Single-cell RNA-sequencing of BK polyomavirus replication in primary human renal proximal tubular epithelial cells identifies specific transcriptome signatures and a novel mitochondrial stress pattern.

Autor:	Weissbach FH; Transplantation & Clinical Virology, Department of Biomedicine, University of Basel, Basel, Switzerland., Follonier OM; Transplantation & Clinical Virology, Department of Biomedicine, University of Basel, Basel, Switzerland.; Biozentrum, University of Basel, Basel, Switzerland.; SIB Swiss Institute of Bioinformatics, Basel, Switzerland., Schmid S; Transplantation & Clinical Virology, Department of Biomedicine, University of Basel, Basel, Switzerland.; Clinical Virology, Laboratory Medicine, University Hospital Basel, Basel, Switzerland., Leuzinger K; Transplantation & Clinical Virology, Department of Biomedicine, University of Basel, Basel, Switzerland.; Clinical Virology, Laboratory Medicine, University Hospital Basel, Basel, Switzerland., Schmid M; Genexa AG, Zürich, Switzerland., Hirsch HH; Transplantation & Clinical Virology, Department of Biomedicine, University of Basel, Basel, Switzerland.
Jazyk:	angličtina
Zdroj:	Journal of virology [J Virol] 2024 Dec 17; Vol. 98 (12), pp. e0138224. Date of Electronic Publication: 2024 Nov 08.
DOI:	10.1128/jvi.01382-24
Abstrakt:	BK polyomavirus (BKPyV) contributes to premature renal failure in 10%-20% of kidney transplant recipients. Current treatment relies on reducing immunosuppression to regain BKPyV-specific immune control. Subsequently, declining allograft function may result from persisting viral cytopathology, BKPyV-specific immune reconstitution, or alloimmunity/rejection, all being poorly distinguishable by current histological or molecular approaches. To reduce the complexity encountered in BKPyV-replicating kidneys, we analyzed differentially expressed genes (DEGs) in primary human renal proximal tubular epithelial cells at 24 and 48 h post-infection (hpi) using single-cell RNA-sequencing (10x-Genomics-3´ kit). At 24 hpi, viral transcript reads predominantly mapped to the early viral gene region ( EVGR ) and shifted to >100-fold higher late viral gene region ( LVGR ) levels at 48 hpi, matching the sequential bi-directional viral protein expression from the circular double-stranded BKPyV-DNA genome. Besides expected coverage "hills" at viral 3´-poly-A sites, unexpected "spike" and "pulse" reads resulted from off-target TSO priming. "Spike" and "pulse" patterns were rare for the mostly unidirectional reads mapping to the circular mitochondrial genome. Bioinformatic curation removed "spikes" and "pulses" and reclassified 10% of DEGs in renal proximal tubular epithelial cells (RPTECs). Up-regulated gene ontologies included S and G2/M phase, double-stranded DNA repair, proximal tubulopathy, and renal tubular dysfunction, whereas allograft rejection, antigen presentation, innate immunity, translation, and autophagy were down-regulated. BKPyV- LVGR expression induced a novel mitochondrial cell stress pattern consisting of discordant up-regulation and down-regulation of mitochondria-encoded and nucleus-encoded mitochondrial genes, respectively. We explored which top-scoring gene sets of late-phase BKPyV-replicating RPTECs can identify BKPyV-associated nephropathy in kidney transplant biopsies. The results should facilitate distinguishing BKPyV-associated pathology from other entities in kidney transplant biopsies.IMPORTANCEBK polyomavirus (BKPyV) infects more than 90% of the general population and then persists in the reno-urinary tract. Subsequently, low-level urinary shedding is seen in 10% of healthy BKPyV-seropositive persons, indicating that BKPyV replication occurs despite the presence of virus-specific cellular and humoral immunity. Notably, transplantation of donor kidneys with low-level BKPyV replication is a risk factor for progression to high-level BKPyV viruria, new-onset BKPyV-DNAemia and biopsy-proven BKPyV nephropathy. Here, we identify a short list of robust up- and down-regulated nucleus-encoded differentially expressed genes potentially allowing to discriminate viral from allograft immune damage. By carefully curating viral and mitochondrial transcriptomes, we identify a novel virus-associated mitochondrial stress pattern of up-regulated mitochondria-encoded and down-regulated nucleus-encoded mitochondrial transcripts which heralds the BKPyV-agnoprotein-mediated immune escape by breakdown of the mitochondrial membrane potential and network and mitophagy. The results may prove useful when assessing the role of BKPyV replication in kidney transplant patients with suspected acute rejection and/or BKPyV nephropathy. Competing Interests: The authors declare no conflict of interest.
Databáze:	MEDLINE
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