CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2026 , Vol. 44 >Issue 3: 328 - 335

DOI: https://doi.org/10.12140/j.issn.1000-7423.2026.03.003

ORIGINAL ARTICLES

Transcriptomic profiling and identification of potential hub molecules during the in vitro bidirectional development of Echinococcus multilocularis protoscoleces

  • SU Xuan ,
  • LAI Xin ,
  • XIE Jun ,
  • XIAO Heyu ,
  • CAO Yilin ,
  • JI Xiaoyao ,
  • CHEN Junhu ,
  • ZHENG Bin ,
  • HU Wei ,
  • ZHOU Xiaonong ,
  • ZHANG Ting
Expand
  • 1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases; NHC Key Laboratory on Parasite and Vector Biology; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology, Shanghai 200025, China
    2 College of Life Sciences, Inner Mongolia University, Hohhot 010020, Inner Mongolia, China
    3 Xizang Center for Disease Control and Prevention, National Health Commission Key Laboratory of Echinococcosis Prevention and Control, Lhasa 850000, Xizang, China

Received date: 2026-02-09

  Revised date: 2026-03-31

  Online published: 2026-05-26

Supported by

National Key Research and Development Program of China(2021YFC2300800);National Key Research and Development Program of China(2021YFC2300804);Special Fund of the Shanghai Municipal Science and Technology Commission(24DZ2203100);Open Project of the National Health Commission Key Laboratory of Echinococcosis Prevention and Control(2024WZK1002)

Fold

Abstract

Objective To develop a bidirectional developmental model for the differentiation of Echinococcus multilocularis protoscoleces into vesicles and for the development of somite, decipher the transcriptomic characteristics, and screen hub regulatory molecules. Methods E. multilocularis protoscoleces were induced into vesicle differentiation and somite development using an in vitro culture system. The early differentiation stage and the stable structure-formation stage were selected as representative time points, and changes in the parasite ultrastructure were observed using scanning electron microscopy. The mature stage of vesicle differentiation and the stage of pseudosomite formation were used as developmental time points. RNA was extracted from parasites for transcriptome sequencing, and differential expression genes (DEGs) were screened with protoscoleces as controls. In addition, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed, and protein-protein interaction (PPI) network was created. Results Swelling of the parasite margin and the visible vesicles were observed on day 9 of differentiation into vesicles, and the protoscolex rostellum degenerated with a reduction in microvilli on day 30 of differentiation into vesicles, with a vesicle formation rate of (34.2 ± 3.4)%. Transcriptomic analysis identified 3 910 DEGs on day 30 of differentiation into vesicles (60.15% of downregulated genes), which was significantly more than that (43.79% of downregulated genes, 621/1 418) on day 9 (χ² = 112.92, P < 0.01). These genes were mainly enriched in membrane receptor signaling and transmembrane transport pathways, and the hub genes were primarily involved in protein synthesis and mitochondrial energy metabolism. Evagination and active motility of protoscoleces was observed on day 3 of development into somite, while obvious neck constriction and posterior pseudosomite enlargement were seen on day 15, with a pseudosomite formation rate of (33.8 ± 2.9)%. Transcriptomic analysis identified 1 520 DEGs (57.83% of upregulated genes) on day 15, which was significantly more than that (51.97% of upregulated genes, 408/785) on day 3 (χ² = 7.19, P < 0.01). These genes were mainly enriched in cell cycle and external factor signaling pathways, and the hub genes were predominantly cell cycle regulatory molecules. Conclusion The bidirectional development of E. multilocularis protoscoleces is regulated by distinct transcriptional programs. Vesicle differentiation is characterized by metabolic and membrane system modulation, whereas somite development is dominated by cell cycle and Wnt signaling, deciphering the molecular regulatory patterns underlying bidirectional development of E. multilocularis protoscoleces. These findings provide the evidence for decoding the regulatory mechanisms of Echinococcus development and for screening potential targets.

Key words： Echinococcus multilocularis; Protoscolex; Transcriptomics; Bidirectional development; Vesicle differentiation; Strobilar development

Cite this article

SU Xuan , LAI Xin , XIE Jun , XIAO Heyu , CAO Yilin , JI Xiaoyao , CHEN Junhu , ZHENG Bin , HU Wei , ZHOU Xiaonong , ZHANG Ting . Transcriptomic profiling and identification of potential hub molecules during the in vitro bidirectional development of Echinococcus multilocularis protoscoleces[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2026 , 44(3) : 328 -335 . DOI: 10.12140/j.issn.1000-7423.2026.03.003

