scholarly journals Impaired Telomere Maintenance and Decreased Canonical WNT Signaling but Normal Ribosome Biogenesis in Induced Pluripotent Stem Cells from X-Linked Dyskeratosis Congenita Patients

PLoS ONE ◽  
2015 ◽  
Vol 10 (5) ◽  
pp. e0127414 ◽  
Author(s):  
Bai-Wei Gu ◽  
Marisa Apicella ◽  
Jason Mills ◽  
Jian-Meng Fan ◽  
Dara A. Reeves ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wnt Signaling ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ribosome Biogenesis ◽  
Dyskeratosis Congenita ◽  
Telomere Maintenance ◽  
Canonical Wnt Signaling ◽  
Induced Pluripotent ◽  
Canonical Wnt

scholarly journals The Expression of Wnt5a Gene Throughout Definitive Endoderm Induction Process in Induced Pluripotent Stem Cells

2020 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Shahrokh Lorzadeh ◽  
Negar Azarpira ◽  
Saeid Ghavami ◽  
Leila Kohan
Keyword(s):  
Stem Cells ◽  
Wnt Signaling ◽  
Induced Pluripotent Stem Cells ◽  
Mrna Expression ◽  
Pluripotent Stem Cells ◽  
Definitive Endoderm ◽  
Expression Level ◽  
Induction Step ◽  
Induced Pluripotent ◽  
Canonical Wnt

Background: Induced pluripotent stem cells (iPSCs) have the ability to proliferate indefinitely and differentiate into three germ layers of ectoderm, mesoderm, and endoderm. Definitive induction is the first and the most delicate stage of differentiation of various iPSC-derived organs. It has been found that the Wnt signaling pathway implicates in embryogenesis, organogenesis, and cell communication. Objectives: In the present study, we aimed to investigate the expression pattern of the Wnt5a gene as an indicator of non-canonical Wnt signaling activity during definitive endoderm induction of iPSCs. Methods: Human iPSCs (RSCB0042) were acquired from Royan stem cell bank of Royan Institute (Tehran, Iran). The iPSCs were cultured on a feeder layer of mitomycin-inactivated mouse embryonic fibroblasts (MEF), and iPSC colonies were collected for embryoid body (EB) generation by suspension culture method. Then endoderm induction step was performed using a series of small molecules. The quantitative real-time PCR was used to assess the mRNA expression of wnt5a, Nanog, OCT4, SOX17, and FOXA2 genes. Results: The production of efficient EBs confirmed by a decrease in Nanog and Oct4 gene expression and the success of DE (definite endoderm) induction step was confirmed by a high expression level of DE specific genes, Sox17, and FoxA2. A significant upregulation of Wnt5a in EB samples and a minor decrease at day 4 was observed. However, the differentiation process followed by an incremental fashion in Wnt5a mRNA expression starting from day 4 of differentiation among the samples of days 6 and 8 (DE stage). Conclusions: Our results suggest that Wnt5a is more activated at the later steps of endoderm induction rather than the early steps, which may be due to the stimulation of canonical Wnt signaling. Finding the expression level of Wnt5a could rise insights for developing more efficient differentiation induction protocols.

Dysregulation of the TGF β Pathway in Induced Pluripotent Stem Cells (iPSCs) Generated from Patients with Diamond Blackfan Anemia (DBA)

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 254-254
Author(s):  
Jingping Ge ◽  
Marisa Apicella ◽  
Jason A. Mills ◽  
Loic Garcon ◽  
Deborah L. French ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ribosome Biogenesis ◽  
Conflicts Of Interest ◽  
Bone Marrow Failure ◽  
Ectopic Expression ◽  
Canonical Pathway ◽  
Hematopoietic Stem ◽  
Induced Pluripotent

