Mem Inst Oswaldo Cruz, Rio de Janeiro, VOLUME 121 | 2026
Research Articles

Comprehensive identification and co-expression analysis of long non-coding RNAs across eight timepoints of Schistosoma mansoni life cycle

Flávia Arêdes-Rocha1, Cristian Chaparro2, Christoph Grunau2, Renata Guerra-Sá3,+

1Universidade Federal de Ouro Preto, Núcleo de Pesquisas em Ciências Biológicas, Programa de Pós-Graduação em Biotecnologia, Ouro Preto, MG, Brasil
2Univ Montpellier, Univ Perpignan Via Domitia, Centre National de la Recherche Scientifique, Institut Français de Recherche pour l’Exploitation de la Mer, Interactions Hôtes-Pathogènes-Environnements, Perpignan, France
3Universidade Federal de Ouro Preto, Instituto de Ciências Exatas e Biológicas, Departamento de Ciências Biológicas, Laboratório de Bioquímica e Biologia Molecular, Ouro Preto, MG, Brasil

DOI: 10.1590/0074-02760250114
448 views 502 downloads
ABSTRACT

BACKGROUND Long non-coding RNAs (lncRNAs) are important regulatory molecules that can be considered potential biomarkers for schistosomiasis. However, the identification and characterisation of these molecules in such complex organism as Schistosoma mansoni, remains very challenging.
OBJECTIVES This study aimed to identify novel lncRNAs in S. mansoni using RNA-seq libraries representative of six developmental stages (eggs, miracidia, sporocysts, cercariae, schistosomula, and juveniles).
METHODS A pipeline for lncRNAs identification was defined and applied to 41 RNA-seq libraries of eggs, miracidia, 1-day, 5-days, 32-days sporocysts, cercariae, schistosomula, and juveniles of S. mansoni. Novel identified lncRNAs and annotated mRNAs were submitted to a weighted co-expression network analysis (WGCNA) to establish lncRNA-mRNAs correlation.
FINDINGS We identified 1,082 novel lncRNAs, mostly presenting stage-specific expression. Co-expression analysis demonstrate that MSTRG.5305.1 can potentially target MEGs and tetraspanin, essential for juvenile worm survival, making it a promising candidate for further functional studies.
MAIN CONCLUSIONS These findings expand the current catalog of S. mansoni lncRNAs and provide new insights into their potential roles in parasite development and host adaptation. Further functional validation could reveal new molecular targets for schistosomiasis control.

