scholarly journals Gene correction of the CLN3 c.175G>A variant in patient‐derived induced pluripotent stem cells prevents pathological changes in retinal organoids

2021 ◽  
Author(s):  
Xiao Zhang ◽  
Dan Zhang ◽  
Jennifer A. Thompson ◽  
Shang‐Chih Chen ◽  
Zhiqin Huang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Pathological Changes ◽  
Gene Correction ◽  
Induced Pluripotent

Modeling Congenital Amegakaryocytic Thrombocytopenia Using Patient-Specific Induced Pluripotent Stem Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 703-703
Author(s):  
Naoya Takayama ◽  
Shinji Hirata ◽  
Ryoko Jono-Ohnishi ◽  
Sou Nakamura ◽  
Sho-ichi Hirose ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Conflicts Of Interest ◽  
Receptor Gene ◽  
Patient Specific ◽  
Gene Correction ◽  
Donor Skin ◽  
Induced Pluripotent

Abstract Abstract 703 Patient-specific, induced pluripotent stem cells (iPSCs) enable us to study disease mechanisms and drug screening. To clarify the phenotypic alterations caused by the loss of c-MPL, the thrombopoietin (TPO) receptor, we established iPSCs derived from skin fibroblasts of a patient who received curative bone marrow transplantation for congenital amegakarycytic thrombocytopenia (CAMT) caused by the loss of the TPO receptor gene, MPL. The resultant CAMT-iPSCs exhibited mutations corresponding to the original donor skin. Then using an in vitro culture system yielding hematopoietic progenitor cells (HPCs), we evaluated the role of MPL on the early and late phases of human hematopoiesis. Although CAMT-iPSCs generated CD34+ HPCs, per se, their colony formation capability was impaired, as compared to control CD34+ HPCs. Intriguingly, both Glycophorin A (GPA)+ erythrocyte development and CD41+ megakaryocyte yields from CAMT-iPSCs were also impaired, suggesting that MPL is indispensable for MEP (megakaryocyte erythrocyte progenitors) development. Prospective analysis along with the hematopoietic hierarchy revealed that, in CAMT-iPSCs but not control iPSCs expressing MPL, mRNA expression and phosphorylation of putative signaling molecules downstream of MPL are severely impaired, as is the transition from CD34+CD43+CD41-GPA- MPP (multipotent progenitors) to CD41+GPA+ MEP. Additional analysis also indicated that c-MPL is required for maintenance of a consistent supply of megakaryocytes and erythrocytes from MEPs. Conversely, complimentary transduction of MPL into CAMT-iPSCs using a retroviral vector restored the defective erythropoiesis and megakaryopoiesis; however, excessive MPL signaling appears to promote aberrant megakaryopoiesis with CD42b (GPIba)-null platelet generation and impaired erythrocyte production. Taken together, our findings demonstrate the usefulness of CAMT-iPSCs for validation of functionality in the human hematopoiesis system. For example, it appears that MPL is not indispensable for the emergence of HPCs, but is indispensible for their maintenance, and for subsequent MEP development. Our results also strongly indicate that an appropriate expression level of an administered gene is necessary to achieve curative gene correction / therapy using patient-derived iPSCs. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Targeted Gene Correction in Osteopetrotic-Induced Pluripotent Stem Cells for the Generation of Functional Osteoclasts

2015 ◽  
Vol 5 (4) ◽  
pp. 558-568 ◽  
Author(s):  
Tui Neri ◽  
Sharon Muggeo ◽  
Marianna Paulis ◽  
Maria Elena Caldana ◽  
Laura Crisafulli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Gene Correction ◽  
Induced Pluripotent

scholarly journals 347. CRISPR/Cas9-Based Gene Correction of Arginase-Deficient Human Induced Pluripotent Stem Cells to Recover Enzyme Function

2016 ◽  
Vol 24 ◽  
pp. S139
Author(s):  
Brian Truong ◽  
Patrick C. Lee ◽  
Agustin Vega-Crespo ◽  
William B. Gilmore ◽  
Kip Hermann ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Enzyme Function ◽  
Gene Correction ◽  
Induced Pluripotent

scholarly journals Targeted gene correction of α1-antitrypsin deficiency in induced pluripotent stem cells

Nature ◽  
2011 ◽  
Vol 478 (7369) ◽  
pp. 391-394 ◽  
Author(s):  
Kosuke Yusa ◽  
S. Tamir Rashid ◽  
Helene Strick-Marchand ◽  
Ignacio Varela ◽  
Pei-Qi Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Gene Correction ◽  
Antitrypsin Deficiency ◽  
Induced Pluripotent

scholarly journals Targeted gene correction of FKRP by CRISPR/Cas9 restores functional glycosylation of α-dystroglycan in cortical neurons derived from human induced pluripotent stem cells

2017 ◽  
Author(s):  
Beatrice Lana ◽  
Jihee Kim ◽  
David Ryan ◽  
Evangelos Konstantinidis ◽  
Sandra Louzada ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Genome Editing ◽  
Gene Mutation ◽  
Pluripotent Stem Cells ◽  
Cortical Neurons ◽  
List Type ◽  
Directed Differentiation ◽  
Gene Correction ◽  
Induced Pluripotent

SummaryMutations in genes required for functional glycosylation of α-dystroglycan cause a group of congenital muscular dystrophies associated with brain malformations, referred to as dystroglycanopathies. The lack of isogenic, physiology-relevant human cellular models has limited our understanding of the cortical abnormalities in dystroglycanopathies. Here we generate induced pluripotent stem cells (iPSCs) from a severe dystroglycanopathy patient with homozygous mutations in the ribitol-5-phosphate transferase gene, FKRP. We carry out targeted gene correction in FKRP-iPSCs using CRISPR/Cas9-mediated genome editing. We characterise the directed differentiation of FKRP- and corrected-iPSCs to neural stem cells, cortical progenitors and cortical neurons. Importantly, we show that targeted gene correction of FKRP restores functional glycosylation of α-dystroglycan in iPSC-derived cortical neurons. We independently validate this result by showing targeted gene mutation of FKRP disrupts functional glycosylation of α-dystroglycan. This work demonstrates the feasibility of using CRISPR/Cas9-engineered human iPSCs for modelling dystroglycanopathies and provides a foundation for therapeutic development.HighlightsGeneration of FKRP-iPSCs for modelling cortical abnormalities in dystroglycanopathiesPrecise gene correction by CRISPR/Cas9-mediated genome editingDirected differentiation of isogenic control and FKRP-iPSC to cortical neuronsFunctional glycosylation of α-dystroglycan is restored in cortical neurons derived from CRISPR/Cas9-corrected iPSCsTargeted gene mutation of FKRP disrupts functional glycosylation of α-dystroglycan in cortical neurons

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