Defining a Stromal Signature that Facilitates Progression of Lethal Cancers

项目摘要/摘要 本研究专家奖申请书中提出的活动符合Dr。 Thea Tlsty(单位主任)S目前由NCI资助的研究项目，该项目目前支持我的 活动/努力：i)了解细胞可塑性和组织对压力反应的动态机制； 和二)探讨组织微环境的作用以及这些多种因素如何促成肿瘤的发生 和进步。在处理最致命的乳房类型时，这些努力尤其值得 癌症、化生性乳腺癌(MBCs)。识别组织成分和信号通路 有助于这些癌症的出现，以便为患者设计更成功的治疗方法 诊断为多发性肌萎缩侧索硬化症确实具有最大的临床意义。这份申请是我个人 Tlsty实验室对乳腺间质-上皮相互作用作用的长期兴趣的“扭曲” 肿瘤发生学。它的核心概念依赖于一个日益受到重视的概念，即癌症的进展需要 一个有益的肿瘤微环境(TME)。产生TME的结果是：获得癌症- 相关的fi成纤维细胞特征，巨噬细胞类型的转变，细胞外基质特征的改变 促进或抵消肿瘤生长、免疫反应受损、血管扩张和渗漏以及 低氧环境与上皮细胞可塑性增强相关。这样的表型是例证 侵袭性肿瘤以促结缔组织间质广泛改变的ECM沉积为特征 以MBCS为代表。因此，MBCS中出现的ECM组成不平衡是广泛的结构性结构的结果 以及由于间质和上皮细胞属性的改变而导致的功能性间质改变。我们的 一种挑衅性的预测，即特定的ECM/间质成分可能会识别结果最差的癌症 在乳腺癌亚型(腔内癌、Her2阳性、基底细胞样癌)中，临床相关性可能不仅限于 MBC和基底细胞样癌，但也适用于晚期复发的腔内乳腺癌。获得有价值的东西 洞察力，我将：i)通过比较确定MBCs进展的间质驱动因素(和抑制因素) 乳腺肿瘤(包括单核细胞)和正常乳腺组织中细胞外基质的蛋白质组学分析 强调翻译后修饰状态；ii)查询基因表达数据库以上调表达 这些ECM蛋白在乳腺癌各亚型(管腔型、Her2阳性、基底型、 Claudin low)通过数据挖掘；iii)测试某些ECM蛋白驱动(或抑制)细胞的能力 3D细胞培养模型的体外可塑性(多系承诺和上皮间充质转化)； 以及iv)在体内证实这些ECM蛋白在小鼠体内的促(或抗)化生/致瘤活性 异种移植模型。我在蛋白质生物化学、细胞生物学和小鼠特性方面的丰富经验 模型结合我最近对数据挖掘和多重免疫组织化学的兴趣，将我置于 最佳的位置，为新的追求ECM变化的基础上的MBCS。

Project Summary/Abstract The activity proposed in this Research Specialist Award application is in line with the long-standing goals of Dr. Thea Tlsty (the Unit Director)'s currently NCI-funded research program which currently supports my activity/effort: i) understand dynamic mechanisms underlying cellular plasticity and tissue response to stress; and ii) explore the role of tissue microenvironment and how these multiple factors contribute to tumor initiation and progression. These efforts are particularly worthwhile when dealing with the most lethal type of breast cancer, metaplastic breast cancers (MBCs). Identifying tissue components and signaling pathways that contribute to the emergence of these cancers in order to design more successful therapies for patients diagnosed with MBCs is indeed of the outmost clinical relevance. This application is the result of my personal ‘twist’ on the long-term interest of the Tlsty laboratory on the role of stromal-epithelial interactions in breast tumorigenesis. Its core concept relies on the increasingly appreciated notion that cancer progression requires an instructive tumor microenvironment (TME). Generation of a TME results from: acquisition of a cancer- associated fibroblast signature, a shift in macrophage type, alterations of ECM characteristics that either facilitate or counteract tumor growth, a corrupted immune response, an expanded and leaky vasculature and as a correlate an hypoxic environment and increased epithelial cell plasticity. Such phenotypes are exemplified in aggressive tumors characterized by a desmoplastic stroma with extensive and altered ECM deposition typified by MBCs. Thus, the imbalance in ECM composition seen in MBCs is the result of extensive structural and functional stromal changes that result from alterations in both stromal and epithelial cell properties. Our provocative prediction that a specific ECM/stromal make-up may identify cancers with the poorest outcome across breast cancer subtypes (luminal, Her2-positive, basal-like) may be of high clinical relevance not only for MBCs and basal-like cancers, but also for luminal breast cancers with late recurrence. To gain valuable insights, I will: i) identify stromal drivers (and repressors) of progression of MBCs through comparative proteomic analysis of ECM from breast tumor (including MBC) and disease-free breast specimens with an emphasis on post-translational modification status; ii) interrogate gene expression databases for upregulation (or down-regulation) of these ECM proteins across breast cancer subtypes (luminal, Her2-positive, basal, claudin low) through data mining; iii) test the ability of some of these ECM proteins to drive (or repress) cell plasticity (multilineage commitment and epithelial mesenchymal transition) in vitro using 3D cell culture models; and iv) confirm in vivo the pro- (or anti-)metaplastic/tumorigenic activity of these ECM proteins in murine xenograft models. My extensive experience in protein biochemistry, cell biology and characterization of mouse models combined with my recent interest in data mining and multiplex immunohistochemistry, place me in an optimal position for the novel pursuit of ECM alterations underlying progression of MBCs.