Intragraft Antiviral-Specific Gene Expression as a Distinctive Transcriptional Signature for Studies in Polyomavirus-Associated Nephropathy.
Intragraft Antiviral-Specific Gene Expression as a Distinctive Transcriptional Signature for Studies in Polyomavirus-Associated Nephropathy.
复制标题
DOI：
10.1097/tp.0000000000001214
复制
发表时间：
2016-10
期刊：
影响因子：
6.2
通讯作者：
Sarwal MM
中科院分区：
文献类型：
--
作者：
Sigdel TK;Bestard O;Salomonis N;Hsieh SC;Torras J;Naesens M;Tran TQ;Roedder S;Sarwal MM
多瘤病毒肾病（PVAN）是同种异体移植肾功能障碍和丢失的常见原因。为了确定 PVAN 特异性基因表达和潜在的分子机制，我们分析了有和没有 PVAN 的肾活检。该研究包括来自 168 名独特同种异体肾移植受者的 168 例移植后同种异体肾活检（T 细胞介导的排斥反应 = 26，PVAN = 10，正常功能移植物 (STA) = 73，间质纤维化/肾小管萎缩 (IF/TA) = 59）。我们进行了基因表达测定和生物信息学分析，以确定一组 PVAN 特异性基因。这些基因子集的有效性和相关性通过 QPCR 和 IHC 进行验证。所有活检组织的无监督层次聚类分析揭示了 PVAN 和 TCMR 基因表达之间的高度相似性。统计严格性的提高分别鉴定了 158 个和 252 个独特的 PVAN 和 TCMR 损伤特异性基因转录本。 TCMR特异性基因绝大多数参与免疫反应共刺激和TCR信号传导，而PVAN特异性基因主要与DNA复制过程、RNA聚合酶组装和病原体识别受体相关。使用这些基因的主成分分析进一步证实了 3 种不同临床表型之间的最佳分离。通过 QPCR 对 4 个 PVAN 特异性基因（RPS15、CFD、LTF 和 NOSIP）进行了验证，并通过免疫组织化学对 2 个具有抗病毒功能的 PVAN 特异性蛋白（LTF 和 IFITM1）进行了确认。总之，尽管 PVAN 和 TCMR 同种异体肾移植物在基因扰动方面有很大相似之处，但 PVAN 特异性基因被鉴定为具有众所周知的抗病毒特性，为辨别 PVAN 和 AR 提供了工具，并为合理的药物设计提供了有吸引力的靶标。
Polyomavirus nephropathy (PVAN) is a common cause of kidney allograft dysfunction and loss. To identify PVAN-specific gene expression and underlying molecular mechanisms we analyzed kidney biopsies with and without PVAN. The study included 168 posttransplant renal allograft biopsies (T cell mediated rejection=26, PVAN=10, normal functioning graft (STA) =73, and interstitial fibrosis/tubular atrophy (IF/TA) =59) from 168 unique kidney allograft recipients. We performed gene expression assays and bioinformatics analysis to identify a set of PVAN-specific genes. Validity and relevance of a subset of these genes are validated by QPCR and IHC. Unsupervised hierarchical clustering analysis of all the biopsies revealed high similarity between PVAN and TCMR gene expression. Increased statistical stringency identified 158 and 252 unique PVAN and TCMR injury-specific gene transcripts respectively. While TCMR-specific genes were overwhelmingly involved in immune response costimulation and TCR signaling, PVAN-specific genes were mainly related to DNA replication process, RNA polymerase assembly and pathogen recognition receptors. A principal component analysis using these genes further confirmed the most optimal separation between the 3 different clinical phenotypes. Validation of 4 PVAN-specific genes (RPS15, CFD, LTF, and NOSIP) by QPCR and confirmation by immunohistochemistry of 2 PVAN-specific proteins with anti-viral function (LTF and IFITM1) was done. In conclusion, even though PVAN and TCMR kidney allografts share great similarities on gene perturbation, PVAN-specific genes were identified with well-known anti-viral properties that provide tools for discerning PVAN and AR as well as attractive targets for rational drug design.