ROLE OF MICRORNAS IN THE MOLECULAR PATHOGENESIS OF HIV/SIV ENTEROPATHY

微生物在 HIV/SIV 肠病分子发病机制中的作用

• 批准号: 8173006
• 负责人: Mahesh Mohan
• 金额: $ 3.94万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173006
• 关键词: Animals; Autopsy; Cells; Chronic; Colitis; Colon; Complex; Computer Retrieval of Information on Scientific Projects Database; Diarrhea; Disease; Epithelium; Funding; Gastrointestinal tract structure; Gene Expression; Grant; Histopathology; Immune system; Immunofluorescence Immunologic; In Situ Hybridization; In Vitro; Infection; Inflammation; Institution; Intestines; Lamina Propria; Macaca; Macaca mulatta; Mass Spectrum Analysis; Messenger RNA; Methods; MicroRNAs; Modeling; Molecular; Nuclear; Nucleotides; Pathogenesis; Regulator Genes; Research; Research Personnel; Resources; Role; Source; Structure; Tissues; Translational Repression; United States National Institutes of Health; Untranslated RNA; improved; intestinal epithelium; locked nucleic acid; macrophage; novel; response; wasting

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Background & Aims: The Gastrointestinal tract (GIT) is a major target of HIV/SIV infection. Although our understanding of HIV/SIV enteropathy has greatly improved, the recent discovery of miRNAs has added yet another novel and complex regulator of gene expression with potential roles in the molecular pathogenesis of this disorder. microRNAs (miRNAs) are genomically transcribed, ~21-23 nucleotide noncoding RNAs that are highly conserved and suppress gene expression by targeting mRNAs for translational repression or degradation. We investigated the contribution of 11 miRNAs to GIT disease and inflammation using the SIV-infected rhesus macaque model. Methods: Colon tissue was collected at necropsy from 10 SIV-infected (G1) with chronic diarrhea and 5 uninfected control macaques (G2). The contributions of miR-21, miR-125b, miR-132, miR-142-3p, miR-142-5p, miR-146a, miR-155, miR-203, miR-212, miR-223 and miR-338, previously known to be associated with inflammation were investigated using QRT-PCR, In situ hybridization/Immunofluorescence, histopathology and mass spectrometry. Results: All G1 macaques had chronic diarrhea, wasting and colitis. Significant to moderate increase in the expression of miR-142-3p, miR-142-5p (5-6-fold), miR-212 (3-fold), miR-21 (2-fold) was observed in the colon of G1 macaques compared to G2 animals. Interestingly, miR-125b and miR-203 were downregulated (2-3 fold) in G1 animals. No change in miR-132, miR-146a, miR-155 , miR-223 and miR-338 expression was observed between the groups. In situ hybridization using Locked-nucleic acid modified miR-212 probes revealed cytoplasmic and nuclear localization in the colonic epithelium and lamina propria cells (macrophages). Using mass spectrometry, we validated several predicted mRNA targets of miR-142-3p and miR-212 following over expression of both miRNAs in in vitro cultured primary intestinal macrophages. Conclusion: Our findings suggest that deregulation in cell/tissue-specific expression of miRNAs occurring in response to HIV/SIV infection may disrupt the functional relationship between the intestinal epithelium and the mucosal immune system causing alterations in intestinal structure and function.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 背景与目标：胃肠道（GIT）是HIV/SIV感染的主要目标。尽管我们对HIV/SIV肠病的理解已大大提高，但最近发现miRNA的发现增加了基因表达的另一种新颖而复杂的调节剂，在该疾病的分子发病机理中潜在作用。 microRNA（miRNA）是基因组转录的，〜21-23个核苷酸非编码RNA，它们是高度保守的，并通过靶向mRNA来抑制转化抑制或降解，从而抑制基因表达。我们使用SIV感染的恒河猕猴模型研究了11个miRNA对GIT疾病和炎症的贡献。方法：从10个SIV感染（G1）的尸检中收集结肠组织，并用慢性腹泻和5种未感染的对照猕猴（G2）收集。 The contributions of miR-21, miR-125b, miR-132, miR-142-3p, miR-142-5p, miR-146a, miR-155, miR-203, miR-212, miR-223 and miR-338, previously known to be associated with inflammation were investigated using QRT-PCR, In situ hybridization/Immunofluorescence, histopathology and mass spectrometry.结果：所有G1猕猴均患有慢性腹泻，浪费和结肠炎。与G2动物相比，在G1猕猴的结肠中观察到miR-142-3p，miR-142-5p（5-6倍），miR-212（3倍），miR-212（3倍），miR-212（3倍），miR-212（3倍）的表达显着到中度增加。有趣的是，在G1动物中，miR-125b和miR-203被下调（2-3倍）。在两组之间观察到miR-132，miR-146a，miR-155，miR-155，miR-155，miR-155和miR-338表达没有变化。使用锁定核酸修饰的miR-212探针的原位杂交揭示了结肠上皮细胞和固有层的胞质和核定位（巨噬细胞）。使用质谱法，我们验证了MiR-142-3p和miR-212的几个预测的mRNA靶标，而在两种miRNA中都在体外培养的原代肠道巨噬细胞中表达。结论：我们的发现表明，在响应HIV/SIV感染时发生的miRNA的细胞/组织特异性表达失调可能会破坏肠上皮和粘膜免疫系统之间的功能关系，从而导致肠结构和功能的改变。