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Identification of Host‐Response in Cerebral Malaria Patients Using Quantitative Proteomic Analysis
Author(s) -
Kumar Manish,
Varun Chakrakodi N.,
Dey Gourav,
Ravikumar Raju,
Mahadevan Anita,
Shankar Susarla Krishna,
Prasad T. S. Keshava
Publication year - 2018
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201600187
Subject(s) - proteome , orbitrap , quantitative proteomics , proteomics , biology , plasmodium falciparum , trypsin , chemistry , biochemistry , mass spectrometry , immunology , chromatography , malaria , gene , enzyme
Purpose The objective of this study was to study the altered proteome in the frontal lobe of patients with CM. Unbiased analysis of differentially abundant proteins could lead to identification of host responses against Plasmodium falciparum infection, which will aid in better understanding of the molecular mechanism of pathophysiology in CM. Experimental design TMT‐based quantitative proteomic analysis using high‐resolution mass spectrometry is employed. In brief, proteins are isolated from frontal lobe samples, which are collected at autopsy from three cases of CM and three control subjects. Equal amounts of protein from each case are digested using trypsin and labeled with different TMT reagents. The pooled sample is fractionated using strong cation exchange chromatography and analyzed on Orbitrap Fusion in triplicates. For accurate quantitation of peptides, the samples are analyzed in MS3 mode. The data is searched against a combined database of human and P. falciparum proteins using Sequest and Mascot search engines. Results A total of 4174 proteins are identified, of which, 107 are found to be differentially abundant in the test samples with significant p ‐value (<0.05). Proteins associated with biological processes such as innate immune response, complement system, coagulation, and platelet activation are found to be elevated in CM cases. In contrast, proteins associated with myelination, oxidative phosphorylation, regulation of reactive oxygen species, and sodium and calcium ions transport are found to be depleted in response to CM. In addition, three P. falciparum proteins exclusively in CM brain samples are also identified. Conclusions and clinical relevance The study signifies neuronal assault due to axonal injury, altered sodium and calcium ion channels, deregulated inflammation and demyelination as a part of host response to CM. Enhanced oxidative stress, repressed oxidative phosphorylation, and demyelination of axons may contribute to the severity of the disease. Further validation of these results on a large cohort can provide leads in the development of neuroprotective therapies for CM.