Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function
The lethal zoonosis alveolar echinococcosis is caused by tumour-like growth of the metacestode stage of the tapeworm Echinococcus multilocularis within host organs. We previously demonstrated that metacestode proliferation is exclusively driven by somatic stem cells (germinative cells), which are th...
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2024.1335946/full |
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| author | Michaela Herz Magdalena Zarowiecki Leonie Wessels Katharina Pätzel Ruth Herrmann Christiane Braun Nancy Holroyd Thomas Huckvale Monika Bergmann Markus Spiliotis Uriel Koziol Uriel Koziol Matthew Berriman Klaus Brehm |
| author_facet | Michaela Herz Magdalena Zarowiecki Leonie Wessels Katharina Pätzel Ruth Herrmann Christiane Braun Nancy Holroyd Thomas Huckvale Monika Bergmann Markus Spiliotis Uriel Koziol Uriel Koziol Matthew Berriman Klaus Brehm |
| author_sort | Michaela Herz |
| collection | DOAJ |
| description | The lethal zoonosis alveolar echinococcosis is caused by tumour-like growth of the metacestode stage of the tapeworm Echinococcus multilocularis within host organs. We previously demonstrated that metacestode proliferation is exclusively driven by somatic stem cells (germinative cells), which are the only mitotically active parasite cells that give rise to all differentiated cell types. The Echinococcus gene repertoire required for germinative cell maintenance and differentiation has not been characterised so far. We herein carried out Illumina sequencing on cDNA from Echinococcus metacestode vesicles, from metacestode tissue depleted of germinative cells, and from Echinococcus primary cell cultures. We identified a set of ~1,180 genes associated with germinative cells, which contained numerous known stem cell markers alongside genes involved in replication, cell cycle regulation, mitosis, meiosis, epigenetic modification, and nucleotide metabolism. Interestingly, we also identified 44 stem cell associated transcription factors that are likely involved in regulating germinative cell differentiation and/or pluripotency. By in situ hybridization and pulse-chase experiments, we also found a new general Echinococcus stem cell marker, EmCIP2Ah, and we provide evidence implying the presence of a slow cycling stem cell sub-population expressing the extracellular matrix factor Emkal1. RNA-Seq analyses on primary cell cultures revealed that metacestode-derived Echinococcus stem cells display an expanded differentiation capability and do not only form differentiated cell types of the metacestode, but also cells expressing genes specific for protoscoleces, adult worms, and oncospheres, including an ortholog of the schistosome praziquantel target, EmTRPMPZQ. Finally, we show that primary cell cultures contain a cell population expressing an ortholog of the tumour necrosis factor α receptor family and that mammalian TNFα accelerates the development of metacestode vesicles from germinative cells. Taken together, our analyses provide a robust and comprehensive characterization of the Echinococcus germinative cell transcriptome, demonstrate expanded differentiation capability of metacestode derived stem cells, and underscore the potential of primary germinative cell cultures to investigate developmental processes of the parasite. These data are relevant for studies into the role of Echinococcus stem cells in parasite development and will facilitate the design of anti-parasitic drugs that specifically act on the parasite germinative cell compartment. |
| first_indexed | 2024-03-08T11:39:45Z |
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| last_indexed | 2024-03-08T11:39:45Z |
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| spelling | doaj.art-bdde70f3053a478896131bb28ac498c52024-01-25T07:20:17ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882024-01-011410.3389/fcimb.2024.13359461335946Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell functionMichaela Herz0Magdalena Zarowiecki1Leonie Wessels2Katharina Pätzel3Ruth Herrmann4Christiane Braun5Nancy Holroyd6Thomas Huckvale7Monika Bergmann8Markus Spiliotis9Uriel Koziol10Uriel Koziol11Matthew Berriman12Klaus Brehm13Consultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyParasite Genomics, Wellcome Sanger Institute, Cambridge, United KingdomConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyParasite Genomics, Wellcome Sanger Institute, Cambridge, United KingdomParasite Genomics, Wellcome Sanger Institute, Cambridge, United KingdomConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanySección Biología Celular, Facultad de Ciencias, Universidad de la República, Montevideo, UruguayParasite Genomics, Wellcome Sanger Institute, Cambridge, United KingdomConsultant Laboratory for Echinococcosis, Institute of Hygiene and Microbiology, University of Würzburg, Würzburg, GermanyThe lethal zoonosis alveolar echinococcosis is caused by tumour-like growth of the metacestode stage of the tapeworm Echinococcus multilocularis within host organs. We previously demonstrated that metacestode proliferation is exclusively driven by somatic stem cells (germinative cells), which are the only mitotically active parasite cells that give rise to all differentiated cell types. The Echinococcus gene repertoire required for germinative cell maintenance and differentiation has not been characterised so far. We herein carried out Illumina sequencing on cDNA from Echinococcus metacestode vesicles, from metacestode tissue depleted of germinative cells, and from Echinococcus primary cell cultures. We identified a set of ~1,180 genes associated with germinative cells, which contained numerous known stem cell markers alongside genes involved in replication, cell cycle regulation, mitosis, meiosis, epigenetic modification, and nucleotide metabolism. Interestingly, we also identified 44 stem cell associated transcription factors that are likely involved in regulating germinative cell differentiation and/or pluripotency. By in situ hybridization and pulse-chase experiments, we also found a new general Echinococcus stem cell marker, EmCIP2Ah, and we provide evidence implying the presence of a slow cycling stem cell sub-population expressing the extracellular matrix factor Emkal1. RNA-Seq analyses on primary cell cultures revealed that metacestode-derived Echinococcus stem cells display an expanded differentiation capability and do not only form differentiated cell types of the metacestode, but also cells expressing genes specific for protoscoleces, adult worms, and oncospheres, including an ortholog of the schistosome praziquantel target, EmTRPMPZQ. Finally, we show that primary cell cultures contain a cell population expressing an ortholog of the tumour necrosis factor α receptor family and that mammalian TNFα accelerates the development of metacestode vesicles from germinative cells. Taken together, our analyses provide a robust and comprehensive characterization of the Echinococcus germinative cell transcriptome, demonstrate expanded differentiation capability of metacestode derived stem cells, and underscore the potential of primary germinative cell cultures to investigate developmental processes of the parasite. These data are relevant for studies into the role of Echinococcus stem cells in parasite development and will facilitate the design of anti-parasitic drugs that specifically act on the parasite germinative cell compartment.https://www.frontiersin.org/articles/10.3389/fcimb.2024.1335946/fullechinococcosistranscriptomegerminative cellsprimary cellsdifferentiationpluripotent |
| spellingShingle | Michaela Herz Magdalena Zarowiecki Leonie Wessels Katharina Pätzel Ruth Herrmann Christiane Braun Nancy Holroyd Thomas Huckvale Monika Bergmann Markus Spiliotis Uriel Koziol Uriel Koziol Matthew Berriman Klaus Brehm Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function Frontiers in Cellular and Infection Microbiology echinococcosis transcriptome germinative cells primary cells differentiation pluripotent |
| title | Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function |
| title_full | Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function |
| title_fullStr | Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function |
| title_full_unstemmed | Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function |
| title_short | Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function |
| title_sort | genome wide transcriptome analysis of echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function |
| topic | echinococcosis transcriptome germinative cells primary cells differentiation pluripotent |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2024.1335946/full |
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