Lymph-Borne Melanoma-Derived Proteins as Determinants of Lymph Node Metastasis

淋巴源性黑色素瘤衍生蛋白作为淋巴结转移的决定因素

• 批准号: 10658869
• 负责人: Haley du Bois
• 金额: $ 1.94万
• 依托单位: NEW YORK UNIVERSITY SCHOOL 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: 2022-07-01 至 2023-10-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10658869
• 关键词: Albumins; Amino Acids; Bathing; Behavior; Benign; Biochemistry; Biology; Blood; Breast; Cancer Biology; Cancer Etiology; Cells; Cessation of life; Chemicals; Chondroitin Sulfate Proteoglycan; Communication; Contralateral; Data; Data Analyses; Dependence; Dermatology; Dermis; Development; Distant; Drainage procedure; Educational process of instructing; Event; Exhibits; Foundations; Future; Goals; Growth; Human; Immune response; Immunosuppression; In Vitro; Institution; Invaded; Label; Length; Lung; Lymph; Lymphangiogenesis; Lymphatic Endothelial Cells; Lymphatic System; Malignant Neoplasms; Mediating; Medicine; Melanoma Cell; Mentors; Metastatic Neoplasm to Lymph Nodes; Methods; Modernization; Molecular; Morbidity - disease rate; Mus; Neoplasm Metastasis; Organ; Pathology; Patient risk; Patients; Pharmacology; Phenotype; Physiological; Play; Population; Pre-Clinical Model; Primary Neoplasm; Prognosis; Prognostic Marker; Proliferating; Proteins; Research; Research Personnel; Role; Serum; Site; Soil; Solid Neoplasm; Source; Stromal Cells; System; Techniques; Testing; Therapeutic Intervention; Tissues; Training; Tumor Escape; Tumor-Derived; Tumor-Secreted Protein; Work; Writing; career; conditioning; data integration; draining lymph node; experimental study; extracellular vesicles; immune function; in vivo; knock-down; lymph nodes; lymphatic vasculature; melanoma; melanomagenesis; multidimensional data; neoplastic cell; novel; novel therapeutic intervention; patient stratification; protein transport; skills; small hairpin RNA; support network; training opportunity; tumor; tumor growth; tumor immunology; uptake; vesicle transport

PROJECT SUMMARY Tissues are primed for metastasis prior to the arrival of tumor cells. Primary tumors drive this transformation by shedding tumor-derived factors (TDFs), including extracellular vesicles and secreted/shed proteins (TSPs) into tumor-associated blood and lymphatic vasculature. Despite being the most common site of metastasis across solid tumor types, we know very little about how the lymph node (LN) is primed for metastasis by TDFs. My preliminary data demonstrates that primary melanomas initiate remodeling in their draining LNs that enhances metastatic outgrowth relative to contralateral, non-draining and naïve LN controls. Furthermore, I have demonstrated that TDFs uniquely activate this pro-metastatic LN phenotype rather than factors derived from benign dermis or mouse albumin. The focus of this field has been on the role tumor-derived extracellular vesicles play in establishing this pre-metastatic niche, however my data further demonstrates TSPs are also sufficient to activate this niche. Using a novel cell-specific labeling strategy termed BONCAT (biorthogonal non-canonical amino acid tagging), I have identified several candidate TSPs transported to the pre-metastatic LN. In this proposal, I will test the hypothesis that TSPs initiate reprogramming events within resident cell populations of the draining LN that support metastatic growth through two specific aims. The first aim will employ a scRNAseq approach to identify cellular networks, TSP-induced crosstalk between the resident LN stroma and seeded tumor cells, that provide trophic support to metastatic cells and the extent to which LN lymphangiogenesis is required for LN metastasis. The second aim will investigate a specific TSP I have identified, CSPG4 (Melanoma- Associated Chondroitin Sulfate Proteoglycan). This protein is shed by melanoma cells, has known melanoma cell-intrinsic roles of invasion and is elevated in human melanoma patients, yet, it is unknown whether this TSP contributes to pre-metastatic niche formation in distant sites. The completion of this work will provide foundational information on TSP-dependent pre-metastatic niche development in the tdLN, and may uncover CSPG4 as a novel driver of niche development. The proposed work will provide me with comprehensive training opportunities integrating new bench techniques with high dimensional data analysis, ultimately preparing me for a modern independent career as a researcher. Other training opportunities proposed will strengthen my professional skills in writing, presenting, teaching/mentoring, and networking, laying the foundation for my long-term goals of leading a research lab at an academic institution. My training objectives are supported by my sponsor (Dr. Amanda Lund; Departments of Dermatology and Pathology) and co-sponsor (Dr. Itai Yanai; Departments of Biochemistry and Molecular Pharmacology and Director of the Institute for Computational Medicine) who bring expertise in cancer biology and immunology, and sequencing data analysis and integration, respectively.

项目概要 在肿瘤细胞到达之前，组织已做好转移的准备。原发性肿瘤通过以下方式驱动这种转变 将肿瘤衍生因子（TDF），包括细胞外囊泡和分泌/脱落蛋白（TSP）释放到 肿瘤相关的血液和淋巴管系统。尽管是最常见的转移部位 对于实体瘤类型，我们对淋巴结 (LN) 如何为 TDF 转移做好准备知之甚少。我的 初步数据表明，原发性黑色素瘤在其引流淋巴结中启动重塑，从而增强 相对于对侧、非引流和初始 LN 对照的转移性生长。此外，我还有 证明 TDF 独特地激活这种促转移 LN 表型，而不是源自 良性真皮或小鼠白蛋白。该领域的重点是肿瘤来源的细胞外囊泡的作用 在建立这种转移前的生态位中发挥着作用，但是我的数据进一步表明 TSP 也足以 激活这个利基市场。使用称为 BONCAT（双正交非规范）的新型细胞特异性标记策略 氨基酸标签），我已经确定了几个转运至转移前 LN 的候选 TSP。在这个 提议，我将测试以下假设：TSP 在常驻细胞群中启动重编程事件 引流液氮通过两个特定目标支持转移生长。第一个目标将采用 scRNAseq 识别细胞网络、TSP 诱导的驻留 LN 基质和种子肿瘤之间串扰的方法 细胞，为转移细胞提供营养支持以及 LN 淋巴管生成所需的程度 用于淋巴结转移。第二个目标将调查我已确定的特定 TSP，CSPG4（黑色素瘤） 相关硫酸软骨素蛋白多糖）。这种蛋白质由黑色素瘤细胞脱落，已知黑色素瘤 侵袭的细胞内在作用，并且在人类黑色素瘤患者中升高，然而，尚不清楚这种 TSP 是否 有助于远处部位转移前生态位的形成。这项工作的完成将为 有关 tdLN 中 TSP 依赖性转移前生态位发育的信息，并可能发现 CSPG4 作为 利基发展的新驱动力。拟议的工作将为我提供全面的培训机会 将新的工作台技术与高维数据分析相结合，最终为我做好了现代准备 作为研究员的独立职业。提出的其他培训机会将增强我的专业技能 在写作、演讲、教学/指导和网络方面，为我的长期目标奠定了基础 在学术机构领导一个研究实验室。我的培训目标得到了我的赞助商（Dr. 阿曼达·隆德；皮肤科和病理科）和共同发起人（Itai Yanai 博士； 生物化学和分子药理学和计算医学研究所所长）谁带来 分别在癌症生物学和免疫学以及测序数据分析和整合方面拥有专业知识。