Obscurin-Deficient Breast Epithelia Generate Secreted Factors that Prime Lung Vascular Smooth Muscle Cell Pre-metastatic Microenvironment Formation

暗蛋白缺陷的乳腺上皮细胞产生分泌因子，促进肺血管平滑肌细胞转移前微环境的形成

• 批准号: 10749467
• 负责人: Matthew Kent Eason
• 金额: $ 4.58万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2026-07-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749467
• 关键词: AKT2 gene; Affect; Automobile Driving; Back; Biochemical; Biological Markers; Breast; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer therapy; Breast Epithelial Cells; Breast cancer metastasis; Cell Adhesion; Cell Line; Cell secretion; Cells; Clone Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen; Collagen Type I; Deposition; Development; Disease; Distant; Down-Regulation; Environment; Evaluation; Extracellular Matrix; Extracellular Matrix Proteins; Fibronectins; Generations; Genes; Goals; Human; In Vitro; Invaded; KRAS2 gene; Knock-out; Knowledge; Link; Lung; MCF10A cells; Mammary Neoplasms; Mediating; Membrane Proteins; Metastatic Neoplasm to the Lung; Modeling; Mus; Muscle; Mutation; Neoplasm Circulating Cells; Neoplasm Metastasis; Oncogene Activation; Oncogenes; Organ; PIK3CG gene; Pathway interactions; Patients; Phenotype; Population; Prevention; Primary Neoplasm; Probability; Process; Production; Proteins; Published Comment; Pulmonary artery structure; Reporting; Research; Risk; Role; Sarcomeres; Smooth Muscle Myocytes; Stromal Cells; Structure of parenchyma of lung; Testing; Therapeutic; Tumor Markers; Tumor Suppressor Proteins; Tumor-Derived; Up-Regulation; Vacuum; Vascular Smooth Muscle; Work; cell motility; diagnostic biomarker; exosome; extracellular; improved; lung colonization; lung metastatic; malignant breast neoplasm; mammary epithelium; metastasis prevention; migration; neoplastic cell; new therapeutic target; novel; novel therapeutics; obscurin; response; restoration; transcription factor; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor microenvironment; tumorigenesis; vascular smooth muscle cell proliferation

Project Summary/Abstract: New therapies blocking metastatic spread are greatly needed to improve breast cancer patient survival. Pre-metastatic niche development has been identified as a key milestone to successful metastasis, generated in response to a growing tumor’s release of secreted factors. Critical to the pre-metastatic niche is the pre- metastatic microenvironment (PMM) composed of extracellular matrix (ECM) proteins, deposited by resident stromal cells, that enhance metastasis. To date, PMM development has been studied solely in the context of highly metastatic tumor models using cell lines rich in tumor suppressor and oncogene mutations, however no direct mechanistic connections between metastatic driver genes and PMM formation have yet been deciphered. Obscurins have recently been pinned as potent metastasis suppressors in breast cancer. Biochemical evidence from our group has mechanistically linked obscurin loss to PI3K/Akt2 activation in breast epithelial cells, driving oncogenesis and metastatic spread. However, no study has yet drawn the connection between the loss of obscurin in breast epithelia and PMM development. This study will test the novel hypothesis that loss of obscurin in breast epithelia primes vascular smooth muscle cell (vSMC)-derived fibronectin and collagen 1a1 deposition in the PMM through secreted factors, potentiating metastasis. The goal of my application is to investigate the potential of obscurin-deficient breast epithelia secreted factors (OBEFs) to drive vSMC ECM fibronectin and collagen 1a1 deposition. OBEFs will be isolated from OBSCN knock-out (KO) MCF10A cell clones, while the human pulmonary artery smooth muscle cell (HPASMC) line will be the representative PMM cell population for the following reasons: 1) Lung PMM development is dependent on phenotypically activated lung vSMC-derived ECM production, and 2) Pre-metastatic lung, vSMC- derived ECM has been shown to enhance metastatic lung seeding in the pre-metastatic lung parenchyma. In particular, I will investigate potential changes in OBEF-driven vSMC-derived ECM fibronectin and collagen 1a1 and their connection to enhanced lung metastatic seeding (Aim 1). Then, I will identify the responsible secreted factors (Aim 2) that drive the subsequent vSMC ECM fibronectin and collagen 1a1 production in the lung PMM. Together, these studies will connect our knowledge of PMM development back to a major suppressor of primary tumor metastasis, indicating the first targetable tumor biomarker linked to the stromal component of PMM generation.

项目摘要/摘要： 新的疗法阻碍转移性扩散以改善乳腺癌患者的生存。 种植前的利基开发已被确定为成功转移的关键里程碑，生成 为了响应肿瘤的释放，分泌因素的释放。对中转移的利基至关重要的是 由居民沉积的细胞外基质（ECM）蛋白组成的转移微环境（PMM） 基质细胞，增强转移。迄今为止，PMM开发仅在 高度转移性肿瘤模型使用富含肿瘤抑制和癌基突变的细胞系，但是否 尚未确定转移驱动基因和PMM形成之间的直接机械连接。 最近已将黑蛋白固定为乳腺癌中潜在的转移补充剂。生化证据 从我们组中，在乳腺上皮细胞中，机械地将近距离的损失与PI3K/AKT2激活相关联，驱动 肿瘤发生和转移扩散。但是，尚未研究损失之间​​的联系 乳房上皮和PMM发育中的遮盖蛋白。这项研究将测试遗体丧失的新假设 在乳房上皮素中，血管平滑肌细胞（VSMC）衍生的纤连蛋白和胶原蛋白1a1沉积 在PMM中通过分泌的因素，潜在的转移。 我应用的目的是调查分泌的模糊缺陷性乳房上皮的潜力 驱动VSMC ECM纤连蛋白和胶原蛋白1A1沉积的因素（肥胖）。肥胖将与 淫秽的敲除（KO）MCF10A细胞克隆，而人类肺动脉平滑肌细胞（HPASMC） 由于以下原因，线将是代表性PMM细胞种群：1）肺PMM发育是 取决于表型激活的肺VSMC衍生的ECM产生，2）肺前肺VSMC- 衍生的ECM已被证明可以增强中转移性肺实质中的转移性肺接种。 特别是，我将调查肥大驱动的VSMC衍生的ECM纤连蛋白和胶原蛋白1A1的潜在变化 以及它们与增强的肺转移播种的联系（AIM 1）。然后，我将确定负责任的分泌 因素（目标2）驱动肺PMM中随后的VSMC ECM纤连蛋白和胶原蛋白1a1的产生。 这些研究将共同​​将我们对PMM开发的了解与主要抑制器联系起来 肿瘤转移，表明与PMM的基质成分相关的第一个靶向肿瘤生物标志物 一代。