Enteric Glia is New Biological Target to Block Drug Resistance in Colon Cancer

肠胶质细胞是阻断结肠癌耐药性的新生物靶点

• 批准号: 10659846
• 负责人: Laurianne Chantal Van Landeghem
• 金额: $ 34.03万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659846
• 关键词: 3-Dimensional; ATM Signaling Pathway; Apoptosis; Automobile Driving; Biological; Cell Line; Cells; Chemoprotection; Chemoprotective Agent; Chemoresistance; Chemotherapy-Oncologic Procedure; Coculture Techniques; Colon; Colon Adenocarcinoma; Colon Carcinoma; Colonic Neoplasms; Complex; Cues; Cytoprotection; DNA Damage; DNA Repair; Development; Dinoprostone; Drug resistance; Drug usage; Embryo; Engraftment; Enteral; Enteric Nervous System; Fluorouracil; Follistatin; Gene Targeting; Genetic; Glial Fibrillary Acidic Protein; Growth; Human; Immunodeficient Mouse; In Vitro; Induction of Apoptosis; Infiltration; Knockout Mice; Ligands; Light; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Molecular; Molecular Profiling; Neuroglia; Operative Surgical Procedures; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Probability; Production; Prognosis; Proliferating; Rattus; Resistance; Signal Transduction; Solid Neoplasm; Survival Rate; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Transgenic Mice; Treatment Efficacy; Tumor Stem Cells; WFDC2 gene; ataxia telangiectasia mutated protein; cancer cell; cancer stem cell; cancer therapy; carcinogenicity; chemotherapy; colon carcinogenesis; design; glial activation; healing; improved; in vivo; mouse model; neoplastic cell; new therapeutic target; novel; novel therapeutics; oxaliplatin; paracrine; pharmacologic; protective effect; public health relevance; resistance factors; response; sensor; single-cell RNA sequencing; stem cell survival; targeted treatment; therapeutic target; tumor; tumor microenvironment

PROJECT SUMMARY/ABSTRACT Resistance to anti-cancer therapies largely explains the abysmal 5-year survival rate of patients with advanced colon cancer. Traditional chemotherapy regimens have been designed to efficiently stop proliferation and initiate apoptosis in cancer cells, but have failed to appreciate the pro-chemoresistance signals emanating from cells surrounding the tumor. We have identified a novel cellular component of the tumor microenvironment: the enteric glial cells (EGC). We and others have shown over the last 15 years that EGC are potent inducers of barrier function and healing in a healthy colon. Recently we have demonstrated that the EGC network substantially infiltrates human colon adenocarcinomas and promotes cancer stem cell tumor-forming abilities via a paracrine PGE2-EP4 pathway. Nevertheless, whether EGC impact colon cancer resistance to chemotherapy remains unknown. Our preliminary studies indicate that EGC protect cancer stem cells against apoptosis induced by chemotherapeutic drugs, allowing for enhanced tumor formation and growth despite the chemotherapy treatment. We also have evidence that this is (1) dependent on activation of the MRN-ATM pathway - a central player in DNA repair- in cancer cells and (2) exacerbated by EGC activation with chemotherapy. Using mass spectrometry analyses, we have identified FSTL3 as a novel EGC-derived factor and generated preliminary results implicating FSTL3 in EGC chemoprotective effects. Therefore, we propose to test the hypothesis that “in response to chemotherapeutic drugs, EGC release larger amounts of FSTL3 in the tumor microenvironment, which enhances cancer stem cell chemoresistance and allows for tumor formation and growth by promoting DNA repair driven by the MRN-ATM pathway”. Specific Aim 1 will determine whether EGC promote cancer stem cell resistance to chemotherapies via the release of FSTL3. Specific Aim 2 will test whether EGC protective effects are mediated by increased DNA repair as a result of activation of the MRN-ATM pathway. Specific Aim 3 will determine whether blocking FSTL3 production in EGC sensitizes colon tumors to chemotherapies in vivo. Studies will use translationally relevant primary cultures of human EGC and cancer cells, 3D co-culture platforms, orthotopic co-engraftment in immunodeficient mice, murine models of colon carcinogenesis, transgenic mice allowing for chemogenetic activation of EGC (GFAP-hM3Dq) and inducible gene targeting in EGC (GFAP- CreERT2), in addition to cutting-edge molecular profiling using single cell RNA seq and mass spectrometry studies to identify the pro-chemoresistance factor(s) (and in particular FSTL3) and pathway(s) involved. These studies will not only improve our understanding of the cellular and molecular mechanisms driving colon cancer chemoresistance but will also demonstrate the therapeutic potential of developing strategies combining targeted therapies against EGC-derived FSTL3 and traditional chemotherapy regimens.

项目总结/摘要 对抗癌治疗的耐药性在很大程度上解释了晚期乳腺癌患者的5年生存率极低。 结肠癌传统的化疗方案已被设计成有效地阻止增殖并启动化疗。 细胞凋亡，但未能认识到从细胞发出的促化学抗性信号 肿瘤周围。我们已经确定了肿瘤微环境的一种新的细胞成分：肠道微环境。 神经胶质细胞（EGC）。在过去的15年里，我们和其他人已经证明EGC是屏障的有效诱导剂， 功能和愈合在一个健康的结肠。最近，我们已经证明，EGC网络实质上 浸润性人结肠腺癌并通过旁分泌促进癌症干细胞肿瘤形成能力 PGE 2-EP 4通路。尽管如此，EGC是否影响结肠癌对化疗的耐药性仍有待研究 未知我们的初步研究表明，EGC保护肿瘤干细胞对抗由 化疗药物，允许增强的肿瘤形成和生长，尽管化疗 治疗我们也有证据表明，这是（1）依赖于激活的MRN-ATM途径-一个中央的 参与DNA修复-在癌细胞和（2）加剧了EGC激活与化疗。利用大众 光谱分析，我们已经确定FSTL 3作为一种新的EGC衍生因子，并产生了初步的 结果表明FSTL 3参与EGC化学保护作用。因此，我们建议测试假设，“在 对化疗药物的反应，EGC在肿瘤微环境中释放更大量的FSTL 3， 它增强了癌症干细胞的化学抗性，并通过促进DNA 由MRN-ATM途径驱动的修复”。特异性目的1将决定EGC是否促进癌症干细胞 通过释放FSTL 3对化疗产生抗性。具体目标2将测试EGC是否具有保护作用 是由MRN-ATM途径激活导致的DNA修复增加介导的。第3章将 确定阻断EGC中FSTL 3的产生是否使结肠肿瘤对体内化疗敏感。 研究将使用与诊断相关的人EGC和癌细胞的原代培养物、3D共培养平台， 免疫缺陷小鼠原位共移植，结肠癌发生的小鼠模型，转基因小鼠 允许EGC的化学发生活化（GFAP-hM 3Dq）和EGC中的诱导型基因靶向（GFAP-hM 3Dq）， CreERT 2），以及使用单细胞RNA测序和质谱的尖端分子分析 研究以鉴定所涉及的前化学抗性因子（并且特别是FSTL 3）和途径。这些 这些研究不仅将提高我们对结肠癌细胞和分子机制的理解， 但也将证明开发策略的治疗潜力， 针对EGC衍生的FSTL 3和传统化疗方案的治疗。