Microbiome-derived regulators of therapy-resistant colorectal tumors

项目总结 结直肠癌(CRC)是美国男性和女性的第三大致命癌症，尽管有效 早期结直肠癌、晚期实体瘤表现为缺氧和/或血管生成不良的治疗方法较多 很难治疗。为了改善这些肿瘤的预后，迫切需要新的治疗方案。 肠道细菌群落(微生物区系)提供了一种可能的解决方案。我们团队的证据也是如此 如其他的，表明通过微生物区系的代谢过程产生的小分子代谢物可以 促进并在某些情况下抑制结直肠癌的发生。这些发现表明， “细菌衍生代谢物疗法(BdMT)”作为耐药结直肠癌的变革性治疗。然而， 我们对这些肿瘤-微生物相互作用的了解还处于初级阶段，还需要做更多的工作 BdMT变成了现实。特别是，我们缺乏关于微生物衍生的代谢产物的身份的知识，这些代谢产物调节 结直肠癌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，我们使用了一个强大的临床前实验模型系统(类肿瘤)来分析 微生物代谢产物对肿瘤细胞的生理作用，结合前沿的多组体 研究肿瘤反应的分子机制和鉴定其小分子的分析工具 监管者。我们的假设是，用从粪便样本中获得的代谢物组分治疗结直肠癌肿瘤 通过多组学分析将揭示微生物组衍生的小分子肿瘤调节因子 可进一步用于治疗应用。我们将通过以下具体目标来检验这一假设： 目的1.建立结直肠癌类肿瘤模型，检测肿瘤细胞对粪便代谢产物组分的反应。 利用Pi Subramanian的专业知识，我们将建立患者来源的CRC肿瘤样，并治疗这些 从结直肠癌患者和健康对照组的粪便样本中分离出代谢物部分。肿瘤细胞 将对每个被测试的组分进行增殖检测；目标2.确定和验证分子调节器和 用多组学方法研究结直肠癌类肿瘤反应的机制。对于引起增殖的代谢物组分 肿瘤细胞反应在AIM 1中，将对肿瘤细胞进行先进的多组体分析来研究 应对机制。粪便样本的元转录分析将确定微生物区系的功能途径 在荷瘤患者中很活跃。肿瘤反应机制和活跃的微生物代谢途径将是 用于预测粪便部分中潜在的小分子调节剂活性。基于质谱学的 代谢组学将确定潜在的小分子肿瘤调节剂。由我们的不同专业知识支持 在癌症微生物学、肿瘤-微生物相互作用和多组体分析方面的团队，我们将发现潜在的新的 进一步研究肿瘤调节代谢物，使BdMT成为现实的关键一步 儿童权利公约的结果。

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.