Li KaShing Institute of Virology, University of Alberta, Edmonton, Alberta, Canada.
Applied Microbiology Research, Department Biomedicine, University of Basel, Basel, Switzerland.
Am J Transplant. 2016 Feb;16(2):650-60. doi: 10.1111/ajt.13464. Epub 2015 Oct 13.
Human (Homo sapiens) micro-RNAs (hsa-miRNAs) regulate virus and host-gene translation, but the biological impact in patients with human cytomegalovirus (hCMV) infection is not well defined in a clinically relevant model. First, we compared hsa-miRNA expression profiles in peripheral blood mononuclear cells from 35 transplant recipients with and without CMV viremia by using a microarray chip covering 847 hsa-miRNAs. This approach demonstrated a set of 142 differentially expressed hsa-miRNAs. Next, we examined the effect of each of these miRNAs on viral growth by using human fibroblasts (human foreskin fibroblast-1) infected with the hCMV Towne strain, identifying a subset of proviral and antiviral hsa-miRNAs. miRNA-target prediction software indicated potential binding sites within the hCMV genome (e.g., hCMV-UL52 and -UL100 [UL = unique long]) and host-genes (e.g., interleukin-1 receptor, IRF1). Luciferase-expressing plasmid constructs and immunoblotting confirmed several predicted miRNA targets. Finally, we determined the expression of selected proviral and antiviral hsa-miRNAs in 242 transplant recipients with hCMV-viremia. We measured hsa-miRNAs before and after antiviral therapy and correlated hsa-miRNA expression levels to hCMV-replication dynamics. One of six antiviral hsa-miRNAs showed a significant increase during treatment, concurrent with viral decline. In contrast, six of eight proviral hsa-miRNAs showed a decrease during viral decline. Our results indicate that a complex and multitargeted hsa-miRNA response occurs during CMV replication in immunosuppressed patients. This study provides mechanistic insight and potential novel biomarkers for CMV replication.
人类（智人）microRNAs（hsa-miRNAs）调节病毒和宿主基因翻译，但在具有人类巨细胞病毒（hCMV）感染的临床相关模型中，其对患者的生物学影响尚不清楚。首先，我们通过使用覆盖 847 个 hsa-miRNAs 的微阵列芯片，比较了 35 例移植受者有无 CMV 病毒血症时外周血单个核细胞中的 hsa-miRNA 表达谱。该方法显示了一组 142 个差异表达的 hsa-miRNAs。接下来，我们通过用 hCMV Towne 株感染人成纤维细胞（人包皮成纤维细胞-1），检查了这些 miRNA 中每一个对病毒生长的影响，鉴定了一组前病毒和抗病毒 hsa-miRNAs。miRNA 靶预测软件表明 hCMV 基因组（例如 hCMV-UL52 和 -UL100[UL = 独特长]）和宿主基因（例如白细胞介素-1 受体，IRF1）内的潜在结合位点。荧光素酶表达质粒构建体和免疫印迹证实了几个预测的 miRNA 靶标。最后，我们在 242 例具有 hCMV 病毒血症的移植受者中测定了选定的前病毒和抗病毒 hsa-miRNAs 的表达。我们在抗病毒治疗前后测量了 hsa-miRNAs，并将 hsa-miRNA 表达水平与 hCMV 复制动力学相关联。六种抗病毒 hsa-miRNAs 中有一种在治疗过程中表达显著增加，与病毒下降同时发生。相比之下，八种前病毒 hsa-miRNAs 中有六种在病毒下降期间表达减少。我们的结果表明，在免疫抑制患者的 CMV 复制过程中，发生了一种复杂的多靶向 hsa-miRNA 反应。该研究为 CMV 复制提供了机制见解和潜在的新型生物标志物。
