Matrix remodeling in tumor growth and metastasis in a murine breast cancer model

小鼠乳腺癌模型中肿瘤生长和转移的基质重塑

• 批准号: 8203343
• 负责人: Lisa L Chang
• 金额: $ 4.84万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8203343
• 关键词: Address; Affect; Binding; Binding Sites; Biological Markers; Breast Cancer Model; Breeding; Cancer Diagnostics; Cancer Etiology; Cancer Patient; Carcinoma; Catalytic Domain; Cell Proliferation; Cessation of life; Characteristics; Cleaved cell; Clinic; Clinical; Collagen; Colon Carcinoma; Data; Development; Diagnosis; Dipeptidyl Peptidases; Disease; Drug resistance; Economics; Endopeptidases; Epithelial; Extracellular Matrix; Family; Fibroblasts; Genes; Human; Integrin Binding; Integrins; Interstitial Collagenase; Knock-in Mouse; Knockout Mice; Lead; Life; Link; Luciferases; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mammary Neoplasms; Mediating; Membrane; Modeling; Molecular Target; Mouse Mammary Tumor Virus; Mouse Strains; Mus; Neoplasm Metastasis; Neoplastic Epithelial Cell; Peptide Hydrolases; Play; Primary Neoplasm; Process; Productivity; Protein Inhibition; Proteolytic Processing; Psyche structure; Reporting; Research; Resources; Role; Serine Protease; Signal Pathway; Signal Transduction; Signaling Molecule; Societies; Specificity; Stromal Cells; Structure; Testing; Therapeutic; Time; Transplantation; Vascularization; Wild Type Mouse; Woman; angiogenesis; base; cancer therapy; collagenase; cost; fibroblast-activating factor; inhibitor/antagonist; malignant breast neoplasm; mortality; mouse model; mutant; mutant mouse model; neoplastic cell; novel; outcome forecast; protein expression; spatial relationship; therapeutic target; tumor; tumor growth; tumor initiation; tumor progression; tumorigenesis; tumorigenic; unpublished works

DESCRIPTION (provided by applicant): My research aims to understand the tumorigenic role of fibroblast activation protein (FAP), a membrane-bound serine protease with dipeptidyl peptidase (DPP) and endopeptidase/collagenase activities. FAP is unique, in that it is expressed in cancer associated fibroblasts (CAFs) in 90% of all human epithelial derived cancers, but not in normal or transformed (malignant/neoplastic) epithelial cells. Importantly, unpublished work from our lab (J.Tchou and E. Puri) shows that FAP can be used as a robust pan-CAF marker in human breast cancer, it being superior to other CAF markers in its selectivity and specificity. CAFs play roles in ECM remodeling, angiogenesis and stromagenesis, and are important players in tumor growth and metastasis. A wide majority of reports on FAP activity describe correlations between FAP expression and poor prognosis, making FAP inhibition an appealing potential target for cancer therapeutics. Our lab recently demonstrated that FAP promotes tumorigenesis in an autochthonous mouse model of mutant K-ras- driven lung cancer and syngeneic transplant models of colon and pancreatic cancer and that the reduction in tumor growth in FAP-deficient mice was associated with enhanced accumulation of disorganized collagen, aberrant integrin signaling and reduced stromagenesis and angiogenesis. However, the mechanisms by which FAP promotes tumorigenesis are yet to be defined and its role in metastasis has not yet been investigated in any tumor type. Our preliminary data indicate that FAP mediates the proteolytic processing of the characteristic 3/4 and 1/4 fragments of collagen generated by collagenase mediated cleavage (eg. MMP-1). A central hypothesis of this proposal is that FAP promotes tumorigenesis through its protease activity by playing a critical role in ECM remodeling. Specifically, I hypothesize that by targeting collagen for degradation and turnover, FAP regulates matrix-dependent signaling of tumor cell proliferation, stromagenesis, angiogenesis and metastasis. To test these hypotheses I will compare tumorigenesis and metastasis of MMTV-Neu driven autochthonous mammary tumors in FAP wild-type mice, FAP-null mice (FAP-knock-ins expressing luciferase in place of the FAP gene) and knock-in mice expressing an enzymatic dead FAP mutant (FAPS624A) or a FAP mutant (FAPA657S) that retains DPP activity but lacks endopeptidase/collagenase activity. Therefore, by making use of these enzymatically defective FAP mice, this study will also examine the mechanism by which FAP acts on ECM remodeling. These novel FAP mouse strains have been produced in the lab and are currently breeding, while the MMTV-Neu mouse is commercially available. ! PUBLIC HEALTH RELEVANCE: Breast cancer is the second leading cause of cancer deaths in women today. The costs of cancer are a financial and mental toll on people diagnosed with cancer, their families, and society as a whole. In 2005 the burden of breast cancer in the U.S. was $12.096 billion in lost time and economic productivity (NCI, 2009/2010), therefore it is imperative to identify new molecular targets for early prognosis, treatment and eradication of this devastating disease. This research aims to better understand the causes and mechanisms underlying tumor initiation, progression and metastasis by studying the relationship between mammary tumor cells and their microenvironment. The research proposed will define the protumorigenic activity of a tumor stromal cell protease and extracellular matrix remodeling in an autochthonous breast cancer model. Results of the proposed studies will reveal novel mechanisms through which this protease regulates tumor development and therefore its value as a target for cancer therapeutics. Further, our studies may lead to an early breast cancer diagnostic biomarker that could be used in the clinic to save time, resources and most importantly, life. !

