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Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines

Renal proximal tubule epithelial cells (RPTECs) are a primary site for kidney injury. We created two RPTEC lines from CD-1 mice immortalized with hTERT (human telomerase reverse transcriptase) or SV40 LgT antigen (Simian Virus 40 Large T antigen). Our hypothesis was that low-level, repeated exposure...

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Autores principales: Merrick, B. Alex, Martin, Negin P., Brooks, Ashley M., Foley, Julie F., Dunlap, Paul E., Ramaiahgari, Sreenivasa, Fannin, Rick D., Gerrish, Kevin E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531624/
https://www.ncbi.nlm.nih.gov/pubmed/37762531
http://dx.doi.org/10.3390/ijms241814228
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author Merrick, B. Alex
Martin, Negin P.
Brooks, Ashley M.
Foley, Julie F.
Dunlap, Paul E.
Ramaiahgari, Sreenivasa
Fannin, Rick D.
Gerrish, Kevin E.
author_facet Merrick, B. Alex
Martin, Negin P.
Brooks, Ashley M.
Foley, Julie F.
Dunlap, Paul E.
Ramaiahgari, Sreenivasa
Fannin, Rick D.
Gerrish, Kevin E.
author_sort Merrick, B. Alex
collection PubMed
description Renal proximal tubule epithelial cells (RPTECs) are a primary site for kidney injury. We created two RPTEC lines from CD-1 mice immortalized with hTERT (human telomerase reverse transcriptase) or SV40 LgT antigen (Simian Virus 40 Large T antigen). Our hypothesis was that low-level, repeated exposure to subcytotoxic levels of 0.25–2.5 μM cisplatin (CisPt) or 12.5–100 μM aflatoxin B1 (AFB1) would activate distinctive genes and pathways in these two differently immortalized cell lines. RNA-seq showed only LgT cells responded to AFB1 with 1139 differentially expressed genes (DEGs) at 72 h. The data suggested that AFB1 had direct nephrotoxic properties on the LgT cells. However, both the cell lines responded to 2.5 μM CisPt from 3 to 96 h expressing 2000–5000 total DEGs. For CisPt, the findings indicated a coordinated transcriptional program of injury signals and repair from the expression of immune receptors with cytokine and chemokine secretion for leukocyte recruitment; robust expression of synaptic and substrate adhesion molecules (SAMs) facilitating the expression of neural and hormonal receptors, ion channels/transporters, and trophic factors; and the expression of nephrogenesis transcription factors. Pathway analysis supported the concept of a renal repair transcriptome. In summary, these cell lines provide in vitro models for the improved understanding of repeated renal injury and repair mechanisms. High-throughput screening against toxicant libraries should provide a wider perspective of their capabilities in nephrotoxicity.
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spelling pubmed-105316242023-09-28 Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines Merrick, B. Alex Martin, Negin P. Brooks, Ashley M. Foley, Julie F. Dunlap, Paul E. Ramaiahgari, Sreenivasa Fannin, Rick D. Gerrish, Kevin E. Int J Mol Sci Article Renal proximal tubule epithelial cells (RPTECs) are a primary site for kidney injury. We created two RPTEC lines from CD-1 mice immortalized with hTERT (human telomerase reverse transcriptase) or SV40 LgT antigen (Simian Virus 40 Large T antigen). Our hypothesis was that low-level, repeated exposure to subcytotoxic levels of 0.25–2.5 μM cisplatin (CisPt) or 12.5–100 μM aflatoxin B1 (AFB1) would activate distinctive genes and pathways in these two differently immortalized cell lines. RNA-seq showed only LgT cells responded to AFB1 with 1139 differentially expressed genes (DEGs) at 72 h. The data suggested that AFB1 had direct nephrotoxic properties on the LgT cells. However, both the cell lines responded to 2.5 μM CisPt from 3 to 96 h expressing 2000–5000 total DEGs. For CisPt, the findings indicated a coordinated transcriptional program of injury signals and repair from the expression of immune receptors with cytokine and chemokine secretion for leukocyte recruitment; robust expression of synaptic and substrate adhesion molecules (SAMs) facilitating the expression of neural and hormonal receptors, ion channels/transporters, and trophic factors; and the expression of nephrogenesis transcription factors. Pathway analysis supported the concept of a renal repair transcriptome. In summary, these cell lines provide in vitro models for the improved understanding of repeated renal injury and repair mechanisms. High-throughput screening against toxicant libraries should provide a wider perspective of their capabilities in nephrotoxicity. MDPI 2023-09-18 /pmc/articles/PMC10531624/ /pubmed/37762531 http://dx.doi.org/10.3390/ijms241814228 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Merrick, B. Alex
Martin, Negin P.
Brooks, Ashley M.
Foley, Julie F.
Dunlap, Paul E.
Ramaiahgari, Sreenivasa
Fannin, Rick D.
Gerrish, Kevin E.
Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines
title Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines
title_full Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines
title_fullStr Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines
title_full_unstemmed Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines
title_short Insights into Repeated Renal Injury Using RNA-Seq with Two New RPTEC Cell Lines
title_sort insights into repeated renal injury using rna-seq with two new rptec cell lines
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531624/
https://www.ncbi.nlm.nih.gov/pubmed/37762531
http://dx.doi.org/10.3390/ijms241814228
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