Telomere dynamics and hematopoietic differentiation of human DKC1-mutant induced pluripotent stem cells

Author(s):Show less -

Donaires, Flavia S. ^[1] ; Alves-Paiva, Raquel M. ^[2] ; Gutierrez-Rodrigues, Fernanda ^[2] ; da Silva, Fernanda Borges ^[2] ; Tellechea, Maria Florencia ^[1] ; Moreira, Lilian Figueiredo ^[2] ; Santana, Barbara A. ^[2] ; Traina, Fabiola ^[2] ; Dunbar, Cynthia E. ^[3] ; Winkler, Thomas ^[3] ; Calado, Rodrigo T. ^[2]

Affiliation

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Genet, Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Med Imaging Hematol & Clin Oncol, Ribeirao Preto, SP - Brazil

^[3] NHLBI, NIH, Bldg 10, Bethesda, MD 20892 - USA

• Document type: Journal article
• Source: STEM CELL RESEARCH; v. 40, OCT 2019.
• Web of Science Citations: 0

Abstract

Telomeropathies are a group of phenotypically heterogeneous diseases molecularly unified by pathogenic mutations in telomere-maintenance genes causing critically short telomeres. X-linked dyskeratosis congenita (DC), the prototypical telomere disease, manifested with ectodermal dysplasia, cancer predisposition, and severe bone marrow failure, is caused by mutations in DKC1, encoding a protein responsible for telomerase holoenzyme complex stability. To investigate the effects of pathogenic DKC1 mutations on telomere repair and hematopoietic development, we derived induced pluripotent stem cells (iPSCs) from fibroblasts of a DC patient carrying the most frequent mutation: DKC1 p.A353V. Telomeres eroded immediately after reprogramming in DKC1-mutant iPSCs but stabilized in later passages. The telomerase activity of mutant iPSCs was comparable to that observed in human embryonic stem cells, and no evidence of alternative lengthening of telomere pathways was detected. Hematopoietic differentiation was carried out in DKC1-mutant iPSC clones that resulted in increased capacity to generate hematopoietic colony-forming units compared to controls. Our study indicates that telomerase-dependent telomere maintenance is defective in pluripotent stem cells harboring DKC1 mutation and unable to elongate telomeres, but sufficient to maintain cell proliferation and self-renewal, as well as to support the primitive hematopoiesis, the program that is recapitulated with our differentiation protocol. (AU)

• FAPESP's process: 12/08119-4 - Induced pluripotent stem (iPS) cells differentiation in hepatocytes, neurons, and blood cells from fibroblasts of patients with telomere diseases
• Grantee: Flávia Sacilotto Donaires Ramos
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 12/18434-4 - Induced pluripotent stem (IPS) cells differentiation in hepatocytes, neurons, and blood cells from fibroblasts of patients with telomere diseases
• Grantee: Flávia Sacilotto Donaires Ramos
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC