Generation of human iPSCs from urine derived cells of a patient with a novel homozygous PAI-1 mutation

Afzal, Muhammad Bilal; Gartz, Melanie; Klyachko, Ekaterina; Khan, Sadiya S.; Shah, Sanjiv J.; Gupta, Sweta; Shapiro, Amy D.; Vaughan, Douglas E.; Strande, Jennifer L.

doi:10.1016/j.scr.2016.11.010

Cited by 3 publications

(3 citation statements)

References 13 publications

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“…As a proof-of-concept study, we used previously characterized iPSC lines to pursue preliminary mechanistic in vitro studies. [7][8][9] Cardiomyocyte differentiation of iPSCs was achieved by following published protocols 10,11 with details outlined in the eMethods in the Supplement from 1 participant homozygous for the c.699_700dupTA SERPINE1 variant and 1 wildtype control participant. Experimental results examining cardiomyocyte response to angiotensin II and metabolic stress are presented as mean (SEM).…”

Section: Stem Cell-derived Cardiomyocytesmentioning

confidence: 99%

Identification of Cardiac Fibrosis in Young Adults With a Homozygous Frameshift Variant in SERPINE1

et al. 2021

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IMPORTANCECardiac fibrosis is exceedingly rare in young adults. Identification of genetic variants that cause early-onset cardiomyopathy may inform novel biological pathways. Experimental models and a single case report have linked genetic deficiency of plasminogen activator inhibitor-1 (PAI-1), a downstream target of cardiac transforming growth factor β, with cardiac fibrosis. OBJECTIVE To perform detailed cardiovascular phenotyping and genotyping in young adults from an Amish family with a frameshift variant (c.699_700dupTA) in SERPINE1, the gene that codes for PAI-1. DESIGN, SETTING, AND PARTICIPANTSThis observational study included participants from 3 related nuclear families from an Amish community in the primary analysis and participants from the extended family in the secondary analysis.

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Section: Stem Cell-derived Cardiomyocytesmentioning

confidence: 99%

Identification of Cardiac Fibrosis in Young Adults With a Homozygous Frameshift Variant in SERPINE1

et al. 2021

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“…The dinucle-otide (TA) insertion in exon 4 of the PAI-1 gene disrupts an endogenous SfcI restriction enzyme digestion site. PCR amplification followed by SfcI digestion allows for detection of wild-type (606 and 147 bp fragments), heterozygous (753, 606 and 147 bp fragments), and homozygous (753 bp fragment) ( Afzal et al, 2016 ) gene products.…”

Section: Methodsmentioning

confidence: 99%

Generation of human iPSCs from urine derived cells of patient with a novel heterozygous PAI-1 mutation

et al. 2017

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“…The ability of stem cells to differentiate into endoderm, mesoderm and ectoderm were tested via embryoid body (EB) formation and stained by Human three germ layer 3-color immunocytochemistry kit (R&D Systems, Minneapolis, MN, USA) 35 .…”

Section: Characterization Of Genetically-modified Ipsc Propertiesmentioning

confidence: 99%

Distinct effects of V617F and exon12-mutated JAK2 expressions on erythropoiesis in a human induced pluripotent stem cell (iPSC)-based model

Nilsri

Jangprasert

Pawinwongchai

et al. 2021

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Activating mutations affecting the JAK-STAT signal transduction is the genetic driver of myeloproliferative neoplasms (MPNs) which comprise polycythemia vera (PV), essential thrombocythemia (ET) and myelofibrosis. The JAK2p.V617F mutation can produce both erythrocytosis in PV and thrombocytosis in ET, while JAK2 exon 12 mutations cause only erythrocytosis. We hypothesized that these two mutations activated different intracellular signals. In this study, the induced pluripotent stem cells (iPSCs) were used to model JAK2-mutated MPNs. Normal iPSCs underwent lentiviral transduction to overexpress JAK2p.V617F or JAK2p.N542_E543del (JAK2exon12) under a doxycycline-inducible system. The modified iPSCs were differentiated into erythroid cells. Compared with JAK2V617F-iPSCs, JAK2exon12-iPSCs yielded more total CD71+GlycophorinA+ erythroid cells, displayed more mature morphology and expressed more adult hemoglobin after doxycycline induction. Capillary Western immunoassay revealed significantly higher phospho-STAT1 but lower phospho-STAT3 and lower Phospho-AKT in JAK2exon12-iPSCs compared with those of JAK2V617F-iPSCs in response to erythropoietin. Furthermore, interferon alpha and arsenic trioxide were tested on these modified iPSCs to explore their potentials for MPN therapy. Both agents preferentially inhibited proliferation and promoted apoptosis of the iPSCs expressing mutant JAK2 compared with those without doxycycline induction. In conclusion, the modified iPSC model can be used to investigate the mechanisms and search for new therapy of MPNs.

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Generation of human iPSCs from urine derived cells of a patient with a novel homozygous PAI-1 mutation

Cited by 3 publications

References 13 publications

Identification of Cardiac Fibrosis in Young Adults With a Homozygous Frameshift Variant in SERPINE1

Identification of Cardiac Fibrosis in Young Adults With a Homozygous Frameshift Variant in SERPINE1

Generation of human iPSCs from urine derived cells of patient with a novel heterozygous PAI-1 mutation

Distinct effects of V617F and exon12-mutated JAK2 expressions on erythropoiesis in a human induced pluripotent stem cell (iPSC)-based model

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