Elsevier

Journal of Proteomics

Volume 127, Part A, 8 September 2015, Pages 103-113
Journal of Proteomics

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

https://doi.org/10.1016/j.jprot.2015.04.032Get rights and content

Highlights

  • P. falciparum infection represents the main reason of malaria associated mortality.

  • First report on proteomic alterations in severe falciparum malaria in Indian population

  • 2D-DIGE and iTRAQ were performed for comparative quantitative proteomic analysis.

  • SAA, HP, and Apo A1 are potential biomarkers for monitoring malaria severity.

  • Vital physiological pathways are modulated adversely in severe falciparum malaria.

Abstract

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 severity 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 abundance 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.

Introduction

Plasmodium falciparum infection represents the major cause of malaria-associated mortality worldwide [1]. This lethal species of malaria 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 considered 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 surrogate markers of infection and disease severity and can provide valuable 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], [11], [12]. However, there is a dearth of similar proteomic analysis of severe falciparum malaria in Indian populations. In an earlier study we have reported the modulations in human serum proteome and various physiological pathways in uncomplicated non-severe falciparum malaria in an adult population from India [13]. In this study, serum samples from adult severe and non-severe falciparum malaria (SFM and NSFM) patients along with healthy community controls from three different endemic regions of India were investigated using 2D-DIGE and iTRAQ-based quantitative proteomics in combination with ESI-Q-TOF and Q-Exactive mass spectrometry. Different hematological and liver function parameters were measured in malaria patients and controls for a comparative statistical analysis. Bioinformatic analysis involving the identified proteins showing altered abundance in the serum samples of falciparum malaria patients revealed the modulation of different vital physiological pathways. This study revealed potential biomarkers for monitoring disease severity of P. falciparum infection and enhanced our understanding regarding pathogenesis of falciparum malaria.

Section snippets

Subject recruitment, blood collection, and serum separation

This proteomics study was conducted involving non-severe and severe falciparum malaria patients and healthy controls from three different endemic regions of India; Mumbai, Kolkata, and Bikaner. This multi-centric analysis was performed with the approval of the institutional ethics committees of Seth GS Medical College and King Edward Memorial Hospital—Mumbai, Medical College Hospital Kolkata — Kolkata, and Malaria Research Center, S.P. Medical College — Bikaner. Written informed consent was

Alterations in clinicopathological parameters in falciparum malaria patients

A total of 250 subjects (HC: n = 146, NSFM: n = 67, SFM: n = 37) were analyzed in this multi-centric study. Platelet counts and Hb levels were found to be significantly lower (p < 0.05) in malaria patients (both NSFM and SFM) as compared to the healthy controls (Table 1 and Fig. S1). Sequential decreases in those two hematological parameters were observed along with the increase in disease severity. Liver function parameters including total bilirubin, serum glutamic oxaloacetic transaminase (SGOT),

Discussion

Specific parasite and host factors, which attribute the complexity of the disease in falciparum malaria have not been clearly deciphered [18]. Hitherto, there is no clear mechanism by which we can comprehend or anticipate the severity of disease at an early stage of the infection [19]. The present study aims to contribute towards unraveling the complexity of severe falciparum malaria pathogenesis. Moreover, the intent of this study was to identify potential predictive and disease monitoring

Competing interests

The authors have declared that no competing interests exist.

Acknowledgments

The active support from Prajakta Gandhe from the Department of Clinical Pharmacology, Seth GS Medical College & KEM Hospital, Mumbai, Sumit Verma from the Medicine Department, Medical College Hospital Kolkata, and Dharmendra Rojh from the Department of Medicine, Malaria Research Center, S.P. Medical College, Bikaner in clinical sample collection process is gratefully acknowledged. We would like to thank Mayuri N. Gandhi and Manali Jadhav from the Centre for Research in Nanotechnology & Science

Reference (48)

  • W. Graninger et al.

    Serum protein concentrations in Plasmodium falciparum malaria

    Acta Trop.

    (1992)
  • P. Balmer et al.

    Experimental erythrocytic malaria infection induces elevated serum amyloid P production in mice

    Immunol. Lett.

    (2000)
  • F.J. Rosales et al.

    Relation of serum retinol to acute phase proteins and malarial morbidity in Papua New Guinea children

    Am. J. Clin. Nutr.

    (2000)
  • R.W. Snow et al.

    The global distribution of clinical episodes of Plasmodium falciparum malaria

    Nature

    (2005)
  • WHO

    WHO World Malaria Report

    (2008)
  • Severe falciparum malaria. World Health Organization, Communicable Diseases Cluster

    Trans. R. Soc. Trop. Med. Hyg.

    (2000)
  • E.A. Winzeler

    Malaria research in the post-genomic era

    Nature

    (2008)
  • S. Ray et al.

    Differential expression of serum/plasma proteins in various infectious diseases: specific or nonspecific signatures

    Proteomics Clin. Appl.

    (2014)
  • S. Ray et al.

    Proteomic technologies for the identification of disease biomarkers in serum: advances and challenges ahead

    Proteomics

    (2011)
  • E.N. Gitau et al.

    Plasma and cerebrospinal proteomes from children with cerebral malaria differ from those of children with other encephalopathies

    J. Infect. Dis.

    (2013)
  • J. Bachmann et al.

    Affinity proteomics reveals elevated muscle proteins in plasma of children with cerebral malaria

    PLoS Pathog.

    (2014)
  • F. Burte et al.

    Severe childhood malaria syndromes defined by plasma proteome profiles

    PLoS One

    (2012)
  • S. Ray et al.

    Proteomic investigation of falciparum and vivax malaria for identification of surrogate protein markers

    PLoS One

    (2012)
  • S. Sharma et al.

    Quantitative proteomic analysis of meningiomas for the identification of surrogate protein markers

    Sci. Rep.

    (2014)
  • Cited by (17)

    • Exploring the role of secretory proteins in the human infectious diseases diagnosis and therapeutics

      2023, Advances in Protein Chemistry and Structural Biology
      Citation Excerpt :

      Up and down-regulation of secretory proteins during mild and severe forms of plasmodial infection along with variations observed in the case of different Plasmodium species have prompted multiple types of research looking into panels of human SPs for potential applications in malarial diagnosis, as disease biomarkers (Ray et al., 2015). Screening of human SPs against healthy control groups has demonstrated alterations in the regulation of 30–40 proteins, namely serum amyloid A with C-reactive protein (CRP), retinol-binding protein-4, apolipoprotein A–I and E, and haptoglobin, alpha 1 antitrypsin, lipopolysaccharide-binding protein, serum paraoxonase, haptoglobin, and carbonic anhydrase (Gillespie et al., 1991; Ray et al., 2012, 2015). Roles of human cytokines and chemokines as immunomodulatory markers in plasmodial infections have been an interesting discussion in the last decades.

    • Calibration-free concentration analysis for an analyte prone to self-association

      2017, Analytical Biochemistry
      Citation Excerpt :

      Calibration-free concentration analysis (CFCA) based on surface plasmon resonance (SPR) extracts the concentration of the active molecules in a mixture of active and inactive (denatured) molecules [1–6]. Recently, the CFCA has been increasingly applied to the studies [7–16]. The CFCA uses the known relation between the diffusion coefficient and the molecular weight of the analyte.

    View all citing articles on Scopus

    This article is part of a Special Issue entitled: Proteomics in India.

    1

    Both authors contributed equally to the preparation of this manuscript.

    View full text