Proteomic analysis of Plasmodium falciparum induced alterations in humans from different endemic regions of India to decipher malaria pathogenesis and identify surrogate markers of severity ☆ Sandipan Ray a,1 , Vipin Kumar a,1 , Amruta Bhave a , Vaidhvi Singh a , Nithya J. Gogtay b , Urmila M. Thatte b , Arunansu Talukdar c , Sanjay K. Kochar d , Swati Patankar a , Sanjeeva Srivastava a, ⁎ a Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India b Department of Clinical Pharmacology, Seth GS Medical College & KEM Hospital, Parel, Mumbai 400012, India c Department of Medicine, Medical College and Hospital Kolkata, 88, College Street, Kolkata 700073, India d Department of Medicine, Malaria Research Center, S.P. Medical College, Bikaner 334003, India abstract article info Article history: Received 26 February 2015 Received in revised form 21 April 2015 Accepted 29 April 2015 Available online xxxx Keywords: India Malaria pathogenesis Plasmodium falciparum Proteomics Severe falciparum malaria Surrogate markers India significantly contributes to the global malaria burden and has the largest population in the world at risk of malaria. This study aims to analyze alterations in the human serum proteome as a consequence of non-severe and severe infections by the malaria parasite Plasmodium falciparum to identify markers related to disease sever- ity and to obtain mechanistic insights about disease pathogenesis and host immune responses. In discovery phase of the study, a comprehensive quantitative proteomic analysis was performed using gel-based (2D-DIGE) and gel-free (iTRAQ) techniques on two independent mass spectrometry platforms (ESI-Q-TOF and Q-Exactive mass spectrometry), and selected targets were validated by ELISA. Proteins showing altered serum abundance in falciparum malaria patients revealed the modulation of different physiological pathways including chemokine and cytokine signaling, IL-12 signaling and production in macrophages, complement cascades, blood coagulation, and protein ubiquitination pathways. Some muscle related and cytoskeletal proteins such as titin and galectin-3- binding protein were found to be up-regulated in severe malaria patients. Hemoglobin levels and platelet counts were also found to be drastically lower in severe malaria patients. Identified proteins including serum amyloid A, C-reactive protein, apolipoprotein E and haptoglobin, which exhibited sequential alterations in their serum abun- dance in different severity levels of malaria, could serve as potential predictive markers for disease severity. To the best of our information, we report here the first comprehensive analysis describing the serum proteomic alterations observed in severe P. falciparum infected patients from different malaria endemic regions of India. This article is part of a Special Issue entitled: Proteomics in India. © 2015 Elsevier B.V. All rights reserved. 1. Introduction Plasmodium falciparum infection represents the major cause of malaria-associated mortality worldwide [1]. This lethal species of ma- laria parasite is responsible for approximately 247 million cases and around one million deaths each year, particularly in the sub-Saharan Africa [2]. India notably contributes to the global malaria burden and has the largest population in the world at risk of malaria [3]. Moreover, due to the extremely variable malaria epidemiology in India, it is consid- ered as an important country for malaria research [4]. Importantly, in recent years there is an increased incidence of P. falciparum compared to Plasmodium vivax malaria in different endemic regions of India [4]. Severe falciparum malaria often leads to fatal and complicated clinical manifestations including hepatic dysfunction, renal dysfunction, severe anemia, hypoglycaemia, acute respiratory distress syndrome (ARDS), cerebral manifestation, and multiple organ involvement [5]. Proteomic techniques pose tremendous potential to provide a wealth of new information to accelerate malaria research [6,7]. In- depth analysis of the differential abundances of serum/plasma proteins during the febrile stage of the infection may help in identification of sur- rogate markers of infection and disease severity and can provide valu- able information regarding disease pathogenesis and host immune responses [8,9]. A few previous studies have investigated the alterations in plasma proteome profiles in cerebral falciparum malaria in children from different endemic and holoendemic regions of Africa [10–12]. However, there is a dearth of similar proteomic analysis of severe Journal of Proteomics xxx (2015) xxx–xxx ☆ This article is part of a Special Issue entitled: Proteomics in India. ⁎ Corresponding author. E-mail address: sanjeeva@iitb.ac.in (S. Srivastava). 1 Both authors contributed equally to the preparation of this manuscript. JPROT-02155; No of Pages 11 http://dx.doi.org/10.1016/j.jprot.2015.04.032 1874-3919/© 2015 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Journal of Proteomics journal homepage: www.elsevier.com/locate/jprot Please cite this article as: S. Ray, et al., Proteomic analysis of Plasmodium falciparum induced alterations in humans from different endemic regions of India..., J Prot (2015), http://dx.doi.org/10.1016/j.jprot.2015.04.032