Deconstructing the collective invasion pack to define Myo10 function

解构集体入侵包定义Myo10功能

• 批准号: 10159870
• 负责人: Adam I. Marcus
• 金额: $ 35.69万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-06 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10159870
• 关键词: 3-Dimensional; Address; Automobile Driving; Back; Cancer Etiology; Cancer Patient; Cell Count; Cell Line; Cells; Cellular biology; Cessation of life; Data; Disease; Disseminated Malignant Neoplasm; Epigenetic Process; Extracellular Matrix; Fibronectins; Filopodia; Genes; Genetic Transcription; Genomics; Heterogeneity; Histologic; Image; Individual; Invaded; Lung; Malignant neoplasm of lung; Mechanics; Mediating; Methylation; Modeling; Molecular; Molecular Analysis; Movement; Neoplasm Metastasis; Pathway interactions; Patients; Phenotype; Population; Prevalence; Publishing; Research; Resolution; Resources; Shapes; Signal Transduction; Slice; Solid Neoplasm; Structure of parenchyma of lung; Supporting Cell; Techniques; Testing; Transcriptional Activation; Western Blotting; Width; Xenograft Model; base; cancer cell; cell population study; cell type; cohesion; epigenomics; ex vivo imaging; fibrillogenesis; genome-wide; genomic platform; genomic profiles; image guided; in vivo; insight; lung cancer cell; lung tumorigenesis; mimicry; mouse model; notch protein; overexpression; programs; promoter; spatiotemporal; therapeutic target; tool; transcriptome; tumor; tumor progression

Project Summary Collective invasion is a major mode of metastasis observed in patients across most solid tumor types. How the collective invasion pack operates, communicates, and navigates as a single cohesive unit remains unclear. To address this, we published on an image-guided genomics platform to isolate any living cell(s) within a collective invasion pack, and expand the population for genomic and molecular analysis, a technique termed Spatiotemporal Cellular & Genomic Analysis (SaGA). We used SaGA to dissect the molecular, epigenetic, and genomic profiles of leader and follower cells invading as a hierarchical cohesive unit. To determine how epigenetic reprogramming drives this phenotypic heterogeneity, we deconstructed the collective invasion pack using SaGA, then integrated genome-wide promoter methylation and transcriptome data to define differentially methylated regions within the leader and follower phenotypes. We observe global epigenomic re-wiring in leader cells supporting an epigenetic basis for the phenotypic heterogeneity within the collective invasion pack. We then identified Myo10 (myosinX) as a top differentially methylated and expressed gene, where the leader cell promoter is hypomethylated, and leaders in several lung cancer lines overexpress Myo10. Myo10 is a canonical modulator of filopodia elongation and we show it drives filopodia elongation, collective invasion, leader cell-driven fibronectin micropatterning (fibrillogenesis), and is transcriptionally activated by Jag1/Notch. We will use this information to test a mechanistic model with the overarching hypothesis that Myo10 activation via promoter hypomethylation in leader cells drives filopodia-based micropatterning of fibronectin to create a leader cell-driven collective invasion path. We propose that this leads to an invasive advantage for lung cancer cells resulting in metastatic disease. In Aim 1 we test the model that Myo10 hypomethylation in leaders allows for Jag1/Notch1- driven transcriptional activation, driving filopodia elongation, and fibronectin micropatterning. In Aim 2 we test how this collective invasion pathway impacts metastasis using in vivo metastasis models and the first patient- derived leader cells. Throughout, we leverage unique resources developed here including SaGA-derived cell lines, ex vivo imaging, and patient-derived lung cancer leader cells. We speculate that these data will provide mechanistic insight into collective invasion and translational value towards understanding lung cancer patient leader cell biology.

项目摘要 集体入侵是大多数实体瘤类型的患者中观察到的一种主要转移方式。如何 集体入侵包在单个凝聚力单元中运行，通信和导航尚不清楚。到 解决这个问题，我们在图像引导的基因组学平台上发表了，以隔离集体中的任何活细胞 入侵包，并扩大种群以进行基因组和分子分析，该技术称为 时空细胞和基因组分析（SAGA）。我们使用传奇来剖析分子，表观遗传和 领导者和追随者细胞的基因组谱作为分层凝聚单元。确定如何 表观遗传重编程驱动了这种表型异质性，我们解构了集体入侵包 然后使用传奇，然后整合全基因组启动子甲基化和转录组数据来定义差异化 领导者和追随者表型内的甲基化区域。我们观察到领导者的全球表观基因组学 支持集体入侵包中表型异质性的表观遗传基础的细胞。然后我们 鉴定为肌脱甲基的甲基化和表达的基因，将myo10（肌动蛋白）识别为领导者细胞 启动子是低甲基化的，并且在几个肺癌系中的领导者过表达myo10。 myo10是一个规范 丝状伸长率调节剂，我们表明它可以驱动丝状延伸，集体入侵，领导者细胞驱动 纤连蛋白微图解（原纤维生成），并被JAG1/NOTCH激活。我们将使用这个 通过总体假设测试机械模型的信息，即Myo10通过启动子激活 领导细胞中的低甲基化驱动基于丝虫的纤连蛋白的微观图案，以创建领导者细胞驱动 集体入侵路径。我们建议这导致肺癌细胞具有侵入性优势，导致 转移性疾病。在AIM 1中，我们测试了Leaders中Myo10甲基化允许JAG1/NOTCH1-的模型。 驱动的转录激活，驱动丝状伸长和纤连蛋白微图案。在AIM 2中我们测试 这种集体入侵途径如何使用体内转移模型和第一个患者影响转移 派生的领导细胞。在整个过程中，我们利用这里开发的独特资源，包括传说中的细胞 线，体内成像和患者来源的肺癌领导细胞。我们推测这些数据将提供 对集体入侵和对了解肺癌患者的转化价值的机械洞察力 领导者细胞生物学。