Skip to main content
Top
Published in: Malaria Journal 1/2018

Open Access 01-12-2018 | Research

Plasmodium falciparum genetic variation of var2csa in the Democratic Republic of the Congo

Authors: Robert Verity, Nicholas J. Hathaway, Andreea Waltmann, Stephanie M. Doctor, Oliver J. Watson, Jaymin C. Patel, Kashamuka Mwandagalirwa, Antoinette K. Tshefu, Jeffrey A. Bailey, Azra C. Ghani, Jonathan J. Juliano, Steven R. Meshnick

Published in: Malaria Journal | Issue 1/2018

Login to get access

Abstract

Background

The Democratic Republic of the Congo (DRC) bears a high burden of malaria, which is exacerbated in pregnant women. The VAR2CSA protein plays a crucial role in pregnancy-associated malaria (PAM), and hence quantifying diversity at the var2csa locus in the DRC is important in understanding the basic epidemiology of PAM, and in developing a robust vaccine against PAM.

Methods

Samples were taken from the 2013–14 Demographic and Health Survey conducted in the DRC, focusing on children under 5 years of age. A short subregion of the var2csa gene was sequenced in 115 spatial clusters, giving country-wide estimates of sequence polymorphism and spatial population structure.

Results

Results indicate that var2csa is highly polymorphic, and that diversity is being maintained through balancing selection, however, there is no clear signal of phylogenetic or geographic structure to this diversity. Linear modelling demonstrates that the number of var2csa variants in a cluster correlates directly with cluster prevalence, but not with other epidemiological factors such as urbanicity.

