Control of Inflammatory Acidity in Mucosal Inflammation

粘膜炎症中炎症酸度的控制

• 批准号: 10512056
• 负责人: Ian Michael Cartwright
• 金额: --
• 依托单位: VA EASTERN COLORADO HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10512056
• 关键词: Acidity; Acidosis; Acids; Acute; Affect; American; Anxiety; Bacteria; Bicarbonates; Biological Assay; Biological Response Modifier Therapy; Buffers; Cell physiology; Cells; Chemicals; Chronic; Colitis; Colonic inflammation; Crohn' s disease; Data; Development; Disease; Disease Outcome; Epithelial Cells; Escherichia coli; Escherichia coli K12; Exposure to; Family; Funding; Gastrointestinal tract structure; Gene Expression Regulation; Generations; Genetic; Genetic Transcription; Germ-Free; Glycolysis; Goals; Home; Hospitalization; Hypochlorous Acid; Ileitis; Immune response; Immunology; In Vitro; Incidence; Indoles; Infiltration; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Intestines; Invaded; Knowledge; Lactic acid; Malignant Neoplasms; Mental Depression; Mentorship; Modeling; Modification; Molds; Monitor; Mucositis; Mucous Membrane; Mus; Pathogenesis; Pathology; Patients; Peroxidases; Phenotype; Post-Traumatic Stress Disorders; Production; Regulation; Reporting; Research Personnel; Resolution; Role; SLC26A3 gene; Severity of illness; Signal Transduction; Site; Source; Surface; Testing; Time; Tissues; Training; Ulcerative Colitis; Veterans; Work; career development; chemically induced colitis; commensal bacteria; cytokine; design; economic impact; effective therapy; experience; extracellular; gastrointestinal; genetic approach; genetic signature; gut inflammation; hospitalization rates; host-microbe interactions; improved; in vivo; inhibitor; intestinal epithelium; intestinal homeostasis; knock-down; member; metabolomics; microbial; microbial host; microbiota; microbiota metabolites; migration; military veteran; mouse model; murine colitis; neutrophil; novel; novel therapeutics; pH Homeostasis; pharmacologic; psychosocial; response; symposium; transcriptome sequencing; wound healing; Acidity; Acidosis; Acids; Acute; Affect; American; Anxiety; Bacteria; Bicarbonates; Biological Assay; Biological Response Modifier Therapy; Buffers; Cell physiology; Cells; Chemicals; Chronic; Colitis; Colonic inflammation; Crohn' s disease; Data; Development; Disease; Disease Outcome; Epithelial Cells; Escherichia coli; Escherichia coli K12; Exposure to; Family; Funding; Gastrointestinal tract structure; Gene Expression Regulation; Generations; Genetic; Genetic Transcription; Germ-Free; Glycolysis; Goals; Home; Hospitalization; Hypochlorous Acid; Ileitis; Immune response; Immunology; In Vitro; Incidence; Indoles; Infiltration; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Intestines; Invaded; Knowledge; Lactic acid; Malignant Neoplasms; Mental Depression; Mentorship; Modeling; Modification; Molds; Monitor; Mucositis; Mucous Membrane; Mus; Pathogenesis; Pathology; Patients; Peroxidases; Phenotype; Post-Traumatic Stress Disorders; Production; Regulation; Reporting; Research Personnel; Resolution; Role; SLC26A3 gene; Severity of illness; Signal Transduction; Site; Source; Surface; Testing; Time; Tissues; Training; Ulcerative Colitis; Veterans; Work; career development; chemically induced colitis; commensal bacteria; cytokine; design; economic impact; effective therapy; experience; extracellular; gastrointestinal; genetic approach; genetic signature; gut inflammation; hospitalization rates; host-microbe interactions; improved; in vivo; inhibitor; intestinal epithelium; intestinal homeostasis; knock-down; member; metabolomics; microbial; microbial host; microbiota; microbiota metabolites; migration; military veteran; mouse model; murine colitis; neutrophil; novel; novel therapeutics; pH Homeostasis; pharmacologic; psychosocial; response; symposium; transcriptome sequencing; wound healing