Abstract DBA is an inherited bone marrow failure syndrome that usually develops in the first year of life with clinical features of red cell aplasia and sometimes variable developmental abnormalities. Most affected patients have heterozygous loss of function mutations of one of the 11 ribosomal proteins (RPs) or mutations in the GATA1 gene which encodes an erythroid specific transcription factor. We have previously demonstrated that induced pluripotent stem cells (iPSCs) from fibroblast of DBA patients with RPS19 or RPL5 mutations recapitulate the pathogenesis of DBA, with the mutant lines showing abnormal ribosome biogenesis and altered erythropoiesis. The mechanism whereby haploinsufficiency for RPs causes failure of erythropoiesis and the other DBA features is still unknown. We investigated the pathways that are affected in these DBA iPSCs using an Affymetrix human exon array, and we observed the striking dysregulation of the TGF β pathway in DBA lines. The TGF β downstream target genes, such as DKK1, BAMBI, FN1, COL3A1, COLA1A1 and PAI-1 significantly increased in the DBA iPSCs. The TGF β signaling is complex and can occur via a canonical pathway or by a number of non-canonical pathways. We measured levels of a number of intermediates in these pathways by western blot, and observed a significant increase in the levels of p-JNK, a mediator of a non-canonical pathway, in the DBA iPSCs. Moreover, when the mutant cells were corrected by ectopic expression of WT RPS19 or RPL5, levels of p-JNK returned to normal. We also investigated the SMAD family, which are mediators of the TGF β canonical pathway and are known to negatively regulate the regeneration of hematopoietic stem cells. We observed a drastic decrease in SMAD4, but no change in p-SMAD2. Again corrected lines showed normal expression levels of SMAD4. Our data suggests that the activation of a non-canonical TGF β pathway in the DBA iPSCs may lead increased expression of the downstream genes; and the decrease of anti-proliferative factor SMAD4 may explain how DBA iPSCs maintain their growth. We conclude that the mutations of RPS19 or RPL5 both affect ribosome biogenesis and TGF β signaling, which can cause the failure of erythropoiesis at the stem cell stage. We further suggest that the suppression of SMAD4 may be used as a therapeutic target for DBA treatment. Disclosures No relevant conflicts of interest to declare.

scholarly journals Stage-Specific Regulation of Reprogramming to Induced Pluripotent Stem Cells by Wnt Signaling and T Cell Factor Proteins

Cell Reports ◽  
2013 ◽  
Vol 3 (6) ◽  
pp. 2113-2126 ◽  
Author(s):  
Ritchie Ho ◽  
Bernadett Papp ◽  
Jackson A. Hoffman ◽  
Bradley J. Merrill ◽  
Kathrin Plath
Keyword(s):  
Stem Cells ◽  
T Cell ◽  
Wnt Signaling ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
T Cell Factor ◽  
Specific Regulation ◽  
Induced Pluripotent

scholarly journals Ribosomal and hematopoietic defects in induced pluripotent stem cells derived from Diamond Blackfan anemia patients

Blood ◽  
2013 ◽  
Vol 122 (6) ◽  
pp. 912-921 ◽  
Author(s):  
Loïc Garçon ◽  
Jingping Ge ◽  
Shwetha H. Manjunath ◽  
Jason A. Mills ◽  
Marisa Apicella ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ribosomal Protein ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ribosome Biogenesis ◽  
Ribosomal Protein Genes ◽  
Diamond Blackfan Anemia ◽  
Key Points ◽  
Induced Pluripotent

Key PointsRibosome biogenesis and hematopoiesis are impaired in iPSCs from DBA patients. The abnormalities of DBA iPSCs are ameliorated by genetic restoration of the defective ribosomal protein genes.

scholarly journals Telomere shortening and loss of self-renewal in dyskeratosis congenita induced pluripotent stem cells

Nature ◽  
2011 ◽  
Vol 474 (7351) ◽  
pp. 399-402 ◽  
Author(s):  
Luis F. Z. Batista ◽  
Matthew F. Pech ◽  
Franklin L. Zhong ◽  
Ha Nam Nguyen ◽  
Kathleen T. Xie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Dyskeratosis Congenita ◽  
Self Renewal ◽  
Telomere Shortening ◽  
Loss Of Self ◽  
Induced Pluripotent

scholarly journals Regulation of WNT Signaling by VSX2 During Optic Vesicle Patterning in Human Induced Pluripotent Stem Cells

Stem Cells ◽  
2016 ◽  
Vol 34 (11) ◽  
pp. 2625-2634 ◽  
Author(s):  
Elizabeth E. Capowski ◽  
Lynda S. Wright ◽  
Kun Liang ◽  
M. Joseph Phillips ◽  
Kyle Wallace ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wnt Signaling ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Optic Vesicle ◽  
Induced Pluripotent
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