Role of WNT-EGFR crosstalk by EVs and exomeres in normal colon and colon cancer

EV 和外泌体 WNT-EGFR 串扰在正常结肠和结肠癌中的作用

• 批准号: 10544807
• 负责人: Robert J. Coffey
• 金额: $ 34.57万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-22 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544807
• 关键词: AREG gene; Apple; Automobile Driving; Biogenesis; Cell Line; Cells; Cetuximab; Colon; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Data; Drug resistance; EGF gene; Epidermal Growth Factor Receptor; Epithelium; Equilibrium; Fluorescence; Genes; Goals; Homeostasis; Human; Intestines; KRAS2 gene; LGR5 gene; LacZ Genes; Large Intestine Carcinoma; Link; Luciferases; MDCK cell; Malignant Neoplasms; Mediating; Methodology; Modeling; Mus; Mutation; Neoplasms; Neoplastic Cell Transformation; Oncogenic; Organoids; Pathway interactions; Patients; Pattern; Play; Population; Preparation; Proteins; Publishing; RNA; Reagent; Regulation; Reporter; Reporting; Resistance; Role; Signal Pathway; Signal Transduction; Sorting; Testing; Text; Tumor-Associated Process; Vesicle; WNT Signaling Pathway; Work; colorectal cancer progression; experimental study; extracellular vesicles; intestinal crypt; mouse model; mutant; nanoparticle; neoplastic; overexpression; stem cell function; stem cell niche; stem cells; tumor; tumor growth; tumor progression; vesicular release

Project 3 Summary (Coffey) The Coffey lab has identified important links between WNT and EGFR signaling in cetuximab (CTX) resistance and intestinal crypt homeostasis. We recently reported a new mode of CTX resistance due to increased WNT signaling mediated by miR-100/-125b. These two miRs are upregulated in extracellular vesicles (EVs) released by CTX-resistant cells and these EVs can transfer CTX resistance. In a unique EGFR and WNT reporter mouse model, we show that activation of WNT signaling in an EGFR-sensitized background dramatically increases both EGFR and non-cell autonomous WNT activity. Based on these findings, we propose a model of opposing gradients of EGFR and WNT activity in the colonic stem cell niche (SCN) that contribute to homeostasis and disruption of the gradient is a feature of neoplastic transformation. We hypothesize that in the normal crypt niche EVs and exomeres released by the EGFR-active and WNT-active compartments reinforce the EGFR-WNT gradient and in CRC these nanoparticles serve to drive tumor growth and define cancer progression due to their oncogenically altered constituents. The model also provides a framework to further examine the role of EVs and exomeres in conferring CTX resistance, at least in part, via increased WNT signaling. To examine this model and to determine how EVs participate in CTX resistance, with the ultimate goal of devising strategies to overcome CTX resistance, we propose three Aims. Aim 1 is to determine the effect of EVs and exomeres isolated from highly informative paired cell lines on EGFR and WNT activity in reporter cell lines. Aim 2 is to test the hypothesis that these EVs and exomeres regulate normal stem cell patterning and tumor progression using our unique EGFR and WNT reporter mouse models and their derived organoids. Aim 3 is to elucidate mechanistic underpinnings of EV participation in resistance to EGFR blockade. This work has the potential to alter our fundamental understanding of normal stem cell function, regulation of tumor growth and processes regulating drug resistance.

项目3摘要（Coffey） 科菲实验室已经确定了西妥昔单抗（CTX）电阻中的Wnt和EGFR信号之间的重要联系 和肠道稳态​​。我们最近报道了由于Wnt增加而导致的一种新的CTX抗性模式 由miR-100/-125b介导的信号传导。这两个miR在释放的细胞外囊泡（EV）中上调 通过耐CTX的细胞和这些电动汽车可以传递CTX耐药性。在独特的EGFR和WNT记者鼠标中 模型，我们表明在EGFR敏感的背景中的Wnt信号的激活大大增加了 EGFR和非电池自动wnt活性。基于这些发现，我们提出了一种对立的模型 在稳态和稳态的结肠干细胞生态位（SCN）中EGFR和WNT活性的梯度 梯度的破坏是肿瘤转化的特征。我们假设在正常地下室 EGFR活性和Wnt-Active室释放的电动汽车和外事物增强了EGFR-WNT 梯度和CRC中，这些纳米颗粒用于驱动肿瘤的生长并确定癌症的进展 肿瘤变化的成分。该模型还提供了一个框架，以进一步研究电动汽车的作用 至少部分通过增加的Wnt信号传导来赋予CTX抗性的外侧。检查这个模型 并确定电动汽车如何参与CTX抵抗，其最终目标是制定策略以克服 CTX抗性，我们提出了三个目标。目的1是确定与与之隔离的电动汽车和外事物的影响 在报告基细胞系中，在EGFR和WNT活性上有足够的信息配对细胞系。目标2是检验假设 这些电动汽车和外体使用我们的独特 EGFR和WNT报告的小鼠模型及其衍生的类器官。目标3是阐明机械的 电动汽车参与对EGFR封锁的阻力的基础。这项工作有可能改变我们的 对正常干细胞功能的基本了解，调节肿瘤生长和调节过程 耐药性。