Decrease in Abundance of apurinic/apyrimidinic Endonuclease
Causes Failure of Base Excision Repair in Culture-Adapted Human
Embryonic Stem Cells

J 2013

Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells

KRUTÁ, Miriama, Lukáš BÁLEK, Renata HEJNOVÁ, Zuzana KUNICKÁ, Lívia EISELLEOVÁ et. al.

Basic information

Original name

Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells

Authors

KRUTÁ, Miriama (703 Slovakia, belonging to the institution), Lukáš BÁLEK (203 Czech Republic, belonging to the institution), Renata HEJNOVÁ (203 Czech Republic, belonging to the institution), Zuzana KUNICKÁ (203 Czech Republic, belonging to the institution), Lívia EISELLEOVÁ (703 Slovakia, belonging to the institution), Kamil MATULKA (203 Czech Republic, belonging to the institution), Tomáš BÁRTA (203 Czech Republic, belonging to the institution), Petr FOJTÍK (203 Czech Republic, belonging to the institution), Jiří FAJKUS (203 Czech Republic, belonging to the institution), Aleš HAMPL (203 Czech Republic, belonging to the institution), Petr DVOŘÁK (203 Czech Republic, belonging to the institution) and Vladimír ROTREKL (203 Czech Republic, guarantor, belonging to the institution)

Edition

Stem Cells, UNITED STATES, WILEY-BLACKWELL, 2013, 1066-5099

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 7.133

RIV identification code

RIV/00216224:14110/13:00065577

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1002/stem.1312

UT WoS

000316624300008

Keywords in English

APE1; base excision repair; human embryonic stem cells; culture adaptation; genome instability

Abstract

V originále

The inevitable accumulation of chromosomal abnormalities in human embryonic stem cells (hESCs) during in vitro expansion represents a considerable obstacle for cell replacement therapies. To determine the source of chromosomal abnormalities, we examined hESCs maintained in culture for over 55 months for defects in telomere maintenance and DNA repair. Although prolonged culture affected neither telomerase activity nor nonhomologous end joining, the efficiency of base excision repair (BER) was significantly decreased and correlated with reduced expression of apurinic/apyrimidinic endonuclease 1 (APE1), the major nuclease required for BER. Interestingly, the expression of other BER enzymes was unchanged. Addition of human recombinant APE1 protein to nuclear extracts from late passage hESCs increased BER efficiency to the level typical of early passage hESCs. The link between BER and double-strand breaks (DSB) was demonstrated by decreased DSB release after downregulation of APE1 in early passage hESCs via siRNA. Correspondingly lower APE1 level in late passage hESC resulted in slower and less intensive but long lasting DSB release upon ionizing radiation (IR). Downregulation of APE1 in early passage hESCs also led to approximately 30% decrease in -H2AX signaling following IR, similar to that in late passage hESCs. We suggest that downregulation of APE1 significantly contributes to the failure of BER during long-term culture of hESCs, and further that BER failure is one of the factors affecting the genomic instability of hESCs by altering BER-dependent DSB release and cell cycle/checkpoint signaling.

Links

GBP302/12/G157, research and development project

Name: Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

Investor: Czech Science Foundation

MSM0021622430, plan (intention)

Name: Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie

Investor: Ministry of Education, Youth and Sports of the CR, Functional and molecular characteristics of cancer and normal stem cells - identification of targets for novel therapeutics and therapeutic strategies

NS10237, research and development project

Name: Identifikace mechanizmů vzniku chromozomálních aberací při dlouhodobé kultivaci lidských embryonálních buněk in vitro

Investor: Ministry of Health of the CR

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Citovat

KRUTÁ, Miriama, Lukáš BÁLEK, Renata HEJNOVÁ, Zuzana KUNICKÁ, Lívia EISELLEOVÁ, Kamil MATULKA, Tomáš BÁRTA, Petr FOJTÍK, Jiří FAJKUS, Aleš HAMPL, Petr DVOŘÁK and Vladimír ROTREKL. Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells. Stem Cells. UNITED STATES: WILEY-BLACKWELL, 2013, vol. 31, No 4, p. 693-702. ISSN 1066-5099. Available from: https://dx.doi.org/10.1002/stem.1312.

