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：
10413032
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/10413032
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 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 Genetic Translation Genomic DNA Goals Growth Human Immune checkpoint inhibitor Immunotherapeutic agent Immunotherapy In Vitro Incidence Individual Interstitial Collagenase Intervention 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 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 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，结肠和直肠的癌症在事件中排名第三，仅次于前列腺和肺部。关于诊断为结肠癌的退伍军人中有三分之一将死于转移性疾病。但是，结肠癌传播基础的分子事件仍然不确定，目前治疗晚期疾病是有限的，仅提供短暂的益处。迫切需要更有效，耐用的治疗方法。和先前的VA值得支持，我们发现M3毒蕈碱受体（M3R）激活促进了结肠癌的进展通过许多不利动作。我们的新发现表明，两个毒蕈碱受体亚型M1R和M3R编码在结肠癌继续和结肠癌干细胞中，CHRM1和CHRM3的过表达（CCSC）驱动肿瘤弹性，继电器和转移。这些发现表明同时针对M1R M3R直接具有巨大的翻译潜力。彼此之间，我们建议检验中心假设有选择地靶向毒蕈碱受体亚型可减轻结肠癌的进展和对治疗的抗性。为此，我们必须填补有关M1R和M3R表达方式如何调节颜色的关键空白肿瘤并确定M1R和M3R在驱动癌症进展中的个体作用。我们的新数据表明随着肿瘤的发展，与M3R表达相比，M1R表达逐渐降低。大多数颜色癌症M3R水平的高度超过了M1R的水平，我们无法在肿瘤侵入性的CCSC中检测到M1R 边缘。值得注意的是，我们确定了M1R表达与两种microRNA的相互关系，mir- 通过计算分析预测的107和miR-103与CHRM1 3'-UTR相互作用。这些发现是与MiR-107/-103一致抑制晚期癌症中的CHRM1 mRNA翻译，从而阻尼M1R 水平。由于miR-107/-103也可以扩增针对ChRM3的β-catenin信号传导，因此这些miRNA可能间接增强M3R水平。 MiR-30-5p的表达降低也可能导致M3R水平升高，这可以与CHRM3 3'-UTR相互作用，与结肠癌中M3R水平有反比关系。有趣的是，我们的新发现还表明，结肠癌还表达了一个圆形（Circ）RNA，CTFRC，能够“ sponging” mir -107 / - 103;因此，通过降低自由miR-107和miR-103的水平，间接调节M1R和M3R水平 - 我们提出利用circrnas的治疗潜力的建议。在这些新发现的基础上，我们建议使用体外，体内和体内模型的体外软化，以对后的细致，重点研究通过这些非编码RNA对M1R和M3R表达的转录调节，并了解如何调节M1R M3R水平会影响重要的临床前终点 - 肿瘤的生长，扩散，对化学和化学的反应免疫疗法和动物生存。为了实现这些目标，我们提出了两个全面的具体目标： AIM 1：严格测试选择性耗尽和失活的M1R和M3R对结肠癌的影响进展，对化学和免疫疗法的反应以及存活。目标2：将miR -107，-103和-30-5p的确切作用定义为M1R和M3R水平的调节剂，以及结肠癌进展的生物标志物以及Circrna海绵的治疗潜力。由于毒蕈碱受体可以被FDA批准的其他适应症的抑制剂选择性地吸毒，因此我们相信在动物模型中调节毒蕈碱受体活性的治疗益处模仿人类疾病将刺激该药物的临床试验，以治疗结肠癌。识别小说与结肠癌进展相关的microRNA/circrna/毒蕈碱受体轴将是一个主要的概念前进我们的新提议的circrna海绵为miR-107/-103表示了治疗诺言。