Proteomic analysis of human plasma in chronic rheumatic mitral stenosis reveals proteins involved in the complement and coagulation cascade.
BACKGROUND
Rheumatic fever in childhood is the most common cause of Mitral Stenosis in developing countries. The disease is characterized by damaged and deformed mitral valves predisposing them to scarring and narrowing (stenosis) that results in left atrial hypertrophy followed by heart failure. Presently, echocardiography is the main imaging technique used to diagnose Mitral Stenosis. Despite the high prevalence and increased morbidity, no biochemical indicators are available for prediction, diagnosis and management of the disease. Adopting a proteomic approach to study Rheumatic Mitral Stenosis may therefore throw some light in this direction. In our study, we undertook plasma proteomics of human subjects suffering from Rheumatic Mitral Stenosis (n = 6) and Control subjects (n = 6). Six plasma samples, three each from the control and patient groups were pooled and subjected to low abundance protein enrichment. Pooled plasma samples (crude and equalized) were then subjected to in-solution trypsin digestion separately. Digests were analyzed using nano LC-MS(E). Data was acquired with the Protein Lynx Global Server v2.5.2 software and searches made against reviewed Homo sapiens database (UniProtKB) for protein identification. Label-free protein quantification was performed in crude plasma only.


RESULTS
A total of 130 proteins spanning 9-192 kDa were identified. Of these 83 proteins were common to both groups and 34 were differentially regulated. Functional annotation of overlapping and differential proteins revealed that more than 50% proteins are involved in inflammation and immune response. This was corroborated by findings from pathway analysis and histopathological studies on excised tissue sections of stenotic mitral valves. Verification of selected protein candidates by immunotechniques in crude plasma corroborated our findings from label-free protein quantification.


CONCLUSIONS
We propose that this protein profile of blood plasma, or any of the individual proteins, could serve as a focal point for future mechanistic studies on Mitral Stenosis. In addition, some of the proteins associated with this disorder may be candidate biomarkers for disease diagnosis and prognosis. Our findings might help to enrich existing knowledge on the molecular mechanisms involved in Mitral Stenosis and improve the current diagnostic tools in the long run.
Cell Biology and Physiology Division, CSIR-Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata, 700032 India.
Somaditya
S
Mukherjee
Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune, 411008 India.
Mashanipalya G
MG
Jagadeeshaprasad
Cell Biology and Physiology Division, CSIR-Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata, 700032 India.
Tanima
T
Banerjee
General Medicine Department, Medical College, Kolkata, India.
Sudip K
SK
Ghosh
Department of Cardiology, Geisinger Community Medical Center & Wright Center for graduate medical education, Scranton, PA 18510 USA.
Monodeep
M
Biswas
Department of Cardio-thoracic and Vascular Surgery, Institute of Post Graduate Medical Education and Research, SSKM Hospital, Kolkata, 700020 India.
Santanu
S
Dutta
Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune, 411008 India.
Mahesh J
MJ
Kulkarni
Rabindranath Tagore International Institute of Cardiac Sciences, Kolkata, 700099 India.
Sanjib
S
Pattari
Cell Biology and Physiology Division, CSIR-Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata, 700032 India.
Arun
A
Bandyopadhyay
Immunotechniques; Inflammation; Mitral Stenosis; Plasma proteomics; Rheumatic fever