Deconvoluting myeloid gene regulation through functional genomics

项目总结 巨噬细胞是肿瘤病变中最大的免疫细胞成分之一，其数量 甚至可以超过肿瘤细胞。尽管MF对组织动态平衡和感染控制很重要，但在 肿瘤MF采用表型状态，使其具有促肿瘤和免疫抑制功能。 正因为如此，MF成为癌症治疗的靶点。然而，调节肿瘤相关的MF已被证明 很难。这在很大程度上是因为我们仍然没有完全了解控制 因此，我们仍然缺乏调节相关MF功能的最佳目标。 这个项目的目标是识别驱动人类免疫效应和抑制免疫活性的基因 Mf.在人类非小细胞肺癌(NSCLC)的单细胞rna-seq分析中，我的赞助实验室发现 与邻近的健康肺组织中的MF相比，肿瘤相关的MF中有大量基因上调。 这包括一些具有免疫抑制功能的基因，但对于大多数已识别的基因来说， 这些功能，特别是可能与免疫力有关的功能，尚不清楚。几乎一半已识别的基因 编码在MF生物学中具有潜在上游调控作用的因子，例如转录因子(Tf)， 信号组件和细胞表面受体。 在这个项目中，我将测试一种假设，即肿瘤相关的MF的免疫抑制活性是由 通过调节因子和信号成分上调肿瘤内的MF。为了检验我的假设，我 我将利用我的赞助实验室开发的一个新的CRISPR基因组学平台，该平台能够实现高维 同时以单细胞分辨率对多个CRISPR基因敲除进行表型鉴定(Wroblewska，Dhainaut 等人的研究。单元格2018)。我将系统地敲除肿瘤相关的MF信号基因，并确定如何 扰动影响细胞的关键表型和功能，包括检查点配体和 调节T细胞活化。通过识别控制MF身份以及效应和抑制功能的基因， 这个项目的结果将为主要细胞之一的分子力学提供新的见解。 在肿瘤微环境中控制免疫反应的方向并提供分子靶点 用于旨在增强抗肿瘤免疫的治疗干预。

PROJECT SUMMARY Macrophages (MF) are one of the largest immune cell components of tumor lesions, where their numbers can even exceed tumor cells. Though MF are important for tissue homeostasis and control of infections, in tumors MF adopt a phenotypic state which confers them with pro-tumorigenic and immunosuppressive functions. Because of this, MF are a target for cancer treatment. However, modulating tumor-associated MF has proved difficult. This is in large part because we still do not have a complete understanding of the genes that control many key aspects of MF biology, and thus we still lack an optimal target for modulating relevant MF functions. The objective of this project is to identify genes that drive effector and suppressor immune activity of human MF. In single cell RNA-seq analysis of human non-small cell lung carcinoma (NSCLC), my sponsor labs identified a large module of genes upregulated in tumor-associated MF compared to MF in adjacent healthy lung tissue. This included a number of genes with immunosuppressive functions, but for a majority of the identified genes, the functions, particularly as they may relate to immunity, are not known. Almost half of the identified genes encode for factors with potential upstream regulatory roles in MF biology, such as transcription factors (TF), signaling components, and cell surface receptors. In this project, I will test the hypothesis that the immunosuppressive activity of tumor-associated MF is driven by the regulatory factors and signaling components upregulated on intratumoral MF. To test my hypothesis, I will utilize a novel CRISPR genomics platform my sponsor lab developed, which enables high-dimensional phenotyping of multiple CRISPR knockouts simultaneously and with single cell resolution (Wroblewska, Dhainaut et al. Cell 2018). I will systematically knockout the tumor-associated MF signature genes and determine how perturbation impacts key phenotypes and functions of the cells, including expression of checkpoint ligands and modulation of T cell activation. By identifying genes that control MF identity and effector and suppressor function, the outcome of this project will provide new insights in to the molecular mechanics of one of the major cells controlling the direction of the immune response in the tumor microenvironment and provide molecular targets for therapeutic interventions aimed at enhancing anti-tumor immunity.