CCL11 as a New Therapeutic Target for Colitis and Colon Cancer

CCL11作为结肠炎和结肠癌的新治疗靶点

• 批准号: 10266036
• 负责人: Lori A Coburn
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10266036
• 关键词: Acute; Admission activity; Adult; Affect; Age; American; Antibodies; Apoptosis; Azoxymethane; Body Weight decreased; Bone Marrow Transplantation; CCL11 gene; Carcinogenicity Tests; Cause of Death; Cells; Chemotactic Factors; Chronic; Colitis; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Crohn' s disease; DNA Damage; Development; Eotaxin; Epithelial; Epithelial Cells; Exhibits; Exposure to; Functional disorder; Goals; Health; Healthcare Systems; Hematopoietic; Histologic; Immune; Immune response; Infiltration; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury; Intervention; Knock-out; Lamina Propria; Lesion; Link; Malignant Neoplasms; Mediating; Mediator of activation protein; Messenger RNA; Modeling; Morbidity - disease rate; Mucous Membrane; Mus; Mutant Strains Mice; Myelogenous; Myeloid Cells; Patients; Permeability; Population; Prevalence; Prevention; Probability; Risk; Role; Serum; Severity of illness; Side; Signal Transduction; Sodium Dextran Sulfate; Testing; Tissues; Tumor Burden; Ulcerative Colitis; Veterans; Wild Type Mouse; Woman; base; carcinogenesis; cell motility; chemokine; cohort; colitis associated cancer; colon carcinogenesis; colon tumorigenesis; cytokine; dextran sulfate sodium induced colitis; disorder control; eosinophil; epithelial injury; fecal microbiome; illness length; immune activation; immune function; injury and repair; insight; macrophage; men; mouse model; mutant; neutralizing antibody; new therapeutic target; novel therapeutics; premalignant; prospective; protective effect; receptor; repair model; response; tumor; tumorigenesis

With more than 1.6 million Americans affected, inflammatory bowel disease (IBD) causes significant morbidity and can progress to colon cancer. There are two main forms of IBD: ulcerative colitis (UC) and Crohn's disease, and the associated abnormal immune response continues to be investigated with the goal of discovering new therapeutics or avenues for intervention. We have previously shown that in a prospectively collected cohort of adult UC patients, CCL11 (also called eotaxin-1), a chemoattractant for eosinophils, was the only analyte increased in both serum and tissue at all levels of disease severity out of 42 cytokines and chemokines assessed, compared to non-IBD control patients. We also observed a significant increase in colonic tissue eosinophils in UC patients vs. controls, and eosinophil counts correlated with tissue CCL11 levels. It has been shown that Ccl11-deficient mice were protected from colitis in an acute dextran sulfate sodium (DSS)-induced colitis model. However, acute colitis studies in mice lacking eosinophils have been mixed, suggesting it is not clearly just decreased tissue eosinophil infiltration leading to improvement. We have now determined that 1) Ccl11 mRNA is expressed in isolated colonic epithelial cells and isolated lamina propria cells in response to the chronic DSS and the azoxymethane (AOM)-DSS colitis-associated carcinogenesis (CAC) models; 2) epithelial cells and macrophages express receptors for CCL11; 3) treatment with an anti-CCL11 antibody leads to protection in an injury and repair model of colitis; 4) Ccl11–/– mice exhibit decreased tumor number, tumor burden, and colonic eosinophils in the AOM-DSS model; 5) bone marrow transplant studies point to loss of both hematopoietic and epithelial-derived CCL11 as key mediators of the protection seen in the AOM-DSS model; 6) CCL11 expression is increased in Apc mutant mouse tumors; and 7) the fecal microbiome is altered in Ccl11–/– mice compared to wild-type mice at baseline. In this proposal, we will dissect the role of CCL11 in modulating epithelial injury and repair, and immune responses in models of chronic colitis, CAC, and sporadic/genetically-mediated colon cancer. Our Hypothesis is: CCL11 is a key mediator of colonic inflammation, epithelial dysfunction, and risk for carcinogenesis. The overarching goal is to establish new insights into the potential benefits of blocking CCL11 signaling in both the treatment of colitis and prevention of colon carcinogenesis. The Specific Aims are: 1. To determine the mechanism of protection in colitis-associated cancer with loss of CCL11. Ccl11–/– mice exhibit protection in the AOM- DSS model associated with altered tissue cytokine levels and decreased tissue eosinophils. However, acute colitis studies in eosinophil-deficient mice suggest this may be an eosinophil independent effect. WT, Ccl11fl/fl, Ccl11∆mye, Ccl11∆GIepi, and Ccl11–/– mice exposed to AOM-DSS will be used to test the hypothesis that protective effects are due to altered immune cell populations other than eosinophils and/or epithelial function when CCL11 is lost by assessing tumor number and size, colonic immune cell composition/function, and epithelial function. 2. To determine if CCL11 is also a modifier in sporadic/genetically-mediated colon tumorigenesis. Loss of CCL11 protects from CAC in murine modeling, and CCL11 expression is increased in Apc mutant tumors. CDX2P-CreERT2Apcfl/fl and CDX2P-CreERT2Apcfl/flCcl11–/– mice that develop colon tumors will be used to test the hypothesis that loss of CCL11 will abrogate tumor development by reducing tumor- promoting inflammation by assessing: tumor number and size, colonic immune cell composition/function, and epithelial function. 3. To determine if targeting CCL11 signaling with neutralizing antibodies protects from colon tumorigenesis. WT mice exposed to AOM-DSS and CDX2P-CreERT2Apcfl/fl mice will be treated with a CCL11 neutralizing antibody (or isotype control) to establish the effects on: tumor number and size, colonic immune cell composition/function, and epithelial function. These studies will provide new mechanistic insights to support the use of treatments based on inhibition of CCL11 in colitis and colon cancer.

