Apoptotic susceptibility to DNA damage of pluripotent stem cells facilitates pharmacologic purging of teratoma risk

Alyson J. Smith, Natalie G. Nelson, Saji Oommen, Katherine A. Hartjes, Clifford Folmes, Andre Terzic, Timothy J Nelson

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Pluripotent stem cells have been the focus of bioengineering efforts designed to generate regenerative products, yet harnessing therapeutic capacity while minimizing risk of dysregulated growth remains a challenge. The risk of residual undifferentiated stem cells within a differentiated progenitor population requires a targeted approach to eliminate contaminating cells prior to delivery. In this study we aimed to validate a toxicity strategy that could selectively purge pluripotent stem cells in response to DNA damage and avoid risk of uncontrolled cell growth upon transplantation. Compared with somatic cell types, embryonic stem cells and induced pluripotent stem cells displayed hypersensitivity to apoptotic induction by genotoxic agents. Notably, hypersensitivity in pluripotent stem cells was stage-specific and consistently lost upon in vitro differentiation, with the mean half-maximal inhibitory concentration increasing nearly 2 orders of magnitude with tissue specification. Quantitative polymerase chain reaction and Western blotting demonstrated that the innate response was mediated through upregulation of the BH3-only protein Puma in both natural and induced pluripotent stem cells. Pretreatment with genotoxic etoposide purged hypersensitive pluripotent stem cells to yield a progenitor population refractory to teratoma formation upon transplantation. Collectively, this study exploits a hypersensitive apoptotic response to DNA damage within pluripotent stem cells to decrease risk of dysregulated growth and augment the safety profile of transplant-ready, bioengineered progenitor cells.

Original languageEnglish (US)
Pages (from-to)709-718
Number of pages10
JournalStem cells translational medicine
Volume1
Issue number10
DOIs
StatePublished - 2012

Fingerprint

Pluripotent Stem Cells
Teratoma
DNA Damage
Induced Pluripotent Stem Cells
Hypersensitivity
Stem Cells
Growth
Transplantation
Puma
Bioengineering
Etoposide
Embryonic Stem Cells
Population
Up-Regulation
Western Blotting
Transplants
Safety
Polymerase Chain Reaction
Proteins

Keywords

  • Apoptosis
  • Pluripotent stem cells
  • Stem cell transplantation
  • Stem cells
  • Tumor cell purging

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology

Cite this

Apoptotic susceptibility to DNA damage of pluripotent stem cells facilitates pharmacologic purging of teratoma risk. / Smith, Alyson J.; Nelson, Natalie G.; Oommen, Saji; Hartjes, Katherine A.; Folmes, Clifford; Terzic, Andre; Nelson, Timothy J.

In: Stem cells translational medicine, Vol. 1, No. 10, 2012, p. 709-718.

Research output: Contribution to journalArticle

@article{a56427bd0daf4d38a524d5fa816f9d7b,
title = "Apoptotic susceptibility to DNA damage of pluripotent stem cells facilitates pharmacologic purging of teratoma risk",
abstract = "Pluripotent stem cells have been the focus of bioengineering efforts designed to generate regenerative products, yet harnessing therapeutic capacity while minimizing risk of dysregulated growth remains a challenge. The risk of residual undifferentiated stem cells within a differentiated progenitor population requires a targeted approach to eliminate contaminating cells prior to delivery. In this study we aimed to validate a toxicity strategy that could selectively purge pluripotent stem cells in response to DNA damage and avoid risk of uncontrolled cell growth upon transplantation. Compared with somatic cell types, embryonic stem cells and induced pluripotent stem cells displayed hypersensitivity to apoptotic induction by genotoxic agents. Notably, hypersensitivity in pluripotent stem cells was stage-specific and consistently lost upon in vitro differentiation, with the mean half-maximal inhibitory concentration increasing nearly 2 orders of magnitude with tissue specification. Quantitative polymerase chain reaction and Western blotting demonstrated that the innate response was mediated through upregulation of the BH3-only protein Puma in both natural and induced pluripotent stem cells. Pretreatment with genotoxic etoposide purged hypersensitive pluripotent stem cells to yield a progenitor population refractory to teratoma formation upon transplantation. Collectively, this study exploits a hypersensitive apoptotic response to DNA damage within pluripotent stem cells to decrease risk of dysregulated growth and augment the safety profile of transplant-ready, bioengineered progenitor cells.",
keywords = "Apoptosis, Pluripotent stem cells, Stem cell transplantation, Stem cells, Tumor cell purging",
author = "Smith, {Alyson J.} and Nelson, {Natalie G.} and Saji Oommen and Hartjes, {Katherine A.} and Clifford Folmes and Andre Terzic and Nelson, {Timothy J}",
year = "2012",
doi = "10.5966/sctm.2012-0066",
language = "English (US)",
volume = "1",
pages = "709--718",
journal = "Stem cells translational medicine",
issn = "2157-6564",
publisher = "AlphaMed Press",
number = "10",

