Comparative Proteomic Analysis of Rapamycin Versus Cyclosporine Combination Treatment in Mouse Podocytes

B. S. Kim, Y. Cho, H. Lee, D. J. Joo, K. H. Huh, M. S. Kim, Y. S. Kim

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background The mechanism of podocyte injury observed with the use of rapamycin (RPM) remains unclear. The conversion from calcineurin inhibitors (CNIs) to RPM in kidney transplant recipients has been associated with a higher incidence of proteinuria and renal injury. In this study, we performed proteomic analyses to investigate the alteration of protein expression in mouse podocytes treated with RPM in comparison with CNI/RPM combination. Methods Immortalized mouse podocytes were treated with 20 nmol/L RPM or 20 nmol/L RPM + 1 μg/mL cyclosporine. Podocyte proteins were separated by 2-dimensional polyacrylamide gel electrophoresis (2DE) and identified by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and peptide fingerprinting. Selected proteins were analyzed by means of Western blot assay. Results We identified 36 differently expressed proteins after isolated RPM or CNI/RPM combination treatment in cultured mouse podocytes. There are 3 distinct patterns of protein expression: (1) potentiated down- or upregulation of proteins by CNI/RPM treatment compared with isolated RPM treatment (n = 4); (2) partial offset of down-regulation by CNI/RPM in comparison with RPM treatment (n = 25); (3) no difference in down-regulation between RPM and CNI/RPM treatment (n = 5). We found a significant interplay between RPM and CNI on the expression of the selected proteins in mouse podocytes. This might explain the higher incidence of proteinuria by CNI/RPM combination in clinical settings. Conclusions Further study is required to elucidate the target protein associated with RPM-induced podocyte injury.

Original languageEnglish
Pages (from-to)1297-1301
Number of pages5
JournalTransplantation Proceedings
Volume48
Issue number4
DOIs
Publication statusPublished - 2016 May 1

Fingerprint

Podocytes
Sirolimus
Proteomics
Cyclosporine
Therapeutics
Proteins
Down-Regulation
Proteinuria
Wounds and Injuries
Kidney
Peptide Mapping
Calcineurin Inhibitors
Incidence

All Science Journal Classification (ASJC) codes

  • Surgery
  • Transplantation

Cite this

Kim, B. S. ; Cho, Y. ; Lee, H. ; Joo, D. J. ; Huh, K. H. ; Kim, M. S. ; Kim, Y. S. / Comparative Proteomic Analysis of Rapamycin Versus Cyclosporine Combination Treatment in Mouse Podocytes. In: Transplantation Proceedings. 2016 ; Vol. 48, No. 4. pp. 1297-1301.
@article{e7ccc12c455945878e3627abd4096000,
title = "Comparative Proteomic Analysis of Rapamycin Versus Cyclosporine Combination Treatment in Mouse Podocytes",
abstract = "Background The mechanism of podocyte injury observed with the use of rapamycin (RPM) remains unclear. The conversion from calcineurin inhibitors (CNIs) to RPM in kidney transplant recipients has been associated with a higher incidence of proteinuria and renal injury. In this study, we performed proteomic analyses to investigate the alteration of protein expression in mouse podocytes treated with RPM in comparison with CNI/RPM combination. Methods Immortalized mouse podocytes were treated with 20 nmol/L RPM or 20 nmol/L RPM + 1 μg/mL cyclosporine. Podocyte proteins were separated by 2-dimensional polyacrylamide gel electrophoresis (2DE) and identified by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and peptide fingerprinting. Selected proteins were analyzed by means of Western blot assay. Results We identified 36 differently expressed proteins after isolated RPM or CNI/RPM combination treatment in cultured mouse podocytes. There are 3 distinct patterns of protein expression: (1) potentiated down- or upregulation of proteins by CNI/RPM treatment compared with isolated RPM treatment (n = 4); (2) partial offset of down-regulation by CNI/RPM in comparison with RPM treatment (n = 25); (3) no difference in down-regulation between RPM and CNI/RPM treatment (n = 5). We found a significant interplay between RPM and CNI on the expression of the selected proteins in mouse podocytes. This might explain the higher incidence of proteinuria by CNI/RPM combination in clinical settings. Conclusions Further study is required to elucidate the target protein associated with RPM-induced podocyte injury.",
author = "Kim, {B. S.} and Y. Cho and H. Lee and Joo, {D. J.} and Huh, {K. H.} and Kim, {M. S.} and Kim, {Y. S.}",
year = "2016",
month = "5",
day = "1",
doi = "10.1016/j.transproceed.2016.01.022",
language = "English",
volume = "48",
pages = "1297--1301",
journal = "Transplantation Proceedings",
issn = "0041-1345",
publisher = "Elsevier USA",
number = "4",

}

Comparative Proteomic Analysis of Rapamycin Versus Cyclosporine Combination Treatment in Mouse Podocytes. / Kim, B. S.; Cho, Y.; Lee, H.; Joo, D. J.; Huh, K. H.; Kim, M. S.; Kim, Y. S.