REFERENCES
01. McManus DP, Dunne DW, Sacko M, Utzinger J, Vennervald BJ, Zhou XN. Schistosomiasis. Nat Rev Dis Primers. 2018; 4(1): 1-19.
02. Ponzo E, Midiri A, Manno A, Pastorello M, Biondo C, Mancuso G. Insights into epidemiology, pathogenesis, and differential diagnosis of schistosomiasis. Eur J Microbiol Immunol. 2024; 4(2): 86-96.
03. Hirai H. Chromosomal differentiation of schistosomes: What is the message? Front Genet. 2014; 5: 1-5.
04. Berriman M, Haas BJ, Loverde PT, Wilson RA, Dillon GP, Cerqueira GC, et al. The genome of the blood fluke Schistosoma mansoni. Nature. 2009; 460(7253): 352-8.
05. WormBase Parasite. Schistosoma mansoni [homepage on the Internet]. Release 19 (WBPS19). 2024 [cited 2025 Nov 3]. Available from: https://parasite.wormbase.org/Schistosoma_mansoni_ prjea36577/Info/Index/.
06. Verjovski-Almeida S, DeMarco R, Martins EAL, Guimarães PEM, Ojopi EPB, Paquola ACM, et al. Transcriptome analysis of the acoelomate human parasite Schistosoma mansoni. Nat Genet. 2003; 35(2): 148-57.
07. Jarroux J, Morillon A, Pinskaya M. History, discovery, and classification of lncRNAs. 2017: 1008: 1-46.
08. Statello L, Guo CJ, Chen LL, Huarte M. Gene regulation by long non-coding RNAs and its biological functions. Nat Rev Mol Cell Biol. 2021; 22(2): 96-118.
09. Silveira GO, Coelho HS, Amaral MS, Verjovski-Almeida S. Long non-coding RNAs as possible therapeutic targets in protozoa, and in Schistosoma and other helminths. Parasitol Res. 2022; 121(4): 1091-115.
10. Mattick JS, Amaral PP, Carninci P, Carpenter S, Chang HY, Chen LL, et al. Long non-coding RNAs: definitions, functions, challenges and recommendations. Nat Rev Mol Cell Biol. 2023; 24(6): 430-47.
11. Maciel LF, Morales-Vicente DA, Silveira GO, Ribeiro RO, Olberg GGO, Pires DS, et al. Weighted gene co-expression analyses point to long non-coding RNA hub genes at different Schistosoma mansoni life-cycle stages. Front Genet. 2019; 10: 823.
12. Oliveira VF, Moares LAG, Mota EA, Jannotti-Passos LK, Coelho PMZ, Mattos ACA, et al. Identification of 170 new long noncoding RNAs in Schistosoma mansoni. Biomed Res Int. 2018; 2018: 1-9.
13. Vasconcelos EJR, Dasilva LF, Pires DS, Lavezzo GM, Pereira ASA, Amaral MS, et al. The Schistosoma mansoni genome encodes thousands of long non-coding RNAs predicted to be functional at different parasite life-cycle stages. Sci Rep. 2017; 7(1): 1-17.
14. Liao Q, Zhang Y, Zhu Y, Chen J, Dong C, Tao Y, et al. Identification of long noncoding RNAs in Schistosoma mansoni and Schistosoma japonicum. Exp Parasitol. 2018; 191: 82-7.
15. Kim HC, Khalil AM, Jolly ER. LncRNAs in molluscan and mammalian stages of parasitic schistosomes are developmentally-regulated and coordinately expressed with protein-coding genes. RNA Biol. 2020; 17(6): 805-15.
16. Buddenborg SK, Lu Z, Sankaranarayan G, Doyle SR, Berriman M. The stage- and sex-specific transcriptome of the human parasite Schistosoma mansoni. Sci Data. 2023; 10(1): 1-8.
17. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014; 30(15): 2114-20.
18. Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013; 29(1): 15-21.
19. Pertea M, Pertea GM, Antonescu CM, Chang TC, Mendell JT, Salzberg SL. StringTie enables improved reconstruction of a transcriptome from RNA-seq reads. Nat Biotechnol. 2015; 33(3): 290-5.
20. Wucher V, Legeai F, Hédan B, Rizk G, Lagoutte L, Leeb T, et al. FEELnc: a tool for long non-coding RNA annotation and its application to the dog transcriptome. Nucleic Acids Res. 2017; 45(8): 1-12.
21. Patro R, Duggal G, Love MI, Irizarry RA, Kingsford C. Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods. 2017; 14(4): 417-9.
22. Langfelder P, Horvath S. WGCNA: an R package for weighted correlation network analysis. BMC Bioinformatics. 2008; 9: 559.
23. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014; 15(12): 550.
24. Oo JA, Brandes RP, Leisegang MS. Long non-coding RNAs: novel regulators of cellular physiology and function. Pflugers Arch Eur J Physiol. 2022; 474(2): 191-204.
25. Olajide JS, Olopade B, Cai J. Functional intricacy and symmetry of long non-coding RNAs in parasitic infections. Front Cell Infect Microbiol. 2021; 11: 1-13.
