Generation of dyskeratosis congenita-like hematopoietic stem cells through the stable inhibition of DKC1

Autor: M. Luz Lozano, Juan A. Bueren, Hidde A. Zittersteijn, Guillermo Guenechea, Cristina Manguan-García, Carlos Carrascoso-Rubio, Leandro Sastre, Beatriz Fernández-Varas, Rosario Perona, Laura Pintado-Berninches
Přispěvatelé: Ministerio de Economía y Competitividad (España), Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Instituto de Salud Carlos III, Fundación Botín, Universidad Autónoma de Madrid
Rok vydání: 2021
Předmět:
0301 basic medicine
congenital
hereditary
and neonatal diseases and abnormalities
Telomerase
medicine.medical_treatment
Short Report
CD34
Medicine (miscellaneous)
Cell Cycle Proteins
Hematopoietic stem cell transplantation
Biochemistry
Genetics and Molecular Biology (miscellaneous)
Dyskeratosis Congenita
Bone marrow failure disorders
lcsh:Biochemistry
Small hairpin RNA
Mice
03 medical and health sciences
0302 clinical medicine
Animals
Humans
Medicine
lcsh:QD415-436
skin and connective tissue diseases
lcsh:R5-920
business.industry
Bone marrow failure
Nuclear Proteins
Lentiviral vectors
Cell Biology
Telomere
Hematopoietic Stem Cells
medicine.disease
Haematopoiesis
030104 developmental biology
Mutation
Cancer research
Molecular Medicine
Stem cell
lcsh:Medicine (General)
Dyskeratosis congenita
DKC1 gene
Short hairpin RNA
business
Hematopoietic stem cells
030215 immunology
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
Stem Cell Research & Therapy
Stem Cell Research & Therapy, Vol 12, Iss 1, Pp 1-8 (2021)
ISSN: 1757-6512
Popis: © The Author(s).
Dyskeratosis congenita (DC) is a rare telomere biology disorder, which results in different clinical manifestations, including severe bone marrow failure. To date, the only curative treatment for the bone marrow failure in DC patients is allogeneic hematopoietic stem cell transplantation. However, due to the toxicity associated to this treatment, improved therapies are recommended for DC patients. Here, we aimed at generating DC-like human hematopoietic stem cells in which the efficacy of innovative therapies could be investigated. Because X-linked DC is the most frequent form of the disease and is associated with an impaired expression of DKC1, we have generated DC-like hematopoietic stem cells based on the stable knock-down of DKC1 in human CD34+ cells with lentiviral vectors encoding for DKC1 short hairpin RNAs. At a molecular level, DKC1-interfered CD34+ cells showed a decreased expression of TERC, as well as a diminished telomerase activity and increased DNA damage, cell senescence, and apoptosis. Moreover, DKC1-interfered human CD34+ cells showed defective clonogenic ability and were incapable of repopulating the hematopoiesis of immunodeficient NSG mice. The development of DC-like hematopoietic stem cells will facilitate the understanding of the molecular and cellular basis of this inherited bone marrow failure syndrome and will serve as a platform to evaluate the efficacy of new hematopoietic therapies for DC.
This work was supported by grants from the “Ministerio de Economía, Comercio y Competitividad and Fondo Europeo de Desarrollo Regional (FEDER)” (SAF2015-68073-R) and from the “Ministerio de Ciencia, Innovación y Universidades and Fondo Europeo de Desarrollo Regional (FEDER)” (RTI2018-097125-B-I00) and P17-01401 from “Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III (FIS-ISCIII)”. The authors also thank the Fundación Botín for promoting translational research at the Hematopoietic Innovative Therapies Division of the CIEMAT. CIBERER is an initiative of the “Instituto de Salud Carlos III” and “FEDER”. CCR was supported by an FPI grant from the Universidad Autónoma de Madrid (UAM).
Databáze: OpenAIRE
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