Novel role of myeloid-derived lymphatic progenitors in induction of breast cancer lymphatics

髓源性淋巴祖细胞在诱导乳腺癌淋巴管中的新作用

• 批准号: 9304980
• 负责人: Sophia Ran
• 金额: $ 33.74万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY SCH OF MED
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9304980
• 关键词: Adoptive Transfer; Blood; Blood Vessels; Bone Marrow; Bone Marrow Transplantation; Breast Cancer Model; Breast Cancer Patient; Breast cancer metastasis; Cancer Patient; Carcinoma; Cell Lineage; Cells; Chimera organism; Clinical; Complement; Data; Detection; Diagnostic; Environment; Generations; Human; ITGAM gene; Immature Bone; In Vitro; Inflammation Mediators; Inflammatory; Ligands; Lymph; Lymphangiogenesis; Lymphatic; Lymphatic Endothelial Cells; Lymphatic Metastasis; Lymphatic Vessel Tumors; Lymphatic vessel; Mammary Neoplasms; Mediating; Metastatic Neoplasm to Lymph Nodes; Metastatic breast cancer; Methods; Modeling; Molecular; Molecular Target; Mus; Myelogenous; Myeloid Cells; NF-kappa B; Neoplasm Metastasis; Organ; Outcome; Outcome Study; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Process; Prognostic Marker; Proteins; Recruitment Activity; Role; Signal Transduction; Stem cells; TLR4 gene; Testing; Therapeutic; Up-Regulation; Validation; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factor Receptor-3; autocrine; base; breast cancer survival; clinically significant; density; factor C; improved; in vivo Model; lymph nodes; lymphatic cancer; malignant breast neoplasm; monocyte; mortality; neoplastic cell; novel; paracrine; podoplanin; prevent; progenitor; receptor; trait; transcription factor; transdifferentiation; tumor; vasculogenesis

PROJECT SUMMARY/ABSTRACT Title: Novel Role of Myeloid-derived Lymphatic Progenitors in Induction of Breast Cancer Lymphatics Metastasis to lymph nodes, a common occurrence in breast cancer (BC), is the most significant prognostic indicator of poor outcome. To reach locoregional lymph nodes, tumor cells exclusively use lymphatic vessels. Not surprisingly, the extent of lymphatic metastasis is directly proportional to the density of tumor lymphatic vessels. It is therefore of high clinical significance to understand the mechanisms of tumor induced lymphangiogenesis, that is, the formation of new lymphatic vessels. It is currently thought that the main mechanism causing the formation of new lymphatic vessels is mediated by a paracrine lymphangiogenic factor VEGF-C that activates its receptor VEGFR-3 expressed in lymphatic endothelial cells. We recently discovered a fundamentally different mechanism of tumor lymphangiogenesis that complements the current views. This mechanism is mediated by tumor-mobilized bone marrow (BM)- derived monocytic progenitors that upon influence of the inflammatory tumor environment differentiate into lymphatic-like cells. These cells dubbed here Monocyte-derived Lymphatic Endothelial Cells Progenitors or M- LECP are characterized by the two main traits: (1) co-expression of myeloid and lymphatic-specific proteins that are typically segregated into distinct lineages; and (2) the ability to integrate into preexisting lymphatic vessels, which is an early prerequisite for lymphatic outgrowth. Using these criteria, we found very high levels of M-LECP in blood and tumors of BC patients as well as in a variety of metastatic orthotopic breast tumors from human and mouse origins. All tumors that contained M- LECP also displayed lymphatic vessels positive for myeloid-specific markers, an established phenomenon indicative of vascular integration of M-LECP that is required for sprouting. Importantly, we recently established that the levels of tumor-recruited M-LECP significantly correlate with tumor lymphatic density and lymph node status in clinical BC patients. Our studies in BC models showed that M-LECP originate from BM-derived CD11b+ cells that are highly positive for a lymphatic marker Podoplanin (Pdpn). Adoptive transfer of a phenotypically distinct BM subset identified by CD11b and Pdpn from metastatic tumor-bearing mice to mice with low-metastatic tumors significantly increased the density of lymphatic vessels and lymphatic metastasis. Preliminary data also show that differentiation of M-LECP can be faithfully reproduced in vitro by activating Toll-like Receptor-4 (TLR4) in human primary normal blood-circulating monocytes. This process is controlled by NF-kB and a transcription factor c-Maf, a newly identified regulator of monocytic-lymphatic reprogramming. TLR4-dependent upregulation of NF-kB and c-Maf leads to activation of the VEGFR-3 pathway which appears to be a critical milestone for acquisition of the lymphatic phenotype. Based on these findings, we hypothesize that tumor lymphatic outgrowth, an essential prerequisite for metastasis, is strongly promoted by M- LECP differentiating from the bone marrow myeloid precursors by TLR4-activating factors. To test this hypothesis we propose the following Specific Aims: (1) Test the hypothesis that M-LECP are directly and significantly relate to tumor-induced lymphangiogenesis and lymphatic metastasis in clinical BC; (2) Delineate the molecular mechanisms that reprogram bone marrow myeloid precursors into M-LECP; and (3) Determine the role of TLR4 in generation of the CD11b+/Pdpn+ bone marrow subset and its direct contribution to the formation of tumor lymphatic vessels and metastasis. Impact and Translational Relevance: We anticipate that these studies will establish a novel mechanism by which BM-derived monocytes recruited by breast tumors promote lymphangiogenesis and metastasis to lymph nodes. Validation of this novel concept will enhance the mechanistic understanding of the formation of tumor lymphatic vessels and suggest new molecular targets for inhibiting differentiation of M-LECP. Additionally, detection of higher levels of tumor- residing and blood-circulating M-LECP might identify BC patients with more aggressive tumors who should be preemptively treated to suppress locoregional spread. Such study outcomes can bolster the current clinical paradigms thus reducing mortality of BC patients.

