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2D-DIGE as a strategy to identify serum protein biomarkers to monitor pharmacological efficacy in dopamine-dictated states of Parkinson’s disease and schizophrenia

Authors Gupta AK, Kumar GK, Rani K, Pokhriyal R, Khan MI, Kumar DR, Goyal V, Tripathi M, Gupta R, Chadda RK, Vanamail P, Mohanty AK, Hariprasad G

Received 17 December 2018

Accepted for publication 19 February 2019

Published 24 April 2019 Volume 2019:15 Pages 1031—1044


Checked for plagiarism Yes

Review by Single anonymous peer review

Peer reviewer comments 2

Editor who approved publication: Dr Roger Pinder

Video abstract presented by Gururao Hariprasad.

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Ashish Kumar Gupta,1 Gaurav Khunger Kumar,1 Komal Rani,1 Ruchika Pokhriyal,1 Mohd Imran Khan,1 Domada Ratna Kumar,1 Vinay Goyal,2 Manjari Tripathi,2 Rishab Gupta,3 Rakesh Kumar Chadda,3 Perumal Vanamail,4 Ashok Kumar Mohanty,5 Gururao Hariprasad1

1Department of Biophysics, All India Institute of Medical Sciences, New Delhi 110029, India; 2Department of Neurology, All India Institute of Medical Sciences, New Delhi 110029, India; 3Department of Psychiatry, All India Institute of Medical Sciences, New Delhi 110029, India; 4Department of Biostatistics, All India Institute of Medical Sciences, New Delhi 110029, India; 5Proteomics Facility, National Diary Research Institute, Karnal, Haryana 132001, India

Objectives: Parkinson’s disease and schizophrenia are clinical scenarios that occur due to dopaminergic deficit and hyperactivity in the midbrain, respectively. Current pharmacological interventions for these two diseases therefore aim to restore normal dopamine levels in the midbrain. But during therapy, there is a overshooting of dopamine concentrations that result in hallucinations in Parkinson’s disease patients and extra-pyramidal symptoms in schizophrenic patients. This causes a lot of inconvenience to the patents and the clinicians. There are no tests currently available to monitor drug efficacy in these two neuropsychiatric diseases.
Materials and methods: Parkinson’s disease and schizophrenic naïve patients were recruited. Serum proteins isolated from these two clinical phenotypes were labeled with fluorescent cyanine dyes and analyzed by two-dimensional difference in gel electrophoresis proteomic experiment. Differentially expressed spots that had consistent expression pattern across five sets of biological replicate gels were trypsin digested and subjected to mass spectrometric analysis for protein identification. Validation experiments were done for the identified proteins using antibody-based assay on a patient cohort that included naïve, treated, and those who had side effects.
Results: Serum α- and β-globin chains were identified as differentially expressed proteins having threefold higher expressions in Parkinson’s patients as compared to schizophrenia. Interestingly, concentrations of these two proteins had an inverse correlation across clinical phenotypes in the dopaminergic spectrum. RBC contamination as a source for these proteins was ruled out.
Conclusion: There is a clear association of free serum globin with dopaminergic clinical states. This lays a platform for protein biomarker–based monitoring of pharmacological efficacy in Parkinson’s disease and schizophrenia.

Keywords: Parkinson’s disease, schizophrenia, gel-based proteomics, biomarkers, dopamine, pharmacological efficacy, difference gel electrophoresis

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