TY - JOUR
T1 - 3D-structured illumination microscopy reveals clustered DNA double-strand break formation in widespread γH2AX foci after high LET heavy-ion particle radiation
AU - Hagiwara, Yoshihiko
AU - Niimi, Atsuko
AU - Isono, Mayu
AU - Yamauchi, Motohiro
AU - Yasuhara, Takaaki
AU - Limsirichaikul, Siripan
AU - Oike, Takahiro
AU - Sato, Hiro
AU - Held, Kathryn D.
AU - Nakano, Takashi
AU - Shibata, Atsushi
N1 - Funding Information:
This work was supported by JSPS KAKENHI Grant Numbers, JP15H02816, JP15K15447 and JP15K15449, and Takeda Science Foundation. This work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan for programs for Leading Graduate Schools, Cultivating Global Leaders in Heavy Ion Therapeutics and Engineering. This work was supported by the Program of the network-type Joint Usage/Research Center for Radiation Disaster Medical Science of Hiroshima University, Nagasaki University, and Fukushima Medical University.
Funding Information:
We thank Yoshimi Omi, Yoko Hayashi, Akiko Shibata and Shiho Nakanishi for assistance with lab work. We thank Drs. Yukari Yoshida and Tatsuaki Kanai for setting up the horizontal carbon-ion irradiation. We thank Dr. Toshiyuki Hatano for setting up the 3D-SIM imaging by OMX. We thank Drs. Ritsuko Watanabe and Kentaro Fujii for fruitful discussion about clustered DSBs formation. This work was done as a part of Research Project with Heavy Ions at GHMC. We would like to thank the GHMC engineering staff for providing support for our heavy-ion experiments. This work was supported by JSPS KAKENHI Grant Numbers, JP15H02816, JP15K15447 and JP15K15449, and Takeda Science Foundation. This work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan for programs for Leading Graduate Schools, Cultivating Global Leaders in Heavy Ion Therapeutics and Engineering. This work was supported by the Program of the network-type Joint Usage/Research Center for Radiation Disaster Medical Science of Hiroshima University, Nagasaki University, and Fukushima Medical University
Publisher Copyright:
© Hagiwara et al.
PY - 2017
Y1 - 2017
N2 - DNA double-strand breaks (DSBs) induced by ionising radiation are considered the major cause of genotoxic mutations and cell death. While DSBs are dispersed throughout chromatin after X-rays or γ-irradiation, multiple types of DNA damage including DSBs, single-strand breaks and base damage can be generated within 1-2 helical DNA turns, defined as a complex DNA lesion, after high Linear Energy Transfer (LET) particle irradiation. In addition to the formation of complex DNA lesions, recent evidence suggests that multiple DSBs can be closely generated along the tracks of high LET particle irradiation. Herein, by using three dimensional (3D)-structured illumination microscopy, we identified the formation of 3D widespread γH2AX foci after high LET carbon-ion irradiation. The large γH2AX foci in G2-phase cells encompassed multiple foci of replication protein A (RPA), a marker of DSBs undergoing resection during homologous recombination. Furthermore, we demonstrated by 3D analysis that the distance between two individual RPA foci within γH2AX foci was approximately 700 nm. Together, our findings suggest that high LET heavy-ion particles induce clustered DSB formation on a scale of approximately 1 μm3. These closely localised DSBs are considered to be a risk for the formation of chromosomal rearrangement after heavy-ion irradiation.
AB - DNA double-strand breaks (DSBs) induced by ionising radiation are considered the major cause of genotoxic mutations and cell death. While DSBs are dispersed throughout chromatin after X-rays or γ-irradiation, multiple types of DNA damage including DSBs, single-strand breaks and base damage can be generated within 1-2 helical DNA turns, defined as a complex DNA lesion, after high Linear Energy Transfer (LET) particle irradiation. In addition to the formation of complex DNA lesions, recent evidence suggests that multiple DSBs can be closely generated along the tracks of high LET particle irradiation. Herein, by using three dimensional (3D)-structured illumination microscopy, we identified the formation of 3D widespread γH2AX foci after high LET carbon-ion irradiation. The large γH2AX foci in G2-phase cells encompassed multiple foci of replication protein A (RPA), a marker of DSBs undergoing resection during homologous recombination. Furthermore, we demonstrated by 3D analysis that the distance between two individual RPA foci within γH2AX foci was approximately 700 nm. Together, our findings suggest that high LET heavy-ion particles induce clustered DSB formation on a scale of approximately 1 μm3. These closely localised DSBs are considered to be a risk for the formation of chromosomal rearrangement after heavy-ion irradiation.
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U2 - 10.18632/oncotarget.22679
DO - 10.18632/oncotarget.22679
M3 - Article
AN - SCOPUS:85037707568
VL - 8
SP - 109370
EP - 109381
JO - Oncotarget
JF - Oncotarget
SN - 1949-2553
IS - 65
ER -