In: Transplantation Proceedings, Vol. 48, No. 4, 01.05.2016, p. 1297-1301.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Comparative Proteomic Analysis of Rapamycin Versus Cyclosporine Combination Treatment in Mouse Podocytes

AU - Kim, B. S.

AU - Cho, Y.

AU - Lee, H.

AU - Joo, D. J.

AU - Huh, K. H.

AU - Kim, M. S.

AU - Kim, Y. S.

PY - 2016/5/1

Y1 - 2016/5/1

N2 - Background The mechanism of podocyte injury observed with the use of rapamycin (RPM) remains unclear. The conversion from calcineurin inhibitors (CNIs) to RPM in kidney transplant recipients has been associated with a higher incidence of proteinuria and renal injury. In this study, we performed proteomic analyses to investigate the alteration of protein expression in mouse podocytes treated with RPM in comparison with CNI/RPM combination. Methods Immortalized mouse podocytes were treated with 20 nmol/L RPM or 20 nmol/L RPM + 1 μg/mL cyclosporine. Podocyte proteins were separated by 2-dimensional polyacrylamide gel electrophoresis (2DE) and identified by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and peptide fingerprinting. Selected proteins were analyzed by means of Western blot assay. Results We identified 36 differently expressed proteins after isolated RPM or CNI/RPM combination treatment in cultured mouse podocytes. There are 3 distinct patterns of protein expression: (1) potentiated down- or upregulation of proteins by CNI/RPM treatment compared with isolated RPM treatment (n = 4); (2) partial offset of down-regulation by CNI/RPM in comparison with RPM treatment (n = 25); (3) no difference in down-regulation between RPM and CNI/RPM treatment (n = 5). We found a significant interplay between RPM and CNI on the expression of the selected proteins in mouse podocytes. This might explain the higher incidence of proteinuria by CNI/RPM combination in clinical settings. Conclusions Further study is required to elucidate the target protein associated with RPM-induced podocyte injury.

AB - Background The mechanism of podocyte injury observed with the use of rapamycin (RPM) remains unclear. The conversion from calcineurin inhibitors (CNIs) to RPM in kidney transplant recipients has been associated with a higher incidence of proteinuria and renal injury. In this study, we performed proteomic analyses to investigate the alteration of protein expression in mouse podocytes treated with RPM in comparison with CNI/RPM combination. Methods Immortalized mouse podocytes were treated with 20 nmol/L RPM or 20 nmol/L RPM + 1 μg/mL cyclosporine. Podocyte proteins were separated by 2-dimensional polyacrylamide gel electrophoresis (2DE) and identified by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and peptide fingerprinting. Selected proteins were analyzed by means of Western blot assay. Results We identified 36 differently expressed proteins after isolated RPM or CNI/RPM combination treatment in cultured mouse podocytes. There are 3 distinct patterns of protein expression: (1) potentiated down- or upregulation of proteins by CNI/RPM treatment compared with isolated RPM treatment (n = 4); (2) partial offset of down-regulation by CNI/RPM in comparison with RPM treatment (n = 25); (3) no difference in down-regulation between RPM and CNI/RPM treatment (n = 5). We found a significant interplay between RPM and CNI on the expression of the selected proteins in mouse podocytes. This might explain the higher incidence of proteinuria by CNI/RPM combination in clinical settings. Conclusions Further study is required to elucidate the target protein associated with RPM-induced podocyte injury.

UR - http://www.scopus.com/inward/record.url?scp=84975147701&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84975147701&partnerID=8YFLogxK

U2 - 10.1016/j.transproceed.2016.01.022

DO - 10.1016/j.transproceed.2016.01.022

M3 - Article

C2 - 27320608

AN - SCOPUS:84975147701

VL - 48

SP - 1297

EP - 1301

JO - Transplantation Proceedings

JF - Transplantation Proceedings

SN - 0041-1345

IS - 4

ER -