Urine exosomal ceruloplasmin: a potential early biomarker of underlying kidney disease

  • Krishnamurthy P. GudehithluEmail author
  • Peter Hart
  • Amit Joshi
  • Ignacio Garcia-Gomez
  • David J. Cimbaluk
  • George Dunea
  • Jose A. L. Arruda
  • Ashok K. Singh
Original article



Previously we found that kidney tissue and urinary exosomes from patients of diabetic kidney disease showed high levels of ceruloplasmin (CP). Because CP is an acute-phase protein of kidney origin, it could be an early marker of many other kidney diseases. To investigate this hypothesis, we first measured urine exosomal and kidney expression of CP in non-diabetic chronic kidney disease (CKD) patients (membranous nephropathy, focal segmental glomerulosclerosis, lupus nephritis and IgA nephropathy) followed by a longitudinal study in rat passive Heymann nephritis (PHN), a model of human membranous nephropathy.


Urinary exosomes were isolated from urine of patients (and rats) by differential centrifugation. The exosomal extracts were used for measuring CP using ELISA. Kidney expression of CP was evaluated by immune-staining biopsy tissues. Similar techniques were applied in rat PHN model (produced by injection of anti-gp600 antiserum) to analyze urine exosomal and kidney CP.


Urine exosomal CP levels were 10–20 times higher in CKD patients than in controls; consistent with this we found high immune-reactive CP localized in tubules and collecting ducts of biopsies of CKD patients. In the PHN model urinary exosomal CP level was significantly higher prior to the onset of proteinuria. Early rise of urine exosomal CP, which preceded proteinuria, correlated with high immunoreactive CP found in rat kidneys at this time.


We propose that urine exosomal CP, observed to increase prior to proteinuria, makes it a potential urinary biomarker to diagnose early kidney disease.


Ceruloplasmin Exosomes Chronic kidney disease Immunohistochemistry Proteinuria Heymann nephritis 



The authors thank Dr. Lev Rappoport and Ms. Anu Hakimiyan for help with the histologic processing of tissue. This work was financially supported by Hektoen Institute for Medicine, Chicago IL (1994–present).

Compliance with ethical standards

Conflict of interest

None declared.

Human subjects ethical approval

The study was approved and conducted according to the ethical standards of the Institutional Review Board of John H. Stroger, Jr. Hospital of Cook County, Chicago IL, and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The IRB approval and the informed patients consent forms are approved by the Institutional Review Board (IRB approved #: 11-210).

Animal ethical approval

All experimental procedures performed in studies involving animals were in accordance with the ethical standards of the Institutional Animal Care Committee of John H. Stroger, Jr. Hospital of Cook County, Chicago IL (IUCAC #: HEK-0018-2015).

Supplementary material

10157_2019_1734_MOESM1_ESM.pptx (3.2 mb)
Supplementary material 1 (PPTX 3316 kb)


