Project 2: Investigating cell intrinsic and extrinsic drivers of prostate cancer bone metastasis

项目摘要/摘要 我们研究的目的是阐明肿瘤细胞产生的内在和外在机制。 以骨转移和明确骨的趋向性。我们研究的基础是NPKYPF小鼠模型，该模型 发生高度穿透性骨转移，与人前列腺癌的骨转移密切相关 癌症。对这些小鼠的分析使研究癌症期间骨转移的进化成为可能 在雄激素完整和去雄激素环境中的自然微环境的进展。我们发现了 MYC和RAS信号的共同激活在骨转移中是必不可少的，而MYC的激活是 在雄激素缺乏的情况下尤其相关。我们的初步研究确定ATAD2是一种 MYC-在骨转移中表达的辅助因子，特别是在雄激素缺乏的情况下，以及 对于骨转移来说是必要的。基于人的全基因组互补体内CRISPR筛选 在前列腺癌异种移植中，我们发现CITED2，另一种MYC辅助因子，是骨的细胞内在驱动因素 转移说明它足以促进体内的骨转移。此外，对原代细胞进行了单细胞测序 NPKYPF小鼠的肿瘤和骨转移导致了识别主调节器(MRS)，这些MRS是 骨转移的候选细胞内在驱动因素，而对非肿瘤成分的分析表明 来自转移性NPKYPF小鼠的原发肿瘤缺乏大多数免疫细胞群，但高度浓缩 用于肿瘤相关的巨噬细胞。 因此，我们将探讨骨转移代表细胞固有的顶峰的假说。 转移骨微环境中的肿瘤细胞外在因素和转移细胞驱动因素。在……里面 目的1、探讨CITED2与MYC协同促进骨特异性的假说。在……里面 同时，我们将研究与RAS途径激活相关的候选主调节因子(MRS)以 阐明它们在骨转移中的潜在作用。在目标2中，我们将研究ATAD2是一种 在雄激素缺乏的情况下，MYC的辅助因素，并阐明雄激素状态与 骨转移的演变和潜在机制。在目标3中，我们将研究细胞的外在机制 在原发瘤和转移性骨的微环境中，鉴定相关细胞群 评估它们在骨转移中的功能作用，并确定骨转移的细胞外在驱动因素 转移。 整合：我们的研究是对肿瘤微环境研究的高度补充 神经内分泌分化(项目3)和肿瘤进展(项目1)。候选监管机构的确认 对于人类的骨转移将需要Core A。此外，我们对单个骨转移的系统分析 单元级，补充项目1和3的工作，而我们从这些数据中互惠互利的能力将是 由核心B的数据管理组件启用。

Project Summary/Abstract The objective of our research is to elucidate tumor cell intrinsic and extrinsic mechanisms that give rise to bone metastasis and specify bone tropism. The foundation for our studies is the NPKYPF mouse model, which develops highly penetrant bone metastasis that is well-conserved with bone metastasis in human prostate cancer. Analyses of these mice enable investigations of the evolution of bone metastases during cancer progression in the native microenvironment in androgen-intact and androgen-deprived contexts. We have found that co-activation of MYC and RAS signaling is essential for bone metastasis, and that MYC activation is particularly relevant in contexts of androgen deprivation. Our preliminary studies have identified ATAD2 as a MYC-co-factor that is expressed in bone metastasis, particularly in contexts of androgen deprivation, and necessary for bone metastasis. In a complementary genome-wide in vivo CRISPR screening based on human prostate cancer xenografts, we identified CITED2, another MYC co-factor, as a cell-intrinsic driver of bone metastasis that it is sufficient to promote bone metastasis in vivo. Furthermore, single cell sequencing of primary tumors and bone metastases from NPKYPF mice has led to the identification master regulators (MRs) that are candidate cell intrinsic drivers of bone metastasis, while analyses of the non-tumor components have shown that primary tumors from metastatic NPKYPF mice are deficient for most immune cell populations, but highly enriched for tumor associated macrophages. Thus, we will investigate the hypothesis that bone metastasis represents the culmination of cell intrinsic drivers from the metastatic cells and tumor cell extrinsic factors in microenvironment of the metastatic bone. In Aim 1, we will investigate the hypothesis that CITED2 collaborates with MYC to promote bone specificity. In parallel, we will investigate candidate master regulators (MRs) associated with RAS pathway activation to elucidate their potential roles in bone metastasis. In Aim 2, we will investigate the hypothesis that ATAD2 is a co-factor for MYC in contexts of androgen deprivation, and elucidate the relationship of androgen status for the evolution and underlying mechanisms of bone metastasis. In Aim 3, we will investigate cell extrinsic mechanisms in the microenvironment of the primary tumor and metastatic bone, to identify relevant cell populations associated with metastasis, to evaluate their functional role for bone metastasis, and to identify cell extrinsic drivers of bone metastasis. Integration: Our studies are highly complementary to investigations of the tumor microenvironment for neuroendocrine differentiation (Project 3) and tumor progression (Project 1). Validation of candidate regulators for human bone metastasis will require Core A. Further, our systematic analysis of bone metastasis at the single cell level, complement efforts in Projects 1 and 3, while our ability to mutually benefit from these data will be enabled by the data management component of Core B.