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: 10535107
• 负责人: Haley du Bois
• 金额: $ 3.9万
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10535107
• 关键词: Albumins; Amino Acids; Behavior; Benign; Biochemistry; Biology; Blood; Breast; CSPG4 gene; 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; Institutes; Institution; Label; Length; Lung; Lymph; Lymphangiogenesis; Lymphatic Endothelial Cells; Lymphatic System; Malignant Neoplasms; Mediating; Medicine; Melanoma Cell; Mentors; Metastatic Neoplasm to Lymph Nodes; Metastatic to; Methods; Modernization; Molecular; Mus; Neoplasm Metastasis; Organ; Pathology; Patient risk; Patients; Pharmacology; Phenotype; Physiological; Play; Population; Pre-Clinical Model; Primary Neoplasm; Prognosis; Prognostic Marker; Proteins; Research; Research Personnel; Role; Seeds; 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; 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.

项目摘要 在肿瘤细胞到达之前，组织针对转移做好准备。原发性肿瘤通过以下方式驱动这种转化： 脱落肿瘤衍生因子（TDFs），包括细胞外囊泡和分泌/脱落蛋白（TSP）， 肿瘤相关的血液和淋巴管系统。尽管是最常见的转移部位， 虽然我们对实体瘤的类型知之甚少，但我们对淋巴结（LN）如何通过TDFs引发转移知之甚少。我 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， 相对于对侧、非引流和初治LN对照的转移性生长。而且我也 表明TDFs独特地激活这种促转移LN表型，而不是来自于 良性真皮或小鼠白蛋白。这一领域的重点一直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作用，然而，我的数据进一步表明TSP也足以 激活这个niche。使用一种新的细胞特异性标记策略，称为BONCAT（双正交非典型 氨基酸标记），我已经鉴定了几种转运到转移前LN的候选TSP。在这 在这个提议中，我将检验TSP在宿主细胞群中启动重编程事件的假设。 引流LN通过两个特定目的支持转移性生长。第一个目标将使用scRNAseq 识别细胞网络的方法，TSP诱导的驻留LN基质和接种肿瘤之间的串扰 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 淋巴结转移。第二个目标是研究我已经确定的一种特定的TSP，CSPG 4（黑色素瘤-黑色素瘤）。 相关硫酸软骨素蛋白聚糖）。这种蛋白质是由黑色素瘤细胞脱落的， 在人类黑色素瘤患者中，这种TSP在细胞内在的侵袭作用中升高，然而，目前尚不清楚这种TSP是否 有助于远处转移前的小生境形成。这项工作的完成将提供基础 tdLN中TSP依赖性转移前小生境发展的信息，并可能揭示CSPG 4作为一种转移因子。 利基发展的新驱动力。建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，最终为我准备了一个现代的 作为一名独立的研究员。建议的其他培训机会将加强我的专业技能 在写作，演示，教学/指导和网络，奠定了我的长期目标的基础， 领导一个学术机构的研究实验室。我的培训目标得到了我的赞助商（博士）的支持。 阿曼达隆德;皮肤科和病理科）和共同申办者（Itai柳井博士; 生物化学和分子药理学和计算医学研究所所长）， 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。