Unique Non-Saccharide Mimetics of Sulfated Glycosaminoglycan Target Colon Cancer Stem Cells

这个协作VA优点应用程序（CMA）利用了成就研究人员的专业知识，他们形成了一个 VA结直肠癌（CRC）细胞基因组学联盟（VA 4C）于2017年5月。癌症干细胞（CSC） 致癌作用的关键介质，并诱导癌症复发，导致不良结果。 糖胺聚糖（GAG），线性多糖，在调节几个标志物中发挥关键作用 与各种细胞表面受体靶点结合，特别是在CSC上。然而，它们的生物学潜能 癌症还没有被发现。基于我们最近非常有趣的观察，G2.2，一种非糖 天然GAG的糖胺聚糖模拟物（NSGM）通过相反定向的免疫抑制选择性抑制CSC。 由于G2.2是一种新型的、选择性的肿瘤干细胞抑制剂，因此我们认为G2.2是一种新型的肿瘤干细胞抑制剂。我们 假设独特NSGM可以通过一种新的 关于生长因子激活的对跖多效性机制。我们提出三个目标。我们 将测试G2.2在50个充分表征的富集原代人结肠直肠CSC中的功效 球状体/类肿瘤以及正常肠类器官，检查全面的CSC表型（自身）， 更新、迁移、侵袭和化疗耐药性），并确定药物的药代动力学特征。 最有前途的NSGMs（目标1）。在目标2中，我们将确定G2.2（及其类似物）的分子机制。我们 将决定如何产生对跖多效性效应，例如，成纤维细胞生长因子受体活化 （FGFR）和抑制胰岛素样生长因子1受体（IGF 1 R），并改变pp 38/pERK信号传导比率， 这有助于CSC表型。最后，在目标3中，我们将确定NSGMs在以下方面的治疗效用： 通过在体内检查晚期干细胞的功效（特别是对CSC）和毒性（对正常干细胞的影响）， 患者来源的异种移植物（PDX）或HT 29原位异种移植物的模型，其单独或与 化疗（5-氟尿嘧啶和奥沙利铂）。建议的研究将大大加强合作， 择优程序具体来说，我们与Mohapatra（Subhra）（坦帕）、Raufman（巴尔的摩） Pisegna（大洛杉矶）在我们的癌症干细胞在病理学和治疗CRC（CSCPT）集群， 以及其他VA 4C合作者，包括Mohapatra/Kelly（坦帕/达勒姆）和Bouvet博士（圣地亚哥） 将允许- a）在新的类肿瘤模型中测试NSGM以进行独立验证（目标1）; B）破译 通过效应与多组学研究的相关性预测NSGM效应的生物标志物;以及 新型动物模型中的NSGM，如Aim 3中的PDX和远端结肠原位异种移植物。同样，我们的 通过该提案获得的专业知识和知识将与Mohapatra博士和Raufman博士（CSC）分享 分析，例如CSC标志物的综合分析。CSC选择性等），我们还将协助其他VA 4C 收集血液、血清和组织样本。预计拟议的研究将有助于 破译新的CSC信号传导，并开发独特的GAG模拟物作为临床使用的原型试剂。

This Collaborative VA Merit Application (CMA) leverages expertise of accomplished researchers who formed a VA Colorectal Cancer (CRC) Cell-Genomics Consortium (VA4C) in May 2017. Cancer stem cells (CSCs) are critical mediators of carcinogenesis and induce cancer relapse resulting in poor outcomes. Glycosaminoglycans (GAGs), linear polysaccharides, that play a critical role in regulation of several hallmarks of cancer, engage various cell surface receptor targets, especially on CSCs. Yet, their biological potential in cancer has not been realized. Based on our highly intriguing recent observations that G2.2, a non-saccharide glycosaminoglycan mimetic (NSGM) of a natural GAG, selectively inhibits CSCs through an oppositely-directed mechanism involving growth factors, we propose that G2.2 is a novel, selective inhibitor of human CSCs. We hypothesize that unique NSGMs can selectively and potently inhibit colorectal CSC growth through a novel mechanism of antipodal pleiotropicity with respect to activation of growth factors. We propose three aims. We will test the efficacy of G2.2 in 50 well characterized CSCs enriched primary human colorectal spheroids/tumoroids as well as normal intestinal organoids, examine comprehensive CSC phenotype (self- renewal, migration, invasion, and chemotherapy resistance), and determine pharmacokinetic profile of the most promising NSGMs (Aim 1). In Aim 2, we will determine molecular mechanism of G2.2 (and analogs). We will determine how antipodal pleiotropic effects arise, e.g., activation of fibroblast growth factor receptor (FGFRs) and inhibition of insulin-like growth factor 1 receptor (IGF1R), and alter pp38/pERK signaling ratio, which contributes to the CSC phenotype. Finally, in Aim 3, we will determine therapeutic utility of NSGMs in vivo by examining efficacy (specifically on CSCs) and toxicity (effect on normal stem cells) in advanced in vivo models of patient-derived xenografts (PDXs) or HT29 orthotopic xenografts either alone or in combination with chemotherapy (5-fluorouracil & oxaliplatin). The proposed studies will be greatly enhanced by collaborative merit process. Specifically, our collaboration with Drs. Mohapatra (Subhra)(Tampa), Raufman (Baltimore) Pisegna (Greater LA) within our cluster of Cancer Stem Cells in Pathology and Treatment of CRCs (CSCPT), as well as other VA4C collaborators including Mohapatra/Kelly (Tampa/Durham), and Dr. Bouvet (San Diego) will allow – a) testing of NSGMs in novel tumoroid models for independent validation (Aim 1); b) decipher predictive biomarkers for NSGM effect through correlation of effect with muti-omics studies; and c) testing of NSGMs in novel animal models such as PDXs and distal colon orthotopic xenografts in Aim 3. Similarly, our expertise and knowledge gained through the proposal will be shared with Drs. Mohapatra and Raufman (CSC assays e.g. comprehensive analyses of CSC markers. CSC selectivity etc.), We will also assist other VA4C investigators in collecting blood, serum and tissue samples. The proposed studies are expected to help decipher novel CSCs signaling and develop unique GAG mimetics as prototypic agents for clinical use.