Microfluidic Techniques for the Molecular and Functional Analysis of Gene Express

用于基因表达分子和功能分析的微流控技术

基本信息

批准号：
7662667
负责人：
STEPHEN R QUAKE
金额：
$ 44.58万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7662667
关键词：
Antibodies Area Automation Behavior Bioinformatics Biological Biological Assay Blood Vessels CD47 gene Cell Count Cell Culture Techniques Cell model Cells Chromatin Chromatin Structure Chromatin Structure Alteration Complement Culture Media DNA Data Data Analyses Devices Diagnosis Disease Environment Epigenetic Process Fibroblasts Gene Expression Gene Expression Regulation Genes Genetic Genomics Growth Growth Factor Heterogeneity Histones Human Human Resources In Vitro Individual Investigation Laboratories Ligands Malignant Neoplasms Measurement Mesenchymal Methylation Microfluidic Microchips Microfluidics Miniaturization Modification Molecular Mus Normal tissue morphology Outcome Pathway Analysis Pathway interactions Patients Play Polycomb Population Post-Translational Protein Processing Proliferating Property Proteins Protocols documentation Radiopharmaceuticals Recruitment Activity Regulation Regulatory Pathway Resistance Role Schedule Screening procedure Sequence Analysis Signal Transduction Pathway Solid Solid Neoplasm Stem cells Stratification Stromal Cells Support System Supporting Cell System Techniques Technology Testing Therapeutic Therapeutic Agents Time Video Microscopy Xenograft procedure base cancer cell cancer stem cell cancer therapy cell behavior cell growth cell motility chromatin immunoprecipitation design high throughput screening in vitro Assay in vitro testing in vivo insight leukemia malignant breast neoplasm matrigel neoplastic cell new therapeutic target research study response self-renewal stem cell population therapeutic target time use tool transcription factor tumor tumor growth tumorigenic

项目摘要

Cancer stem cells in solid tumors have been recently isolated and appear to be primarily responsible for the growth and spread of the disease. The presence of a stem cell population in a tumor has implications for the diagnosis and treatment of cancer, as it is these cancer stem cells that must be targeted to achieve a cure. Preliminary evidence demonstrates that there is a host of genes differentially expressed by the cancer stem cells and their non-tumorigenic progeny. Many of these genes are thought to play a role in essential cancer functions including proliferation, survival, self renewal and resistance to standard therapeutics. Uncovering the true functional stratification of the superficially uniform population of stem cells in leukemia, breast cancer and other solid cancers is a challenge which requires new kinds of measurements at the single-cell level. Based on recent discoveries by the Clarke and Weissman groups, we believe that gene expression in normal stem cells, as well as cancer stem cells is significantly regulated at the epigenetic level. Epigenetic regulation of gene expression is in part controlled by the posttranslational modification of histone proteins, as well as methylation of DMA, both of which result in the alteration of chromatin structure. These modifications are examined using Chromatin immunoprecipitation (ChIP) assays. Epigenetic analysis is a necessary complement to gene expression analysis in order to understand the control of normal and cancer stem cell self-renewal and discover new therapeutic targets. The microfluidic tools developed by the Quake lab for single cell gene expression (aim1), chip (aim 2) and high throughput in vitro cell culture (aim 3) will allow the Clarke and Weissman laboratories to perform gene expression and epigenetic analyses on rare, purified cells from model mice and primary human cancer or cancer xenografts. These assays will all use microfluidic platforms which have already been developed and validated in the Quake laboratory or are commercially available. New systems will also be designed as needed in the course of this project. Microfluidic epigenetic and genetic assays will allow the study of highly purified, homogeneous, rare cell populations that were previously inaccessible with the standard techniques. Microfluidic cell culture platforms will allow finding appropriate conditions to culture cancer stem cells in vitro and test new therapeutic targets

实体瘤中的癌症干细胞最近被分离出来，似乎主要是造成疾病的生长和传播。肿瘤中干细胞种群的存在对癌症的诊断和治疗，因为这些癌症干细胞必须针对治愈。初步证据表明，癌症茎有差异表达的基因细胞及其非肿瘤后代。这些基因中的许多被认为在基本癌症中起作用功能包括增殖，生存，自我更新和对标准治疗剂的抵抗力。发现白血病，乳腺癌中干细胞表面均匀种群的真正功能分层其他固体癌症是一个挑战，需要在单细胞水平上进行新的测量。根据克拉克和魏斯曼群体的最新发现，我们相信基因在正常干细胞以及癌症干细胞在表观遗传水平上受到显着调节。表观遗传学基因表达的调节部分由组蛋白的翻译后修饰作为控制以及DMA的甲基化，这两者都会改变染色质结构。这些修改使用染色质免疫沉淀（CHIP）测定进行检查。表观遗传分析是必要的补充基因表达分析以了解正常和癌干细胞的控制自我更新并发现新的治疗靶标。地震实验室开发的微流体工具单细胞基因表达（AIM1），CHIP（AIM 2）和高通量体外细胞培养（AIM 3）将允许克拉克（Clarke）和魏斯曼（Weissman）实验室对稀有，纯化的基因表达和表观遗传分析进行来自模型小鼠和原发性人类癌症或癌症异种移植的细胞。这些测定都将使用微流体已经在地震实验室开发和验证的平台或商业上是可用的。在此项目过程中，新系统也将根据需要设计。微流体表观遗传学遗传测定将允许研究高度纯化的，均匀的，稀有的细胞群体以前与标准技术无法访问。微流体细胞培养平台将允许查找适当的培养癌症干细胞体外和测试新治疗靶标的条件