超过160万美国人受到影响，炎症性肠病（IBD）导致显著的发病率 并可能发展为结肠癌IBD有两种主要形式：溃疡性结肠炎（UC）和克罗恩病 疾病，以及相关的异常免疫反应继续进行研究，目标是 发现新的治疗方法或干预途径。我们以前已经表明，在一个前瞻性的 收集的成年UC患者队列中，CCL 11（也称为嗜酸性粒细胞趋化因子-1）是嗜酸性粒细胞的化学引诱物， 在42种细胞因子中，只有分析物在所有疾病严重程度水平的血清和组织中增加， 与非IBD对照患者相比，评估趋化因子。我们还观察到， UC患者与对照组的结肠组织嗜酸性粒细胞，嗜酸性粒细胞计数与组织CCL 11相关 程度.已经表明，Ccl 11缺陷小鼠在急性硫酸葡聚糖治疗中免受结肠炎的影响。 钠（DSS）诱导结肠炎模型。然而，在缺乏嗜酸性粒细胞的小鼠中进行的急性结肠炎研究已经被证实是一种新的治疗方法。 混合，表明它不是明确的只是减少组织嗜酸性粒细胞浸润导致改善。我们有 现在确定1）Ccl 11 mRNA在分离的结肠上皮细胞和分离的板层中表达 固有细胞对慢性DSS和氧化偶氮甲烷（AOM）-DSS结肠炎相关 癌发生（CAC）模型; 2）上皮细胞和巨噬细胞表达CCL 11的受体; 3）治疗 与抗-CCL 11抗体一起使用导致在结肠炎的损伤和修复模型中的保护; 4）Ccl 11-/-小鼠表现出 AOM-DSS模型中肿瘤数量、肿瘤负荷和结肠嗜酸性粒细胞减少; 5）骨髓 移植研究指出，造血和上皮来源的CCL 11的丢失是移植过程中的关键介质。 6）CCL 11表达在Apc突变小鼠肿瘤中增加;和 7)基线时，与野生型小鼠相比，Ccl 11-/-小鼠的粪便微生物组发生了改变。在本提案中，我们 将剖析CCL 11在调节上皮损伤和修复中的作用，以及免疫反应在模型中的作用。 慢性结肠炎、CAC和散发性/遗传介导的结肠癌。我们的假设是：CCL 11是一个关键 结肠炎症介质、上皮功能障碍和致癌风险。总体 我们的目标是建立新的见解，以了解阻断CCL 11信号传导在治疗 结肠炎和预防结肠癌发生。具体目标是：1。为了确定 保护结肠炎相关癌症与CCL 11的损失。Ccl 11-/-小鼠在AOM中表现出保护作用- DSS模型与组织细胞因子水平改变和组织嗜酸性粒细胞减少相关。然而，急性 在嗜酸性粒细胞缺乏小鼠中的结肠炎研究表明这可能是嗜酸性粒细胞非依赖性效应。WT，Ccl11fl/fl， 将使用暴露于AOM-DSS的Ccl 11 Myc、Ccl 11 Gleepi和Ccl 11-/-小鼠来检验以下假设： 保护作用是由于除了嗜酸性粒细胞和/或上皮功能以外的免疫细胞群的改变 当通过评估肿瘤数量和大小、结肠免疫细胞组成/功能， 上皮功能2.确定CCL 11是否也是散发性/遗传介导的结肠炎中的修饰剂 肿瘤发生在小鼠模型中，CCL 11的缺失可保护小鼠免受CAC的影响，并且在小鼠模型中CCL 11的表达增加。 APC突变肿瘤。发生结肠肿瘤的CDX 2 P-CreERT 2Apcfl/fl和CDX 2 P-CreERT 2Apcfl/flCcl 11-/-小鼠 将被用于检验CCL 11的缺失将通过减少肿瘤- 通过评估肿瘤数量和大小、结肠免疫细胞组成/功能促进炎症，以及 上皮功能3.为了确定用中和抗体靶向CCL 11信号传导是否能保护 从结肠肿瘤的形成。将处理暴露于AOM-DSS的WT小鼠和CDX 2 P-CreERT 2Apcfl/fl小鼠 与CCL 11中和抗体（或同种型对照）接触，以确定对肿瘤数量和大小的影响， 结肠免疫细胞组成/功能和上皮功能。这些研究将提供新的机制 支持使用基于抑制CCL 11的治疗方法治疗结肠炎和结肠癌。