Conclusions

Results suggest that the DRC fits within the global pattern of high var2csa diversity and little genetic differentiation between regions. A broad multivalent VAR2CSA vaccine candidate could benefit from targeting stable regions and common variants to address the substantial genetic diversity.
Appendix
Available only for authorised users
Literature
1.
go back to reference Steketee RW, Nahlen BL, Parise ME, Menendez C. The burden of malaria in pregnancy in malaria-endemic areas. Am J Trop Med Hyg. 2001;64:28–35.CrossRefPubMed Steketee RW, Nahlen BL, Parise ME, Menendez C. The burden of malaria in pregnancy in malaria-endemic areas. Am J Trop Med Hyg. 2001;64:28–35.CrossRefPubMed
2.
go back to reference Desai M, ter Kuile FO, Nosten F, McGready R, Asamoa K, Brabin B, et al. Epidemiology and burden of malaria in pregnancy. Lancet Infect Dis. 2007;7:93–104.CrossRefPubMed Desai M, ter Kuile FO, Nosten F, McGready R, Asamoa K, Brabin B, et al. Epidemiology and burden of malaria in pregnancy. Lancet Infect Dis. 2007;7:93–104.CrossRefPubMed
3.
go back to reference Van Geertruyden JP, Thomas F, Erhart A, D’Alessandro U. The contribution of malaria in pregnancy to perinatal mortality. Am J Trop Med Hyg. 2004;71(2_suppl):35–40.PubMed Van Geertruyden JP, Thomas F, Erhart A, D’Alessandro U. The contribution of malaria in pregnancy to perinatal mortality. Am J Trop Med Hyg. 2004;71(2_suppl):35–40.PubMed
4.
go back to reference Salanti A, Dahlbäck M, Turner L, Nielsen MA, Barfod L, Magistrado P, et al. Evidence for the involvement of VAR2CSA in pregnancy-associated malaria. J Exp Med. 2004;200:1197–203.CrossRefPubMedPubMedCentral Salanti A, Dahlbäck M, Turner L, Nielsen MA, Barfod L, Magistrado P, et al. Evidence for the involvement of VAR2CSA in pregnancy-associated malaria. J Exp Med. 2004;200:1197–203.CrossRefPubMedPubMedCentral
5.
go back to reference Pereira MA, Clausen TM, Pehrson C, Mao Y, Resende M, Daugaard M, et al. Placental sequestration of Plasmodium falciparum malaria parasites is mediated by the interaction between VAR2CSA and chondroitin sulfate A on syndecan-1. PLoS Pathog. 2016;12:e1005831.CrossRef Pereira MA, Clausen TM, Pehrson C, Mao Y, Resende M, Daugaard M, et al. Placental sequestration of Plasmodium falciparum malaria parasites is mediated by the interaction between VAR2CSA and chondroitin sulfate A on syndecan-1. PLoS Pathog. 2016;12:e1005831.CrossRef
7.
go back to reference Eisele TP, Larsen DA, Anglewicz PA, Keating J, Yukich J, Bennett A, et al. Malaria prevention in pregnancy, birthweight, and neonatal mortality: a meta-analysis of 32 national cross-sectional datasets in Africa. Lancet Infect Dis. 2012;12:942–9.CrossRefPubMed Eisele TP, Larsen DA, Anglewicz PA, Keating J, Yukich J, Bennett A, et al. Malaria prevention in pregnancy, birthweight, and neonatal mortality: a meta-analysis of 32 national cross-sectional datasets in Africa. Lancet Infect Dis. 2012;12:942–9.CrossRefPubMed
8.
go back to reference Chêne A, Houard S, Nielsen MA, Hundt S, D’Alessio F, Sirima SB, et al. Clinical development of placental malaria vaccines and immunoassays harmonization: a workshop report. Malar J. 2016;15:476.CrossRefPubMedPubMedCentral Chêne A, Houard S, Nielsen MA, Hundt S, D’Alessio F, Sirima SB, et al. Clinical development of placental malaria vaccines and immunoassays harmonization: a workshop report. Malar J. 2016;15:476.CrossRefPubMedPubMedCentral
11.
