻译 有价值的动物模型的发现有望促进验证 VA患者的癌症风险评估，化学预防和治疗的新策略。