RNA-sequencing of human pluripotent stem-cell derived endothelial cells under control and Wnt-activating conditions
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE173206 | GEO | 2021/11/10
REPOSITORIES: GEO
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Project description:Canonical Wnt signaling in endothelial cells (ECs) is required for vascularization of the central nervous system (CNS) and for formation and maintenance of barrier properties unique to CNS vasculature. Gpr124 is an orphan member of the adhesion G-protein-coupled receptor family that is expressed in ECs and is essential for CNS angiogenesis and barrier formation via an unknown mechanism. Using canonical Wnt signaling assays in cell culture and genetic loss- and gain-of-function experiments in mice, we show that Gpr124 functions as a co-activator of Wnt7a- and Wnt7b-stimulated canonical Wnt signaling via a Frizzled receptor and Lrp co-receptor, and that Gpr124-stimulated signaling functions in concert with Norrin/Frizzled4 signaling to control CNS vascular development. These experiments identify Gpr124 as a ligand-specific co-activator of canonical Wnt signaling. Total mRNA from HEK-293/STF cells was subjected to RNAseq
2014-08-20 | E-GEOD-60529 | biostudies-arrayexpress
Project description:Canonical Wnt signaling in endothelial cells (ECs) is required for vascularization of the central nervous system (CNS) and for formation and maintenance of barrier properties unique to CNS vasculature. Gpr124 is an orphan member of the adhesion G-protein-coupled receptor family that is expressed in ECs and is essential for CNS angiogenesis and barrier formation via an unknown mechanism. Using canonical Wnt signaling assays in cell culture and genetic loss- and gain-of-function experiments in mice, we show that Gpr124 functions as a co-activator of Wnt7a- and Wnt7b-stimulated canonical Wnt signaling via a Frizzled receptor and Lrp co-receptor, and that Gpr124-stimulated signaling functions in concert with Norrin/Frizzled4 signaling to control CNS vascular development. These experiments identify Gpr124 as a ligand-specific co-activator of canonical Wnt signaling.
2014-08-20 | GSE60529 | GEO
Project description:Roles of reactive perivascular astrocytes in dysregulation of the blood-brain barrier (BBB) function and cerebral perfusion are not well defined. Here, we investigated transformation of reactive astrocyte function for vascular repair after ischemic stroke by targeted deletion of astrocytic Na+/H+ exchanger isoform 1 in Gfap-CreERT2+/-;Nhe1f/f (Nhe1 Astro-KO) mice. Control Gfap-CreERT2+/-;Nhe1f/f (wild-type) mice displayed BBB damage (elevated endothelial transcytosis and intracellular vesicles) and persistent cerebral hypoperfusion in an ischemic stroke model (transient middle cerebral artery occlusion). In contrast, Nhe1 Astro-KO mice exhibited significantly less endothelial transcytosis and vesicle formation, and increased angiogenesis and cerebral blood perfusion (CBF) post-stroke. Bulk RNA-sequencing transcriptome analysis of isolated GFAP+ reactive astrocytes from wild-type and Nhe1 Astro-KO ischemic brains revealed that ~177 genes were differentially upregulated, with the Wnt7a mRNA among the top upregulated genes, along with additional Wnt pathway genes (Wnt7b, Fzd9, Fzd10, and Ndp). Abundant Wnt7a/b and β-catenin protein expression was detected in cerebral vessels of Nhe1 Astro-KO ischemic brains but not in the wild-type brains. Selective activation of Wnt/β-catenin pathway in cerebral vessels of Nhe1 Astro-KO ischemic brains was further validated using the Wnt reporter line TCF/LEF::H2B-eGFP; Gfap-CreERT2+/-;Nhe1f/f mice. Lastly, the role of Wnt/β-catenin pathway in resistance of Nhe1 Astro-KO mice to stroke-mediated BBB damage was tested by administration of Wnt/β-catenin inhibitor XAV-939. Taken together, we report that transforming reactive astrocyte function by upregulating astrocytic Wnt/β-catenin signaling activity is a novel mechanism to reduce the BBB endothelial damage, stimulate vascular repair, and restore cerebral blood flow after ischemic stroke.
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Project description:Each organ of the human body requires locally-adapted blood vessels1–3. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model, we reveal a molecular mechanism for brain-specific angiogenesis that operates under the control of Wnt7a/b ligands, well-known blood-brain barrier maturation signals4–6. The control mechanism relies on Wnt7a/b-dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI-anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane which lines the brain surface, and whose distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically, Mmp25 confers brain invasive competence by cleaving the pial basement membrane-enriched Col4a5/6 within a short non-collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition, the Wnt/β-catenin-dependent organotypic control of brain angiogenesis is lost, resulting in a properly patterned, yet blood-brain barrier-defective cerebrovasculature. This work reveals an organ-specific angiogenesis mechanism, sheds light on tip cell mechanistic angiodiversity, and thereby illustrates how organs, by imposing local constraints on angiogenic tip cells, can select vessels matching their distinctive physiological requirements.
2024-02-08 | PXD042613 | Pride
Project description:A small molecule, AT7867, promotes proliferation of human pluripotent stem cell-derived pancreatic progenitor cells (PPCs), yet detailed mechanisms of its proliferative effect were not fully understood. Here, we performed cell-based siRNA screening on a subset of genes to reveal that WNT7B is a growth factor of human PPCs as previously shown in mouse pancreas development. Although recombinant proteins had no in vitro activity, feeder cell lines stably expressing Wnt7a or Wnt7b enhanced PPC proliferation. Further analyses showed that canonical Wnt signaling pathway was not affected by AT7867 treatment as well as Wnt7a and Wnt7b. Knockdown of a non-canonical Wnt pathway component, PKCα, and its downstream substrate, MARCKS, blocked PPC proliferation suggesting a requirement of this pathway. Phosphoproteome analysis revealed that AT7867 inhibited Yin Yang 1 (YY1) phosphorylation at Ser118, which directly or indirectly regulated expression of WNT7B in PPCs. Taken together, non-canonical Wnt signaling mediated by WNT7B plays a critical role for the expansion of human PPC population that is a promising alternative cell source to generate β cells for the cure of diabetes.
