Roles for Adenomatous polyposis coli in colon injury prevention and wound healing

腺瘤性大肠杆菌在预防结肠损伤和伤口愈合中的作用

• 批准号: 10707443
• 负责人: KRISTI L NEUFELD
• 金额: $ 37.15万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-22 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707443
• 关键词: APC gene; APC mutation; Acute; Address; American; Anti-Inflammatory Agents; Automobile Driving; Binding; Biological Assay; Candidate Disease Gene; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Cells; ChIP-seq; Chemotaxis; Chronic; Clinical; Clinical Treatment; Colitis; Colon; Colon Injury; Complex; Cytoplasm; Data; Development; Diagnosis; Distal; Epithelial Cells; Epithelium; Etiology; Future; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Goals; Goblet Cells; Homeostasis; Human; Immune response; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory Bowel Diseases; Injury; Interleukin-1; Intervention; Intestines; Invaded; Irritants; Knock-in; Knowledge; Link; Lymphoid Follicle; Mediator; Mucin-2 Staining Method; Mucous Membrane; Mucous body substance; Mus; Mutation; Natural regeneration; Nuclear; Nuclear Import; Pathologic; Patients; Penetration; Phenotype; Predisposition; Prevention; Process; Production; Proliferating; Publishing; Recovery; Recurrence; Reporter; Reporting; Research; Role; Signal Pathway; Signaling Protein; Small Interfering RNA; Sodium Dextran Sulfate; Stains; Testing; Therapeutic; Thick; Time; Tissue Sample; Tissues; Transcriptional Regulation; Translating; Ulcer; Ulcerative Colitis; WNT Signaling Pathway; Wild Type Mouse; Work; cell mediated immune response; chemokine; cytokine; design; epithelial injury; epithelial repair; epithelial wound; gut microbes; healing; human tissue; in vivo; injury and repair; injury prevention; intestinal homeostasis; microbial; microbial composition; microbiota; mouse model; neutrophil; novel; novel strategies; novel therapeutic intervention; novel therapeutics; prevent; promoter; response; restoration; single-cell RNA sequencing; symptom management; tissue injury; tissue repair; transcriptome sequencing; treatment response; wound; wound healing

PROJECT SUMMARY Over 900,000 Americans are currently diagnosed with Ulcerative colitis (UC), an incurable inflammatory bowel disease of complex etiology. UC is multifactorial, with genetics, microbiota, and uncontrolled immune response leading to recurring colon ulcers and inflammation. With no defined cause, symptom management is critical for patients with this chronic condition. A major endpoint for clinical treatment of UC and other inflammatory bowel diseases is mucosal healing. Current UC therapeutics are predominated by anti- inflammatories. To expand this arsenal, it is important to establish exactly how ulceration prevention and wound healing occur such that these features can be enhanced in a targeted manner. Since our early published discovery that Adenomatous Polyposis Coli (APC) protein has both cytoplasmic and nuclear functions, our research has focused on mechanisms by which nuclear APC maintains normal intestinal homeostasis. To enable our analysis, we generated mice with mutations in APC which compromise nuclear import. These Apc-mNLS mice displayed decreased numbers of mucus-secreting goblet cells (GCs) and increased inflammation, chemokine expression, and tissue damage in response to treatment with the colon irritant DSS compared to their DSS-treated wild-type (WT) littermates. More recently, we published that in WT mice, DSS-treatment could induce a subset of distal colon GCs to express elevated APC levels. Similarly, colon tissue from human CD patients also displayed elevated APC protein level in most GCs, a phenotype not observed in unaffected tissue. Based on these striking results, we wondered whether the elevated APC in UC colon might be a response to ulceration in an attempt to restore normal colonic tissue homeostasis. As such, we sought to determine the mechanistic basis for and cellular consequences of higher APC protein levels in UC tissue. RNA-seq analysis of normal colon cells depleted for APC revealed a set of genes significantly downregulated. Cross-referencing this list with APC ChIP-seq data as well as genes whose expression is altered in UC, we identified 7 genes with roles in wound repair and mucus barrier production that are candidates for transcriptional regulation by nuclear APC. We hypothesize that nuclear APC promotes wound repair and resolution of UC through driving transcription of the major mucus barrier component MUC2 as well as specific wound repair mediators. This hypothesis will be tested by addressing the following questions: Does nuclear APC 1) regulate transcription of modulators of inflammation and wound repair (AIM1)? 2) promote colon wound healing (AIM2)? 3) promote mucus layer generation and thereby impede microbial penetration, alter microbial composition and prevent inflammation (AIM3)? Answering these questions is expected to establish nuclear APC as a key contributor to wound repair, a finding that could ultimately be translated into new therapies for UC and other IBDs.

项目摘要 目前有超过90万美国人被诊断患有溃疡性结肠炎（UC），这是一种无法治愈的炎症性疾病。 病因复杂的肠道疾病。UC是多因素的，遗传学，微生物群和不受控制的免疫 反应导致复发性结肠溃疡和炎症。没有明确的原因，症状管理是 对患有这种慢性疾病的患者至关重要。UC和其他疾病临床治疗的主要终点 炎症性肠病是粘膜愈合。目前的UC治疗主要是抗- 炎症为了扩大这一武库，重要的是要确定究竟如何溃疡预防和伤口 愈合发生，使得这些特征可以以有针对性的方式增强。自从我们的早期出版 发现腺瘤性结肠息肉病（APC）蛋白具有细胞质和细胞核功能，我们 研究集中在核APC维持正常肠内稳态的机制上。使 在我们的分析中，我们产生了具有APC突变的小鼠，该突变损害了核输入。这些Apc-mNLS 小鼠表现出分泌粘液的杯状细胞（GC）数量减少和炎症增加， 趋化因子的表达，以及对结肠刺激剂DSS治疗的反应， DSS处理的野生型（WT）同窝仔。最近，我们发表了在WT小鼠中，DSS治疗可以 诱导远端结肠GC亚组表达升高的APC水平。类似地，来自人CD的结肠组织 在大多数GC中，患者还显示APC蛋白水平升高，这是在未受影响的组织中未观察到的表型。 基于这些惊人的结果，我们想知道UC结肠中APC的升高是否是一种反应， 以恢复正常的结肠组织稳态。因此，我们试图确定 UC组织中较高APC蛋白水平的机制基础和细胞后果。RNA测序分析 耗尽APC的正常结肠细胞显示一组基因显著下调。交叉比对这个 列出APC ChIP-seq数据以及在UC中表达改变的基因，我们鉴定了7个基因， 在创伤修复和粘液屏障产生中的作用，其是通过核APC进行转录调节的候选者。 我们假设核APC通过驱动伤口修复和UC的解决来促进伤口修复和UC的解决。 主要粘液屏障成分MUC 2以及特异性伤口修复介质的转录。 这一假设将通过解决以下问题来检验：核APC 1）调节转录吗 炎症和伤口修复调节剂（AIM 1）？2)促进结肠伤口愈合（AIM 2）？3)促进 粘液层的产生，从而阻止微生物的渗透，改变微生物的组成， 炎症（AIM 3）？解决这些问题有望使核APC成为 伤口修复，这一发现最终可能转化为UC和其他IBD的新疗法。