  1. 1.
    Pisitkun T, Shen R-F, Knepper MA. Identification and proteomic profiling of exosomes in human urine. Proc Natl Acad Sci USA. 2004;101:13368–73.CrossRefGoogle Scholar
  2. 2.
    Moon P-G, You S, Lee J-E, Hwang D, Baek M-C. Urinary exosomes and proteomics. Mass Spectrom Rev. 2011;30:1185–202.CrossRefGoogle Scholar
  3. 3.
    Gudehithlu KP, Garcia-Gomez I, Vernik J, Brecklin C, Kraus M, Cimbaluk DJ, Hart P, Dunea G, Arruda JAL, Singh AK. In diabetic kidney disease urinary exosomes better represent kidney specific protein alterations than whole urine. Am J Nephrol. 2015;42:418–24.CrossRefGoogle Scholar
  4. 4.
    Chun-Yan L, Zi-Yi Z, Tian-Lin Y, Yi-Li W, Bao L, Jiao L, Wei-Jun D. Liquid biopsy biomarkers of renal interstitial fibrosis based on urinary exosomes. Exp Mol Pathol. 2018;105:223–8.CrossRefGoogle Scholar
  5. 5.
    Makker SP, Singh AK. Characterization of the antigen gp600 of Heymann Nephritis. Lab Invest. 1984;30:287–96.Google Scholar
  6. 6.
    Berradi H, Bertho J-M, Dudoignon N, Mazur A, Grandcolas L, Baudelin C, Grison S, Voisin P, Gourmelon P, Dublineau I. Renal anemia induced by chronic ingestion of depleted uranium in rats. Toxicol Sci. 2008;103:397–408.CrossRefGoogle Scholar
  7. 7.
    Huang Q, Dunn RT, Jayadev S, DiSorbo O, Pack FD, Farr SB, Stoll RE, Blanchard KT. Assessment of cisplatin-induced nephrotoxicity by microarray technology. Toxicol Sci. 2001;63:196–207.CrossRefGoogle Scholar
  8. 8.
    Kondo C, Minowa Y, Uehara T, Okuno Y, Nakatsu N, Ono A, Maruyama T, Kato I, Yamate J, Yamada H, Ohno Y, Urushidani T. Identification of genomic biomarkers for concurrent diagnosis of drug-induced renal tubular injury using a large-scale toxicogenomics database. Toxicology. 2009;265:15–26.CrossRefGoogle Scholar
  9. 9.
    Luhe A, Hildebrand H, Bach U, Dingermann T, Ahr H-J. A new approach to studying ochratoxin A (OTA)-induced nephrotoxicity: expression profiling in vivo and in vitro employing cDNA microarrays. Toxicol Sci. 2003;73:315–28.CrossRefGoogle Scholar
  10. 10.
    Moon PG, Lee J-E, You S, Kim T-K, Cho J-H, Kim I-S, Kwon T-H, Kim C-D, Park S-H, Hwang D, Kim Y-L, Baek M-C. Proteomic analysis of urinary exosomes from patients of early IgA nephropathy and thin basement membrane nephropathy. Proteomics. 2011;11:2459–75.CrossRefGoogle Scholar
  11. 11.
    Osaki S, Johnson DA, Frieden E. The possible significance of the ferrous oxidase activity of ceruloplasmin in normal human serum. J Biol Sci. 1966;241:2746–51.Google Scholar
  12. 12.
    Morita H, Ikeda S-I, Yamamoto K, Morita S, Yoshida K, Nomoto S, Kato M, Yanagisawa N. Hereditary ceruloplasmin deficiency with hemosiderosis: a clinicopathological study of a Japanese family. Ann Neurol. 1995;37:646–56.CrossRefGoogle Scholar
  13. 13.
    Hellman NE, Gitlin JD. Ceruloplasmin metabolism and function. Annu Rev Nutr. 2002;22:439–58.CrossRefGoogle Scholar
  14. 14.
    Patel BN, David S. A novel glycosylphosphatidylinositol-anchored form of ceruloplasmin is expressed by mammalian astrocytes. J Biol Chem. 1997;272:20185–90.CrossRefGoogle Scholar
  15. 15.
    Jeong SY, David S. Glycosylphosphatidylinositol-anchored ceruloplasmin is required for iron efflux from cells in the central nervous system. J Biol Chem. 2003;278:27144–8.CrossRefGoogle Scholar

Copyright information

© Japanese Society of Nephrology 2019

Authors and Affiliations

  • Krishnamurthy P. Gudehithlu
    • 1
    Email author
  • Peter Hart
    • 1
    • 2
  • Amit Joshi
    • 1
    • 2
  • Ignacio Garcia-Gomez
    • 3
    • 4
  • David J. Cimbaluk
    • 2
  • George Dunea
    • 1
    • 4
  • Jose A. L. Arruda
    • 1
    • 3
  • Ashok K. Singh
    • 1
    • 3
    • 4
  1. 1.Division of NephrologyJohn H. Stroger, Jr. Hospital of Cook CountyChicagoUSA
  2. 2.Department of Internal MedicineRush University Medical CollegeChicagoUSA
  3. 3.Section of NephrologyUniversity of Illinois at ChicagoChicagoUSA
  4. 4.Hektoen Institute of MedicineChicagoUSA

Personalised recommendations