@article{1081531,
   author = {Krutá, Miriama and Bálek, Lukáš and Hejnová, Renata and Kunická, Zuzana and Eiselleová, Lívia and Matulka, Kamil and Bárta, Tomáš and Fojtík, Petr and Fajkus, Jiří and Hampl, Aleš and Dvořák, Petr and Rotrekl, Vladimír},
   article_location = {UNITED STATES},
   article_number = {4},
   doi = {http://dx.doi.org/10.1002/stem.1312},
   keywords = {APE1; base excision repair; human embryonic stem cells; culture adaptation; genome instability},
   language = {eng},
   issn = {1066-5099},
   journal = {Stem Cells},
   note = {Projekt LSHG-CT-2006-018739 (PLATFORMS FOR BIOMEDICAL DISCOVERY WITH HUMAN ES CELLS) za ČR: AV ČR},
   title = {Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells},
   url = {http://onlinelibrary.wiley.com/doi/10.1002/stem.1312/citedby},
   volume = {31},
   year = {2013}
}

TY  - JOUR
ID  - 1081531
AU  - Krutá, Miriama - Bálek, Lukáš - Hejnová, Renata - Kunická, Zuzana - Eiselleová, Lívia - Matulka, Kamil - Bárta, Tomáš - Fojtík, Petr - Fajkus, Jiří - Hampl, Aleš - Dvořák, Petr - Rotrekl, Vladimír
PY  - 2013
TI  - Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells
JF  - Stem Cells
VL  - 31
IS  - 4
SP  - 693-702
EP  - 693-702
PB  - WILEY-BLACKWELL
SN  - 10665099
N1  - Projekt LSHG-CT-2006-018739 (PLATFORMS FOR BIOMEDICAL DISCOVERY WITH HUMAN ES CELLS) za ČR: AV ČR
KW  - APE1
KW  - base excision repair
KW  - human embryonic stem cells
KW  - culture adaptation
KW  - genome instability
UR  - http://onlinelibrary.wiley.com/doi/10.1002/stem.1312/citedby
N2  - The inevitable accumulation of chromosomal abnormalities in human embryonic stem cells (hESCs) during in vitro expansion represents a considerable obstacle for cell replacement therapies. To determine the source of chromosomal abnormalities, we examined hESCs maintained in culture for over 55 months for defects in telomere maintenance and DNA repair. Although prolonged culture affected neither telomerase activity nor nonhomologous end joining, the efficiency of base excision repair (BER) was significantly decreased and correlated with reduced expression of apurinic/apyrimidinic endonuclease 1 (APE1), the major nuclease required for BER. Interestingly, the expression of other BER enzymes was unchanged. Addition of human recombinant APE1 protein to nuclear extracts from late passage hESCs increased BER efficiency to the level typical of early passage hESCs. The link between BER and double-strand breaks (DSB) was demonstrated by decreased DSB release after downregulation of APE1 in early passage hESCs via siRNA. Correspondingly lower APE1 level in late passage hESC resulted in slower and less intensive but long lasting DSB release upon ionizing radiation (IR). Downregulation of APE1 in early passage hESCs also led to approximately 30% decrease in -H2AX signaling following IR, similar to that in late passage hESCs. We suggest that downregulation of APE1 significantly contributes to the failure of BER during long-term culture of hESCs, and further that BER failure is one of the factors affecting the genomic instability of hESCs by altering BER-dependent DSB release and cell cycle/checkpoint signaling.
ER  -

KRUTÁ, Miriama, Lukáš BÁLEK, Renata HEJNOVÁ, Zuzana KUNICKÁ, Lívia EISELLEOVÁ, Kamil MATULKA, Tomáš BÁRTA, Petr FOJTÍK, Jiří FAJKUS, Aleš HAMPL, Petr DVOŘÁK and Vladimír ROTREKL. Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells. \textit{Stem Cells}. UNITED STATES: WILEY-BLACKWELL, 2013, vol.~31, No~4, p.~693-702. ISSN~1066-5099. Available from: https://dx.doi.org/10.1002/stem.1312.

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