(PQ7)Multi-scale Analysis of Tumor Microenvironment Heterogeneity

摘要 肿瘤发生与先天性和获得性免疫效应器的异质性进化变化有关 肿瘤微环境。记录、识别和量化这些细胞和分子变化 可能会改变我们对肿瘤发生的理解。我们和其他人已经证明，癌基因，如 MYC增选和/或颠覆这些免疫效应器，从而逃避免疫检测并促进肿瘤的发生 促进肿瘤生长的微环境；然而，这似乎发生在一种必然和 可以预见的是，肿瘤极易受到急性癌基因失活的影响，从而使免疫效应器 被激活，从而导致显著的肿瘤消退，这种现象称为“癌基因成瘾”。因此， 我们假设，这些不同的宿主免疫效应器的识别、量化和定位 在肿瘤发生和肿瘤消退期间的肿瘤微环境中，将识别出 将预测癌基因失活的治疗反应。我们的方法将是采用TET系统 MYC诱导的肝细胞癌的调控模型和MYC/Twist1诱导的肝癌模型 转移性肝癌。然后，利用：FACS/CyTOF、Codex/MIBI、IVM/BLI和基因表达/CiberSort 识别细胞效应物和标志性基因和代谢物。首先，我们的MYC转基因小鼠模型- 诱导的肝细胞癌已经被我们和其他许多人广泛使用。使用TET系统，我们的模型展示 MYC癌基因表达的精确可逆和可滴定控制。肿瘤的形成 随着时间的推移，癌基因依赖会导致高度依赖免疫的肿瘤退化。 回应。此外，最近我们制作了一个未发表的小鼠模型，戏剧性地说明了MYC- 联合转基因Twist1诱导的肝肿瘤经血流快速转移 表情。Twist1以前被认为与转移有关。我们已经取得了初步的结果， 显示转移与巨噬细胞的募集有关，并且似乎需要巨噬细胞的募集。最后，我们发现 我们可以使用可用的人类TCGA数据来识别与 人肝细胞癌的发病机制。因此，肿瘤发生的所有阶段似乎都依赖于间质和 免疫反应(初始、进展、转移、消退)，是评估肿瘤的独特工具- 微环境异质性。我们有三个目标：第一，剖析适应性 先天免疫系统促进肝癌的进展、转移和消退；第二，确定 应用体内(IVM和BLI)成像对上述已鉴定细胞群的动力学和定位 技术，特别强调细胞与细胞之间的相互作用，第三，评估这些发现的能力 对人类肝细胞癌的临床行为和预后进行预测。

ABSTRACT Tumorigenesis is associated with heterogeneous evolving changes of innate and adaptive immune effectors in the tumor microenvironment. Recording, identifying and quantifying these cellular and molecular changes could transform our understanding of tumorigenesis. We and others have shown that oncogenes, such as MYC co-opt and/or subvert these immune effectors, thereby evading immune detection and promoting a tumor microenvironment that fuels tumor growth; however, this appears to occur in a manner that necessarily and predictably leaves tumors highly vulnerable to acute oncogene inactivation, whereby immune effectors become activated, thereby resulting in dramatic tumor regression, a phenomenon called “oncogene addiction”. Hence, we hypothesize that the identification，quantification and localization of these different host immune effectors in the tumor microenvironment during tumorigenesis and tumor regression will identify cellular biomarkers that will predict therapeutic response to oncogene inactivation. Our approach will be to employ the Tet system regulated model of MYC-induced hepatocellular carcinoma (HCC) and MYC/Twist1-induced model of metastatic HCC. Then, to utilize: FACS/CyTOF, CODEX/MIBI, IVM/BLI, and Gene expression/CIBERSORT to identify cellular effectors and hallmark genes and metabolites. First, our transgenic mouse model of MYC- induced HCC has been widely utilized by us and many others. Employing the Tet System, our model exhibit precise reversible and titrable control of the gene expression of the MYC oncogene. Tumor formation occurs slowly over time, oncogene-dependency results in tumor regression that is highly dependent on the immune response. Further, recently we generated a not published mouse model dramatically illustrating that MYC- induced liver tumors rapidly metastasize through the blood stream when combined with transgenic Twist1 expression. Twist1 has previously been associated with metastasis. We have obtained preliminary results that show metastasis is associated with and appears to require recruitment of macrophages. Finally, we have found that we can use human TCGA data available to identify potential cellular effectors associated with the pathogenesis of human HCC. Thus, all stages of tumorigenesis appear to be dependent on the stromal and immune response (initial, progressed, metastatic, regressed) and represent a unique tool to assess tumor- microenvironment heterogeneity. We have three aims: first, to dissect the mechanism by which the adaptive and innate immune system facilitates HCC progression, metastasis, and regression; second, to determine the kinetics and localizations of the above identified cell populations using in vivo (IVM and BLI) imaging technologies with special emphasis on cell-cell interactions and, third, to assess the ability of these findings to make predictions regarding the clinical behavior and prognosis of human HCC.