2021-07-01 | PXD018221 | JPOST Repository
Project description:A small molecule, AT7867, promotes proliferation of human pluripotent stem cell-derived pancreatic progenitor cells (PPCs), yet detailed mechanisms of its proliferative effect were not fully understood. Here, we performed cell-based siRNA screening on a subset of genes to reveal that WNT7B is a growth factor of human PPCs as previously shown in mouse pancreas development. Although recombinant proteins had no in vitro activity, feeder cell lines stably expressing Wnt7a or Wnt7b enhanced PPC proliferation. Further analyses showed that canonical Wnt signaling pathway was not affected by AT7867 treatment as well as Wnt7a and Wnt7b. Knockdown of a non-canonical Wnt pathway component, PKCα, and its downstream substrate, MARCKS, blocked PPC proliferation suggesting a requirement of this pathway. Phosphoproteome analysis revealed that AT7867 inhibited Yin Yang 1 (YY1) phosphorylation at Ser118, which directly or indirectly regulated expression of WNT7B in PPCs. Taken together, non-canonical Wnt signaling mediated by WNT7B plays a critical role for the expansion of human PPC population that is a promising alternative cell source to generate β cells for the cure of diabetes.
2022-03-27 | PXD018222 | JPOST Repository
Project description:A limited number of accessible and representative models of human trophoblast cells currently exist for disease modeling and improving our understanding of placentation. Current stem cell models involve either a transition through a naïve stem cell fate or precise dynamic control of multiple interacting growth factor and small molecule cues. Here, we show that simple exposure of hiPSCs to two small molecules, retinoic acid (RA) and the Wnt signaling agonist CHIR 99021 (CHIR), results in rapid, synergistic upregulation of CDX2, an important transcription factor for trophectoderm cell induction and maintenance of the trophoblast cell population. RA+CHIR-treated cells have a transcriptional profile with high similarity to that of primary trophectoderm cells, and they can also be further differentiated into cells with features of syncytiotrophoblasts and extravillous trophoblasts. We assessed RA+CHIR-treated cells for the established set of criteria defining a trophectoderm cell model, and the cells possess all of the features necessary to be considered a valid model. Taken together, our data demonstrate the development of a simple method for the generation of functional trophoblast-like cells that can be used to better understand mechanisms and interactions involving the placenta.
2024-08-15 | GSE232692 | GEO
Project description:Wnt signalling is a critical determinant of cell lineage development. This study used Wnt dose-dependent induction programs to gain insights into molecular regulation of stem cell differentiation. We performed single cell RNA-sequencing of cells responding to a dose-escalation protocol with Wnt agonist CHIR-99021 during the exit from pluripotency to identify cell types and genetic activity driven by Wnt stimulation. Results of activated gene sets and cell types were then used to build a multiple regression model that predicts the efficiency of cardiomyocyte differentiation. Cross-referencing Wnt-associated gene expression profiles to the Connectivity Map database, we identified a novel property of the small molecule drug, tranilast. We found that tranilast synergistically activates Wnt signalling to promote cardiac lineage differentiation which we validate in vitro and in vivo. Our study provides an integrated workflow that links experimental datasets, prediction models and small molecule databases to identify novel drug-like compounds that control cell differentiation.
2024-01-19 | GSE246080 | GEO
Project description:Wnt signalling is a critical determinant of cell lineage development. This study used Wnt dose-dependent induction programs to gain insights into molecular regulation of stem cell differentiation. We performed single cell RNA-sequencing of cells responding to a dose-escalation protocol with Wnt agonist CHIR-99021 during the exit from pluripotency to identify cell types and genetic activity driven by Wnt stimulation. Results of activated gene sets and cell types were then used to build a multiple regression model that predicts the efficiency of cardiomyocyte differentiation. Cross-referencing Wnt-associated gene expression profiles to the Connectivity Map database, we identified a novel property of the small molecule drug, tranilast. We found that tranilast synergistically activates Wnt signalling to promote cardiac lineage differentiation which we validate in vitro and in vivo. Our study provides an integrated workflow that links experimental datasets, prediction models and small molecule databases to identify novel drug-like compounds that control cell differentiation.
2024-01-19 | GSE246079 | GEO
Project description:Wnt signalling is a critical determinant of cell lineage development. This study used Wnt dose-dependent induction programs to gain insights into molecular regulation of stem cell differentiation. We performed single cell RNA-sequencing of cells responding to a dose-escalation protocol with Wnt agonist CHIR-99021 during the exit from pluripotency to identify cell types and genetic activity driven by Wnt stimulation. Results of activated gene sets and cell types were then used to build a multiple regression model that predicts the efficiency of cardiomyocyte differentiation. Cross-referencing Wnt-associated gene expression profiles to the Connectivity Map database, we identified a novel property of the small molecule drug, tranilast. We found that tranilast synergistically activates Wnt signalling to promote cardiac lineage differentiation which we validate in vitro and in vivo. Our study provides an integrated workflow that links experimental datasets, prediction models and small molecule databases to identify novel drug-like compounds that control cell differentiation.
2024-01-19 | GSE246078 | GEO