go back to reference Clausen TM, Christoffersen S, Dahlbäck M, Langkilde AE, Jensen KE, Resende M, et al. Structural and functional insight into how the Plasmodium falciparum VAR2CSA protein mediates binding to chondroitin sulfate A in placental malaria. J Biol Chem. 2012;287:23332–45.CrossRefPubMedPubMedCentral Clausen TM, Christoffersen S, Dahlbäck M, Langkilde AE, Jensen KE, Resende M, et al. Structural and functional insight into how the Plasmodium falciparum VAR2CSA protein mediates binding to chondroitin sulfate A in placental malaria. J Biol Chem. 2012;287:23332–45.CrossRefPubMedPubMedCentral
12.
go back to reference Bordbar B, Tuikue-Ndam N, Bigey P, Doritchamou J, Scherman D, Deloron P. Identification of Id1-DBL2X of VAR2CSA as a key domain inducing highly inhibitory and cross-reactive antibodies. Vaccine. 2012;30:1343–8.CrossRefPubMed Bordbar B, Tuikue-Ndam N, Bigey P, Doritchamou J, Scherman D, Deloron P. Identification of Id1-DBL2X of VAR2CSA as a key domain inducing highly inhibitory and cross-reactive antibodies. Vaccine. 2012;30:1343–8.CrossRefPubMed
13.
go back to reference Trimnell AR, Kraemer SM, Mukherjee S, Phippard DJ, Janes JH, Flamoe E, et al. Global genetic diversity and evolution of var genes associated with placental and severe childhood malaria. Mol Biochem Parasitol. 2006;148:169–80.CrossRefPubMed Trimnell AR, Kraemer SM, Mukherjee S, Phippard DJ, Janes JH, Flamoe E, et al. Global genetic diversity and evolution of var genes associated with placental and severe childhood malaria. Mol Biochem Parasitol. 2006;148:169–80.CrossRefPubMed
14.
go back to reference Bockhorst J, Lu F, Janes JH, Keebler J, Gamain B, Awadalla P, et al. Structural polymorphism and diversifying selection on the pregnancy malaria vaccine candidate VAR2CSA. Mol Biochem Parasitol. 2007;155:103–12.CrossRefPubMed Bockhorst J, Lu F, Janes JH, Keebler J, Gamain B, Awadalla P, et al. Structural polymorphism and diversifying selection on the pregnancy malaria vaccine candidate VAR2CSA. Mol Biochem Parasitol. 2007;155:103–12.CrossRefPubMed
15.
go back to reference Bordbar B, Ndam NT, Renard E, Jafari-Guemouri S, Tavul L, Jennison C, et al. Genetic diversity of VAR2CSA ID1-DBL2Xb in worldwide Plasmodium falciparum populations: impact on vaccine design for placental malaria. Infect Genet Evol. 2014;25:81–92.CrossRefPubMed Bordbar B, Ndam NT, Renard E, Jafari-Guemouri S, Tavul L, Jennison C, et al. Genetic diversity of VAR2CSA ID1-DBL2Xb in worldwide Plasmodium falciparum populations: impact on vaccine design for placental malaria. Infect Genet Evol. 2014;25:81–92.CrossRefPubMed
16.
go back to reference Doritchamou J, Sabbagh A, Jespersen JS, Renard E, Salanti A, Nielsen MA, et al. Identification of a major dimorphic region in the functionally critical N-Terminal ID1 domain of VAR2CSA. PLoS ONE. 2015;10:e0137695.CrossRefPubMedPubMedCentral Doritchamou J, Sabbagh A, Jespersen JS, Renard E, Salanti A, Nielsen MA, et al. Identification of a major dimorphic region in the functionally critical N-Terminal ID1 domain of VAR2CSA. PLoS ONE. 2015;10:e0137695.CrossRefPubMedPubMedCentral
17.
go back to reference Patel JC, Hathaway NJ, Parobek CM, Thwai KL, Madanitsa M, Khairallah C, et al. Increased risk of low birth weight in women with placental malaria associated with P. falciparum VAR2CSA clade. Sci Rep. 2017;7:7768.CrossRefPubMedPubMedCentral Patel JC, Hathaway NJ, Parobek CM, Thwai KL, Madanitsa M, Khairallah C, et al. Increased risk of low birth weight in women with placental malaria associated with P. falciparum VAR2CSA clade. Sci Rep. 2017;7:7768.CrossRefPubMedPubMedCentral
18.
