GATA6-AS1 Regulates GATA6 Expression to Modulate Human Endoderm Differentiation

Yang, Jie; Lu, Pei Jung; Li, Mao; Yan, Chenchao; Zhang, Tianzhe; Jiang, Wei

doi:10.1016/j.stemcr.2020.07.014

Cited by 23 publications

(35 citation statements)

References 51 publications

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“…CHIR99021 (CHIR), also known as GSK3 inhibitor, could induce GATA6 expression for hepatic differentiation from human embryonic stem cells [ 37 ]. Also, down-regulated GSK3 could induce upregulation of GATA6 in cancer cells [ 38 ].…”

Section: Resultsmentioning

confidence: 99%

Efficient hepatic differentiation and regeneration potential under xeno-free conditions using mass-producible amnion-derived mesenchymal stem cells

et al. 2021

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Background Amnion-derived mesenchymal stem cells (AM-MSCs) are an attractive source of stem cell therapy for patients with irreversible liver disease. However, there are obstacles to their use due to low efficiency and xeno-contamination for hepatic differentiation. Methods We established an efficient protocol for differentiating AM-MSCs into hepatic progenitor cells (HPCs) by analyzing transcriptome-sequencing data. Furthermore, to generate the xeno-free conditioned differentiation protocol, we replaced fetal bovine serum (FBS) with polyvinyl alcohol (PVA). We investigated the hepatocyte functions with the expression of mRNA and protein, secretion of albumin, and activity of CYP3A4. Finally, to test the transplantable potential of HPCs, we transferred AM-MSCs along with hepatic progenitors after differentiated days 11, 12, and 13 based on the expression of hepatocyte-related genes and mitochondrial function. Further, we established a mouse model of acute liver failure using a thioacetamide (TAA) and cyclophosphamide monohydrate (CTX) and transplanted AM-HPCs in the mouse model through splenic injection. Results We analyzed gene expression from RNA sequencing data in AM-MSCs and detected downregulation of hepatic development-associated genes including GATA6, KIT, AFP, c-MET, FGF2, EGF, and c-JUN, and upregulation of GSK3. Based on this result, we established an efficient hepatic differentiation protocol using the GSK3 inhibitor, CHIR99021. Replacing FBS with PVA resulted in improved differentiation ability, such as upregulation of hepatic maturation markers. The differentiated hepatocyte-like cells (HLCs) not only synthesized and secreted albumin, but also metabolized drugs by the CYP3A4 enzyme. The best time for translation of AM-HPCs was 12 days from the start of differentiation. When the AM-HPCs were transplanted into the liver failure mouse model, they settled in the damaged livers and differentiated into hepatocytes. Conclusion This study offers an efficient and xeno-free conditioned hepatic differentiation protocol and shows that AM-HPCs could be used as transplantable therapeutic materials. Thus, we suggest that AM-MSC-derived HPCs are promising cells for treating liver disease.

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Section: Resultsmentioning

confidence: 99%

Efficient hepatic differentiation and regeneration potential under xeno-free conditions using mass-producible amnion-derived mesenchymal stem cells

et al. 2021

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“…Moreover, mRNA expression analysis indicated that both knockout and wild-type EBs highly expressed lineage markers, while the pluripotent markers were downregulated compared to undifferentiated ESCs ( Figure 5D ). We further performed directed definitive endoderm differentiation assay ( Figure 5E ; Yang et al, 2020 ). After 3 days of treatment with Activin A, KDM6-knockout ESCs could be induced to FOXA2-positive and OCT-negative endoderm cells by immunofluorescence examination ( Figure 5F ); flow cytometric analysis based on endoderm markers CXCR4 and SOX17 indicated that KDM6-knockout ESCs kept the endoderm differentiation capacity, although much lower than wild-type ESCs ( Figure 5G ), which was consistent with a previous study ( Jiang et al, 2013 ).…”

Section: Resultsmentioning

confidence: 99%

“…For definitive endoderm differentiation, the endoderm differentiation medium [50% IMDM and 50% F12, supplemented with 0.5% B27 without vitamin A, 0.2% BSA (YEASEN, #B57370), and 100 ng/ml activin A (PeproTech, # 120-14P)] was applied for 3 days ( Yang et al, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Depletion of Demethylase KDM6 Enhances Early Neuroectoderm Commitment of Human PSCs

Meng

Zhang

Zheng

et al. 2021

Front. Cell Dev. Biol.