描述（由申请人提供）：我的研究旨在了解成纤维细胞激活蛋白（FAP）的致瘤作用，FAP是一种具有二肽基肽酶（DPP）和内肽酶/胶原酶活性的膜结合丝氨酸蛋白酶。 FAP 是独特的，因为它在 90% 的人类上皮衍生癌症中的癌症相关成纤维细胞 (CAF) 中表达，但在正常或转化（恶性/肿瘤）上皮细胞中不表达。重要的是，我们实验室（J.Tchou 和 E. Puri）未发表的工作表明，FAP 可用作人类乳腺癌中强大的泛 CAF 标记物，其选择性和特异性优于其他 CAF 标记物。 CAF 在 ECM 重塑、血管生成和基质生成中发挥作用，并且在肿瘤生长和转移中发挥重要作用。大多数关于 FAP 活性的报告描述了 FAP 表达与不良预后之间的相关性，这使得 FAP 抑制成为癌症治疗中有吸引力的潜在目标。我们的实验室最近证明，FAP 可促进突变 K-ras 驱动的肺癌的本地小鼠模型以及结肠癌和胰腺癌的同基因移植模型中的肿瘤发生，并且 FAP 缺陷小鼠中肿瘤生长的减少与紊乱的细胞积累的增加有关。胶原蛋白、异常的整合素信号传导以及基质生成和血管生成减少。然而，FAP 促进肿瘤发生的机制尚未明确，并且尚未在任何肿瘤类型中研究其在转移中的作用。我们的初步数据表明，FAP 介导由胶原酶介导的裂解产生的特征性 3/4 和 1/4 胶原片段（例如 MMP-1）的蛋白水解加工。该提议的一个中心假设是，FAP 在 ECM 重塑中发挥关键作用，通过其蛋白酶活性促进肿瘤发生。具体来说，我假设通过靶向胶原蛋白的降解和周转，FAP 调节肿瘤细胞增殖、基质生成、血管生成和转移的基质依赖性信号传导。为了检验这些假设，我将比较 FAP 野生型小鼠、FAP 缺失小鼠（表达荧光素酶代替 FAP 基因的 FAP 敲入小鼠）和表达荧光素酶的敲入小鼠中 MMTV-Neu 驱动的本地乳腺肿瘤的肿瘤发生和转移。酶促死亡的 FAP 突变体 (FAPS624A) 或保留 DPP 活性但缺乏 DPP 活性的 FAP 突变体 (FAPA657S)内肽酶/胶原酶活性。因此，通过利用这些酶缺陷的 FAP 小鼠，本研究还将研究 FAP 作用于 ECM 重塑的机制。这些新型 FAP 小鼠品系已在实验室生产，目前正在育种，而 MMTV-Neu 小鼠已上市。 ！ 公共卫生相关性：乳腺癌是当今女性癌症死亡的第二大原因。癌症造成的代价是对癌症患者、他们的家人和整个社会造成的经济和精神损失。 2005 年，美国乳腺癌造成的时间和经济生产力损失为 120.96 亿美元（NCI，2009/2010），因此必须确定新的分子靶点来早期预测、治疗和根除这种破坏性疾病。 本研究旨在通过研究乳腺肿瘤细胞与其微环境之间的关系，更好地了解肿瘤发生、进展和转移的原因和机制。拟议的研究将定义肿瘤基质细胞蛋白酶的促肿瘤活性和原生乳腺癌模型中的细胞外基质重塑。拟议研究的结果将揭示这种蛋白酶调节肿瘤发展的新机制，从而揭示其作为癌症治疗靶点的价值。此外，我们的研究可能会产生一种早期乳腺癌诊断生物标志物，可用于临床以节省时间、资源，最重要的是，节省生命。 ！