Robust reprogramming of Ataxia-Telangiectasia patient and carrier erythroid cells to induced pluripotent stem cells

Stem Cell Res. 2016 Sep;17(2):296-305. doi: 10.1016/j.scr.2016.08.006. Epub 2016 Aug 12.

Abstract

Biallelic mutations in ATM result in the neurodegenerative syndrome Ataxia-Telangiectasia, while ATM haploinsufficiency increases the risk of cancer and other diseases. Previous studies revealed low reprogramming efficiency from A-T and carrier fibroblasts, a barrier to iPS cell-based modeling and regeneration. Here, we tested the feasibility of employing circulating erythroid cells, a compartment no or minimally affected in A-T, for the generation of A-T and carrier iPS cells. Our results indicate that episomal expression of Yamanaka factors plus BCL-xL in erythroid cells results in highly efficient iPS cell production in feeder-free, xeno-free conditions. Moreover, A-T iPS cells generated with this protocol maintain long-term replicative potential, stable karyotypes, re-elongated telomeres and capability to differentiate along the neural lineage in vitro and to form teratomas in vivo. Finally, we find that haploinsufficiency for ATM does not limit reprogramming from human erythroid cells or in vivo teratoma formation in the mouse.

Keywords: ATM; Ataxia-Telangiectasia; induced pluripotent stem cells; radiation; telomere; teratoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Animals
  • Ataxia Telangiectasia / genetics
  • Ataxia Telangiectasia / metabolism
  • Ataxia Telangiectasia / pathology*
  • Ataxia Telangiectasia Mutated Proteins / genetics
  • Base Sequence
  • Cell Differentiation
  • Cell Line
  • Cell Lineage
  • Cellular Reprogramming*
  • DNA Mutational Analysis
  • Erythroid Cells / cytology*
  • Erythroid Cells / metabolism
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / transplantation
  • Karyotype
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism
  • Telomere / metabolism
  • Telomere Shortening
  • Teratoma / metabolism
  • Teratoma / pathology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Transcription Factors
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins