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Forschungsdatenbank PMU-SQQUID

LC-MS/MS identification of doublecortin as abundant beta cell-selective protein discharged by damaged beta cells in vitro.
Jiang, L; Brackeva, B; Stangé, G; Verhaeghen, K; Costa, O; Couillard-Després, S; Rotheneichner, P; Aigner, L; Van Schravendijk, C; Pipeleers, D; Ling, Z; Gorus, F; Martens, GA;
J Proteomics. 2013; 80:26-80
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Aigner Ludwig
Couillard-Després Sébastien
Rotheneichner Peter

Abstract

There is a clinical need for plasma tests that can directly detect injury to pancreatic beta cells in type 1 diabetes. Such tests require biomarkers that are abundantly and selectively released into plasma by damaged beta cells. We combined LC-MS/MS proteomics and tissue-comparative transcriptomics of FACS-purified beta cells for bottom-up identification of candidate markers. Less than 10% of 467 proteins detected in beta cells showed endocrine-enriched expression. One surprising candidate was the neuronal migration marker doublecortin: in situ analysis revealed uniform doublecortin expression in the cytoplasm of all beta cells. Western blotting and real-time PCR confirmed its strong beta cell-selectivity outside the brain and its high molar abundance, indicating promising biomarker properties in comparison to GAD65, a more established marker of beta cell injury. DCX potential was validated in vitro: chemically-induced necrosis of rat and human beta cells led to a discharge of intracellular doublecortin into the extracellular space, proportionate to the amount of injured cells, and similar to GAD65. In vivo, recombinant DCX showed favorable pharmacokinetic properties, with a half-life in plasma of around 3h. Combined, our findings provide first proof-of-principle for doublecortin as biomarker for beta cell injury in vitro, advocating its further validation as biomarker in vivo.


Useful keywords (using NLM MeSH Indexing)

Animals

Biological Markers/analysis*

Glutamate Decarboxylase/blood

Humans

Insulin-Secreting Cells/metabolism

Insulin-Secreting Cells/pathology*

Microtubule-Associated Proteins/immunology

Microtubule-Associated Proteins/isolation*

purification

Microtubule-Associated Proteins/metabolism*

Neuropeptides/immunology

Neuropeptides/isolation*

purification

Neuropeptides/metabolism*

Proteomics

Rats

Transcriptome


Find related publications in this database (Keywords)

Insulin-secreting cells
Biological markers
Cell death
Organ specificity
Diagnostic techniques and procedures
Proteomics