Modulation of mitochondrial respiration to treat colitis

调节线粒体呼吸来治疗结肠炎

• 批准号: 10560494
• 负责人: Terrence A. Barrett
• 金额: --
• 依托单位: VA MEDICAL CENTER - LEXINGTON, KY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-10-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560494
• 关键词: 3-Dimensional; 3-nitrotyrosine; Acceleration; Antioxidants; Auranofin; Bioenergetics; Biogenesis; Biopsy; Cell Survival; Cells; Childhood; Chronic; Clinical; Clinical Data; Colitis; Colon; Crohn' s disease; Data; Disease; Disease Outcome; Disease remission; Distant; Electron Transport; Epithelial Cells; Event; Failure; Gene Expression; Genes; Glutathione; Goals; Gold; Gold Compounds; Healthcare Systems; Human; Hydrogen Peroxide; Immunosuppression; In Vitro; Incubated; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestines; Messenger RNA; Mitochondria; Modeling; Morbidity - disease rate; Mucositis; Mucous Membrane; Mus; Oral; Organ; Organoids; Outcome; Oxidative Phosphorylation; Oxidative Stress; Oxygen Consumption; PPAR gamma; Patients; Pharmaceutical Preparations; Phosphatidylinositols; Phosphotransferases; Production; Progress Reports; Proteins; Reactive Oxygen Species; Refractory; Research; Respiration; Respiratory Chain; Rheumatoid Arthritis; Risk; Role; SOD2 gene; Secondary to; Signal Induction; Signal Transduction; Small Intestines; Superoxide Dismutase; TNF gene; Testing; Therapeutic; Therapeutic Effect; Therapeutic Studies; Tissues; Toxicology; Ulcer; Ulcerative Colitis; Veterans; Work; antioxidant enzyme; catalase; chronic ulcer; cytokine; dextran sulfate sodium induced colitis; enzyme activity; healing; improved; in vivo; innovation; intestinal crypt; intestinal epithelium; mtTF1 transcription factor; murine colitis; novel; novel therapeutics; nuclear factor-erythroid 2; oxidant stress; preclinical study; prospective; protein expression; repaired; response; restoration; synergism; uptake

The importance of mucosal healing in inflammatory bowel disease (IBD) derives from clinical data that demonstrate it to be a predictor of remission. Unfortunately, many IBD patients remain refractory or lose responsiveness to therapies leading to persistent mucosal ulceration. Prospective pediatric studies show that reduced mitochondrial gene expression predict unfavorable outcomes in ulcerative colitis (UC). Consistently, increased expression of mitochondrial respiratory chain genes has been shown to predict favorable Crohn’s disease (CD) outcomes. Based on these results and our own data, we hypothesize that a critical driver of chronic ulceration in IBD is the failure of intestinal epithelial cells (IEC) to increase mitochondrial respiration. In studies leading up to this submission we found that mitochondrial deficiency caused poor ulcer healing due to inadequate crypt fissioning occurring at ulcer edges (Progress Report). These findings led us to develop a novel gold (Au)- based drug based on the current safe and effective gold therapy for Rheumatoid arthritis (Auranofin). This new drug, AuPhos, localizes to mitochondria where it increases mitochondrial respiration and mitochondrial reactive oxygen species (mtROS) production. Induction of low levels of “endogenous” mtROS are known to promote healthy adaptive responses in cells including anti-oxidant enzyme activities (Background). In preliminary studies, we found that oral AuPhos 1) enhances mucosal repair in DSS colitis, 2) increases IEC mitochondrial oxygen consumption rate (OCR) and oxidative phosphorylation (OXPHOS), 3) mtROS and H2O2 production in IEC as well as 4) phosphoinositides-3 kinase (PI3K) and nuclear factor erythroid 2-related factor (Nrf2) signaling with 5) improved expression of proteins that induce mitochondrial biogenesis [peroxisome proliferator activated receptor-γ coactivator 1 alpha (PGC1α) and transcription factor A, mitochondrial (TFAM). Studies proposed in Aim 1 interrogate mitochondrial function under baseline conditions to allow assessment of AuPhos-induced changes in mitochondrial mass, activity (mtROS) and OCR, ATP turnover, electron transport chain (ETC) activity, induction of adaptive responses [anti-oxidant catalase, MnSOD (superoxide dismutase) and glutathione (reduced/oxidized)] as well as assess changes in tissue oxidant stress (4HNE, protein carbonylation, nitrotyrosine IHC). Aim 2 will build upon these studies by evaluating AuPhos effects in TNF-stimulated small bowel (SB) and colonic organoids from veterans (normal and IBD). Studies in 3D organoids will allow for mechanistic testing of AuPhos-induced crypt budding and IEC gene expression. Results of Aim 2 will instruct studies in Aim 3A that study therapeutic effects of AuPhos on IEC gene expression and crypt fissioning that heal ulcers in DSS colitis. In Aim 3B, scRNAseq analysis of human biopsies incubated with AuPhos will allow us to examine induction of mitochondrial gene expression in discrete IEC clusters. Proposed studies are driven by a novel hypothesis that restoration of IEC mitochondrial respiration is a key step in mucosal healing. Together, these “preclinical” studies with a VA-sponsored new drug (AuPhos) have the potential of transitioning this agent to the therapy of IBD in veterans.

粘膜愈合在炎症性肠病（IBD）中的重要性来自于临床数据 证明它是缓解的预测指标。不幸的是，许多IBD患者仍然难治或失去 对疗法的反应能力导致持续性粘膜溃疡。前瞻性小儿研究表明 线粒体基因表达降低预测溃疡性结肠炎（UC）的不利结果。一贯 线粒体呼吸链基因的表达增加已被证明可以预测有利的克罗恩 疾病（CD）结果。基于这些结果和我们自己的数据，我们假设是慢性的关键驱动力 IBD溃疡是肠上皮细胞（IEC）增加线粒体呼吸的失败。在研究中 在此提交之前，我们发现线粒体缺陷导致溃疡愈合不足，因此不足 隐窝裂变发生在溃疡边缘（进度报告）。这些发现使我们开发了一种新颖的黄金（AU） - 基于当前用于类风湿关节炎（Auranofin）的安全有效金疗法的药物。这个新 药物，auphos，将其定位于线粒体增加线粒体呼吸和线粒体反应性 氧（MTROS）生产。已知诱导低水平的“内源性” MTRO会促进 包括抗氧化酶活性的细胞中健康的适应性反应（背景）。在初步 研究，我们发现口服Auphos 1）增强DSS结肠炎的粘膜修复，2）增加IEC线粒体 氧消耗率（OCR）和氧化磷酸化（OXPHOS），3）MTROS和H2O2的产生 IEC以及4）磷酸肌醇-3激酶（PI3K）和核因子2相关因子（NRF2）信号传导 5）改善了诱导线粒体生物发生的蛋白质表达[过氧化物组增殖剂激活 受体-γ共激活因子1α（PGC1α）和转录因子A，线粒体（TFAM）。研究提出了 AIM 1在基线条件下询问线粒体功能，以评估AUPHOS诱导的 线粒体质量，活性（MTROS）和OCR，ATP周转率，电子传输链（ETC）活性的变化， 诱导自适应反应[抗氧化剂过氧化氢酶，MNSOD（超氧化物歧化酶）和谷胱甘肽 （降低/氧化）]以及组织氧化应激的评估变化（4HNE，蛋白羰基化， 硝基酪氨酸IHC）。 AIM 2将通过评估TNF刺激的小的AUPHOS效应来基于这些研究的基础 从退伍军人（正常和IBD）的肠道（SB）和结肠类器官。 3D类器官的研究将允许 AUPHOS诱导的隐窝萌芽和IEC基因表达的机械测试。 AIM 2的结果将指示 AIM 3A的研究研究了Auphos对IEC基因表达和隐窝裂变的治疗作用 在DSS结肠炎中治愈溃疡。在AIM 3B中，与Auphos一起孵化的人类活检的SCRNASEQ分析将允许 我们检查离散IEC簇中线粒体基因表达的诱导。提出的研究是驱动的 通过一个新的假设，即恢复IEC线粒体呼吸是粘膜愈合的关键步骤。一起， 这些使用VA赞助的新药（Auphos）的“临床前”研究有可能过渡该试剂 在退伍军人中对IBD的治疗。