Fibroblast lineage tracing in metastatic and pre-metastatic microenvironments

转移和转移前微环境中的成纤维细胞谱系追踪

• 批准号: 10224666
• 负责人: Jamie L Null
• 金额: $ 3.47万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224666
• 关键词: 3-Dimensional; Ablation; Area; Automobile Driving; Biological Assay; Biomechanics; Blood Vessels; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Cancer Etiology; Cause of Death; Cells; Characteristics; Chronic; Coculture Techniques; Collagen; Collagen Fiber; Complex; Data; Deposition; Development; Disease; Disease Progression; Disseminated Malignant Neoplasm; Distant; Extracellular Matrix; Fibroblasts; Fibrosis; Generations; Genetic; Heart Injuries; Human; Image; Immune; Impairment; In Vitro; Inflammation; Inflammatory; Invaded; Knowledge; Label; Lead; Lung; MDA MB 231; Malignant Neoplasms; Mammary Neoplasms; Measures; Metastatic Neoplasm to the Lung; Microscopy; Modeling; Molecular; Mus; Myofibroblast; Neoplasm Metastasis; Patient-Focused Outcomes; Phenotype; Play; Population Heterogeneity; Primary Neoplasm; Property; Proteins; Reporter; Role; Scaffolding Protein; Seeds; Site; Small Interfering RNA; Source; Sum; Testing; Tissues; Traction Force Microscopy; Transforming Growth Factor beta; Tumor Cell Invasion; Tumor Cell Migration; Tumor stage; Tumor-Associated Process; Variant; Woman; acute wound; cancer cell; cancer type; cell motility; cell type; crosslink; experimental study; fibrillogenesis; improved; improved outcome; in vivo; knock-down; lymph nodes; malignant breast neoplasm; mechanical properties; mechanotransduction; melanoma; migration; mortality; neoplastic cell; periostin; response; second harmonic; single-cell RNA sequencing; tool; tumor; tumor growth; tumor microenvironment; tumor progression; wound healing

Abstract Breast cancer is one of the leading causes of cancer-related mortality among women worldwide, and the overwhelming majority of breast cancer patient deaths are caused by metastatic disease rather than primary tumor growth. However, the complex process of tumor cell dissemination and colonization of distant tissues is incompletely understood and is largely incurable using existing therapies. Metastatic disease is particularly intractable because metastasis is not solely driven by tumor cell-intrinsic properties but is instead a consequence of dynamic, heterotypic interactions between cancer cells and other cell types in the tumor microenvironment (TME) including cancer-associated fibroblasts (CAFs). CAFs are a prominent component of the breast tumor microenvironment and are important during multiple stages of tumor development, including metastasis. However, CAFs have been shown to display paradoxical functions across different cancer types, suggesting that heterogeneous CAF subpopulations may play diverse or opposing roles in the tumor microenvironment. It was recently shown that periostin, a TGFβ-induced matricellular protein that serves as a protein scaffold to promote collagen cross-linking and extracellular matrix stiffening, can distinguish CAF subtypes in breast cancer. This specific CAF subpopulation is not well characterized, and its source, functional properties, and contribution to disease progression have not been studied in vivo. Thus, I propose to use in vivo genetic labelling/targeting of periostin+ subpopulations and their progeny to better delineate their expansion and function during breast tumor growth and metastasis. My preliminary data indicates that ZSGreen-labelled periostin-expressing CAFs are more abundant in the primary tumors and metastases of high-metastatic breast tumor cells compared to their low-metastatic counterparts. I also observed that the collagen organization of high-metastatic tumors containing abundant periostin-expressing CAFs is altered, with high-metastatic tumors displaying long, aligned collagen fibers and low-metastatic tumors displaying disorganized collagen. Accordingly, I found that knocking down periostin in primary human CAFs inhibits tumor cell invasion through collagen in a CAF/tumor cell co-culture spheroid assay, indicating that periostin- expressing CAFs may remodel the extracellular matrix via collagen cross-linking, making it more conducive to cell migration and invasion. In addition, I observed that ZSGreen+ cells in highly metastatic tumors are larger and more spindle-shaped, and that periostin knockdown in primary CAFs reduces their cellular area and ability to migrate, suggesting that periostin is associated with biomechanical properties of CAFs that function in cell spreading and motility. I propose that high-metastatic cancer cells have an enhanced ability to activate periostin-expressing CAFs, driving their proliferation and expansion in the TME and subsequently promoting collagen remodeling and collective cell migration of CAFs and cancer cells, resulting in metastasis.

抽象的 乳腺癌是全世界女性癌症相关死亡的主要原因之一， 绝大多数乳腺癌患者死亡是由转移性疾病而非原发性疾病引起 然而，肿瘤细胞向远处组织播散和定植的过程非常复杂。 转移性疾病尤其不完全了解，并且使用现有疗法基本上无法治愈。 这是棘手的，因为转移不仅是由肿瘤细胞的内在特性驱动的，而且是一种 癌细胞与肿瘤中其他细胞类型之间动态、异型相互作用的结果 微环境（TME），包括癌症相关成纤维细胞（CAF）是其重要组成部分。 乳腺肿瘤微环境在肿瘤发展的多个阶段都很重要，包括 然而，CAF 已被证明在不同的癌症类型中表现出矛盾的功能， 表明异质 CAF 亚群可能在肿瘤中发挥不同或相反的作用 最近表明，骨膜素是一种 TGFβ 诱导的基质细胞蛋白，可作为一种微环境。 蛋白质支架促进胶原交联和细胞外基质硬化，可区分CAF 该特定 CAF 亚群的特征及其来源、功能尚不明确。 性质以及对疾病进展的贡献尚未在体内研究过，因此，我建议在体内使用。 对骨膜素+亚群及其后代进行体内遗传标记/靶向，以更好地描述其 我的初步数据表明，乳腺肿瘤生长和转移过程中的扩张和功能。 ZSGreen 标记的表达骨膜素的 CAF 在原发肿瘤和转移瘤中更为丰富 我还观察到，高转移性乳腺肿瘤细胞与其低转移性盟友相比。 含有丰富表达骨膜素的 CAF 的高转移肿瘤的胶原组织发生改变， 高转移性肿瘤显示长且排列的胶原纤维，低转移性肿瘤显示 因此，我发现敲除原代人 CAF 中的骨膜素可以抑制肿瘤。 在 CAF/肿瘤细胞共培养球体测定中通过胶原蛋白进行细胞侵袭，表明骨膜素 表达 CAF 可能通过胶原蛋白交联重塑细胞外基质，使其更有利于 此外，我观察到高度转移性肿瘤中的 ZSGreen+ 细胞更大。 和更多的纺锤形，并且原代 CAF 中的骨膜蛋白敲低减少了它们的细胞面积和能力 迁移，表明骨膜素与在细胞中发挥作用的 CAF 的生物力学特性有关 我认为高转移性癌细胞的激活能力增强。 表达周期蛋白的 CAF，驱动其在 TME 中增殖和扩张，随后促进 胶原蛋白重塑以及 CAF 和癌细胞的集体细胞迁移，导致转移。