go back to reference Taylor SM, Van Eijk AM, Hand CC, Mwandagalirwa K, Messina JP, Tshefu AK, et al. Quantification of the burden and consequences of pregnancy-associated malaria in the Democratic Republic of the Congo. J Infect Dis. 2011;204:1762–71.CrossRefPubMedPubMedCentral Taylor SM, Van Eijk AM, Hand CC, Mwandagalirwa K, Messina JP, Tshefu AK, et al. Quantification of the burden and consequences of pregnancy-associated malaria in the Democratic Republic of the Congo. J Infect Dis. 2011;204:1762–71.CrossRefPubMedPubMedCentral
20.
go back to reference Taylor SM, Antonia AL, Parobek CM, Juliano JJ, Janko M, Emch M, et al. Plasmodium falciparum sulfadoxine resistance is geographically and genetically clustered within the DR Congo. Sci Rep. 2013;3:1165.CrossRefPubMedPubMedCentral Taylor SM, Antonia AL, Parobek CM, Juliano JJ, Janko M, Emch M, et al. Plasmodium falciparum sulfadoxine resistance is geographically and genetically clustered within the DR Congo. Sci Rep. 2013;3:1165.CrossRefPubMedPubMedCentral
21.
go back to reference Taylor SM, Antonia AL, Harrington WE, Goheen MM, Mwapasa V, Chaluluka E, et al. Independent lineages of highly sulfadoxine-resistant Plasmodium falciparum haplotypes, eastern Africa. Emerg Infect Dis. 2014;20:1140.CrossRefPubMedPubMedCentral Taylor SM, Antonia AL, Harrington WE, Goheen MM, Mwapasa V, Chaluluka E, et al. Independent lineages of highly sulfadoxine-resistant Plasmodium falciparum haplotypes, eastern Africa. Emerg Infect Dis. 2014;20:1140.CrossRefPubMedPubMedCentral
22.
go back to reference Carrel M, Patel J, Taylor SM, Janko M, Mwandagalirwa MK, Tshefu AK, et al. The geography of malaria genetics in the Democratic Republic of Congo: a complex and fragmented landscape. Soc Sci Med. 2015;133:233–41.CrossRefPubMed Carrel M, Patel J, Taylor SM, Janko M, Mwandagalirwa MK, Tshefu AK, et al. The geography of malaria genetics in the Democratic Republic of Congo: a complex and fragmented landscape. Soc Sci Med. 2015;133:233–41.CrossRefPubMed
23.
go back to reference Miller RH, Hathaway NJ, Kharabora O, Mwandagalirwa K, Tshefu A, Meshnick SR, et al. A deep sequencing approach to estimate Plasmodium falciparum complexity of infection (COI) and explore apical membrane antigen 1 diversity. Malar J. 2017;16:490.CrossRefPubMedPubMedCentral Miller RH, Hathaway NJ, Kharabora O, Mwandagalirwa K, Tshefu A, Meshnick SR, et al. A deep sequencing approach to estimate Plasmodium falciparum complexity of infection (COI) and explore apical membrane antigen 1 diversity. Malar J. 2017;16:490.CrossRefPubMedPubMedCentral
24.
go back to reference Doctor SM, Liu Y, Whitesell A, Thwai KL, Taylor SM, Janko M, et al. Malaria surveillance in the Democratic Republic of the Congo: comparison of microscopy, PCR, and rapid diagnostic test. Diagn Microbiol Infect Dis. 2016;85:16–8.CrossRefPubMedPubMedCentral Doctor SM, Liu Y, Whitesell A, Thwai KL, Taylor SM, Janko M, et al. Malaria surveillance in the Democratic Republic of the Congo: comparison of microscopy, PCR, and rapid diagnostic test. Diagn Microbiol Infect Dis. 2016;85:16–8.CrossRefPubMedPubMedCentral
25.
go back to reference Plowe CV, Djimde A, Bouare M, Doumbo O, Wellems TE. Pyrimethamine and proguanil resistance-conferring mutations in Plasmodium falciparum dihydrofolate reductase: polymerase chain reaction methods for surveillance in Africa. Am J Trop Med Hyg. 1995;52:565–8.CrossRefPubMed Plowe CV, Djimde A, Bouare M, Doumbo O, Wellems TE. Pyrimethamine and proguanil resistance-conferring mutations in Plasmodium falciparum dihydrofolate reductase: polymerase chain reaction methods for surveillance in Africa. Am J Trop Med Hyg. 1995;52:565–8.CrossRefPubMed
26.
