Cultured Renal Proximal Tubular Epithelial Cells Resemble a Stressed/Damaged Kidney While Supporting BK Virus Infection.
Cultured Renal Proximal Tubular Epithelial Cells Resemble a Stressed/Damaged Kidney While Supporting BK Virus Infection.
复制标题
培养的肾近端肾小管上皮细胞类似于受压/受损的肾脏，同时支持 BK 病毒感染。
DOI：
10.1128/jvi.00343-23
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发表时间：
2023
影响因子：
5.4
通讯作者：
Pipas,JamesM
中科院分区：
文献类型：
--
作者：
An,Ping;SáenzRobles,MariaTeresa;Cantalupo,PaulG;Naik,AbhijitS;Sealfon,Rachel;Imperiale,MichaelJ;Pipas,JamesM
BK病毒（BKV；人类多瘤病毒1）感染在大多数个体中是无症状的，并且该病毒终生存在而无危害。然而，BKV对移植患者和免疫抑制性疾病患者是一种威胁。在这种情况下，病毒可以在近端小管上皮细胞（PT）中大量复制。培养的肾近端小管上皮细胞（RPTE）允许BKV感染，并已广泛用于表征BKV感染的不同方面。最近，htert永生的RPTE菌株已经出现，初步研究表明它们也支持BKV感染。我们的研究结果表明，BKV感染在原发性和永生化RPTE中导致相似的反应。此外，我们检测了原代和永生化RPTE的整体基因表达模式，并将其与未培养的新鲜从人肾脏分离的PT进行了比较。正如预期的那样，从健康肾脏中分离出来的PT表达了许多与肾功能相关的分化特异性基因。然而，在培养的RPTE中，大多数这些基因的表达不存在或被抑制。相反，培养的RPTE表现出一种基因表达谱，表明肾脏受到应激或损伤。用BKV接种培养的RPTE可抑制许多与肾应激相关的基因。总之，本研究表明，原发性和永生化RPTE之间的基因表达模式和对BKV感染的反应相似。此外，大量转录组测序（RNA-seq）和SCT实验的结果显示，与未培养的新鲜分离的人肾脏PT相比，原发性RPTE中不同的转录组特征代表细胞损伤和应激。培养的原代人细胞为研究病毒感染周期和宿主病毒相互作用提供了强有力的工具。在bkv相关肾病的病例中，病毒复制主要发生在肾脏近端小管上皮。因此，培养的原代和永生化肾近端小管上皮细胞（RPTE）被广泛用于研究BKV感染。在这项工作中，我们使用大量和单细胞转录组学，发现原代和永生化RPTE对BKV感染的反应相似。然而，未感染的原代和永生化RPTE的基因表达谱与直接从人肾中分离的健康近小管上皮明显不同。培养的RPTE处于基因表达状态，表明肾脏受损或应激。这些结果提出了BKV在肾病期间在损伤或应激的肾上皮细胞中优先复制的可能性。
BK virus (BKV; human polyomavirus 1) infections are asymptomatic in most individuals, and the virus persists throughout life without harm. However, BKV is a threat to transplant patients and those with immunosuppressive disorders. Under these circumstances, the virus can replicate robustly in proximal tubule epithelial cells (PT). Cultured renal proximal tubule epithelial cells (RPTE) are permissive to BKV and have been used extensively to characterize different aspects of BKV infection. Recently, lines of hTERT-immortalized RPTE have become available, and preliminary studies indicate they support BKV infection as well. Our results indicate that BKV infection leads to a similar response in primary and immortalized RPTE. In addition, we examined the patterns of global gene expression of primary and immortalized RPTE and compared them with uncultured PT freshly dissociated from human kidney. As expected, PT isolated from the healthy kidney express a number of differentiation-specific genes that are associated with kidney function. However, the expression of most of these genes is absent or repressed in cultured RPTE. Rather, cultured RPTE exhibit a gene expression profile indicative of a stressed or injured kidney. Inoculation of cultured RPTE with BKV results in the suppression of many genes associated with kidney stress. In summary, this study demonstrated similar global gene expression patterns and responses to BKV infection between primary and immortalized RPTE. Moreover, results from bulk transcriptome sequencing (RNA-seq) and SCT experiments revealed distinct transcriptomic signatures representing cell injury and stress in primary RPTE in contrast to the uncultured, freshly dissociated PT from human kidney.IMPORTANCECultured primary human cells provide powerful tools for the study of viral infectious cycles and host virus interactions. In the case of BKV-associated nephropathy, viral replication occurs primarily in the proximal tubule epithelia in the kidney. Consequently, cultured primary and immortalized renal proximal tubule epithelial cells (RPTE) are widely used to study BKV infection. In this work, using bulk and single-cell transcriptomics, we found that primary and immortalized RPTE responded similarly to BKV infection. However, both uninfected primary and immortalized RPTE have gene expression profiles that are markedly different from healthy proximal tubule epithelia isolated directly from human kidney without culture. Cultured RPTE are in a gene expression state indicative of an injured or stressed kidney. These results raise the possibility that BKV replicates preferentially in injured or stressed kidney epithelial cells during nephropathy.