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

此协作退伍军人奖励应用程序(CMA)利用了经验丰富的研究人员的专业知识，这些研究人员形成了 VA结直肠癌(CRC)细胞基因组学联盟(VA4C)于2017年5月成立。癌症干细胞(CSCs) 致癌的关键介质，并导致癌症复发，从而导致不良的结果。 糖胺多聚糖(GAG)是一种线性多糖，在几个特征的调节中发挥关键作用 在癌症中，与各种细胞表面受体靶点接触，特别是在肿瘤干细胞上。然而，它们在生物方面的潜力 癌症还没有被认识到。基于我们最近非常耐人寻味的观察，G2.2，一种非糖类 天然GAG的糖胺多糖模拟物(NSGM)，通过反向定向选择性抑制CSCs 在涉及生长因子的作用机制方面，我们认为G2.2是一种新型的选择性抑制人CSCs的药物。我们 假设独特的NSGM可以选择性和有效地通过一种新的 关于激活生长因子的反足多向作用机制。我们提出了三个目标。我们 将在50个特性良好的CSCs富集人原代结直肠癌中测试G2.2的疗效 球体/肿瘤样体以及正常肠道器官样体，检查综合CSC表型(自身 更新、迁移、侵袭和化疗耐药)，并确定其药代动力学 最有前途的NSGM(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 将确定反足多效性是如何产生的，例如成纤维细胞生长因子受体的激活 (FGFRs)和抑制胰岛素样生长因子1受体(IGF1R)，并改变pp38/perk信号转导比， 这对CSC的表型有贡献。最后，在目标3中，我们将确定NSGMs在 通过检测晚期体内干细胞的有效性(特别是对CSCs)和毒性(对正常干细胞的影响) 患者来源的异种移植物(PDX)或HT29原位异种移植物模型 化疗(5-氟尿嘧啶和奥沙利铂)。拟议的研究将通过合作大大加强 择优过程。具体地说，我们与Mohapatra博士(Subhra)(Tampa)，Raufman博士(巴尔的摩)合作 Pisegna(Greater LA)在我们的癌症病理和治疗干细胞集群(CSCPT)中， 以及其他VA4C合作者，包括莫哈帕特拉/凯利(坦帕/达勒姆)和布韦博士(圣地亚哥) 将允许-a)在新的肿瘤样模型中测试NSGM以进行独立验证(目标1)；b)解密 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c)测试 新型动物模型中的NSGMs，如AIM 3中的PDX和远端结肠原位异种移植。类似地，我们的 通过该提案获得的专业知识将与Mohapatra博士和Raufman博士(CSC 检测，如CSC标记的综合分析。CSC选择性等)，我们还将协助其他VA4C 调查人员采集血液、血清和组织样本。预计拟议的研究将有所帮助。 破译新的CSCs信号并开发独特的GAG模拟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.