Connexin 43 is involved in early differentiation of human embryonic stem cells

Peng, Qian; Yue, C. Patrick; Chen, Andy Chun Hang; Lee, Kai Chuen; Fong, Sze Wan; Yeung, William Shu Biu; Lee, Yin Lau

doi:10.1016/j.diff.2018.12.003

Cited by 17 publications

(13 citation statements)

References 47 publications

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“…These results suggest that yet-to-be-identified signals exchanged through Cx43 GJ channels between differentiating ESCs must positively affect developmental decisions in ESCs. Our conclusions are also in agreement with those of previous studies showing that the combined genetic ablation of Cx43 and Cx45 in mouse ESC results in their failure to form primitive endoderm representing a key inductive stage for further differentiating into downstream cell lineages (Nishii et al., 2014, Wörsdörfer et al., 2017), while other studies have also suggested an important dynamic modulation of Cx43 during hESC differentiation (Galat et al., 2012, Peng et al., 2019).…”

Section: Discussionsupporting

confidence: 93%

Connexin 43 Functions as a Positive Regulator of Stem Cell Differentiation into Definitive Endoderm and Pancreatic Progenitors

et al. 2019

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Summary Efficient stem cell differentiation into pancreatic islet cells is of critical importance for the development of cell replacement therapies for diabetes. Here, we identify the expression pattern of connexin 43 (Cx43), a gap junction (GJ) channel protein, in human embryonic stem cell (hESC)-derived definitive endoderm (DE) and primitive gut tube cells, representing early lineages for posterior foregut (PF), pancreatic progenitors (PP), pancreatic endocrine progenitors (PE), and islet cells. As the function of GJ channels is dependent on their gating status, we tested the impact of supplementing hESC-derived PP cell cultures with AAP10, a peptide that promotes Cx43 GJ channel opening. We found that this treatment promotes the expression of DE markers FoxA2 and Sox17, leads to a more efficient derivation of DE, and improves the yield of PF, PP, and PE cells. These results demonstrate a functional involvement of GJ channels in the differentiation of embryonic stem cells into pancreatic cell lineages.

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Section: Discussionsupporting

confidence: 93%

Connexin 43 Functions as a Positive Regulator of Stem Cell Differentiation into Definitive Endoderm and Pancreatic Progenitors

et al. 2019

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“…CX43 is present in human embryonic stem cells, and its reduced expression is a necessity for the differentiation of the stem cells into trophoblast cells [148]. CX43 is also present in the CTBs, STB [143,149], GCs, and in the decidual stroma [150].…”

Section: Gap Junctionsmentioning

confidence: 99%

The Involvement of Cell Adhesion Molecules, Tight Junctions, and Gap Junctions in Human Placentation

et al. 2020

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Placentation is a major determinant of the success of pregnancy. It is regulated by several factors such as cell adhesion molecules, tight junctions, and gap junctions. The cell adhesion molecules are integrins, cadherins, immunoglobulins, nectins, and selectins. The tight junctions are composed of claudins, occludin, and junction adhesion molecule proteins while the gap junctions are composed of connexins of varying molecular weights. During placentation, some of these molecules regulate trophoblast proliferation, trophoblast fusion, trophoblast migration, trophoblast invasion, trophoblastendothelium adhesion, glandular remodeling, and spiral artery remodeling. There is a dysregulated placental expression of some of these molecules during obstetric complications. We have, hereby, indicated the expression patterns of the subunits of each of these molecules in the various trophoblast subtypes and in the decidua, and have highlighted their involvement in physiological and pathological placentation. The available evidence points to the relevance of these molecules as distinguishing markers of the various trophoblast lineages and as potential therapeutic targets in the management of malplacentation-mediated diseases.

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“…In line with the broad expression of Cx43 throughout the developing embryo, we found comparable Cx43 transcript expression across control iPSCs and all three embryonic germ lineages ( Figure 1 ). Due to the widely reported role of Cx43 in stem cell differentiation [ 12 , 20 , 22 , 27 , 29 ], along with our findings that Cx43 transcripts are similarly expressed during iPSC lineage commitment, we focused on Cx43 protein expression and function across cells of the three germ lineages. Confocal microscopy and Western blot analyses of lineage-specific proteins demonstrate the successful differentiation of iPSCs to ectoderm (PAX6 or Nestin), endoderm (SOX17), and mesoderm (Brachyury) ( Figure 2 ).…”

Section: Resultsmentioning

confidence: 99%

“…Cx43 is reportedly upregulated during human ESC differentiation toward endoderm, and continued Cx43 activity promotes differentiation to endoderm-derived pancreatic precursors [27]. However, this reported role of Cx43 in endoderm differentiation is contrasted by the work of Peng and colleagues, wherein Cx43 knockdown impaired mesoderm formation while endoderm formation remained unaltered [22]. Therefore, questions remain as to how connexin-mediated intercellular communication influences lineage specification.…”

Section: Introductionmentioning

confidence: 98%

“…Human ESCs also express transcripts for eighteen out of the twenty-one identified connexin isoforms [ 13 , 21 ]. Inhibiting GJIC significantly decreases iPSC reprogramming efficiency, while pharmacological GJIC inhibition leads to human iPSC cell death [ 20 , 22 ]. While a variety of Cxs are expressed in pluripotent stem cells, it is unclear how these Cx channels affect cell fate decisions and subsequent tissue development.…”

Section: Introductionmentioning

confidence: 99%

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Connexin 43 Gene Ablation Does Not Alter Human Pluripotent Stem Cell Germ Lineage Specification

2021

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During embryonic germ layer development, cells communicate with each other and their environment to ensure proper lineage specification and tissue development. Connexin (Cx) proteins facilitate direct cell–cell communication through gap junction channels. While previous reports suggest that gap junctional intercellular communication may contribute to germ layer formation, there have been limited comprehensive expression analyses or genetic ablation studies on Cxs during human pluripotent stem cell (PSC) germ lineage specification. We screened the mRNA profile and protein expression patterns of select human Cx isoforms in undifferentiated human induced pluripotent stem cells (iPSCs), and after directed differentiation into the three embryonic germ lineages: ectoderm, definitive endoderm, and mesoderm. Transcript analyses by qPCR revealed upregulation of Cx45 and Cx62 in iPSC-derived ectoderm; Cx45 in mesoderm; and Cx30.3, Cx31, Cx32, Cx36, Cx37, and Cx40 in endoderm relative to control human iPSCs. Generated Cx43 (GJA1) CRISPR-Cas9 knockout iPSCs successfully differentiated into cells of all three germ layers, suggesting that Cx43 is dispensable during directed iPSC lineage specification. Furthermore, qPCR screening of select Cx transcripts in our GJA1-/- iPSCs showed no significant Cx upregulation in response to the loss of Cx43 protein. Future studies will reveal possible compensation by additional Cxs, suggesting targets for future CRISPR-Cas9 ablation studies in human iPSC lineage specification.

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Connexin 43 is involved in early differentiation of human embryonic stem cells

Cited by 17 publications

References 47 publications

Connexin 43 Functions as a Positive Regulator of Stem Cell Differentiation into Definitive Endoderm and Pancreatic Progenitors

Connexin 43 Functions as a Positive Regulator of Stem Cell Differentiation into Definitive Endoderm and Pancreatic Progenitors

The Involvement of Cell Adhesion Molecules, Tight Junctions, and Gap Junctions in Human Placentation

Connexin 43 Gene Ablation Does Not Alter Human Pluripotent Stem Cell Germ Lineage Specification

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