TY - JOUR
T1 - Quantitative proteomics for identifying biomarkers for tuberculous meningitis
AU - Kumar, Ghantasala S.Sameer
AU - Venugopal, Abhilash K.
AU - Mahadevan, Anita
AU - Renuse, Santosh
AU - Harsha, H. C.
AU - Sahasrabuddhe, Nandini A.
AU - Pawar, Harsh
AU - Sharma, Rakesh
AU - Kumar, Praveen
AU - Rajagopalan, Sudha
AU - Waddell, Keith
AU - Ramachandra, Yarappa L.
AU - Satishchandra, Parthasarathy
AU - Chaerkady, Raghothama
AU - Prasad, T. S.Keshava
AU - Shankar, K.
AU - Pandey, Akhilesh
N1 - Funding Information:
This study was supported by “DBT Programme Support on Neuroproteomics for Proteomic Investigation of Neurological Disorders.” Nandini Sahasrabuddhe and Harsh Pawar are recipients of Senior Research fellowship award from the Council for Scientific and Industrial Research (CSIR) of the Government of India. Santosh Renuse is a recipient of Senior Research fellowship award from the University Grants Commission (UGC) of the Government of India. Rakesh Sharma is a Research associate supported by DBT. T. S. Keshava Prasad is supported by a research grant on “Establishment of a National Database on Tuberculosis” and “Development of Infrastructure and a Computational Framework for Analysis of Proteomic Data” from DBT. Harsha Gowda is a Wellcome Trust-DBT India Alliance Early Career Fellow. Human brain tissues for the study were obtained from the Human Brain Tissue Repository, a national research facility in the Department of Neuropathology, National Institute of Mental Health and Neuro Sciences, Bangalore, India. Secretarial assistance of Mrs. Manjula Madan is acknowledged.
PY - 2012
Y1 - 2012
N2 - Introduction: Tuberculous meningitis is a frequent extrapulmonary disease caused by Mycobacterium tuberculosis and is associated with high mortality rates and severe neurological sequelae. In an earlier study employing DNA microarrays, we had identified genes that were differentially expressed at the transcript level in human brain tissue from cases of tuberculous meningitis. In the current study, we used a quantitative proteomics approach to discover protein biomarkers for tuberculous meningitis. Methods: To compare brain tissues from confirmed cased of tuberculous meningitis with uninfected brain tissue, we carried out quantitative protein expression profiling using iTRAQ labeling and LC-MS/MS analysis of SCX fractionated peptides on Agilent's accurate mass QTOF mass spectrometer. Results and conclusions: Through this approach, we identified both known and novel differentially regulated molecules. Those described previously included signal-regulatory protein alpha (SIRPA) and protein disulfide isomerase family A, member 6 (PDIA6), which have been shown to be overexpressed at the mRNA level in tuberculous meningitis. The novel overexpressed proteins identified in our study included amphiphysin (AMPH) and neurofascin (NFASC) while ferritin light chain (FTL) was found to be downregulated in TBM. We validated amphiphysin, neurofascin and ferritin light chain using immunohistochemistry which confirmed their differential expression in tuberculous meningitis. Overall, our data provides insights into the host response in tuberculous meningitis at the molecular level in addition to providing candidate diagnostic biomarkers for tuberculous meningitis.
AB - Introduction: Tuberculous meningitis is a frequent extrapulmonary disease caused by Mycobacterium tuberculosis and is associated with high mortality rates and severe neurological sequelae. In an earlier study employing DNA microarrays, we had identified genes that were differentially expressed at the transcript level in human brain tissue from cases of tuberculous meningitis. In the current study, we used a quantitative proteomics approach to discover protein biomarkers for tuberculous meningitis. Methods: To compare brain tissues from confirmed cased of tuberculous meningitis with uninfected brain tissue, we carried out quantitative protein expression profiling using iTRAQ labeling and LC-MS/MS analysis of SCX fractionated peptides on Agilent's accurate mass QTOF mass spectrometer. Results and conclusions: Through this approach, we identified both known and novel differentially regulated molecules. Those described previously included signal-regulatory protein alpha (SIRPA) and protein disulfide isomerase family A, member 6 (PDIA6), which have been shown to be overexpressed at the mRNA level in tuberculous meningitis. The novel overexpressed proteins identified in our study included amphiphysin (AMPH) and neurofascin (NFASC) while ferritin light chain (FTL) was found to be downregulated in TBM. We validated amphiphysin, neurofascin and ferritin light chain using immunohistochemistry which confirmed their differential expression in tuberculous meningitis. Overall, our data provides insights into the host response in tuberculous meningitis at the molecular level in addition to providing candidate diagnostic biomarkers for tuberculous meningitis.
KW - Cerebrospinal fluid
KW - Early diagnosis
KW - Histopathology
KW - Relative quantitation
KW - Tuberculosis
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U2 - 10.1186/1559-0275-9-12
DO - 10.1186/1559-0275-9-12
M3 - Article
AN - SCOPUS:84879606270
SN - 1542-6416
VL - 9
JO - Clinical Proteomics
JF - Clinical Proteomics
IS - 1
M1 - 12
ER -