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Conference Paper: YAP1 knockout impaired human primordial germ cell induction from human expanded potential stem cells
| Title | YAP1 knockout impaired human primordial germ cell induction from human expanded potential stem cells |
|---|---|
| Authors | |
| Issue Date | 1-Jul-2023 |
| Abstract | The Hippo signaling pathway is critical for stem cell self-renewal and differentiation during embryogenesis. The expression of YAP1, one of the Hippo pathway genes, is initiated in the outer cells of human morula. YAP1 is an essential modulator for first-lineage segregation. Previous studies reported that Yap1 modulates differentiation of mouse primordial germ cells (PGC) in vitro. The germline competence is compromised in Yap1-null mouse epiblast stem cells due to the reduced response to Wnt pathway, a critical pathway for mouse epiblast- PGC transition. However, the role of YAP1 in human germ cell lineage development is not fully elucidated. To address this question, we first re-analyzed the published dataset of human fetal PGC development. Interestingly, expression of YAP1 was detected from mitotic to meiotic stages of human PGC. We then utilized our novel human expanded potential stem cells derived from human embryos (hEPSC-em) as an in-vitro model for studying the role of YAP1 in early PGC development. Following the published protocol, hEPSC-em were first differentiated into pre-mesendoderm (pre-ME) stage, followed by further differentiation into PGC-like cell (PGCLC) in six days. Gene expression analyses revealed significant induction of markers of pre-ME (Emoes, Mixl1) and PGCLC (Sox17, Nanos3, Tfap2c) at their respective stages, showing the efficient induction of germ cell lineage from hEPSC-em. In addition, decreased Dnmt3b and enhanced Tet2 expression were found during PGCLC differentiation, which was concordant with the reported global DNA demethylation. YAP1-knockout hEPSC-em generated by CRISPR/Cas9 approach was further used to confirm the role of YAP1 in PGC development. Our results showed that YAP1 depletion led to retention of markers of pre-ME but reduced expressions of germ-cell related markers during PGCLC differentiation. On the other hand, the expressions of DNA methylation-related genes were unaffected upon YAP1 knockout. Our findings demonstrated the importance of YAP1 during early PGC development in human. |
| Persistent Identifier | http://hdl.handle.net/10722/340153 |
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.author | Chen, Chun Hang | - |
| dc.contributor.author | Wu, Minju | - |
| dc.contributor.author | Fong, Sze Wan | - |
| dc.contributor.author | Yeung, William Shu Biu | - |
| dc.contributor.author | Li, Raymond Hang Wun | - |
| dc.contributor.author | Liu, Kui | - |
| dc.contributor.author | Lee, Cherie Yin Lau | - |
| dc.date.accessioned | 2024-03-11T10:42:03Z | - |
| dc.date.available | 2024-03-11T10:42:03Z | - |
| dc.date.issued | 2023-07-01 | - |
| dc.identifier.uri | http://hdl.handle.net/10722/340153 | - |
| dc.description.abstract | <p>The Hippo signaling pathway is critical for stem cell self-renewal and differentiation during embryogenesis. The expression of <em>YAP1</em>, one of the Hippo pathway genes, is initiated in the outer cells of human morula. <em>YAP1</em> is an essential modulator for first-lineage segregation. Previous studies reported that <em>Yap1</em> modulates differentiation of mouse primordial germ cells (PGC) <em>in vitro</em>. The germline competence is compromised in Yap1-null mouse epiblast stem cells due to the reduced response to Wnt pathway, a critical pathway for mouse epiblast- PGC transition. However, the role of <em>YAP1</em> in human germ cell lineage development is not fully elucidated. To address this question, we first re-analyzed the published dataset of human fetal PGC development. Interestingly, expression of <em>YAP1</em> was detected from mitotic to meiotic stages of human PGC. We then utilized our novel human expanded potential stem cells derived from human embryos (hEPSC-em) as an <em>in-vitro</em> model for studying the role of YAP1 in early PGC development. Following the published protocol, hEPSC-em were first differentiated into pre-mesendoderm (pre-ME) stage, followed by further differentiation into PGC-like cell (PGCLC) in six days. Gene expression analyses revealed significant induction of markers of pre-ME (<em>Emoes, Mixl1</em>) and PGCLC (<em>Sox17, Nanos3, Tfap2c</em>) at their respective stages, showing the efficient induction of germ cell lineage from hEPSC-em. In addition, decreased <em>Dnmt3b</em> and enhanced <em>Tet2 </em>expression were found during PGCLC differentiation, which was concordant with the reported global DNA demethylation. <em>YAP1</em>-knockout hEPSC-em generated by CRISPR/Cas9 approach was further used to confirm the role of <em>YAP1</em> in PGC development. Our results showed that <em>YAP1</em> depletion led to retention of markers of pre-ME but reduced expressions of germ-cell related markers during PGCLC differentiation. On the other hand, the expressions of DNA methylation-related genes were unaffected upon <em>YAP1</em> knockout. Our findings demonstrated the importance of YAP1 during early PGC development in human.</p> | - |
| dc.language | eng | - |
| dc.relation.ispartof | International Society for Stem Cell Research Annual Meeting 2023 (14/06/2023-17/06/2023, Boston) | - |
| dc.title | YAP1 knockout impaired human primordial germ cell induction from human expanded potential stem cells | - |
| dc.type | Conference_Paper | - |