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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要目前有超过 900,000 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病因复杂的肠道疾病。 UC 是多因素影响的，包括遗传、微生物群和不受控制的免疫导致复发性结肠溃疡和炎症的反应。在没有明确原因的情况下，症状管理是对于患有这种慢性病的患者来说至关重要。 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。为了扩大这一武器库，重要的是要准确地确定如何预防溃疡和伤口愈合的发生使得这些功能可以有针对性地增强。自从我们早期发表发现结肠腺瘤性息肉病 (APC) 蛋白同时具有细胞质和核功能，我们研究重点是核 APC 维持正常肠道稳态的机制。启用根据我们的分析，我们生成了 APC 突变的小鼠，该突变会损害核输入。这些 Apc-mNLS 小鼠的粘液分泌杯状细胞（GC）数量减少，炎症增加，与结肠刺激剂 DSS 治疗相比，趋化因子表达和组织损伤 DSS 处理的野生型 (WT) 同窝小鼠。最近，我们发表了在 WT 小鼠中，DSS 治疗可以诱导远端结肠 GC 的子集表达升高的 APC 水平。同样，来自人类 CD 的结肠组织患者在大多数 GC 中也表现出 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 升高是否可能是一种反应溃疡以试图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-seq 分析去除 APC 的正常结肠细胞显示一组基因显着下调。交叉引用这个列出 APC ChIP-seq 数据以及 UC 中表达发生改变的基因，我们确定了 7 个具有作用的基因伤口修复和粘液屏障生成，是核 APC 转录调控的候选者。我们假设核 APC 通过驱动促进伤口修复和 UC 解决主要粘液屏障成分 MUC2 以及特定伤口修复介质的转录。该假设将通过解决以下问题来检验：核 APC 1) 是否调节转录炎症和伤口修复调节剂（AIM1）？ 2）促进结肠伤口愈合（AIM2）？ 3）推广粘液层的产生，从而阻碍微生物渗透，改变微生物组成并防止炎症（AIM3）？回答这些问题有望将核 APC 确立为关键贡献者伤口修复，这一发现最终可能转化为治疗 UC 和其他 IBD 的新疗法。