Within the gastrointestinal (GI) tract the primary function of the mucosal surface is to provide a selective barrier to the outside. It is widely understood that the GI tract exists in a state of low-grade inflammation, as a result of constant processing of luminal antigenic material. The GI tract is home to trillions of commensal bacteria which contribute significantly to GI homeostasis, but can also initiate and drive the progression of mucosal diseases. Acute intestinal inflammation involves early accumulation of neutrophils (PMN) followed by either resolution or progression to chronic inflammation. Without efficient PMN clearance at sites of infiltration, PMN can accumulate and contribute to chronic inflammatory conditions, including inflammatory bowel disease. The ongoing studies outline in this proposal have revealed that PMN transepithelial migration (TEM) results in a significant decrease in extracellular pH, deemed inflammatory acidification. Extensions of these studies have shown that this acidic microenvironment impacts barrier function and gene transcription in intestinal epithelial cells (IEC). Guided by an unbiased RNAseq of IEC exposed to low pH in vitro and tissue metabolomics in vivo, we identified a gene signature that includes induction of both TNF and members of the DUSP and NR4A families. We also identified a role of IEC GPR31 in sensing extracellular pH and the loss of microbial-derived indoles during active inflammation. Extending these studies, we observed significant improvement in IEC barrier function under acidic conditions in T84 cells which had GPR31 knocked down. Additionally, we observed that indole treatment significantly decreased inflammatory acidification following PMN TEM and in K12 E. coli acidic conditions induce the production of indole. Based on these observations, we hypothesize that inflammatory acidification promotes inflammatory signaling in IEC and that the microbiota-derived indoles have a significant role in promoting pH homeostasis at sites of inflammation. In vitro functional assays will define the impact of extracellular acidosis and pH sensing on PMN TEM, IEC barrier function, and IEC adaptation to chronic extracellular acidosis. These studies will be extended to examine the influence of microbial/host interactions on IEC responses to extracellular acidification. Finally, utilizing in vivo murine colitis and ileitis models we will profile inflammatory acidification. We will extend these studies to investigate the impact of the microbiota and microbiota-derived metabolites on inflammation-associated acidosis. The career development and training plan outlined in this application is designed to guide and promote my transition to independence as a VA funded researcher. The proposed combination of didactic courses, professional conferences, and mentorship committee have been designed to strengthen my knowledge of mucosal immunology and mucosal inflammation. Technically, I will gain experience in the generation of murine intestinal enteroids, genetic modification of enteroids, both chemically induced and spontaneous murine models of ileitis and colitis, and expand my knowledge of microbial/host response and technical experience in manipulating the microbiota. Upon completion of this proposal I will be well prepared to establish myself as an expert in pH regulation and inflammation-associated acidification. Furthermore, it is our hope that the studies outline in this proposal will lay the groundwork for the identification of novel therapeutic avenues in IBD, directly benefiting the veteran population.

在胃肠道（GI）中，粘膜表面的主要功能是提供选择性屏障 到外面。众所周知，由于胃肠道存在于低度炎症状态 腔内抗原材料的持续加工。胃肠道是数万亿个共生细菌的家园 对胃肠道稳态做出了重大贡献，但也可以启动和推动粘膜疾病的进展。 急性肠道炎症涉及嗜中性粒细胞（PMN）的早期积累，然后分辨率或 发展为慢性炎症。没有有效的PMN渗透部位，PMN可以 积累并导致慢性炎症状况，包括炎症性肠病。 该提案中正在进行的研究大纲表明，PMN跨性别迁移（TEM）导致 细胞外pH值显着降低，被认为是炎性酸化。这些研究的扩展已有 表明这种酸性微环境会影响肠上皮的屏障功能和基因转录 细胞（IEC）。在体外暴露于低pH值的IEC的无偏RNASEQ的指导下，体内的组织代谢组学 我们确定了一个基因特征，其中包括诱导TNF和DUSP和NR4A成员 家庭。我们还确定了IEC GPR31在感测细胞外pH和微生物衍生的丧失中的作用 活性炎症期间的吲哚。扩展了这些研究，我们观察到IEC的显着改善 T84细胞中GPR31撞倒的T84细胞中的屏障功能。另外，我们 观察到，吲哚治疗在PMN TEM和IN IN后显着降低了炎症性酸化 K12大肠杆菌酸性条件诱导吲哚的产生。基于这些观察，我们假设 炎性酸化促进了IEC中的炎症信号，并且微生物群衍生的吲 在促进炎症部位促进pH稳态方面发挥着重要作用。体外功能分析将 定义细胞外酸中毒和pH感应对PMN TEM，IEC屏障功能和IEC的影响 适应慢性细胞外酸中毒。这些研究将扩展到检查 IEC对细胞外酸化反应的微生物/宿主相互作用。最后，使用体内鼠结肠炎 和回肠炎模型我们将介绍炎性酸化。我们将扩展这些研究以调查 微生物群和微生物群的代谢产物对炎症相关酸中毒的影响。职业 本申请中概述的制定和培训计划旨在指导和促进我的过渡到 作为VA资助​​的研究人员的独立性。教学课程的拟议组合，专业 会议和精神委员会旨在增强我对粘膜的了解 免疫学和粘膜感染。从技术上讲，我将获得鼠肠的一代经验 肠to，肠道遗传的遗传修饰，无论是化学诱导的肠炎和赞助鼠模型 和结肠炎，并扩大我对微生物/宿主反应的了解以及操纵的技术经验 微生物群。完成此提案后，我将做好充分的准备，以确立自己的pH专家 调节和与炎症相关的酸化。此外，我们希望研究概述这一点 提案将为识别IBD的新型治疗途径奠定基础，直接受益 退伍军人人口。