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Proteomic analysis of inhibitory protein profiles in the urine of children with nephrolithiasis: implication for disease prevention

  • Urology - Original Paper
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Abstract

Purpose

In this study we aimed to screen for the presence of biomarkers that are downregulated in children with nephrolithiasis (RS) compared to healthy controls (HC) using a proteomic approach. We hypothesized that RS and HC would display unique inhibitory protein profiles that could be used for comparative pathway analysis.

Methods

This is a prospective, controlled, pilot study of pooled urine from RS (N = 30, 24 females, mean age 12.95 ± 4.03 years) versus age- and gender-matched HC, using liquid chromatography-mass spectrometry. The criteria for protein selection were: (1) patient/control abundance ratio of < 0.5; and (2) ≤ 0.05 p-value for the Fisher’s Exact Test. Results were confirmed by ELISA testing in individual samples.

Results

67 proteins were downregulated in RS group, and 17 of those were significantly different compared to controls. Of those seventeen proteins, five (two actins, annexin A5, keratin 6B, and serpin B4) were completely absent in the urine of stone patients but were found in controls. The remaining twelve proteins were significantly less abundant in the patient’s urine compared to healthy controls. Protein–protein interaction modeling of significant proteins identified syndecan-1 as the key node, a protein associated with adhesion pathways. ELISA analysis by subgroups showed statistically significant difference in the urinary excretion of osteopontin (5.1 ± 3.22 ng/mg creatinine vs 14.1 ± 9.5 ng/mg creatinine, p = 0.046) between stone patients with hypocitraturia and controls. Urinary osteopontin concentration was positively correlated with urinary citrate excretion (r = 0.417, p = 0.03).

Conclusions

Children with RS have a different urinary inhibitory polypeptide profile compared to HC. Decreased urinary excretion of these proteins indicates their potential inhibitory role in renal stone formation, especially of the adhesion phase. Lower concentration of urinary osteopontin in children with nephrolithiasis and hypocitraturia suggests its potential involvement in the pathogenesis of this disease. Further characterization of these proteins in a larger sample is imperative.

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Funding

This study was funded by Children’s Hospital of Michigan Foundation (grant number R2-2014-31).

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Authors and Affiliations

  1. Department of Pediatric Urology, Children’s Hospital of Michigan, Michigan State University, 3901 Beaubien Blvd, Detroit, MI, 48201, USA

    Larisa Kovacevic, Natalija Kovacevic & Yegappan Lakshmanan

  2. Vattikuti Urology Institute, Henri Ford Hospital, Detroit, MI, USA

    Natalija Kovacevic

Authors
  1. Larisa Kovacevic
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  2. Natalija Kovacevic
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  3. Yegappan Lakshmanan
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Corresponding author

Correspondence to Larisa Kovacevic.

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Conflict of interest

Author Larisa Kovacevic declares that she has no conflict of interest. Author Natalija Kovacevic declares that she has no conflict of interest. Author Yegappan Lakshmanan declares that he has no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.

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Informed consent was obtained from all individual participants included in the study.

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Kovacevic, L., Kovacevic, N. & Lakshmanan, Y. Proteomic analysis of inhibitory protein profiles in the urine of children with nephrolithiasis: implication for disease prevention. Int Urol Nephrol 54, 2783–2788 (2022). https://doi.org/10.1007/s11255-022-03310-5

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  • DOI: https://doi.org/10.1007/s11255-022-03310-5

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