References

[1] Citterio CV, Obber F, Trevisiol K, et al. Echinococcus multilocularis and other cestodes in red foxes (Vulpes vulpes) of northeast Italy, 2012-2018[J]. Parasites Vectors, 2021, 14(1): 29.
[2] Wang X, Kui Y, Xue CZ, et al. Past, present and future epidemiology of echinococcosis in China based on nationwide surveillance data 2004-2022[J]. J Infect, 2025, 90(3): 106445.
[3] Autier B, Gottstein B, Millon L, et al. Alveolar echinococcosis in immunocompromised hosts[J]. Clin Microbiol Infect, 2023, 29(5): 593-599.
[4] Casulli A, Barth TFE, Tamarozzi F. Echinococcus multilocularis[J]. Trends Parasitol, 2019, 35(9): 738-739.
[5] Kaethner M, Epping K, Bernthaler P, et al. Transforming growth factor-β signalling regulates Protoscolex formation in the Echinococcus multilocularis metacestode[J]. Front Cell Infect Microbiol, 2023, 13: 1153117.
[6] Thompson RCA. Biology and systematics of Echinococcus[M]// Echinococcus and Echinococcosis, Part A. Amsterdam: Elsevier, 2017: 65-109.
[7] Tsai IJ, Zarowiecki M, Holroyd N, et al. The genomes of four tapeworm species reveal adaptations to parasitism[J]. Nature, 2013, 496(7443): 57-63.
[8] Korhonen PK, Kinkar L, Young ND, et al. Chromosome-scale Echinococcus granulosus (genotype G1) genome reveals the Eg95 gene family and conservation of the EG95-vaccine molecule[J]. Commun Biol, 2022, 5: 199.
[9] Gao HJ, Huo LL, Mo XJ, et al. Suppressive effect of pseudolaric acid B on Echinococcus multilocularis involving regulation of TGF-β1 signaling in vitro and in vivo[J]. Front Microbiol, 2022, 13: 1008274.
[10] Bai Y, Zhang ZZ, Jin L, et al. Dynamic changes in the global transcriptome and microRNAome reveal complex miRNA-mRNA regulation in early stages of the bi-directional development of Echinococcus granulosus protoscoleces[J]. Front Microbiol, 2020, 11: 654.
[11] Wang H, Li J, Guo BP, et al. In vitro culture of Echinococcus multilocularis producing protoscoleces and mouse infection with the cultured vesicles[J]. Parasites Vectors, 2016, 9(1): 411.
[12] Chen SF. Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp[J]. iMeta, 2023, 2(2): e107.
[13] Kim D, Langmead B, Salzberg SL. HISAT: a fast spliced aligner with low memory requirements[J]. Nat Meth, 2015, 12(4): 357-360.
[14] Srinivasan KA, Virdee SK, McArthur AG. Strandedness during cDNA synthesis, the stranded parameter in htseq-count and analysis of RNA-Seq data[J]. Brief Funct Genom, 2020, 19(5/6): 339-342.
[15] Zhang YG, Wang SL, Xiao JC, et al. Bioinformatics analysis to identify the key genes affecting the progression and prognosis of hepatocellular carcinoma[J]. Biosci Rep, 2019, 39(2): BSR20181845.
[16] Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing[J]. J R Stat Soc Ser B Stat Methodol, 1995, 57(1): 289-300.
[17] Eisen MB, Spellman PT, Brown PO, et al. Cluster analysis and display of genome-wide expression patterns[J]. Proc Natl Acad Sci U S A, 1998, 95(25): 14863-14868.
[18] The Gene Ontology Consortium. The gene ontology resource: 20 years and still GOing strong[J]. Nucleic Acids Res, 2019, 47(D1): D330-D338.
[19] Kanehisa M, Furumichi M, Sato Y, et al. KEGG for taxonomy-based analysis of pathways and genomes[J]. Nucleic Acids Res, 2023, 51(D1): D587-D592.
[20] Szklarczyk D, Kirsch R, Koutrouli M, et al. The STRING database in 2023: protein-protein association networks and functional enrichment analyses for any sequenced genome of interest[J]. Nucleic Acids Res, 2023, 51(D1): D638-D646.
[21] Chin CH, Chen SH, Wu HH, et al. cytoHubba: Identifying hub objects and sub-networks from complex interactome[J]. BMC Syst Biol, 2014, 8(4): S11.
[22] Manterola C, Rivadeneira J, Pogue SD, et al. Morphology of Echinococcus granulosus protoscolex[J]. Int J Morphol, 2023, 41(2): 646-653.
[23] Müller J, Zumkehr B, Heller M, et al. Host proteins in Echinococcus multilocularis metacestodes[J]. Int J Mol Sci, 2025, 26(7): 3266.
[24] Debarba JA, Sehabiague MPC, Monteiro KM, et al. Transcriptomic analysis of the early strobilar development of Echinococcus granulosus[J]. Pathogens, 2020, 9(6): 465.
[25] Herz M, Zarowiecki M, Wessels L, et al. Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function[J]. Front Cell Infect Microbiol, 2024, 14: 1335946.
[26] 王正荣, 张艳艳, 薄新文, 等. 6个wnt基因家族基因在细粒棘球绦虫原头节和成虫的差异表达分析[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(6): 552-557.
  Wang ZR, Zhang YY, Bo XW, et al. Differential expression of six Wnt gene family members in Echinococcus granulosus protoscoleces and adult worms[J]. Chin J Parasitol Parasit Dis, 2016, 34(6): 552-557. (in Chinese)
[27] Aoki K, Higuchi T, Akieda Y, et al. Mechano-gradients drive morphogen-noise correction to ensure robust patterning[J]. Sci Adv, 2024, 10(46): eadp2357.
[28] Lemmens B, Hegarat N, Akopyan K, et al. DNA replication determines timing of mitosis by restricting CDK1 and PLK1 activation[J]. Mol Cell, 2018, 71(1): 117-128.e3.
[29] Ambrozkiewicz MC, Lorenz S. Understanding ubiquitination in neurodevelopment by integrating insights across space and time[J]. Nat Struct Mol Biol, 2025, 32(1): 14-22.
[30] Stadelmann B, Aeschbacher D, Huber C, et al. Profound activity of the anti-cancer drug bortezomib against Echinococcus multilocularis metacestodes identifies the proteasome as a novel drug target for cestodes[J]. PLoS Negl Trop Dis, 2014, 8(12): e3352.
Options
Outlines

/

〈 〉