项目概要/摘要 标题：髓源性淋巴祖细胞在乳腺癌淋巴管诱导中的新作用 淋巴结转移是乳腺癌 (BC) 中常见的情况，是最重要的预后因素 不良结果的指标。为了到达局部淋巴结，肿瘤细胞专门使用淋巴管。 毫不奇怪，淋巴转移的程度与肿瘤淋巴管的密度成正比。 船只。因此，了解肿瘤发生的机制具有重要的临床意义。 淋巴管生成，即新淋巴管的形成。 目前认为引起新淋巴管形成的主要机制是介导 一种旁分泌淋巴管生成因子 VEGF-C，可激活淋巴管中表达的受体 VEGFR-3 内皮细胞。我们最近发现了一种根本不同的肿瘤淋巴管生成机制 这补充了当前的观点。该机制是由肿瘤动员的骨髓 (BM) 介导的 - 衍生的单核细胞祖细胞在炎症肿瘤环境的影响下分化为 淋巴样细胞。这些细胞在这里被称为单核细胞源性淋巴内皮细胞祖细胞或 M- LECP具有两个主要特征：（1）骨髓和淋巴特异性蛋白的共表达 通常分为不同的谱系； (2) 融入已有淋巴管的能力 血管，这是淋巴管生长的早期先决条件。 使用这些标准，我们发现 BC 患者的血液和肿瘤以及 来自人类和小鼠的多种转移性原位乳腺肿瘤。所有含有 M- 的肿瘤 LECP 还显示淋巴管对骨髓特异性标记物呈阳性，这是一种既定现象 表明发芽所需的 M-LECP 的血管整合。重要的是，我们最近成立了 肿瘤招募的 M-LECP 水平与肿瘤淋巴密度和淋巴结显着相关 临床 BC 患者的状况。我们对 BC 模型的研究表明，M-LECP 源自 BM 衍生的 CD11b+ 细胞对淋巴标记 Podoplanin (Pdpn) 呈高度阳性。过继转让 CD11b 和 Pdpn 鉴定出从转移性肿瘤小鼠到小鼠的表型不同的 BM 子集 低转移性肿瘤的淋巴管密度和淋巴转移明显增加。 初步数据还表明，通过激活 M-LECP 的分化可以在体外忠实地复制。 人类原代正常血液循环单核细胞中的 Toll 样受体 4 (TLR4)。这个过程是受控的 由 NF-kB 和转录因子 c-Maf 组成，c-Maf 是一种新发现的单核细胞淋巴重编程调节因子。 TLR4 依赖性 NF-kB 和 c-Maf 上调导致 VEGFR-3 通路激活 成为获得淋巴表型的一个重要里程碑。根据这些发现，我们假设 M-强烈促进肿瘤淋巴管生长，这是转移的重要先决条件 LECP 通过 TLR4 激活因子与骨髓前体细胞分化。 为了检验这一假设，我们提出以下具体目标： (1) 检验M-LECP与肿瘤诱导的淋巴管生成直接且显着相关的假设 以及临床BC中的淋巴转移； (2) 描绘骨髓重编程的分子机制 骨髓前体细胞转化为 M-LECP； (3) 确定 TLR4 在 CD11b+/Pdpn+ 骨生成中的作用 骨髓亚群及其对肿瘤淋巴管形成和转移的直接贡献。 影响力和转化相关性： 我们预计这些研究将建立一种新的机制，通过该机制招募骨髓来源的单核细胞 乳腺肿瘤促进淋巴管生成并转移至淋巴结。验证这个新颖的概念 将增强对肿瘤淋巴管形成机制的理解并提出新的建议 抑制 M-LECP 分化的分子靶标。此外，检测到更高水平的肿瘤 居住的和血液循环的 M-LECP 可能会识别出患有更具侵袭性肿瘤的 BC 患者，这些患者应该接受 抢先治疗以抑制局部扩散。此类研究结果可以支持当前的临床 从而降低 BC 患者的死亡率。