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Comparative microRNA expression profiles of Schistosoma japonicum from SCID mouse and BALB/c mouse

ABSTRACT: Background: In immunodeficient mice, the growth and reproduction of schistosome were significantly retarded, and their virulence on the host was also weakened. Some studies have suggested that host factors including endocrine and immune signals can modulate the parasite’s development and maturation. However, it is still not clear for the molecular mechanisms underlying the growth and maturation regulation of schistosome. Methods: We isolated the total RNAs of female and male schistosome from immunodeficient SCID mice and immunocompetent BALB/c mice (labeled as S_F, S_M, B_F and B_M) obtained at five weeks post infection with cercariae, and constructed cDNA libraries of small RNAs using small RNA sample pre kit for small RNA deep sequencing on an Illumina (Solexa) platform. Results: Of the 122 (75 known and 47 novel) identified S. japonicum-specific miRNAs, statistical analysis revealed that 33 miRNAs (17 miRNAs were up-regulated and 16 were down-regulated, P<0.05) in female schistosome and 33 miRNAs (13 miRNAs were up-regulated and 20 were down-regulated, P<0.05) in male schistosome from SCID mice were differentially expressed when compared with those from BALB/c mice. The Venn diagram of the differentially expressed miRNAs shows 21 miRNAs were common between S_F vs B_ F and S_M vs B_M, 12 miRNAs were specific in female comparison, and other 12 were specific in male comparison. Conclusions: This study provides a comparative microRNAs expression profiles of schistosomes derived from SCID mice and immunocompetent BALB/c mice. A number of microRNAs were identified to potentially participate in regulating the development of schistosomes. These findings lay the foundation for schistosomiasis research in regulation of the development of schistosomes and provide a valuable resource for screening antischistosomal intervention targets or vaccine candidates.

ORGANISM(S): Schistosoma japonicum

PROVIDER: GSE122318 | GEO | 2020/11/06

REPOSITORIES: GEO

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Project description:Background: In immunodeficient mice, the growth and egg laying of schistosome were significantly retarded or inhibited, and their virulence on the host was also weakened. Some studies have suggested that host factors including endocrine and immune signals can modulate the parasite’s development and maturation. However, it is still not clear for the molecular mechanisms underlying the growth and maturation regulation of schistosome. Methodology/Principal results: We isolated the total RNAs of female and male schistosome from immunodeficient SCID mice and immunocompetent BALB/c mice (labeled as S_F, S_M, B_F and B_M) obtained at five weeks post infection with cercariae, and constructed cDNA libraries for RNA deep sequencing on an Illumina (Solexa) platform. In the results, the mRNAs were distinguished from long non-coding RNAs after blast to the reference Schistosoma japonicum genome, assemble and assessment of coding potential of the clean reads between S_F and B_F, and between S_M and B_M. The RNA-Seq yielded 90 to 142 million raw reads and 84 to 134 million clear reads per sample for the four schistosome samples. There are 56% to 65% of the clean reads mapped to the reference S. japonicum genome data. Given P-value <0.05, 298 differentially expressed genes (DEGs, mRNAs) for pairwise comparison of the above schistosome samples were identified. In detailed, 240 DEGs were identified in S_F vs B_F, of which 152 DEGs were up-regulated (147 DEGs were present in S_F worms but absent or undetected technically in B_F worms) and 88 were down-regulated (83 DEGs were present in B_F worms but absent or undetected technically in S_F worms). 62 DEGs were identified in S_M vs B_M, of which 39 were up-regulated (29 DEGs were present in S_M worms but absent or undetected technically in B_M worms) and 23 were down-regulated (22 DEGs were present in B_M worms but absent or undetected technically in S_M worms). Four differentially expressed mRNAs were shared among the two comparisons (present both in female and male worms), which were Sjp_0117620 (26S proteasome subunit P28-related), Sjp_0127870 (leucine zipper-EF-hand-containing transmembrane protein 1, mitochondrial precursor), Sjp_0130040 (putative dynein heavy chain) and Sjp_0094540 (hypothetical protein containing proteinase inhibitor I25, cystatin domain，SJCHGC07500 protein, partial). There were other 236 DEG specific in the comparison of S_F and B_F, and 58 DEG specific in the comparison of S_M and B_M. GO analysis was performed to summarize and explore the major GO categories of the DEGs which may be susceptible to host environments. The above DEGs of female comparison and male comparison were annotated with GO terms in three independent categories: biological processes (400 GO terms were hit for annotation of DEGs of female comparison and 287 GO terms were hit for annotation of DEGs of male comparison), molecular functions (231 GO terms were hit for annotation of DEGs of female comparison and 163 GO terms were hit for annotation of DEGs of male comparison), and cellular components (88 GO terms were hit for annotation of DEGs of female comparison and 75 GO terms were hit for annotation of DEGs of male comparison). The biological processes analysis revealed the predominant DEGs of female worms were involved in response to microtubule-based process, while predominant DEGs of male worms were involved in response to response to DNA damage stimulus, oxidation-reduction process and cellular response to stress. The molecular functions, some DEGs of female and male worms were annotated with calcium-dependent phospholipid binding activities in common. Notably, genes coding for microtubule associated complex, cytoskeletal part, cytoskeleton and microtubule cytoskeleton accounted for large portion of annotated DEGs of female worms in the cellular components analysis, while gene coding for nucleoplasmic THO complex of male worms were identified in the cellular components analysis. The KEGG pathway analysis showed many of the DEGs of female worms were involved in purine metabolism, RNA polymerase, fatty acid biosynthesis, fatty acid degradation and pyrimidine metabolism. Many of the DEGs of male worms were involved in oxidative phosphorylation, riboflavin metabolism and tyrosine metabolism. Conclusions/Significance: This study provides a comparative gene expression profiles of schistosomes derived from SCID mice and immunocompetent BALB/c mice. A number of genes potentially regulating the development of schistosomes were identified. These findings lay the foundation for schistosomiasis research in parasite-host interaction at the transcriptional level and provide a valuable resource for screening antischistosomal intervention targets or vaccine candidates.

