Microbiome-derived regulators of therapy-resistant colorectal tumors

项目摘要 结直肠癌（CRC）是美国男性和女性的第三大致命癌症。 对于早期CRC、表现出缺氧和/或血管形成不良的晚期实体瘤的治疗更多地是 为了改善这些肿瘤的预后，迫切需要新的CRC治疗方案。 肠道细菌群落（微生物群）提供了一种可能的解决方案。我们小组的证据也是 和其他人一样，表明通过微生物群的代谢过程产生的小分子代谢物可以 促进并且在某些情况下抑制CRC肿瘤发生。这些发现表明， “细菌衍生代谢物疗法（BdMT）”作为耐药CRC肿瘤的变革性治疗。然而，在这方面， 我们对这些肿瘤-微生物相互作用的理解还处于起步阶段，需要做更多的工作， BdMT成为现实。特别是，我们缺乏对微生物衍生的代谢物的身份的知识， CRC肿瘤，以及这些分子调节肿瘤细胞的潜在分子机制。 为了填补这些空白，我们使用强大的临床前实验模型系统（类肿瘤）进行测定， 微生物衍生代谢物对肿瘤细胞的生理作用，加上尖端的多组学 分析工具，以研究肿瘤反应的分子机制，并确定其小分子 监管部门我们的假设是，用从粪便样品中获得的代谢物组分治疗的CRC类肿瘤 通过多组学分析CRC患者将揭示微生物组来源的小分子肿瘤调节剂 其可进一步用于治疗应用。我们将通过这些具体目标来检验这一假设： 目标1.建立CRC类肿瘤模型并测试肿瘤细胞对粪便代谢物组分的反应。 利用PI Subramanian的专业知识，我们将建立患者源性CRC类肿瘤，并治疗这些肿瘤。 用从CRC患者和健康对照的粪便样品中分离的代谢物级分。肿瘤细胞 将测定每个测试级分的增殖;目的2.识别和验证分子调节剂， 使用多组学的CRC类肿瘤反应机制。对于引起增殖的代谢物组分， 目标1中的肿瘤细胞反应，将对肿瘤细胞进行高级多组学分析以研究 反应机制。粪便样本的元转录组学分析将确定微生物群功能途径 在肿瘤患者中有效。肿瘤反应机制和活跃的微生物代谢途径将在 用于预测在粪便组分中具有活性的潜在小分子调节剂。基于质谱的 代谢组学将确定潜在的小分子肿瘤调节剂。通过我们的各种专业知识， 癌症微生物学，肿瘤-微生物相互作用和多组学分析团队，我们将确定潜在的新的 肿瘤调节代谢物进行进一步研究，这是使BdMT成为现实的关键一步， CRC的结果。

PROJECT SUMMARY Colorectal cancer (CRC) is the third most deadly cancer for both men and women in the U.S. Despite effective treatments for early-stage CRC, late-stage solid tumors exhibiting hypoxic and/or poor vascularization are more difficult to treat. To improve the prognosis for these tumors, new treatment regimes for CRC are urgently needed. Intestinal tract bacterial communities (microbiota) present a possible solution. Evidence from our team, as well as others, indicates small molecule metabolites produced via metabolic processes of the microbiota can promote, and in some cases inhibit, CRC tumorigenesis. These findings have suggested the possibility of “Bacterial-derived Metabolite Therapy (BdMT)” as a transformative treatment of resistant CRC tumors. However, our understanding of these tumor-microbe interactions is only in its infancy, and more work is needed to make BdMT a reality. In particular, we lack knowledge on the identity of microbe-derived metabolites that regulate CRC tumors, and also the underlying molecular mechanisms by which these molecules regulate the tumor cells. In order to fill in these gaps, we use a powerful preclinical experimental model system (tumoroids) for assaying physiological effects of microbe-derived metabolites on tumor cells, coupled with cutting-edge multi-omic analysis tools to investigate molecular mechanisms of tumor response and identify their small molecular regulators. Our hypothesis is that CRC tumoroids treated with metabolite fractions obtained from fecal samples of CRC patients and analyzed via multi-omics will reveal microbiome-derived small molecule tumor regulators which can be further leveraged for therapeutic applications. We will test this hypothesis via these Specific Aims: Aim 1. Establish a CRC tumoroid model and test tumor cell response to fecal metabolite fractions. Leveraging the expertise of PI Subramanian, we will establish patient-derived CRC tumoroids, and treat these with metabolite fractions isolated from fecal samples of CRC patients and healthy controls. Tumor cell proliferation will be assayed for each fraction tested; Aim 2. Identify and validate molecular regulators and mechanisms of CRC tumoroid response using multi-omics. For metabolite fractions eliciting proliferative tumor cell response in Aim 1, advanced multi-omic analysis will be performed on the tumor cells to investigate response mechanisms. Metatranscriptomic analysis of fecal samples will identify microbiota functional pathways active in tumor-bearing patients. Tumor response mechanisms and active microbial metabolic pathways will be used to predict potential small molecule regulators active in the fecal fractions. Mass spectrometry-based metabolomics will identify potential small molecule tumor regulators. Enabled by the diverse expertise of our team in cancer microbiology, tumor-microbe interactions and multi-omic analysis, we will identify potential new tumor-regulating metabolites for further investigation, a critical step towards making BdMT a reality for improving outcomes of CRC.