go back to reference Taylor SM, Juliano JJ, Trottman PA, Griffin JB, Landis SH, Kitsa P, et al. High-throughput pooling and real-time PCR-based strategy for malaria detection. J Clin Microbiol. 2010;48:512–9.CrossRefPubMed Taylor SM, Juliano JJ, Trottman PA, Griffin JB, Landis SH, Kitsa P, et al. High-throughput pooling and real-time PCR-based strategy for malaria detection. J Clin Microbiol. 2010;48:512–9.CrossRefPubMed
27.
go back to reference Beshir KB, Hallett RL, Eziefula AC, Bailey R, Watson J, Wright SG, et al. Measuring the efficacy of anti-malarial drugs in vivo: quantitative PCR measurement of parasite clearance. Malar J. 2010;9:312.CrossRefPubMedPubMedCentral Beshir KB, Hallett RL, Eziefula AC, Bailey R, Watson J, Wright SG, et al. Measuring the efficacy of anti-malarial drugs in vivo: quantitative PCR measurement of parasite clearance. Malar J. 2010;9:312.CrossRefPubMedPubMedCentral
30.
go back to reference Ndam NT, Bischoff E, Proux C, Lavstsen T, Salanti A, Guitard J, et al. Plasmodium falciparum transcriptome analysis reveals pregnancy malaria associated gene expression. PLoS ONE. 2008;3:e1855.CrossRef Ndam NT, Bischoff E, Proux C, Lavstsen T, Salanti A, Guitard J, et al. Plasmodium falciparum transcriptome analysis reveals pregnancy malaria associated gene expression. PLoS ONE. 2008;3:e1855.CrossRef
31.
go back to reference Sander AF, Salanti A, Lavstsen T, Nielsen MA, Magistrado P, Lusingu J, et al. Multiple var2csa-type PfEMP1 genes located at different chromosomal loci occur in many Plasmodium falciparum isolates. PLoS ONE. 2009;4:e6667.CrossRefPubMedPubMedCentral Sander AF, Salanti A, Lavstsen T, Nielsen MA, Magistrado P, Lusingu J, et al. Multiple var2csa-type PfEMP1 genes located at different chromosomal loci occur in many Plasmodium falciparum isolates. PLoS ONE. 2009;4:e6667.CrossRefPubMedPubMedCentral
32.
go back to reference Bahl A, Brunk B, Crabtree J, Fraunholz MJ, Gajria B, Grant GR, et al. PlasmoDB: the Plasmodium genome resource. A database integrating experimental and computational data. Nucleic Acids Res. 2003;31:212–5.CrossRefPubMedPubMedCentral Bahl A, Brunk B, Crabtree J, Fraunholz MJ, Gajria B, Grant GR, et al. PlasmoDB: the Plasmodium genome resource. A database integrating experimental and computational data. Nucleic Acids Res. 2003;31:212–5.CrossRefPubMedPubMedCentral
33.
go back to reference Bodenhofer U, Bonatesta E, Horejš-Kainrath C, Hochreiter S. msa: an R package for multiple sequence alignment. Bioinformatics. 2015;31:3997–9.PubMed Bodenhofer U, Bonatesta E, Horejš-Kainrath C, Hochreiter S. msa: an R package for multiple sequence alignment. Bioinformatics. 2015;31:3997–9.PubMed
35.
go back to reference Kumar S, Stecher G, Tamura K. MEGA7: molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33:1870–4.CrossRefPubMed Kumar S, Stecher G, Tamura K. MEGA7: molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33:1870–4.CrossRefPubMed
36.
go back to reference Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, et al. Clustal W and Clustal X version 2.0. Bioinformatics. 2007;23:2947–8.CrossRefPubMed Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, et al. Clustal W and Clustal X version 2.0. Bioinformatics. 2007;23:2947–8.CrossRefPubMed
37.
go back to reference Tajima F. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics. 1989;123:585–95.PubMedPubMedCentral Tajima F. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics. 1989;123:585–95.PubMedPubMedCentral
38.
go back to reference Nei M, Gojobori T. Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. Mol Biol Evol. 1986;3:418–26.PubMed Nei M, Gojobori T. Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. Mol Biol Evol. 1986;3:418–26.PubMed
40.
41.