Project description:Microtus fortis (M. fortis) is the only mammal in which the growth, development and maturation of schistosomes (Schistosoma japonicum) is prevented, resulting in the failure of the parasite to mature and complete its life cycle. MicroRNAs (miRNAs) are a class of endogenous, non-coding small RNAs, has been found to introduce a whole new layer of gene regulation in eukaryotes. The anti-schistosomiasis mechanosm of M. fortis may require the participation of miRNA-mediated gene expression. In the present study, the difference pathological change of different tissue such as liver, spleen and lung of M. fortis were observed by using haematoxylin-eosin staining. Also, the miRNA expression in different tissue of M. fortis and mice before challenge and 10 days post-infection with schistosomes were first compared using microRNA microarray analysis. Histological analyses showed that S. japonicum infection in M. fortis resulted in more intensive inflammatory response and pathologic change than mice. The microarray investigations showed that 388 miRNAs detected common expressed in the two species, and 11 miRNAs in liver, 25 miRNAs in spleen and 28 miRNAs in lung differentially expressed in non-permissive M. fortis while increased, decreased or nearly fixed in mice. Further studies of the differentially expressed miRNAs demonstrated that many important signal pathway were triggered after the S. japonicum infection in M. fortis rather than the mouse, such as the metabolism of some nutrient material such as fatty-acid, cholesterol, lipid, insulin, and carbohydrate; immune response such as B and T cell differentiation, monocyte differentiation, the inflammation, NF-kappaB binding, even the in innate immune system; Cell differentiation and apoptosis such as erythrocytic differentiation and targeting proapoptotic and antiapoptotic proteins. These results may provide new insights into general mechanisms of regulation in non-permissive M. fortis, exploit the potential miRNA regulatory networks and the interaction between parasites and different hosts, which provide valuable new information on schistosome biology and valuable information for the better understanding of schistosome development and host-parasite interactions. We collected liver, spleen and lung from control and 10 days post-infection with schistosomes of M. fortis, mice and rat, respectively.

Project description:STUDY THE EFFECT OF TGF beta ON SCHISTOSOME ADULT WORMSGoal: Primary: Assess the effects of TGF beta on transcription of adult male and female schistosomes. Secondary: Identify the pathways regulated by TGF beta in worm pairs, mature males, mature females, immature males and immature females.METHODS:Parasite: Schistosoma mansoni employed is the NMRI strain that is maintained in a Puerto Rican Biomphalaria glabrata and Golden Syrian hamsters. The life cycle is maintained in the LoVerde lab.Cercariae obtained from infected snails were used to infect experimental animals. The cercariae wre shed from snails exposed to multiple miracidia for mixed infections or from snails exposed to a single miracidia for single-sex infections. Schistosome worms were recovered from hamsters by perfusion (Duvall and DeWitt, 1967) 45 days post infection. Freshly perfused adult worms containing mixed-sex or single-sex infections were collected from infected golden hamsters, washed and incubated for overnight at 37C, 5% CO2 in MEM medium supplemented with 1 mM sodium pyruvate, 1 X non-essential amino acid solution, 1 X amino acid solution, 100 U ml penicillin, 100 ug ml streptomycin 1 ug ml amphotericin B (antibiotic antimycotic solution) and 2 mM Glutamax-I. Recombinant human TGF- beta1 was added individually to about 25 pairs or 30 individual schistosomes to a final concentrations of 1.0 nM and incubation was continued for another 24 hr. The controls were treated as above except no TGF-beta1 was added. The worms were placed in RNAlater and sent to WTSI (Anna) for RNA extraction and sequencing.Duvall RH, Dewitt WD. An improved perfusion technique for recovering adult schistosomes from laboratory animals. Am J Trop Med Hyg 1967;16:483-486.. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/

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Comparative microRNA expression profiles of Schistosoma japonicum from SCID mouse and BALB/c mouse

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