26. Graham ML, Li M, Gong AY, Deng S, Jin K, Wang S, et al. Cryptosporidium parvum hijacks a host’s long noncoding RNA U90926 to evade intestinal epithelial cell-autonomous antiparasitic defense. Front Immunol. 2023; 14: 1-14.
27. Nian X, Li L, Ma X, Li X, Li W, Zhang N, et al. Understanding pathogen-host interplay by expression profiles of lncRNA and mRNA in the liver of Echinococcus multilocularis-infected mice. PLoS Negl Trop Dis. 2022; 16(5): 1-26.
28. Morales-Vicente DA, Zhao L, Silveira GO, Tahira AC, Amaral MS, Collins JJ, et al. Single-cell RNA-seq analyses show that long non-coding RNAs are conspicuously expressed in Schistosoma mansoni gamete and tegument progenitor cell populations. Front Genet. 2022; 13: 1-19.
29. Maciel LF, Morales-Vicente DA, Verjovski-Almeida S. Dynamic expression of long non-coding RNAs throughout parasite sexual and neural maturation in Schistosoma japonicum. Non-coding RNA. 2020; 6(2): 1-18.
30. Mota EA, de Oliveira VF, do Patrocínio AB, Rodrigues V, Guerra- Sá R. Detection of Schistosoma mansoni long non-coding RNAs in the infected C57BL/6 mouse liver. Exp Parasitol. 2021; 222: 108062.
31. Chen L, Chen Y, Zhang D, Hou M, Yang B, Zhang F, et al. Protection and immunological study on two tetraspanin-derived vaccine candidates against schistosomiasis japonicum. Parasite Immunol. 2016; 38(10): 589-98.
32. Mekonnen GG, Tedla BA, Pearson MS, Becker L, Field M, Amoah AS, et al. Characterisation of tetraspanins from Schistosoma haematobium and evaluation of their potential as novel diagnostic markers. PLoS Negl Trop Dis. 2022; 16(1): 1-21.
33. Tran MH, Pearson MS, Bethony JM, Smyth DJ, Jones MK, Duke M, et al. Tetraspanins on the surface of Schistosoma mansoni are protective antigens against schistosomiasis. Nat Med. 2006; 12(7): 835-40.
34. Tran MH, Freitas TC, Cooper L, Gaze S, Gatton ML, Jones MK, et al. Suppression of mRNAs encoding tegument tetraspanins from Schistosoma mansoni results in impaired tegument turnover. PLoS Pathog. 2010; 6(4): e1000840.
35. Keitel WA, Potter GE, Diemert D, Bethony J, Sahly HM El, Kennedy JK, et al. A phase 1 study of the safety, reactogenicity, and immunogenicity of a Schistosoma mansoni vaccine with or without glucopyranosyl lipid A aqueous formulation (GLA-AF) in healthy adults from a non-endemic area. Vaccine. 2019; 37(43): 6500-9.
36. Martins VP, Morais SB, Pinheiro CS, Assis NRG, Figueiredo BCP, Ricci ND, et al. Sm10.3, a member of the micro-exon gene 4 (MEG-4) family, induces erythrocyte agglutination in vitro and partially protects vaccinated mice against Schistosoma mansoni infection. PLoS Negl Trop Dis. 2014; 8(3): e2701.
37. Li XH, DeMarco R, Neves LX, James SR, Newling K, Ashton PD, et al. Microexon gene transcriptional profiles and evolution provide insights into blood processing by the Schistosoma japonicum esophagus. PLoS Negl Trop Dis. 2018; 12(2): e0006275.
38. Mambelli FS, Figueiredo BC, Morais SB, Assis NRG, Fonseca CT, Oliveira SC. Recombinant micro-exon gene 3 (MEG-3) antigens from Schistosoma mansoni failed to induce protection against infection but show potential for serological diagnosis. Acta Trop. 2020; 204: 105356.
39. Farias LP, Vance GM, Coulson PS, Vitoriano-Souza J, da Silva Neto AP, Wangwiwatsin A, et al. Epitope mapping of exposed tegument and alimentary tract proteins identifies putative antigenic targets of the attenuated schistosome vaccine. Front Immunol. 2021; 11: 624613.

Financial support: CNPq, CAPES (Finance Code 001), FAPEMIG (APQ-03158-24).
+ Corresponding author: rguerra@ufop.edu.br | ORCID https://orcid.org/0000-0003-2486-0534
Received 07 May 2025
Accepted 30 October 2025

HOW TO CITE
Arêdes-Rocha F, Chaparro C, Grunau C, Guerra-Sá R. Comprehensive identification and co-expression analysis of long non-coding RNAs across eight timepoints of Schistosoma mansoni life cycle. Mem Inst Oswaldo Cruz. 2025. 120: e250114.

HANDLING EDITOR
Adeilton Alves Brandão | ORCID https://orcid.org/0000-0001-5877-607X

Our Location

Memórias do Instituto Oswaldo Cruz

Av. Brasil 4365, Castelo Mourisco 
sala 201, Manguinhos, 21040-900 
Rio de Janeiro, RJ, Brazil

Tel.: +55-21-2562-1222

This email address is being protected from spambots. You need JavaScript enabled to view it.

Support Program

logo iocb

logo governo federal03h 
faperj   cnpq capes