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iPSC-Derived Intestinal Organoids from Cystic Fibrosis Patients Acquire CFTR Activity upon TALEN-Mediated Repair of the p.F508del Mutation
- Source :
- Molecular Therapy: Methods & Clinical Development, Vol 17, Iss, Pp 858-870 (2020), Addi. Archivo Digital para la Docencia y la Investigación, instname, Digital.CSIC. Repositorio Institucional del CSIC, Molecular Therapy. Methods & Clinical Development
- Publication Year :
- 2020
- Publisher :
- Cell Press, 2020.
-
Abstract
- Cystic fibrosis (CF) is the main genetic cause of death among the Caucasian population. The disease is characterized by abnormal fluid and electrolyte mobility across secretory epithelia. The first manifestations occur within hours of birth (meconium ileus), later extending to other organs, generally affecting the respiratory tract. It is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR encodes a cyclic adenosine monophosphate (cAMP)-dependent, phosphorylation-regulated chloride channel required for transport of chloride and other ions through cell membranes. There are more than 2,000 mutations described in the CFTR gene, but one of them, phenylalanine residue at amino acid position 508 (p.F508del), a recessive allele, is responsible for the vast majority of CF cases worldwide. Here, we present the results of the application of genome-editing techniques to the restoration of CFTR activity in p.F508del patient-derived induced pluripotent stem cells (iPSCs). Gene-edited iPSCs were subsequently used to produce intestinal organoids on which the physiological activity of the restored gene was tested in forskolin-induced swelling tests. The seamless restoration of the p.F508del mutation resulted in normal expression of the mature CFTR glycoprotein, full recovery of CFTR activity, and a normal response of the repaired organoids to treatment with two approved CF therapies: VX-770 and VX-809.<br />Graphical Abstract<br />Cystic Fibrosis (CF) is the main genetic cause of death among the Caucasian population. Here, we present the results of the application of genome-editing techniques to repair the CF defect in patient-derived cells and organoids. This protocol could be the foundation for future treatments based on cell repair and transplantation.
- Subjects :
- 0301 basic medicine
lcsh:QH426-470
Biology
medicine.disease_cause
FIS assay
Cystic fibrosis
Article
gene targeting
cystic fibrosis
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
TALEN
Genetics
Organoid
medicine
Cyclic adenosine monophosphate
lcsh:QH573-671
Induced pluripotent stem cell
Molecular Biology
Mutation
iPSC
lcsh:Cytology
Gene targeting
medicine.disease
Molecular biology
Cystic fibrosis transmembrane conductance regulator
3. Good health
lcsh:Genetics
030104 developmental biology
chemistry
intestinal organoids
030220 oncology & carcinogenesis
Chloride channel
biology.protein
Molecular Medicine
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Molecular Therapy: Methods & Clinical Development, Vol 17, Iss, Pp 858-870 (2020), Addi. Archivo Digital para la Docencia y la Investigación, instname, Digital.CSIC. Repositorio Institucional del CSIC, Molecular Therapy. Methods & Clinical Development
- Accession number :
- edsair.doi.dedup.....9f7bc20059a564dc32e43fd7355dcf0f