Project 2: Contribution of the Stromal Microenvironment to Early Dissemination

项目 2：基质微环境对早期传播的贡献

• 批准号: 10705081
• 负责人: Mayumi Ito
• 金额: $ 34.68万
• 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: 2022-09-15 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705081
• 关键词: ATAC-seq; Address; Adjuvant Therapy; Behavior; Benign; Cancer Control; Cancerous; Cell Communication; Cell Lineage; Cells; ChIP-seq; Characteristics; Chromatin; Clinical; Clinical Data; Congenic Mice; Cytokine Signaling; Data; Dermis; Development; Diagnosis; Disease; Disseminated Malignant Neoplasm; Distal; Environment; Epidermis; Epigenetic Process; Extracellular Matrix; Fibroblasts; Gene Cluster; Genes; Genetic Engineering; Genetic Transcription; Growth; Growth Factor; Heterogeneity; Histones; Human; Human Pathology; IGF1 gene; IGF1R gene; Immune; Immune system; Immunofluorescence Microscopy; Immunohistochemistry; Invaded; Ligands; Link; Lymphatic; Malignant Neoplasms; Maps; Melanoma Cell; Metastatic Melanoma; Metastatic Neoplasm to Lymph Nodes; Modeling; Molecular; Mus; Neoplasm Metastasis; Organ; Pathologic; Pathway Analysis; Pathway interactions; Patient Selection; Patients; Pattern; Phenotype; Play; Population; Primary Neoplasm; Process; Prognostic Marker; Regulation; Regulator Genes; Regulatory Element; Research; Resolution; Resources; Role; Signal Pathway; Signal Transduction; Solid Neoplasm; Spatial Distribution; Specimen; Stromal Cells; Technology; Testing; Time; Tissues; Tomatoes; cancer cell; cancer type; cell type; chemokine; clinically relevant; cytokine; differential expression; draining lymph node; genetic manipulation; in vivo; insight; melanocyte; melanoma; metastatic process; migration; molecular marker; morphogens; mouse model; neoplastic cell; pancreatic cancer model; prevent; prognostic value; programs; receptor; single-cell RNA sequencing; stem cells; synergism; targeted treatment; therapeutic target; tool; transcription factor; transcriptome; transcriptomics; tumor; tumor heterogeneity; tumor initiation; tumor progression; tumorigenesis

PROJECT 2 – SUMMARY Metastasis initiates early in melanoma, and coincides with phenotypic and functional changes in tumor cells, surrounding stromal cells, and extracellular matrix components. Amongst the stromal cell types, cancer associated fibroblasts (CAFs) are emerging as a particularly relevant cell population. Although they are genetically stable, CAFs are phenotypically and functionally diverse and distinct subsets of CAFs were found to support or inhibit cancerous growth in pancreatic cancer models. Although these findings suggest functionally diverse sets of CAFs could also exist in other cancers, they have yet to be defined in most other cancer types. Furthermore, because specific CAFs might be valuable prognostic markers or therapeutic targets, it is becoming increasingly important to better understand their development and regulation. Here, we propose to investigate how signals between melanoma cells and fibroblasts create heterogeneity within tumors and how this heterogeneity unleashes metastatic behaviors in a subset of melanoma cells. To test this hypothesis, we developed genetically engineered congenic mouse models that allow us to genetically manipulate melanocyte stem cells to initiate tumors that faithfully recapitulate the pathology of human melanoma and to independently inhibit specific genes in melanoma cells or CAFs. These models will uniquely enable us to investigate how specific signaling mechanisms influence CAF functions in melanoma development and metastasis. We propose to couple our mouse models with single-cell RNA sequencing, spatial transcriptomics, ATAC-seq, and ChIP-seq approaches to transcriptionally define distinctive CAF states. We will also use them along with melanoma specimens from patients with and without metastatic progression to map specific CAFs and their proximity to other cell types with highly multiplexed immuno-histochemistry (Aim 1). Next, we will use these tools and data to discover cell state specific receptor-ligand interactions and study how IGF1-IGF1R signaling between melanoma cells and CAFs –one model pathway our preliminary studies identified – influences intratumor heterogeneity and metastatic progression in vivo (Aim 2). Project 2 will leverage the pathological, technological, and analytical resources of Cores B and C. It will synergize with Project 1 to determine whether and how melanoma cells interact with distinctive CAFs and how these interactions allow for metastatic dissemination. Project 2 will also enhance Project 3 by shedding new light onto CAF-secreted cytokines that modulate tumor draining lymph nodes and make immune cells tolerant to metastasizing melanoma cells. Once we understand these processes on a molecular level and link them to clinical characteristics, we can begin to inform patient selection for adjuvant therapy in early-stage melanoma and catalyze a rational development of targeted therapies that prevent or treat metastatic dissemination in early- stage melanoma and other cancer types.

项目2--摘要 转移始于黑色素瘤的早期，与肿瘤细胞的表型和功能变化相一致。 周围的基质细胞和细胞外基质成分。在基质细胞类型中，癌症 相关成纤维细胞(CAF)正在成为一个特别相关的细胞群。尽管他们是 CAF在遗传上是稳定的，是表型和功能不同的，CAF的不同亚集被发现 支持或抑制胰腺癌模型中的癌细胞生长。尽管这些发现在功能上表明 在其他癌症中也可能存在不同的CAF组，但在大多数其他癌症类型中尚未定义它们。 此外，由于特定的CAF可能是有价值的预后标记物或治疗靶点，它正在成为 更好地了解它们的发展和监管变得越来越重要。在这里，我们建议调查 黑色素瘤细胞和成纤维细胞之间的信号如何在肿瘤内产生异质性，以及这是如何 异质性释放了黑色素瘤细胞亚群的转移行为。为了检验这一假设，我们 开发了基因工程的同种小鼠模型，允许我们从基因上操纵黑素细胞 干细胞启动肿瘤，忠实地概括人类黑色素瘤的病理，并独立 抑制黑色素瘤细胞或CAF中的特定基因。这些模型将独特地使我们能够调查 特定的信号机制影响CAF在黑色素瘤发生和转移中的功能。我们建议 将我们的小鼠模型与单细胞rna测序、空间转录、atac-seq和chip-seq相结合 从转录上定义不同的CAF状态的方法。我们也会将它们与黑色素瘤一起使用 从有和无转移进展的患者样本中定位特定的CAF及其与 其他细胞类型采用高度多元化的免疫组织化学(目标1)。接下来，我们将使用这些工具和数据来 发现细胞状态特异性受体-配体的相互作用，并研究IGF1-IGF1R信号在黑色素瘤之间 细胞和CAF--我们初步研究确定的一个模型途径--影响肿瘤内的异质性和 体内转移进展(目标2)。 项目2将利用核心B和C的病理、技术和分析资源。它将 与项目1协同，以确定黑色素瘤细胞是否以及如何与独特的CAF相互作用以及如何相互作用 这些相互作用允许转移性传播。项目2还将通过提供新的亮点来增强项目3 CAF分泌的细胞因子调节肿瘤引流淋巴结并使免疫细胞耐受 转移性黑色素瘤细胞。一旦我们在分子水平上了解了这些过程并将它们与 临床特征，我们可以开始通知患者选择早期黑色素瘤的辅助治疗 并促进靶向治疗的合理发展，以防止或治疗早期转移扩散- 分期黑色素瘤和其他癌症类型。