Identification of factors involved in somatic stem cells in the Drosophila testis
Ontology highlight
ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE148230 | GEO | 2021/03/28
REPOSITORIES: GEO
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Project description:After sex specification during development, inappropriate sex maintenance in adults can lead to numerous defects and pathologies across many species. The molecular and cellular signals involved remain unknown. In the adult Drosophila testis, removing the JAK-STAT downstream effector Chinmo from adult somatic cyst stem cells (CySCs) causes their feminization, such that the sex-concordant interactions between male germ cells and the adjacent somatic cells are disrupted, causing gradual formation of testis germ cell tumors (GCT). Here we use single cell RNA-Seq to characterize cell-type specific transcriptional profiles for chinmo mutant testes undergoing sex transformation. This revealed both expected and novel mutant-specific cell populations and the full repertoire of transcriptomic changes accompanying sex transformation. Cell-cell communication networks were computed among all major cell types, revealing that feminized CySCs have reduced outgoing signaling to themselves and hub cells but elevated outgoing signaling including insulin signaling to germline stem cells (GSCs) /spermatogonia. Using Chinmo CUT & Tag, we identified Chinmo’s direct targets including the sex determination factor dsx and members of the insulin signaling pathway. We also found that Chinmo mainly functions as an activator in CySCs. By functional assays, we found ectopic expression of insulin pathway members in GSCs/spermatogonia phenocopies feminized chinmo mutant testes. In addition, knocking down insulin signaling in germ cells partially rescues the chinmo mutant phenotype. Altogether, we uncovered that Chinmo directly regulates canonical sex determination signals, as well as insulin signaling, to drive sex conversion of CySCs and germline tumorigenesis.
2024-08-28 | GSE201673 | GEO
Project description:After sex specification during development, inappropriate sex maintenance in adults can lead to numerous defects and pathologies across many species. The molecular and cellular signals involved remain unknown. In the adult Drosophila testis, removing the JAK-STAT downstream effector Chinmo from adult somatic cyst stem cells (CySCs) causes their feminization, such that the sex-concordant interactions between male germ cells and the adjacent somatic cells are disrupted, causing gradual formation of testis germ cell tumors (GCT). Here we use single cell RNA-Seq to characterize cell-type specific transcriptional profiles for chinmo mutant testes undergoing sex transformation. This revealed both expected and novel mutant-specific cell populations and the full repertoire of transcriptomic changes accompanying sex transformation. Cell-cell communication networks were computed among all major cell types, revealing that feminized CySCs have reduced outgoing signaling to themselves and hub cells but elevated outgoing signaling including insulin signaling to germline stem cells (GSCs) /spermatogonia. Using Chinmo CUT & Tag, we identified Chinmo’s direct targets including the sex determination factor dsx and members of the insulin signaling pathway. We also found that Chinmo mainly functions as an activator in CySCs. By functional assays, we found ectopic expression of insulin pathway members in GSCs/spermatogonia phenocopies feminized chinmo mutant testes. In addition, knocking down insulin signaling in germ cells partially rescues the chinmo mutant phenotype. Altogether, we uncovered that Chinmo directly regulates canonical sex determination signals, as well as insulin signaling, to drive sex conversion of CySCs and germline tumorigenesis.
