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The proteome of cblC defect: in vivo elucidation of altered cellular pathways in humans

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Methylmalonic acidemia with homocystinuria, cobalamin deficiency type C (cblC) (MMACHC) is the most common inborn error of cobalamin metabolism. Despite a multidrug treatment, the long-term follow-up of early-onset patients is often unsatisfactory, with progression of neurological and ocular impairment. Here, the in-vivo proteome of control and MMACHC lymphocytes (obtained from patients under standard treatment with OHCbl, betaine, folate and L-carnitine) was quantitatively examined by two dimensional differential in-gel electrophoresis (2D-DIGE) and mass spectrometry. Twenty three proteins were found up-regulated and 38 proteins were down-regulated. Consistent with in vivo studies showing disturbance of glutathione metabolism, a deregulation in proteins involved in cellular detoxification, especially in glutathione metabolism was found. In addition, relevant changes were observed in the expression levels of proteins involved in intracellular trafficking and protein folding, energy metabolism, cytoskeleton organization and assembly. This study demonstrates relevant changes in the proteome profile of circulating lymphocytes isolated from treated cblC patients. Some results confirm previous observations in vivo on fibroblast, thus concluding that some dysregulation is ubiquitous. On the other hand, new findings could be tissue-specific. These observations expand our current understanding of the cblC disease and may ignite new research and therapeutic strategies to treat this disorder.

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Abbreviations

AdoCbl:

adenosylcobalamin

cblC:

cobalamin deficiency type C

MS:

mass spectrometry

meCbl:

methylcobalamin

MMACHC:

Methylmalonicaciduria with homocystinuria type C

OHCbl:

hydroxicobalamin

2D-DIGE:

two dimensional differential in-gel electrophoresis

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Acknowledgments

The financial support of the Mariani Foundation of Milan (Grant n. R-12-92), of the association “la Vita è unDono”, and of the Italian Ministry of Health (RicercaCorrente) are gratefully acknowledged. This work has been supported by POR Campania FSE 2007-2013 project CRÈME to MR.

Compliance with ethics guidelines

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Conflict of interest

None.

Informed consent

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000. Informed consent was obtained from all patients for being included in the study.

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    Authors and Affiliations

    1. Fondazione SDN-IRCCS, Naples, Italy

      Marianna Caterino & Esther Imperlini

    2. Division of Metabolic Diseases, Department of Pediatric Medicine, Bambino Gesù Children’s Hospital, IRCCS, Rome, Italy

      Anna Pastore, Gianna Di Giovamberardino, Sara Boenzi, Ferdinando Ceravolo, Diego Martinelli & Carlo Dionisi-Vici

    3. Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli, “Federico II”, Naples, Italy

      Maria Grazia Strozziero & Margherita Ruoppolo

    4. CEINGE Biotecnologie Avanzate, Naples, Italy

      Emanuela Scolamiero, Laura Ingenito & Margherita Ruoppolo

    5. Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Via S. Pansini, 5, 80131, Naples, Italy

      Margherita Ruoppolo

    Authors
    1. Marianna Caterino
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    2. Anna Pastore
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    3. Maria Grazia Strozziero
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    Corresponding author

    Correspondence to Margherita Ruoppolo.

    Additional information

    Communicated by: Brian Fowler

    Marianna Caterino and Anna Pastore equally contributed

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    Table 1S

    Supplemental material. Details of tandem mass identification (XLS 33 kb)

    Table 2S

    Supplemental material. Details of ‘peptide mass fingerprint’ identification (XLS 73 kb)

    Figure 1S

    Supplemental material. 2D DIGEanalitical gels (GIF 382 kb)

    High resolution image (TIFF 2572 kb)

    Figure 2S

    Supplemental material. IPA cluster of differentially expressed proteins in cblcpatients lymphocytes (GIF 160 kb)

    High resolution image (TIFF 526 kb)

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    Caterino, M., Pastore, A., Strozziero, M.G. et al. The proteome of cblC defect: in vivo elucidation of altered cellular pathways in humans. J Inherit Metab Dis 38, 969–979 (2015). https://doi.org/10.1007/s10545-014-9806-4

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    • DOI: https://doi.org/10.1007/s10545-014-9806-4

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