Mechanisms of linkage of stem and invasive phenotypes during metastatic colonization

转移定植过程中干细胞和侵袭表型的联系机制

• 批准号: 10659164
• 负责人: Maja Hrzenjak Oktay
• 金额: $ 39.07万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-05 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659164
• 关键词: Actins; Affect; Biology; Biosensor; Blood Circulation; Blood Vessels; Blood capillaries; Bone Marrow; Brain; Cell membrane; Cells; Cessation of life; Chemoresistance; Circulation; Data; Development; Diagnosis; Disease; Disseminated Malignant Neoplasm; Distant; Distant Metastasis; Endothelial Cells; Endothelium; Event; Excision; Extravasation; Future; Hypoxia; Life; Light; Link; Lung; Macrophage; Malignant Neoplasms; Mammary Neoplasms; Mediating; Metastatic Neoplasm to the Lung; Metastatic breast cancer; Molecular Target; NF-kappa B; Neoplasm Metastasis; Organ; Patient-Focused Outcomes; Patients; Phenotype; Phosphatidylinositols; Population; Primary Neoplasm; Process; Prognosis; Protein Isoforms; Publishing; Regulation; Relapse; Resolution; Signal Transduction; Site; Tissues; Tumor Biology; Tumor-associated macrophages; Woman; base; cancer cell; cancer stem cell; chemotherapy; curative treatments; genetic regulatory protein; improved; improved outcome; in vivo; in vivo imaging; insight; intravital imaging; lung colonization; lung imaging; malignant breast neoplasm; migration; mortality; neoplastic cell; new technology; novel; programs; stem; stem cells; stemness; transcription factor; tumor; tumor growth; tumor initiation; tumor microenvironment

ABSTRACT Metastasis, the primary cause of breast cancer-related mortality, is a multistep process culminating with the formation of tumor foci within distant organs. However, only a subpopulation of cancer cells within the primary tumor microenvironment is capable of completing the entire metastatic cascade, which includes intravasation, survival in circulation, extravasation, and tumor growth at distant sites. We identified a population of highly invasive, non-proliferating, non-apoptotic, chemo-resistant cancer cells capable of intravasation (published) and extravasation (preliminary results). These cells express high levels of MenaINV, a pro-metastatic isoform of the actin-regulatory protein Mena required for maturation of invasive protrusions called invadopodia, which enable cancer cells to cross endothelium and disseminate. MenaINV localization to the cell membrane, which is required for its function, may be regulated by PI3Kβ through phosphoinositide signaling. We found that MenaINV expression (published) and a stem cell program (preliminary results) are induced in tumor cells by direct contact with tumor-associated macrophages. Interestingly, we also found that chemotherapy co-induces MenaINV and the stem program in a macrophage-dependent manner. The emergence of MenaINV expressing stem cells may be one of the crucial steps to metastasis because these cells are not only transendothelial migration-competent but also have tumor-initiating capability. In primary breast tumors, cancer cells expressing high levels of MenaINV are able to enter blood vessels through Tumor Microenvironments of Metastasis (TMEM) doorways. These tightly controlled transient openings in capillary walls were first described by our group and are composed of macrophages, endothelial cells and Mena-expressing tumor cells in direct physical contact. TMEM doorways and cancer cell re-dissemination from lung metastases are also observed in lung metastases, but it is currently unknown if stem program is required for cancer cell dissemination from this secondary site. Interestingly, we and others found that chemotherapy in a macrophage-dependent manner increases the proportion of cancer cells co-expressing high levels of MenaINV and stem cell transcription factor SOX9. Thus, we hypothesize that induction of the transendothelial migration-competent phenotype in cancer cells (identified by the expression of MenaINV) is mechanistically linked to the stem program, and that the induction of this transendothelial migration- competent stem phenotype is potentiated by chemotherapy. We will use high resolution intravital imaging of the lungs in combination with stem and Mena biosensors to determine in vivo how the interplay of stemness and MenaINV regulate extravasation and metastatic colonization of the lungs and determine the mechanism by which chemotherapy induced hypoxia and macrophage influx affect cancer phenotype in metastatic foci in the lungs. The successful completion of this project will increase our understanding of the mechanisms involved in development of distant metastases, and provide a base for development of anti-metastatic therapies needed for improvement of survival in patients with metastatic breast cancer.

抽象的 转移是与乳腺癌相关死亡率的主要原因，是一个多步过程 远处器官内肿瘤焦点的形成。但是，仅在原发性癌细胞中进行亚群 肿瘤微环境能够完成整个转移性级联反应，包括侵入， 在辨别部位的循环，渗出和肿瘤生长中的生存。我们确定了高度的人口 侵入性，非增殖，非凋亡，化学抗性的癌细胞（已发表）和 渗出（初步结果）。这些细胞表达高水平的MenaInv，这是一种促近代的同工型 肌动蛋白调节蛋白欧洲现代是入侵蛋白的成熟，称为Invadopodia，这使得 癌细胞越过内皮并传播。 Menainv定位到细胞膜，这是必需的 对于其功能，可以通过PI3Kβ通过磷酸肌醇信号传导调节。我们发现Menainv 通过直接接触在肿瘤细胞中诱导表达（已发布）和干细胞程序（初步结果） 与肿瘤相关的巨噬细胞。有趣的是，我们还发现化学疗法共同诱导了Menainv和 STEM程序以巨噬细胞依赖性方式。表达干细胞的Menainv的出现可能 是转移的关键步骤之一，因为这些细胞不仅是跨内皮迁移 但也具有肿瘤发射能力。在原发性乳腺肿瘤中，表达高水平MENAINV的癌细胞 能够通过转移（TMEM）门的肿瘤微环境进入血管。这些紧紧 毛细管壁中的受控瞬态开口首先由我们的组描述，由 巨噬细胞，内皮细胞和ernaa肿瘤细胞直接接触。 TMEM门 在肺转移中也观察到了肺转移中的癌细胞再隔离，但目前是 未知是否需要从这个次要部位传播癌细胞的STEM程序。有趣的是，我们和 其他人发现，以巨噬细胞依赖性方式化疗增加了癌细胞的比例 共表达高水平的MENAINV和干细胞转录因子SOX9。那我们假设 诱导癌细胞中跨内皮迁移竞争性表型（通过表达 Menainv）机械地与STEM程序有关，并且这种跨性皮迁移的诱导 - 合理的茎表型是通过化学疗法潜在的。我们将使用高分辨率的浸润成像 肺与STEM和MENA生物传感器结合，以确定体内的肺 MENAINV调节肺的渗出和转移性定植，并确定其机制 化学疗法诱导的缺氧和巨噬细胞影响会影响肺转移灶中的癌症表型。 该项目的成功完成将增加我们对涉及机制的理解 远处转移的发展，并为开发抗转移性疗法的发展提供了基础 转移性乳腺癌患者的存活率提高。