Project 2: Profiling Gene Expression and Mechanophenotype in Circulating Tumor Cells Ex Vivo

项目 2：离体循环肿瘤细胞的基因表达和机械表型分析

基本信息

批准号：
10681243
负责人：
Ian Y Wong
金额：
$ 22.58万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681243
关键词：
3-Dimensional Affect Biochemistry Biocompatible Materials Bioinformatics Biological Assay Biomedical Engineering Blood Circulation Bone Marrow Stem Cell Breast Cancer Cell Cell Communication Cells Cellular Spheroids Centers of Research Excellence Coculture Techniques Computational Biology Confocal Microscopy Cultured Tumor Cells Data Disease Progression Distant Drug Screening Drug resistance Engineering Epithelium Estrogen receptor positive Exhibits Extracellular Matrix Family suidae Fibroblasts Fibroins Gene Expression Gene Expression Profile Gene Expression Profiling Genetic Transcription Genomics Goals Heterogeneity Homing Human Individual Invaded Liver Lung Malignant Neoplasms Maps Mechanics Mentors Mesenchymal Metastatic Neoplasm to the Bone Microfluidics Micrometastasis Modeling Morphology National Institute of Biomedical Imaging and Bioengineering Neoplasm Circulating Cells Neoplasm Metastasis Organ Outcome Patients Pattern Pharmaceutical Preparations Phenotype Primary Neoplasm Proliferating Research Research Personnel Research Project Grants Sampling Silk Site Sorting Stimulus Stromal Cells Technology Therapeutic Tissue constructs Tissues Traction Validation Xenograft procedure bone cancer cell cell motility cell type colonization resistance high throughput screening human disease inhibitor malignant breast neoplasm mechanical stimulus mimetics neoplastic cell pre-clinical programs response self assembly single cell analysis spatiotemporal therapy resistant transcriptome sequencing transcriptomic profiling tumor microenvironment

项目摘要

PROJECT SUMMARY Primary tumors shed circulating tumor cells (CTCs) into the bloodstream that metastasize preferentially to distant organs, resulting in 90% of cancer related fatalities. For example, estrogen receptor positive (ER+) breast cancers exhibit high rates of metastasis to bone, with decreased rates to liver and lung. CTCs exhibit heterogeneous gene expression programs and functional phenotypes, which are selected by soluble and mechanical interactions within each metastatic “niche.” A critical challenge is to predict how patient-specific CTCs disseminate throughout the body and respond to therapeutic treatments. An exciting strategy is to culture CTCs ex vivo for drug screening informed by genomic and transcriptional profiling. We seek to elucidate how CTCs respond to different features of the metastatic niche by engineering controlled interactions with tissue-specific extracellular matrix (ECM) and with human primary stromal cells, which may recapitulate disease progression and therapeutic resistance in these microenvironmental contexts. Our long-term goal is to establish preclinical assays for patient CTCs to predict metastatic disease progression and screen targeted inhibitors. Realization of this goal involves several technical challenges, including: 1) Tissue-mimetic matrix with tunable biochemistry and mechanics, 2) Multicellular tissue constructs with controlled size and cellular composition, 3) Gene expression profiling of CTCs in response to soluble and mechanical stimuli, 4) Spatiotemporal analysis of phenotypic heterogeneity, including the epithelial- mesenchymal transition (EMT), and 5) Validation with human patient samples. Thus, our overall objective is to understand how CTC gene expression and mechanophenotype is regulated by matrix or stromal interactions in tissue-mimetic microenvironments. This discovery-based approach can deconstruct patterns of gene expression driven by decellularized extracellular matrix or the secretome of human stromal cells. Such bioinformatics analyses can identify possible therapeutic strategies based on existing patient, xenograft, and high-throughput screens. PI: Wong is a New Investigator with expertise in cancer cell migration, biomaterials, and microfluidics. Mentor: Reichner is an expert on directed cell migration and mechanobiology and Mentor: Bertone is an expert on single cell analyses in cancer. We investigate the relative contributions of microenvironmental stimuli using three aims: AIM 1 will elucidate how individual breast cancer cells interact with tissue-mimetic matrix and AIM 2 will engineer co-cultured multicellular spheroids with stromal cells.

项目概要原发性肿瘤将循环肿瘤细胞 (CTC) 释放到血流中并发生转移优先影响远处器官，导致 90% 的癌症相关死亡，例如雌激素。受体阳性 (ER+) 乳腺癌骨转移率较高，而肝转移率较低和肺的 CTC 表现出异质的基因表达程序和功能表型。通过每个转移“生态位”内的可溶性和机械相互作用进行选择。预测患者特异性 CTC 如何在全身传播以及对治疗的反应。令人兴奋的策略是离体培养 CTC，通过基因组和转录谱进行药物筛选。我们试图通过工程控制来阐明 CTC 如何响应转移生态位的不同特征与组织特异性细胞外基质（ECM）和人类原代基质细胞的相互作用，这可能概括这些微环境背景下的疾病进展和治疗耐药性。我们的长期目标是建立患者 CTC 的临床前检测以预测转移性疾病进展和筛选靶向抑制剂的实现涉及几个技术挑战，包括：1) 具有可调节生物化学和力学功能的组织模拟基质，2) 多细胞组织构建体具有受控的大小和细胞组成，3) CTC 响应可溶性和机械刺激，4）表型异质性的时空分析，包括上皮细胞间充质转化 (EMT)，以及 5) 使用人类患者样本进行验证因此，我们的总体目标是。了解基质或基质相互作用如何调节 CTC 基因表达和机械表型这种基于发现的方法可以解构基因的模式。由去细胞化的细胞外基质或人类基质细胞的分泌组驱动的表达。生物信息学分析可以根据现有患者、异种移植物和高通量屏幕。 PI：Wong 是一位新研究员，在癌细胞迁移、生物材料和微流体领域具有专业知识。 Reichner 是定向细胞迁移和机械生物学方面的专家，导师：Bertone 是定向细胞迁移和机械生物学方面的专家我们利用单细胞分析研究微环境刺激的相关贡献。三个目标：AIM 1 将阐明个体乳腺癌细胞如何与组织模拟基质和 AIM 2 相互作用将设计与基质细胞共培养的多细胞球体。