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Muscarinic Receptors Regulate Colon Cancer Stem Cell Function and Invasiveness

毒蕈碱受体调节结肠癌干细胞功能和侵袭性

基本信息

批准号：
10664886
负责人：
JEAN-PIERRE RAUFMAN
金额：
--
依托单位：
BALTIMORE VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10664886
关键词：
3&apos Untranslated Regions 3-Dimensional Advanced Malignant Neoplasm Affect Animal Model Animals Attenuated Automobile Driving Binding Sites Biological Assay Biological Markers Bioluminescence CHRM1 gene CHRM3 gene Cell Culture Techniques Cells Clinical Trials Colon Colon Carcinoma Colonic Neoplasms Computer Analysis Computer Models Data Diagnosis Disease Equilibrium Event Exonuclease FDA approved Fluorouracil Genetic Transcription Genomic DNA Goals Growth Human Immune checkpoint inhibitor Immunotherapeutic agent Immunotherapy In Vitro Incidence Individual Interstitial Collagenase Intervention Invaded Investigation Knowledge Learning Link Luciferases Lung Malignant Neoplasms Messenger RNA MicroRNAs Molecular Mothers Mus Muscarinic Acetylcholine Receptor Muscarinic M3 Receptor Neoplasm Metastasis Neoplasms Neurons Organoids Pattern Persons Porifera Post-Transcriptional Regulation Property Prostate RNA Receptor Activation Rectal Cancer Regulation Relapse Reporter Reporting Research Resistance Role Signal Transduction Testing Therapeutic Transfection Translations Untranslated RNA Untranslated Regions Veterans Work Xenograft Model Xenograft procedure advanced disease beta catenin cancer cell cancer stem cell chemotherapy colon cancer progression colon cancer treatment design drug repurposing established cell line human disease in vivo in vivo Model inhibitor innovation intervention effect mRNA Translation novel overexpression oxaliplatin pre-clinical prognostic programmed cell death protein 1 promoter receptor receptor expression resilience response stem cell differentiation stem cell function stem cell proliferation therapy resistant translational potential treatment response tumor tumor growth tumor progression

项目摘要

In Veterans, cancers of the colon and rectum rank third in incidence behind those of the prostate and lung; about one-third of Veterans diagnosed with colon cancer will die, primarily from metastatic disease. However, the molecular events underlying the spread of colon cancer remain uncertain and current treatments for advanced disease are limited and provide only transient benefit. More effective, durable treatments are urgently needed. With previous VA Merit support, we found M3 muscarinic receptor (M3R) activation promotes colon cancer progression by many adverse actions. Our new findings reveal that two muscarinic receptor subtypes, M1R and M3R encoded by CHRM1 and CHRM3, are overexpressed early in the colon cancer continuum and in colon cancer stem cells (CCSCs) which drive tumor resilience, relapse, and metastasis. These findings suggest concurrently targeting M1R and M3R directly may have great translational potential. Accordingly, we propose to test the central hypothesis that selectively targeting muscarinic receptor subtypes attenuates colon cancer progression and resistance to therapy. To do so, we must fill key gaps in knowledge regarding how M1R and M3R expression is regulated in colon neoplasia and identify the individual roles of M1R and M3R in driving cancer progression. Our new data reveal that as neoplasia progresses, M1R expression is progressively reduced compared to M3R expression; in most colon cancers M3R levels greatly exceed those of M1R and we could not detect M1R in CCSCs at the tumor invasive edge. Notably, we identified a reciprocal relationship between M1R expression and that of two microRNAs, miR- 107 and miR-103, predicted by computational analysis to interact with the CHRM1 3’-UTR. These findings are consistent with miR-107/-103 suppressing CHRM1 mRNA translation in advanced cancer, thereby damping M1R levels. As miR-107/-103 can also amplify β-catenin signaling, which targets CHRM3, these miRNAs may indirectly augment M3R levels. Increased levels of M3R may also result from reduced expression of miR-30-5p which can interact with the CHRM3 3’-UTR and has an inverse relationship with M3R levels in colon cancer. Intriguingly, our new findings also show colon cancers also express a circular (circ)RNA, cTFRC, capable of ‘sponging’ miR-107 / - 103; and, thus indirectly regulating M1R and M3R levels by reducing levels of free miR-107 and miR-103 – we propose to leverage the therapeutic potential of circRNAs. Building on these new findings, we propose to use sophisticated in vitro, ex vivo, and in vivo models to perform a meticulous, focused investigation of the post- transcriptional regulation of M1R and M3R expression by these non-coding RNAs and learn how modulating M1R and M3R levels impacts important preclinical endpoints – tumor growth, spread, response to chemo- and immunotherapy, and animal survival. To accomplish these goals, we propose two comprehensive Specific Aims: Aim 1: Rigorously test the effects of selectively depleting and inactivating M1R and M3R on colon cancer progression, responses to chemo- and immunotherapy, and survival. Aim 2: Define the exact roles of miR-107, -103, and -30-5p as regulators of M1R and M3R levels and biomarkers for colon cancer progression, and the therapeutic potential of circRNA sponges. As muscarinic receptors are selectively druggable by inhibitors already FDA-approved for other indications, we believe demonstrating the therapeutic benefits of modulating muscarinic receptor activity in animal models that mimic human disease will spur clinical trials of such agents repurposed to treat colon cancer. Identifying a novel microRNA/circRNA/muscarinic receptor axis linked to colon cancer progression will be a major conceptual advance whose therapeutic promise is exemplified by our newly proposed circRNA sponges for miR-107/-103.