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Epigenetic modifications play a crucial role in neurogenesis, learning, and memory, but the study of their role in early neuroectoderm commitment from pluripotent inner cell mass is relatively lacking. Here we utilized the system of directed neuroectoderm differentiation from human embryonic stem cells and identified that KDM6B, an enzyme responsible to erase H3K27me3, was the most upregulated enzyme of histone methylation during neuroectoderm differentiation by transcriptome analysis. We then constructed KDM6B-null embryonic stem cells and found strikingly that the pluripotent stem cells with KDM6B knockout exhibited much higher neuroectoderm induction efficiency. Furthermore, we constructed a series of embryonic stem cell lines knocking out the other H3K27 demethylase KDM6A, and depleting both KDM6A and KDM6B, respectively. These cell lines together confirmed that KDM6 impeded early neuroectoderm commitment. By RNA-seq, we found that the expression levels of a panel of WNT genes were significantly affected upon depletion of KDM6. Importantly, the result that WNT agonist and antagonist could abolish the differential neuroectoderm induction due to manipulating KDM6 further demonstrated that WNT was the major downstream of KDM6 during early neural induction. Moreover, we found that the chemical GSK-J1, an inhibitor of KDM6, could enhance neuroectoderm induction from both embryonic stem cells and induced pluripotent stem cells. Taken together, our findings not only illustrated the important role of the histone methylation modifier KDM6 in early neurogenesis, providing insights into the precise epigenetic regulation in cell fate determination, but also showed that the inhibitor of KDM6 could facilitate neuroectoderm differentiation from human pluripotent stem cells.

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“…To further investigate the functional role of CHD8 in lineage determination of human ESCs, we first performed the definitive endoderm differentiation [ 34 ]. After differentiation, we analyzed the expression levels of endoderm markers ( FOXA2, SOX17 , and GATA6 ) and pluripotent markers ( OCT4 and SOX2 ) by RT-qPCR.…”

Section: Resultsmentioning

confidence: 99%

CHD8 safeguards early neuroectoderm differentiation in human ESCs and protects from apoptosis during neurogenesis

et al. 2021

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The chromatin remodeler CHD8, which belongs to the ATP-dependent chromatin remodelers CHD family, is one of the most high-risk mutated genes in autism spectrum disorders. However, the role of CHD8 in neural differentiation and the mechanism of CHD8 in autism remains unclear, despite there are a few studies based on the CHD8 haploinsufficient models. Here, we generate the CHD8 knockout human ESCs by CRISPR/Cas9 technology and characterize the effect of loss-of-function of CHD8 on pluripotency maintenance and lineage determination by utilizing efficient directed differentiation protocols. The results show loss-of-function of CHD8 does not affect human ESC maintenance although having slight effect on proliferation and cell cycle. Interestingly, CHD8 depletion results in defective neuroectoderm differentiation, along with severe cell death in neural progenitor stage. Transcriptome analysis also indicates CHD8 does not alter the expression of pluripotent genes in ESC stage, but in neural progenitor cells depletion of CHD8 induces the abnormal expression of the apoptosis genes and suppresses neuroectoderm-related genes. These results provide the evidence that CHD8 plays an essential role in the pluripotency exit and neuroectoderm differentiation as well as the regulation of apoptosis during neurogenesis.

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GATA6-AS1 Regulates GATA6 Expression to Modulate Human Endoderm Differentiation

Cited by 23 publications

References 51 publications

Efficient hepatic differentiation and regeneration potential under xeno-free conditions using mass-producible amnion-derived mesenchymal stem cells

Efficient hepatic differentiation and regeneration potential under xeno-free conditions using mass-producible amnion-derived mesenchymal stem cells

Depletion of Demethylase KDM6 Enhances Early Neuroectoderm Commitment of Human PSCs

CHD8 safeguards early neuroectoderm differentiation in human ESCs and protects from apoptosis during neurogenesis

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