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Transcriptome of bovine trophoblast stem cells (TSC) - in self-renewal and differentiation

ABSTRACT: In this study, we examine the transcriptome of bovine TSCs under self-renewal that is sustained by inhibiting RhoA-Rock pathway. Using systems biology of spontaneous differentiation, we identify that TGFB1 induces differentiation of bovine TSCs. We analyse the transcriptome of TSCs under TGFB1 induced differentiation. We also analyze pathways of differentiation that are dependent and independent of RhoA-Rock signaling through analysis of the transcriptome of TSCs exposed to both TGFB1 and ROCK inhibition.

ORGANISM(S): Bos taurus

PROVIDER: GSE181252 | GEO | 2022/03/11

REPOSITORIES: GEO

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Project description:The small GTPase RhoA regulates a variety of cellular processes, including cell motility, proliferation, survival and permeability. In addition, there are reports suggesting that the RhoA-ROCK axis plays a role in VEGF-mediated angiogenesis, whereas other work has shown opposite effects. To elucidate this conflicting data, we examined HUVEC and HCAEC after stable overexpression (lentiviral transduction) of constitutively active (G14V/Q63L), dominant-negative (T19N), or wild-type RhoA using a variety of in vitro angiogenesis assays (proliferation, migration, tube formation, angiogenic sprouting, endothelial cell viability) and a HUVEC xenograft assay in immune incompetent NSGTM mice in vivo. We observed that expression of active as well as wild-type RhoA but not expression of dominant-negative RhoA significantly increased endothelial cell death as well as inhibited endothelial cell proliferation, migration, tube formation and angiogenic sprouting of endothelial cells in vitro and reduced HUVEC-related angiogenesis in vivo. Inhibition of RhoA by C3 transferase antagonized inhibitory RhoA effects and strongly enhanced VEGF-induced angiogenic sprouting in control-treated cells. In contrast, inhibition of RhoA effectors ROCK1/2 and LIMK1/2 had no significant effect on RhoA-related effects, but again increased angiogenic sprouting and migration of control-treated cells. In line with these data, VEGF did not activate RhoA in HUVEC as measured by a FRET-based biosensor. Furthermore, global transcriptome and subsequent bioinformatic gene ontology (GO) enrichment analyses revealed that constitutively active RhoA induces a differentially expressed gene pattern that is enriched for GO biological process terms such as mitotic nuclear division, cell proliferation, cell motility and cell adhesion and includes a significant decrease in VEGFR-2 and NOS3 expression. Thus, our data demonstrate that increased RhoA activity has the potential to trigger endothelial dysfunction and anti-angiogenic effects independently of its well-characterized downstream effectors ROCK and LIMK.

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Transcriptome of bovine trophoblast stem cells (TSC) - in self-renewal and differentiation

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