在退伍军人中，结肠癌和直肠癌的发病率排名第三，仅次于前列腺癌和肺癌; 三分之一被诊断患有结肠癌的退伍军人会死亡，主要是死于转移性疾病。但结肠癌扩散背后的分子事件仍然不确定，疾病是有限的，只能提供短暂的好处。迫切需要更有效、更持久的治疗方法。与在之前的VA Merit支持下，我们发现M3毒蕈碱受体（M3 R）激活促进结肠癌进展许多不利的行动。我们的新发现揭示了两种毒蕈碱受体亚型，M1 R和M3 R编码在结肠癌连续体和结肠癌干细胞中早期过表达（CCSC），其驱动肿瘤复原力、复发和转移。这些发现表明同时靶向M1 R 而M3 R可能直接具有很大的翻译潜力。因此，我们建议测试中心假设，选择性靶向毒蕈碱受体亚型减弱了结肠癌的进展和对治疗的抗性。要做到这一点，我们必须填补有关结肠中M1 R和M3 R表达如何调节的知识空白。肿瘤，并确定M1 R和M3 R在推动癌症进展中的各自作用。我们的新数据显示，随着肿瘤的进展，M1 R表达与M3 R表达相比逐渐减少;在大多数结肠中，肿瘤细胞M3 R水平大大超过M1 R水平，我们在肿瘤浸润的CCSC中不能检测到M1 R。边缘值得注意的是，我们确定了M1 R表达与两种microRNA，miR- 107和miR-103，通过计算分析预测与CHRM 1 3 '-UTR相互作用。这些发现与miR-107/-103抑制晚期癌症中CHRM 1 mRNA翻译一致，从而抑制M1 R 程度.由于miR-107/-103也可以放大靶向CHRM 3的β-catenin信号传导，因此这些miRNAs可以间接地增加M3 R水平。M3 R水平的增加也可能是由于miR-30- 5 p表达的减少，在结肠癌中，M3 R与CHRM 3 3 '-UTR相互作用，并与M3 R水平呈反比关系。有趣的是，我们新的发现还表明，结肠癌也表达一种环状（circ）RNA，cTFRC，能够“吸收”miR-107 / - 因此，通过降低游离miR-107和miR-103的水平来间接调节M1 R和M3 R水平-我们发现，提出利用circRNA的治疗潜力。在这些新发现的基础上，我们建议使用复杂的体外、离体和体内模型，以进行细致的、有重点的研究，这些非编码RNA对M1 R和M3 R表达的转录调控，并了解如何调节M1 R 和M3 R水平影响重要的临床前终点-肿瘤生长，扩散，对化疗的反应-和免疫疗法和动物存活率。为了实现这些目标，我们提出了两个全面的具体目标：目的1：严格测试选择性消耗和失活M1 R和M3 R对结肠癌的影响进展、对化疗和免疫治疗的反应以及存活率。目的2：确定miR-107、-103和-30-5p作为M1 R和M3 R水平调节剂的确切作用，结肠癌进展的生物标志物，以及circRNA海绵的治疗潜力。由于毒蕈碱受体可被FDA批准用于其他适应症的抑制剂选择性药物化，我们相信在动物模型中证明调节毒蕈碱受体活性的治疗益处，模拟人类疾病将刺激临床试验，这些药物将被重新用于治疗结肠癌。识别一本小说与结肠癌进展相关的microRNA/circRNA/毒蕈碱受体轴将是一个重大的概念性进展其治疗前景由我们新提出的针对miR-107/-103的circRNA海绵例证。