go back to reference Piry S, Chapuis MP, Gauffre B, Papaïx J, Cruaud A, Berthier K. Mapping Averaged Pairwise Information (MAPI): a new exploratory tool to uncover spatial structure. Methods Ecol Evol. 2016;7:1463–75.CrossRef Piry S, Chapuis MP, Gauffre B, Papaïx J, Cruaud A, Berthier K. Mapping Averaged Pairwise Information (MAPI): a new exploratory tool to uncover spatial structure. Methods Ecol Evol. 2016;7:1463–75.CrossRef
43.
44.
go back to reference Hamilton WL, Claessens A, Otto TD, Kekre M, Fairhurst RM, Rayner JC, et al. Extreme mutation bias and high AT content in Plasmodium falciparum. Nucleic Acids Res. 2016;45:1889–901.PubMedCentral Hamilton WL, Claessens A, Otto TD, Kekre M, Fairhurst RM, Rayner JC, et al. Extreme mutation bias and high AT content in Plasmodium falciparum. Nucleic Acids Res. 2016;45:1889–901.PubMedCentral
45.
go back to reference Conway DJ, Roper C, Oduola AM, Arnot DE, Kremsner PG, Grobusch MP, et al. High recombination rate in natural populations of Plasmodium falciparum. Proc Natl Acad Sci USA. 1999;96:4506–11.CrossRefPubMedPubMedCentral Conway DJ, Roper C, Oduola AM, Arnot DE, Kremsner PG, Grobusch MP, et al. High recombination rate in natural populations of Plasmodium falciparum. Proc Natl Acad Sci USA. 1999;96:4506–11.CrossRefPubMedPubMedCentral
46.
go back to reference Anderson TJ, Haubold B, Williams JT, Estrada-Franco JG, Richardson L, Mollinedo R, et al. Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum. Mol Biol Evol. 2000;17:1467–82.CrossRefPubMed Anderson TJ, Haubold B, Williams JT, Estrada-Franco JG, Richardson L, Mollinedo R, et al. Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum. Mol Biol Evol. 2000;17:1467–82.CrossRefPubMed
47.
go back to reference Nkhoma SC, Nair S, Al-Saai S, Ashley E, McGready R, Phyo AP, et al. Population genetic correlates of declining transmission in a human pathogen. Mol Ecol. 2013;22:273–85.CrossRefPubMed Nkhoma SC, Nair S, Al-Saai S, Ashley E, McGready R, Phyo AP, et al. Population genetic correlates of declining transmission in a human pathogen. Mol Ecol. 2013;22:273–85.CrossRefPubMed
48.
go back to reference Daubersies P, Sallenave-Sales S, Magne S, Trape JF, Contamin H, Fandeur T, et al. Rapid turnover of Plasmodium falciparum populations in asymptomatic individuals living in a high transmission area. Am J Trop Med Hyg. 1996;54:18–26.CrossRefPubMed Daubersies P, Sallenave-Sales S, Magne S, Trape JF, Contamin H, Fandeur T, et al. Rapid turnover of Plasmodium falciparum populations in asymptomatic individuals living in a high transmission area. Am J Trop Med Hyg. 1996;54:18–26.CrossRefPubMed
49.
go back to reference Gratepanche S, Gamain B, Smith JD, Robinson BA, Saul A, Miller LH. Induction of crossreactive antibodies against the Plasmodium falciparum variant protein. Proc Natl Acad Sci USA. 2003;100:13007–12.CrossRefPubMedPubMedCentral Gratepanche S, Gamain B, Smith JD, Robinson BA, Saul A, Miller LH. Induction of crossreactive antibodies against the Plasmodium falciparum variant protein. Proc Natl Acad Sci USA. 2003;100:13007–12.CrossRefPubMedPubMedCentral
Metadata
Title
Plasmodium falciparum genetic variation of var2csa in the Democratic Republic of the Congo
Authors
Robert Verity
Nicholas J. Hathaway
Andreea Waltmann
Stephanie M. Doctor
Oliver J. Watson
Jaymin C. Patel
Kashamuka Mwandagalirwa
Antoinette K. Tshefu
Jeffrey A. Bailey
Azra C. Ghani
Jonathan J. Juliano
Steven R. Meshnick
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2018
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-018-2193-9

Other articles of this Issue 1/2018

Malaria Journal 1/2018 Go to the issue