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

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 胃肠道（GIT）是HIV/SIV感染的主要目标。虽然我们对HIV/SIV肠病的理解已经有了很大的提高，但最近发现的miRNAs又增加了另一种新的复杂的基因表达调控因子，在这种疾病的分子发病机制中具有潜在的作用。microRNA（miRNAs）是基因组转录的约21-23个核苷酸的非编码RNA，其高度保守并通过靶向mRNA进行翻译抑制或降解来抑制基因表达。我们使用SIV感染的恒河猴模型研究了miRNA对GIT疾病和炎症的贡献。在尸检时从8只急性SIV感染的猕猴（G1）（n=3，SIV感染后8天和n=5，SIV感染后13-30天）和5只未感染的对照猕猴（G2）收集结肠组织。使用TaqMan低密度阵列（TLDA）、QRT-PCR、原位杂交/免疫荧光和组织病理学研究miRNA对GIT疾病和炎症的贡献。卡A上的377个miRNA中的大约70%和卡B上的377个miRNA中的50%与恒河猴交叉反应。在使用STATMINER进行数据分析后，我们鉴定了总共25个卡片A和12个卡片B miRNA在响应SIV感染时差异表达（p 0.05）。重要的差异表达卡片A miRNA包括miR-1、-26 a、-29 c、-34 a、-125 b、-126、-130 a、-133 b、-135 a、-140-5 p、-let 7 b、-let 7 c、-222、-224、-383。相似地，显著差异表达的卡片B miRNA包括-190 b、-550、-1247。 目前正在进行QRT-PCR以进一步确认TLDA表达谱和原位杂交研究以鉴定表达这些miRNA的特定细胞类型。 我们的研究结果表明，在响应HIV/SIV感染发生的细胞/组织特异性表达的miRNA的失调可能会破坏肠上皮细胞和粘膜免疫系统之间的功能关系，导致肠道结构和功能的改变。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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 miRNAs to GIT disease and inflammation using the SIV-infected rhesus macaque model. Colon tissue was collected at necropsy from 8 acutely SIV-infected (G1) (n=3, 8 days post SIV infection and n=5, 13-30 days post SIV infection) and 5 uninfected control macaques (G2). The contribution of miRNAs to GIT disease and inflammation was investigated using TaqMan Low Density Array (TLDA), QRT-PCR, In situ hybridization/Immunofluorescence and histopathology. Approximately 70% of the 377 miRNAs on card A and 50% of the 377 miRNAs on card B cross-reacted with the rhesus macaque. Following data analysis using STATMINER we identified a total of 25 card A and 12 card B miRNAs to be differentially expressed (p 0.05) in response to SIV infection. Important differentially expressed card A miRNAs included miR-1, -26a, -29c, -34a, -125b, -126, -130a, -133b, -135a, -140-5p, -let7b, -let7c, -222, -224, -383. Similarly, notable differentially expressed card B miRNAs included -190b, -550, -1247. QRT-PCR to further confirm TLDA expression profiles and in situ hybridization studies to identify specific cell types expressing these miRNAs are currently in progress. 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.