}

TY - JOUR

T1 - Apoptotic susceptibility to DNA damage of pluripotent stem cells facilitates pharmacologic purging of teratoma risk

AU - Smith, Alyson J.

AU - Nelson, Natalie G.

AU - Oommen, Saji

AU - Hartjes, Katherine A.

AU - Folmes, Clifford

AU - Terzic, Andre

AU - Nelson, Timothy J

PY - 2012

Y1 - 2012

N2 - Pluripotent stem cells have been the focus of bioengineering efforts designed to generate regenerative products, yet harnessing therapeutic capacity while minimizing risk of dysregulated growth remains a challenge. The risk of residual undifferentiated stem cells within a differentiated progenitor population requires a targeted approach to eliminate contaminating cells prior to delivery. In this study we aimed to validate a toxicity strategy that could selectively purge pluripotent stem cells in response to DNA damage and avoid risk of uncontrolled cell growth upon transplantation. Compared with somatic cell types, embryonic stem cells and induced pluripotent stem cells displayed hypersensitivity to apoptotic induction by genotoxic agents. Notably, hypersensitivity in pluripotent stem cells was stage-specific and consistently lost upon in vitro differentiation, with the mean half-maximal inhibitory concentration increasing nearly 2 orders of magnitude with tissue specification. Quantitative polymerase chain reaction and Western blotting demonstrated that the innate response was mediated through upregulation of the BH3-only protein Puma in both natural and induced pluripotent stem cells. Pretreatment with genotoxic etoposide purged hypersensitive pluripotent stem cells to yield a progenitor population refractory to teratoma formation upon transplantation. Collectively, this study exploits a hypersensitive apoptotic response to DNA damage within pluripotent stem cells to decrease risk of dysregulated growth and augment the safety profile of transplant-ready, bioengineered progenitor cells.

AB - Pluripotent stem cells have been the focus of bioengineering efforts designed to generate regenerative products, yet harnessing therapeutic capacity while minimizing risk of dysregulated growth remains a challenge. The risk of residual undifferentiated stem cells within a differentiated progenitor population requires a targeted approach to eliminate contaminating cells prior to delivery. In this study we aimed to validate a toxicity strategy that could selectively purge pluripotent stem cells in response to DNA damage and avoid risk of uncontrolled cell growth upon transplantation. Compared with somatic cell types, embryonic stem cells and induced pluripotent stem cells displayed hypersensitivity to apoptotic induction by genotoxic agents. Notably, hypersensitivity in pluripotent stem cells was stage-specific and consistently lost upon in vitro differentiation, with the mean half-maximal inhibitory concentration increasing nearly 2 orders of magnitude with tissue specification. Quantitative polymerase chain reaction and Western blotting demonstrated that the innate response was mediated through upregulation of the BH3-only protein Puma in both natural and induced pluripotent stem cells. Pretreatment with genotoxic etoposide purged hypersensitive pluripotent stem cells to yield a progenitor population refractory to teratoma formation upon transplantation. Collectively, this study exploits a hypersensitive apoptotic response to DNA damage within pluripotent stem cells to decrease risk of dysregulated growth and augment the safety profile of transplant-ready, bioengineered progenitor cells.

KW - Apoptosis

KW - Pluripotent stem cells

KW - Stem cell transplantation

KW - Stem cells

KW - Tumor cell purging

UR - http://www.scopus.com/inward/record.url?scp=84876510977&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84876510977&partnerID=8YFLogxK

U2 - 10.5966/sctm.2012-0066

DO - 10.5966/sctm.2012-0066

M3 - Article

VL - 1

SP - 709

EP - 718

JO - Stem cells translational medicine

JF - Stem cells translational medicine

SN - 2157-6564

IS - 10

ER -