Chromosomal Instability and Molecular Defects in Induced Pluripotent Stem Cells from Nijmegen Breakage Syndrome Patients

Tomer Halevy; Shira Akov; Martina Bohndorf; Barbara Mlody; James Adjaye; Nissim Benvenisty; Michal Goldberg

doi:10.1016/j.celrep.2016.07.071

Chromosomal Instability and Molecular Defects in Induced Pluripotent Stem Cells from Nijmegen Breakage Syndrome Patients

Cell Rep. 2016 Aug 30;16(9):2499-511. doi: 10.1016/j.celrep.2016.07.071. Epub 2016 Aug 18.

Authors

Tomer Halevy¹, Shira Akov¹, Martina Bohndorf², Barbara Mlody², James Adjaye², Nissim Benvenisty³, Michal Goldberg⁴

Affiliations

¹ The Azrieli Center for Stem Cells and Genetic Research, The Hebrew University, Givat-Ram, Jerusalem 91904, Israel; Department of Genetics, Institute of Life Sciences, The Hebrew University, Givat-Ram, Jerusalem 91904, Israel.
² Institute for Stem Cell Research and Regenerative Medicine, Medical Faculty, Heinrich-Heine-University Duesseldorf, Moorenstrasse 5, 40225 Duesseldorf, Germany.
³ The Azrieli Center for Stem Cells and Genetic Research, The Hebrew University, Givat-Ram, Jerusalem 91904, Israel; Department of Genetics, Institute of Life Sciences, The Hebrew University, Givat-Ram, Jerusalem 91904, Israel. Electronic address: [email protected].
⁴ Department of Genetics, Institute of Life Sciences, The Hebrew University, Givat-Ram, Jerusalem 91904, Israel. Electronic address: [email protected].

PMID: 27545893
DOI: 10.1016/j.celrep.2016.07.071

Abstract

Nijmegen breakage syndrome (NBS) results from the absence of the NBS1 protein, responsible for detection of DNA double-strand breaks (DSBs). NBS is characterized by microcephaly, growth retardation, immunodeficiency, and cancer predisposition. Here, we show successful reprogramming of NBS fibroblasts into induced pluripotent stem cells (NBS-iPSCs). Our data suggest a strong selection for karyotypically normal fibroblasts to go through the reprogramming process. NBS-iPSCs then acquire numerous chromosomal aberrations and show a delayed response to DSB induction. Furthermore, NBS-iPSCs display slower growth, mitotic inhibition, a reduced apoptotic response to stress, and abnormal cell-cycle-related gene expression. Importantly, NBS neural progenitor cells (NBS-NPCs) show downregulation of neural developmental genes, which seems to be mediated by P53. Our results demonstrate the importance of NBS1 in early human development, shed light on the molecular mechanisms underlying this severe syndrome, and further expand our knowledge of the genomic stress cells experience during the reprogramming process.

MeSH terms

Acid Anhydride Hydrolases
Base Sequence
Cell Cycle Proteins / genetics*
Cell Cycle Proteins / metabolism
Cell Proliferation
Cellular Reprogramming
Chromosomal Instability*
DNA Breaks, Double-Stranded
DNA Repair
DNA Repair Enzymes / genetics
DNA Repair Enzymes / metabolism
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Fibroblasts / metabolism*
Fibroblasts / pathology
Gene Expression Regulation
Humans
Induced Pluripotent Stem Cells / metabolism*
Induced Pluripotent Stem Cells / pathology
Karyotyping
MRE11 Homologue Protein
Mitosis
Mutation
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism
Neural Stem Cells / metabolism*
Neural Stem Cells / pathology
Nijmegen Breakage Syndrome / genetics*
Nijmegen Breakage Syndrome / metabolism
Nijmegen Breakage Syndrome / pathology
Nuclear Proteins / genetics*
Nuclear Proteins / metabolism
Primary Cell Culture
Signal Transduction
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism

Substances

Cell Cycle Proteins
DNA-Binding Proteins
MRE11 protein, human
NBN protein, human
Nerve Tissue Proteins
Nuclear Proteins
Tumor Suppressor Protein p53
MRE11 Homologue Protein
Acid Anhydride Hydrolases
RAD50 protein, human
DNA Repair Enzymes