2024-08-28 | GSE201579 | GEO
Project description:JAK/STAT pathway plays important roles in controlling Drosophila intestinal homeostasis and regulating the ISC proliferation and differentiation. However,the downstream targets of its transcription factor-STAT92E remain largely unknown.To further identify the regualtory mechanisms of the JAK/STAT pathway in controlling intestinal homeostasis,we performed the ChIP-Seq assay with mouse raised STAT92E antibody using JAK/STAT signaling highly activated adult intestines.Through the ChIP assay, we have identified over 1000 significant peaks (p<0.01) around the putative targets.The well-characterized JAK/STAT downstream targets including Domeless,Socs36E,STAT92E and chinmo were identified in our ChIP assay,indicating that our experiment is workable to identify novel JAK/STAT downstream targets in adult intestines.This work will provide insights into our understanding of regulatory mechanisms of JAK/STAT signaling during Drosophila intestinal development. Identify the ChIP peaks of STAT92E antibody using JAK/STAT signaling highly actived Drosophila adult intestines, compared with input libaray as the control
2015-05-12 | E-GEOD-67346 | biostudies-arrayexpress
Project description:JAK/STAT pathway plays important roles in controlling Drosophila intestinal homeostasis and regulating the ISC proliferation and differentiation. However,the downstream targets of its transcription factor-STAT92E remain largely unknown.To further identify the regualtory mechanisms of the JAK/STAT pathway in controlling intestinal homeostasis,we performed the ChIP-Seq assay with mouse raised STAT92E antibody using JAK/STAT signaling highly activated adult intestines.Through the ChIP assay, we have identified over 1000 significant peaks (p<0.01) around the putative targets.The well-characterized JAK/STAT downstream targets including Domeless,Socs36E,STAT92E and chinmo were identified in our ChIP assay,indicating that our experiment is workable to identify novel JAK/STAT downstream targets in adult intestines.This work will provide insights into our understanding of regulatory mechanisms of JAK/STAT signaling during Drosophila intestinal development.
2015-05-12 | GSE67346 | GEO
Project description:Although the JAK/STAT pathway regulates numerous processes in vertebrates and invertebrates through modulating transcription, its functionally-relevant transcriptional targets remain largely unknown. With one jak and one stat (stat92E), Drosophila provides a powerful system for finding new JAK/STAT target genes. Genome-wide expression profiling on eye discs in which Stat92E is hyperactivated, revealed 584 differentially-regulated genes, including known targets domeless, socs36E and wingless. Other differentially-regulated genes (chinmo, lama, Mo25, Imp-L2, Serrate, Delta) were validated and may represent new Stat92E targets. Genetic experiments revealed that Stat92E cell-autonomously represses Serrate, which encodes a Notch ligand. Loss of Stat92E led to de-repression of Serrate in the dorsal eye, resulting in ectopic Notch signaling and aberrant eye growth there. Thus, our micro-array documents a new Stat92E target gene and a previously-unidentified inhibitory action of Stat92E on Notch signaling. These data suggest that this study will be a useful resource for the identification of additional Stat92E targets. Identification of the JAK/STAT pathway target genes in the Drosophila eye disc Keywords: Genotype comparison Gene expression profiles from five biological replicates of eye discs with yw (control) and GMR-upd (overexpressing JAK/STAT ligand unpaired) were compared using genome wide mRNA expression profiling by Affymetrix genechip arrays (Drosophila 2.0) and key targets were validated by clonal analysis, in situ hybridization, immunohistochemical staining and quantitative real-time PCR.
2009-08-01 | E-GEOD-15868 | biostudies-arrayexpress
Project description:We report the transcriptional profiles obtained by next-generation sequencing (NGS) of FACS-purified cells from the dpp-domain of wing imaginal discs from mid-third instar Drosophila melanogaster larvae. The goals of this study are to compare NGS-derived transcriptome profiling (RNA-seq) of dpp-domain cells that have elevated JAK/STAT signaling or elevated Myc expression compared to control cells.
2019-06-21 | GSE130993 | GEO
Project description:Developing targeted therapy for cutaneous T cell lymphoma (CTCL) patients still requires actionable mutated genes and deregulated pathways to be identified. There is increasing evidence that activating mutations in JAK genes and deregulated JAK/STAT signaling are important mechanisms involved in multiple B and T cell malignancies, including CTCL. Therefore, in this study we focused on studying the mutational status of JAK1, JAK2 and JAK3 genes in a series of human CTCL lesions and cell lines using next-generation sequencing (NGS). We found that 7 of 48 (14.7%) of the analyzed cases harbored mutations in the JAK1 and JAK3 genes that mainly affected the pseudokinase domain of the corresponding proteins. On the basis of these results, we used a specific JAK inhibitor (INCB018424) in a series of CTCL cell lines with deregulated JAK/STAT activity. Treatment of CTCL cells with INCB018424 resulted in dose-dependent reduction of activated STAT expression, diminished cell viability, and increased apoptosis. We also studied global changes in gene expression in cells with mutated JAK1 and JAK3 proteins treated with INCB018424 and identified multiple genes that were differentially regulated by JAK/STAT signaling, such as FGF20 (upregulated) and EGR1 (downregulated). Thus, our results show that the detection of deregulated JAK/STAT signaling in CTCL lesions via JAK mutations or other surrogate markers may serve to indicate the clinical use of JAK/STAT inhibitors. 3 replicates of cells treated with DMSO or JAKi during 30 min and 3h
2015-10-31 | E-GEOD-68562 | biostudies-arrayexpress
Project description:Although the JAK/STAT pathway regulates numerous processes in vertebrates and invertebrates through modulating transcription, its functionally-relevant transcriptional targets remain largely unknown. With one jak and one stat (stat92E), Drosophila provides a powerful system for finding new JAK/STAT target genes. Genome-wide expression profiling on eye discs in which Stat92E is hyperactivated, revealed 584 differentially-regulated genes, including known targets domeless, socs36E and wingless. Other differentially-regulated genes (chinmo, lama, Mo25, Imp-L2, Serrate, Delta) were validated and may represent new Stat92E targets. Genetic experiments revealed that Stat92E cell-autonomously represses Serrate, which encodes a Notch ligand. Loss of Stat92E led to de-repression of Serrate in the dorsal eye, resulting in ectopic Notch signaling and aberrant eye growth there. Thus, our micro-array documents a new Stat92E target gene and a previously-unidentified inhibitory action of Stat92E on Notch signaling. These data suggest that this study will be a useful resource for the identification of additional Stat92E targets. Identification of the JAK/STAT pathway target genes in the Drosophila eye disc Keywords: Genotype comparison
2009-08-01 | GSE15868 | GEO
Project description:Chronically activated JAK/STAT signaling leads to several lung diseases, including lung cancer, asthma, and chronic obstructive pulmonary disease. We used the fruit fly Drosophila melanogaster to elucidate the molecular processes which transform elevated JAK/STAT signaling in airway epithelial cells into pathology.
2022-08-31 | GSE212132 | GEO
Project description:Developing targeted therapy for cutaneous T cell lymphoma (CTCL) patients still requires actionable mutated genes and deregulated pathways to be identified. There is increasing evidence that activating mutations in JAK genes and deregulated JAK/STAT signaling are important mechanisms involved in multiple B and T cell malignancies, including CTCL. Therefore, in this study we focused on studying the mutational status of JAK1, JAK2 and JAK3 genes in a series of human CTCL lesions and cell lines using next-generation sequencing (NGS). We found that 7 of 48 (14.7%) of the analyzed cases harbored mutations in the JAK1 and JAK3 genes that mainly affected the pseudokinase domain of the corresponding proteins. On the basis of these results, we used a specific JAK inhibitor (INCB018424) in a series of CTCL cell lines with deregulated JAK/STAT activity. Treatment of CTCL cells with INCB018424 resulted in dose-dependent reduction of activated STAT expression, diminished cell viability, and increased apoptosis. We also studied global changes in gene expression in cells with mutated JAK1 and JAK3 proteins treated with INCB018424 and identified multiple genes that were differentially regulated by JAK/STAT signaling, such as FGF20 (upregulated) and EGR1 (downregulated). Thus, our results show that the detection of deregulated JAK/STAT signaling in CTCL lesions via JAK mutations or other surrogate markers may serve to indicate the clinical use of JAK/STAT inhibitors.
2015-10-31 | GSE68562 | GEO