Use of Micro-RNA Arrays to Identify MDSC Functional Pathways and Markers

使用 Micro-RNA 阵列识别 MDSC 功能途径和标记

• 批准号: 8601921
• 负责人: Timothy L. Ratliff
• 金额: $ 15.86万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-02 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8601921
• 关键词: Acute; Arginine; Basic Amino Acid Transport Systems; Biological; Biological Process; Cell physiology; Cells; Chronic; Data; Development; Down-Regulation; Environment; Enzymes; Figs - dietary; Folate; Foundations; Gene Expression Profiling; Genes; Genetic Transcription; Human; ITGAM gene; Immune; Immunity; Inflammation; Inflammatory; Investigation; Knowledge; Link; Liver neoplasms; Malignant Neoplasms; MicroRNAs; Molecular; Molecular Profiling; Mus; Myelogenous; Myeloproliferative disease; NOS2A gene; Na(+)-K(+)-Exchanging ATPase; Names; Pathway interactions; Pattern; Phenotype; Population; Process; Proteins; RNA analysis; Role; Site; Spleen; Supplementation; Suppressor-Effector T-Lymphocytes; System; T cell response; T-Lymphocyte; Tumor Cell Invasion; Tumor Immunity; Tumor-Derived; Up-Regulation; Vitamin D; Vitamin D3 Receptor; arginase; base; human NOS2A protein; insight; network models; novel; public health relevance; receptor; tumor; tumor growth; uptake

DESCRIPTION (provided by applicant): Myeloid derived suppressor cells (MDSC) are a heterogeneous, ill-defined population of immune regulatory cells that are a critical cell population in suppressing tumor immunity and controlling inflammatory processes. Currently, MDSC function is based on studies of phenotypically defined cell populations that contain both suppressive and non-suppressive cells. Markers clearly defining functional MDSC are not available nor have functional pathways unique to MDSC been described. We recently showed that spleen Gr1+CD11b+ cells in mice, the prototypical MDSC phenotype, are precursor MDSC and that active regulatory MDSC are present only within a tumor or at the site of inflammation. Only tumor- or inflammation-derived MDSC to express Arginase I (Arg-I) and inducible nitric oxide synthase (iNOS) and suppress T cell responses, clearly localizing functional MDSC to active inflammatory sites. Thus, array analyses on MDSC both from spleen and tumor were performed to identify gene and micro-RNA (miRNA) expression patterns restricted to tumor-derived (functional) MDSC. Gene array analysis showed 150 genes to be upregulated at least 5-fold, including Arg-I (129-fold) and iNOS (33-fold), in tumor-derived (functional) MDSC compared to spleen (precursor) MDSC. Among the novel genes differentially expressed, to date two, slc7a2 and vitamin D receptor, were observed to regulate MDSC T cell suppressor activity. Thus we hypothesize that identifying miRNAs linked to MDSC function will identify new targets that will enable identification and modulation of active MDSC. Additionally, integrating the miRNA analysis with the gene array data will enhance pathway discovery unique to functional MDSC. Differential miRNA arrays were performed on the same spleen- and tumor-derived MDSC that were used for the gene array analysis. The miRNA arrays show a statistically significant 2-fold or greater up-regulation of 46 miRNAs and similarly a 2-fold or more down- regulation of 40 miRNAs. We propose to utilize the differential miRNA arrays in conjunction with the gene arrays to identify novel MDSC-specific genes and pathways that are associated with MDSC function. The identification of regulatory miRNAs and associated pathways will provide the foundation for better understanding the molecular basis for MDSC function and also will provide opportunity to better target MDSC and/or identify the presence of active MDSC. Two specific aims are proposed: (1) Identify genes, gene sets, pathways, and network modules that are significantly related to differentiating between functional and precursor MDSC to gain global insights of the biological process and its relevance to cancer. Hypothesis: functional MDSC utilize distinct functional pathways from precursor MDSC that will distinguish the two populations. The differential pathway expression linked to function can be detected at the systems level through pathway/network modeling and integrative expression profiling and (2) Validate the biological relevance of select miRNA identified via the analyses performed in Aim 1.

描述（由申请人提供）：髓样衍生的抑制细胞（MDSC）是免疫调节细胞的异质性，不确定的人群，在抑制肿瘤免疫和控制炎症过程中是关键细胞群体。目前，MDSC功能基于对包含抑制性和非抑制细胞的表型定义细胞群的研究。明显定义功能性MDSC的标记尚无可用，也没有描述MDSC独特的功能途径。我们最近表明，脾脏GR1+ CD11b+细胞在小鼠中，原型MDSC表型是前体MDSC，而主动调节MDSC仅存在于肿瘤或炎症部位。仅肿瘤或炎症衍生的MDSC表达精氨酸酶I（ARG-I）和诱导型一氧化氮合酶（INOS）并抑制T细胞反应，显然将功能性MDSC定位在活性炎症位点上。因此，对来自脾脏和肿瘤的MDSC进行了阵列分析，以鉴定仅限于肿瘤衍生（功能）MDSC的基因和微RNA（miRNA）表达模式。基因阵列分析表明，与脾脏（Prestors）MDSC相比，在肿瘤衍生的（功能性）MDSC中，150个基因至少上调了5倍，包括Arg-I（129倍）和Inos（33倍）。在差异表达的新型基因中，迄今为止，SLC7A2和维生素D受体被观察到调节MDSC T细胞抑制活性。因此，我们假设识别与MDSC函数相关的miRNA将确定可以识别和调节活动MDSC的新目标。此外，将miRNA分析与基因阵列数据相结合将增强功能MDSC独有的途径发现。差异miRNA阵列在用于基因阵列分析的同一脾脏和肿瘤衍生的MDSC上进行。 miRNA阵列显示出46个miRNA的统计学意义2倍或更高的上调，同样，有40个miRNA的2倍或更多下调。我们建议利用差异miRNA阵列与基因阵列一起识别与MDSC功能相关的新型MDSC特异性基因和途径。调节性miRNA和相关途径的鉴定将为更好地理解MDSC功能的分子基础提供基础，并将为更好地靶向MDSC和/或确定活性MDSC的存在提供机会。提出了两个具体目的：（1）确定与区分功能和前体MDSC显着相关的基因，基因集，途径和网络模块，以获得对生物学过程及其与癌症相关的全球见解。假设：功能性MDSC利用了与前体MDSC不同的功能途径，该途径将区分这两个人群。可以通过途径/网络建模和集成表达分析在系统级别检测到链接到功能的差分途径表达，并且（2）通过在AIM 1中执行的分析鉴定的选择miRNA的生物学相关性。