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Insights into novel cellular injury mechanisms by gene expression profiling in nephropathic cystinosis

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Nephropathic cystinosis is a rare, inherited metabolic disease caused by functional defects of cystinosin associated with mutations in the CTNS gene. The mechanisms underlying the phenotypic alterations associated with this disease are not well known. In this study, gene expression profiles in peripheral blood of nephropathic cystinosis patients (N = 7) were compared with controls (N = 7) using microarray technology. In unsupervised hierarchical clustering analysis, cystinosis samples co-clustered, and 1,604 genes were significantly differentially expressed between both groups. Gene ontology analysis revealed that differentially expressed genes in cystinosis were enriched in cell organelles such as mitochondria, lysosomes, and endoplasmic reticulum (p ≤ 0.030). The majority of the differentially regulated genes were involved in oxidative phosphorylation, apoptosis, mitochondrial dysfunction, endoplasmic reticulum stress, antigen processing and presentation, B-cell-receptor signaling, and oxidative stress (p ≤ 0.003). Validation of selected genes involved in apoptosis and oxidative phosphorylation was performed by quantitative real-time polymerase chain reaction (PCR). Electron microscopy and confocal imaging of cystinotic renal proximal tubular epithelial cells further confirmed anomalies in the cellular organelles and pathways identified by microarray analysis. Further analysis of these genes and pathways may offer critical insights into the clinical spectrum of cystinosis patients and ultimately lead to novel links for targeted therapy.

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Abbreviations

FDR:

False discovery rate

CTNS :

Cystinosin

SAM:

Significance analysis of microarray

PAM:

Prediction analysis of microarray

LSD:

Lysosomal storage disorders

RPTE:

Renal proximal tubular epithelial

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Acknowledgements

This work was supported by grants from the Cystinosis Foundation, Ireland, and the Intramural Research Program of the National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA. We are indebted to Dr. L.C. Racusen (Department of Pathology, Johns Hopkins University) for generating an incredible resource of cystinotic renal proximal tubular epithelial cells; to Dr. Benedict Yen (Department of Pathology, University of California, San Francisco) for support with electron microscopy experiments, and to Dr. William A. Gahl (Section on Human Biochemical Genetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health), for providing us with cystinotic RPTE cells and for helpful advice and review of the manuscript.

Funding Sources

The work was supported by grants from the Cystinosis Foundation Ireland and the Intramural Research Program of the National Human Genome Research Institute, National Institute of Health.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, USA

    Poonam Sansanwal, Li Li, Szu-Chuan Hsieh & Minnie M. Sarwal

  2. Department of Pediatrics, G306, 300 Pasteur Drive, Stanford, CA, 94304, USA

    Minnie M. Sarwal

  3. Department of Pediatrics, S378, 300 Pasteur Drive, Stanford, CA, 94304, USA

    Poonam Sansanwal

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  1. Poonam Sansanwal
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  2. Li Li
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  3. Szu-Chuan Hsieh
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  4. Minnie M. Sarwal
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Correspondence to Poonam Sansanwal or Minnie M. Sarwal.

Additional information

Communicated by: Jean-Marie Saudubray

Competing interest:

None declared.

Poonam Sansanwal and Li Li contributed equally to this study.

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Sansanwal, P., Li, L., Hsieh, SC. et al. Insights into novel cellular injury mechanisms by gene expression profiling in nephropathic cystinosis. J Inherit Metab Dis 33, 775–786 (2010). https://doi.org/10.1007/s10545-010-9203-6

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  • DOI: https://doi.org/10.1007/s10545-010-9203-6

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