Project II: Immune regulatory circuits in primary colon cancer and lymph node and liver metastases

项目二.原发性结肠癌、淋巴结和肝脏的免疫调节回路 转移 实验负责人：Rudensky 计算主管：Leslie 实验联合负责人：Ganesh 项目摘要 结直肠癌（CRC）是美国和全球第三大常见癌症。尽管主要 通过免疫疗法提供的某些癌症的治疗进展，错配修复熟练（pMMR）CRC， 占绝大多数（85%）的病例，5年生存率约为15%。我们 假设这种失败是由于免疫和基质的CRC促进局部回路的活性， 辅助细胞，其中抑制性调节性T（Treg）细胞和非炎性组织支持性Th17 细胞是重要的组成部分。我们还提出，这些T细胞的结构和细胞组成 细胞回路在原发性CRC和近端淋巴结和远端肝转移中是不同的。在这个项目中， 我们将剖析主要的免疫调节细胞状态，包括Treg和Th17细胞，以及它们的动态变化。 与原发性肿瘤和淋巴结中的细胞组分（“免疫调节邻域”）的相互作用 在相同的pMMR CRC患者中的肝转移以及一种新的自发性肝转移的小鼠遗传模型， 转移性pMMR CRC（目的1）。我们的扰动研究将阐明这些关键的免疫调节作用， 细胞及其伙伴在转移性疾病进展和免疫治疗抵抗（目的2），并检查 关键免疫细胞衍生因子对人类原发性和转移性CRC类器官中癌细胞的影响 (Aim 3）。我们将采用尖端的“组学”分析在一个单一的细胞分辨率结合空间 转录组学和高度多路复用的光学成像和显微镜和流式细胞术， 实验扰动迭代与新的计算建模和分析解构原型 CRC中基于T细胞的细胞回路及其细胞的保守染色质状态和基因调控程序 选民。这些研究将有助于更好地了解促转移细胞的出现机制， 在pMMR CRC中的状态，并为此提供正交合理组合免疫疗法的开发 疾病，并阐明靶向免疫上皮电路在预防和 治疗晚期CRC患者的转移。 .

Project II. Immune regulatory circuits in primary colon cancer and lymph node and liver metastases Experimental Lead: Rudensky Computational Lead: Leslie Experimental Co-Lead: Ganesh PROJECT SUMMARY Colorectal cancer (CRC) represents the third most common cancer in the US and worldwide. Despite major advances in treatment of some cancer afforded by immunotherapy, mismatch repair-proficient (pMMR) CRC, which accounts for the vast majority (85%) of cases, fails to respond with ~15% 5-year survival rate. We hypothesize that this failure is due to the activity of CRC-promoting local circuits of immune and stromal accessory cells, in which suppressive regulatory T (Treg) cells and non-inflammatory tissue-supporting Th17 cells serve as essential components. We also propose that the architecture and cellular composition of these T cell circuits are distinct in primary CRC and in proximal lymph node and distal liver metastases. In this project, we will dissect the dynamics of the main immunomodulatory cells states including Treg and Th17 cell and their interactions with cellular components (“immunomodulatory neighborhoods”) in primary tumors and lymph node and liver metastases in the same pMMR CRC patients alongside a novel mouse genetic model of spontaneously metastasizing pMMR CRC (Aim 1). Our perturbation studies will elucidate roles of these key immunomodulatory cells and their partners in metastatic disease progression and resistance to immunotherapy (Aim 2) and examine the impact of key immune cell-derived factors on cancer cells in human primary and metastatic CRC organoids (Aim 3). We will employ cutting edge “omics” analyses at a single cell resolution combined with spatial transcriptomics and highly multiplexed optical imaging and microscopy and flow cytometry coupled to experimental perturbations iteratively with novel computational modeling and analyses to deconstruct archetypal T cell-based cell circuits in CRC and conserved chromatin states and gene regulatory programs of their cellular constituents. These studies will enable better mechanistic understanding of emergence of pro-metastatic cell states in pMMR CRC and inform development of orthogonal rational combination immunotherapies for this disease and elucidate the therapeutic actionability of targeting immune-epithelial circuits in preventing and treating metastasis in patients with advanced CRC. .