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BMP7 in melanoma niche morphogenesis and homeostasis

BMP7 在黑色素瘤生态位形态发生和稳态中的作用

基本信息

批准号：
8634038
负责人：
MEI-YU HSU
金额：
$ 31.96万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-04 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8634038
关键词：
Angiogenic Factor Apoptosis Area Blood Circulation Blood Vessels Brain Neoplasms Cell Fraction Cell Line Cells Complex Coupled Data Dependency Development Ensure Environment Equilibrium Evaluation Studies Genomics Goals Growth Heterogeneity Homeostasis Human Immunofluorescence Immunologic In Situ In Vitro Knowledge Label Maintenance Malignant Neoplasms Mediating Melanoma Cell Mesenchymal Metabolic Microcirculation Microspheres Modeling Molecular Morphogenesis Neoplasm Metastasis Nutrient Pattern Perfusion Pericytes Population Property Proteomics RNA Interference Resistance Signal Pathway Signal Transduction Staging Stem cells Structure Testing Therapeutic Time Tissues Treatment Failure Up-Regulation Validation Vascular Endothelial Growth Factors Virulence Virulent Xenograft Model angiogenesis autocrine base bone morphogenetic protein 7 cadherin 5 cancer therapy density gain of function in vivo laser capture microdissection loss of function melanoma mimicry novel novel therapeutics paracrine reconstitution self-renewal spatial relationship tumor tumor growth tumor microenvironment tumor progression

项目摘要

Summary Melanoma is arguably the most virulent among human cancers, in part due to its propensity to metastasize, and its resistance to conventional anti-cancer therapies. One key factor responsible for treatment failure relates to tumor heterogeneity, particularly subpopulations that possess stem cell-like properties, known as melanoma initiating cells (MICs). Our long-term goal is to understand the molecular mechanisms, whereby MICs and their stroma collaborate to promote growth and progression, as a means to develop effective therapeutic strategies. In the current project, we focus on defining the microenvironmental "niche", of CD133+/ABCB5+ MICs as a gateway to elucidate the complex cellular and molecular interplay that maintains the metabolic and replicative integrity of MICs. Our preliminary data indicate that : 1) the CD133+/ABCB5+ MICs are spatially arranged in a pattern identical to that of "vasculogenic mimicry (VM)"; 2) the vessel-like channels so formed in VM are themselves CD144 (VE-cadherin)+ and intimately associated with authentic endothelial structures, in keeping with so-called "perivascular niches"; 3) melanoma cell expression of bone morphogenetic protein 7 (BMP7) and its antagonist Noggin is associated with tumor progression and also co-localizes to areas of VM; and 4) BMP7 upregulates the angiogenic factor VEGF in the stromal microenvironment, but at the same time induces selective apoptosis in Noggin-deficient melanoma cells. Collectively, our findings support the central hypothesis that CD133+/ABCB5+ MIC-associated BMP7 may not only support the maintenance of MICs by facilitating "niche" morphogenesis (through coordinated VM and angiogenesis), but also balance metabolic demands (through stimulating VEGF-dependent angiogenesis and inducing selective apoptosis in the competitive, Noggin-deficient, non-initiating population). Using multi-label immunofluorescence and immuno- guided laser capture microdissection followed by qRT-PCR, together with functional circulation analysis, we propose to further delineate the temporal/spatial relationship between melanoma cell CD133, ABCB5, BMP7 and CD144 expression, and the types/ patterns of microcirculation (e.g. melanoma VM channel formation vs. angiogenesis) in vivo (Aim 1a). The dynamics and functional impacts of CD133+ melanoma subsets in "niche" formation (Aim 1b) will be tested in an orthotopic xenograft model using inducible RNAi-mediated knockdown at varying tempos. Finally, to dissect the CD133+/ ABCB5+ MIC-associated BMP7 molecular signals in "niche" morphogenesis (Aim 2a) and tumor heterogeneity/ homeostasis (Aim 2b), we will employ loss- and gain-of- function approaches to assess the BMP7-VEGF/ BMP7-Noggin axes, in a tissue context in unique three- dimensional (3D) organotypic cultures in vitro and melanoma xenograft models in vivo. Defining mechanisms through which MIC-associated BMP7 signals contribute to "niche" development and maintenance offers a novel opportunity to therapeutically eliminate MICs directly or indirectly by targeting their stromal dependency.

总结黑色素瘤可以说是人类癌症中最致命的，部分原因是它有转移的倾向，以及它对传统抗癌疗法的抵抗力。治疗失败的一个关键因素是肿瘤异质性，特别是具有干细胞样特性的亚群，称为黑色素瘤起始细胞（MIC）。我们的长期目标是了解MIC和它们的基质协作以促进生长和进展，作为开发有效治疗的手段。战略布局在目前的项目中，我们专注于定义微环境的“生态位”，CD 133 +/ABCB 5 + MIC作为阐明维持代谢和代谢的复杂细胞和分子相互作用的门户，我们的初步数据表明：1）CD 133 +/ABCB 5 + MIC在空间上是以与“血管生成拟态（VM）"相同的模式排列; 2）在血管生成拟态中如此形成的血管样通道， VM本身是CD 144（VE-钙粘蛋白）+，与真实的内皮结构密切相关，符合所谓的“血管周围龛”; 3）黑色素瘤细胞表达骨形态发生蛋白7 （BMP 7）及其拮抗剂Noggin与肿瘤进展相关，并且还共定位于VM区域; 4）BMP 7上调基质微环境中的血管生成因子VEGF，但同时诱导Noggin缺陷型黑素瘤细胞的选择性凋亡。总的来说，我们的研究结果支持中央假设CD 133 +/ABCB 5 + MIC相关的BMP 7可能不仅通过促进“生态位”形态发生（通过协调VM和血管生成），但也平衡代谢需求（通过刺激VEGF依赖的血管生成和诱导选择性凋亡）。竞争性、Noggin缺陷、非起始群体）。使用多标记免疫荧光和免疫- 引导激光捕获显微切割，然后进行qRT-PCR，结合功能循环分析，我们进一步阐明黑色素瘤细胞CD 133、ABCB 5、BMP 7之间的时空关系和CD 144表达，以及微循环的类型/模式（例如黑素瘤VM通道形成与血管生成）（目的1a）。CD 133+黑色素瘤亚群在“生态位”中的动态变化及其功能影响将在原位异种移植模型中使用诱导型RNAi介导的敲低来测试Aim 1b 在不同的节奏。最后，在“小生境”中分析CD 133 +/ABCB 5 + MIC相关的BMP 7分子信号，形态发生（目标2a）和肿瘤异质性/稳态（目标2b），我们将采用功能的方法来评估BMP 7-VEGF/BMP 7-Noggin轴，在组织背景下，在独特的三个- 体外三维（3D）器官型培养物和体内黑素瘤异种移植模型。界定机制通过MIC相关的BMP 7信号有助于“利基”发展和维护，通过靶向其基质依赖性直接或间接治疗性消除MIC的新机会。