Repositioning disulfiram as a radiosensitizer against atypical teratoid/rhabdoid tumor.
BACKGROUND
Atypical teratoid/rhabdoid tumor (AT/RT) is one of the most common malignant brain tumors in infants. Although cancer stem cells of AT/RT express aldehyde dehydrogenase (ALDH), effective chemotherapies against AT/RT have not been established. Here, we examined radiosensitizing effects of disulfiram (DSF), an irreversible inhibitor of ALDH against AT/RT for a novel therapeutic method.


METHODS
Patient-derived primary cultured AT/RT cells (SNU.AT/RT-5 and SNU.AT/RT-6) and established AT/RT cell lines (BT-12 and BT-16) were used to assess therapeutic effects of combining DSF with radiation treatment (RT). Survival fraction by clonogenic assay, protein expression, immunofluorescence, and autophagy analysis were evaluated in vitro. Antitumor effects of combining DSF with RT were verified by bioluminescence imaging, tumor volume, and survival analysis in vivo.


RESULTS
The results demonstrated that DSF at low concentration enhanced the radiosensitivity of AT/RT cells with reduction of survival fraction to 1.21‒1.58. DSF increased DNA double-strand break (γ-H2AX, p-DNA-PKcs, and p-ATM), apoptosis (cleaved caspase-3), autophagy (LC3B), and cell cycle arrest (p21) in irradiated AT/RT cells, while it decreased anti-apoptosis (nuclear factor-kappaB, Survivin, and B-cell lymphoma 2 [Bcl2]). In vivo, DSF and RT combined treatment significantly reduced tumor volumes and prolonged the survival of AT/RT mouse models compared with single treatments. The combined treatment also increased γ-H2AX, cleaved caspase-3, and LC3B expression and decreased ALDH1, Survivin, and Bcl2 expression in vivo.


CONCLUSIONS
DSF and RT combination therapy has additive therapeutic effects on AT/RT by potentiating programmed cell death, including apoptosis and autophagy of AT/RT cells. We suggest that DSF can be applied as a radiosensitizer in AT/RT treatment.
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Young Eun
YE
Lee
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Seung Ah
SA
Choi
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Pil Ae
PA
Kwack
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Hak Jae
HJ
Kim
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Il Han
IH
Kim
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Kyu-Chang
KC
Wang
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Ji Hoon
JH
Phi
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Ji Yeoun
JY
Lee
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Sangjoon
S
Chong
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Sung-Hye
SH
Park
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Kyung Duk
KD
Park
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Do Won
DW
Hwang
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Kyeung Min
KM
Joo
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, South Korea; Adolescent Cancer Center, Seoul National University Cancer Hospital, Seoul, South Korea; Department of Radiation Oncology, Seoul National University Hospital, Seoul, South Korea; Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea; Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea; Department of Pediatrics, Seoul National University Children's Hospital, Seoul, South Korea; Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea; Department of Health Science and Technology, Samsung Advanced Institute for Health Sciences and Technology, SungKyunKwan University, Seoul, South Korea; Department of Anatomy and Cell Biology, SungKyunKwan University School of Medicine, Suwon, South Korea; Single Cell Network Research Center, Sungkyunkwan University School of Medicine, Suwon, South Korea.
Seung-Ki
SK
Kim
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000473:pathology; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004221:Disulfiram / Q000494:pharmacology*; D006801:Humans; D007223:Infant; D008297:Male; D051379:Mice; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D011836:Radiation Tolerance / Q000187:drug effects; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D018335:Rhabdoid Tumor / Q000188:drug therapy* / Q000378:metabolism; D013724:Teratoma / Q000188:drug therapy*; D023041:Xenograft Model Antitumor Assays / Q000379:methods
D011838:Radiation-Sensitizing Agents; D004221:Disulfiram
AT/RT; DNA damage; DSF; radiation therapy; radiosensitizer