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: 8451031
• 负责人: Timothy L. Ratliff
• 金额: $ 21.05万
• 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 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8451031
• 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）：骨髓源性抑制细胞（MDSC）是异质的、不明确的免疫调节细胞群，是抑制肿瘤免疫和控制炎症过程的关键细胞群。目前，MDSC 功能基于对包含抑制性细胞和非抑制性细胞的表型定义的细胞群的研究。没有明确定义功能性 MDSC 的标记，也没有描述 MDSC 独特的功能途径。我们最近表明，小鼠脾脏 Gr1+CD11b+ 细胞（典型的 MDSC 表型）是前体 MDSC，并且活性调节性 MDSC 仅存在于肿瘤内或炎症部位。只有肿瘤或炎症来源的 MDSC 才能表达精氨酸酶 I (Arg-I) 和诱导型一氧化氮合酶 (iNOS) 并抑制 T 细胞反应，从而将功能性 MDSC 明确定位于活跃的炎症部位。因此，对来自脾脏和肿瘤的 MDSC 进行阵列分析，以确定仅限于肿瘤来源（功能性）MDSC 的基因和微 RNA (miRNA) 表达模式。基因阵列分析显示，与脾脏（前体）MDSC 相比，肿瘤源性（功能性）MDSC 中的 150 个基因上调至少 5 倍，包括 Arg-I（129 倍）和 iNOS（33 倍）。在差异表达的新基因中，迄今为止观察到 slc7a2 和维生素 D 受体这两个基因可调节 MDSC T 细胞抑制活性。因此，我们假设识别与 MDSC 功能相关的 miRNA 将识别出新的靶标，从而能够识别和调节活性 MDSC。此外，将 miRNA 分析与基因阵列数据相结合将增强功能性 MDSC 独特的通路发现。在用于基因阵列分析的相同脾脏和肿瘤来源的 MDSC 上进行差异 miRNA 阵列。 miRNA 阵列显示 46 个 miRNA 具有统计学显着性的 2 倍或以上上调，类似地，40 个 miRNA 具有 2 倍或以上下调。我们建议利用差异 miRNA 阵列与基因阵列相结合来识别与 MDSC 功能相关的新型 MDSC 特异性基因和通路。调节性 miRNA 和相关通路的鉴定将为更好地理解 MDSC 功能的分子基础奠定基础，也将为更好地靶向 MDSC 和/或鉴定活性 MDSC 的存在提供机会。提出了两个具体目标：（1）识别与区分功能性 MDSC 和前体 MDSC 显着相关的基因、基因集、通路和网络模块，以获得生物过程及其与癌症相关性的全局见解。假设：功能性 MDSC 利用与前体 MDSC 不同的功能途径来区分这两个群体。通过通路/网络建模和综合表达谱分析，可以在系统水平上检测与功能相关的差异通路表达；(2) 验证通过目标 1 中进行的分析确定的选定 miRNA 的生物学相关性。