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： 为了提高乳腺癌患者的存活率，迫切需要阻止转移扩散的新疗法。 转移前的生态位发展被认为是成功转移的关键里程碑，产生了 以应对不断增长的肿瘤释放的分泌因子。对转移前利基至关重要的是前- 由细胞外基质(ECM)蛋白组成的转移微环境(PMM)，由居民沉积 间质细胞，增强转移能力。到目前为止，PMM的发展仅在以下背景下进行研究 使用富含抑癌基因和癌基因突变的细胞系建立高转移肿瘤模型，但没有 转移驱动基因和PMM形成之间的直接机制联系尚未破译。 Obscurins最近被认为是乳腺癌的有效转移抑制因子。生化证据 从机制上讲，乳房上皮细胞中暗蛋白丢失与PI3K/Akt2激活有关，推动 肿瘤发生和转移扩散。然而，目前还没有研究得出这种丧失之间的联系 乳房上皮细胞中的遮盖素与PMM发育。这项研究将检验这一新的假设，即遮挡消失 乳腺上皮中血管平滑肌细胞(VSMC)来源的纤维连接蛋白和胶原1a1的沉积 在PMM中通过分泌因子，增强转移。 我的应用程序的目标是研究隐匿蛋白缺陷的乳腺上皮分泌的可能性。 促进vSMC、ECM、纤维连接蛋白和胶原1a1沉积的因素(OBEF)。OBEF将从以下位置隔离 OBSCN基因敲除(KO)的MCF10A细胞克隆，而人肺动脉平滑肌细胞(HPASMC) 将具有代表性的PMM细胞群体分为以下几个原因：1)肺PMM发育是 依赖于表型激活的肺vSMC来源的ECM的产生，以及2)转移前肺，vSMC- 衍生的ECM已被证明能增强转移前肺实质内的转移肺种植。在……里面 特别是，我将研究OBEF驱动的vSMC来源的ECM纤维连接蛋白和胶原1a1的潜在变化 以及它们与增强的肺转移种植的联系(目标1)。然后，我会找出责任人的秘密 推动肺PMM中随后的vSMC、ECM、纤维连接蛋白和胶原1a1产生的因素(目标2)。 综上所述，这些研究将把我们对PMM发展的了解与初级疾病的主要抑制因素联系起来。 肿瘤转移，表明第一个与PMM间质成分相关的靶向肿瘤生物标志物 一代。