Next Article in Journal
Immunotherapy in Bladder Cancer: Current Methods and Future Perspectives
Next Article in Special Issue
Combined PARP Inhibition and Immune Checkpoint Therapy in Solid Tumors
Previous Article in Journal
On-Treatment Changes in FIB-4 and 1-Year FIB-4 Values Help Identify Patients with Chronic Hepatitis B Receiving Entecavir Therapy Who Have the Lowest Risk of Hepatocellular Carcinoma
Previous Article in Special Issue
ATR Inhibition Potentiates PARP Inhibitor Cytotoxicity in High Risk Neuroblastoma Cell Lines by Multiple Mechanisms
Article

Targeting Nuclear NAD+ Synthesis Inhibits DNA Repair, Impairs Metabolic Adaptation and Increases Chemosensitivity of U-2OS Osteosarcoma Cells

1
Department of Medical Chemistry, Faculty of Medicine, University of Debrecen, H-4032 Debrecen, Hungary, [email protected] (A.K.)
2
Doctoral School of Molecular Medicine, University of Debrecen, H-4032 Debrecen, Hungary
3
Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, H-4032 Debrecen, Hungary, [email protected]
4
Drug Radiation Research Department, National Center for Radiation Research and Technology, Atomic Energy Authority, 113701 Cairo, Egypt
5
MTA-DE Cell Biology and Signaling Research Group, H-4032 Debrecen, Hungary
*
Authors to whom correspondence should be addressed.
Cancers 2020, 12(5), 1180; https://doi.org/10.3390/cancers12051180
Received: 11 February 2020 / Revised: 1 May 2020 / Accepted: 4 May 2020 / Published: 7 May 2020
(This article belongs to the Special Issue PARPs, PAR and NAD Metabolism and Their Inhibitors in Cancer)
Osteosarcoma (OS) is the most common bone tumor in children and adolescents. Modern OS treatment, based on the combination of neoadjuvant chemotherapy (cisplatin + doxorubicin + methotrexate) with subsequent surgical removal of the primary tumor and metastases, has dramatically improved overall survival of OS patients. However, further research is needed to identify new therapeutic targets. Here we report that expression level of the nuclear NAD synthesis enzyme, nicotinamide mononucleotide adenylyltransferase-1 (NMNAT1), increases in U-2OS cells upon exposure to DNA damaging agents, suggesting the involvement of the enzyme in the DNA damage response. Moreover, genetic inactivation of NMNAT1 sensitizes U-2OS osteosarcoma cells to cisplatin, doxorubicin, or a combination of these two treatments. Increased cisplatin-induced cell death of NMNAT1−/− cells showed features of both apoptosis and necroptosis, as indicated by the protective effect of the caspase-3 inhibitor z-DEVD-FMK and the necroptosis inhibitor necrostatin-1. Activation of the DNA damage sensor enzyme poly(ADP-ribose) polymerase 1 (PARP1), a major consumer of NAD+ in the nucleus, was fully blocked by NMNAT1 inactivation, leading to increased DNA damage (phospho-H2AX foci). The PARP inhibitor, olaparib, sensitized wild type but not NMNAT1−/− cells to cisplatin-induced anti-clonogenic effects, suggesting that impaired PARP1 activity is important for chemosensitization. Cisplatin-induced cell death of NMNAT1−/− cells was also characterized by a marked drop in cellular ATP levels and impaired mitochondrial respiratory reserve capacity, highlighting the central role of compromised cellular bioenergetics in chemosensitization by NMNAT1 inactivation. Moreover, NMNAT1 cells also displayed markedly higher sensitivity to cisplatin when grown as spheroids in 3D culture. In summary, our work provides the first evidence that NMNAT1 is a promising therapeutic target for osteosarcoma and possibly other tumors as well. View Full-Text
Keywords: NAD+; NMNAT1; cisplatin; chemotherapy; apoptosis; PARP1; osteosarcoma; cancer NAD+; NMNAT1; cisplatin; chemotherapy; apoptosis; PARP1; osteosarcoma; cancer
Show Figures

Figure 1

MDPI and ACS Style

Kiss, A.; Ráduly, A.P.; Regdon, Z.; Polgár, Z.; Tarapcsák, S.; Sturniolo, I.; El‐Hamoly, T.; Virág, L.; Hegedűs, C. Targeting Nuclear NAD+ Synthesis Inhibits DNA Repair, Impairs Metabolic Adaptation and Increases Chemosensitivity of U-2OS Osteosarcoma Cells. Cancers 2020, 12, 1180. https://doi.org/10.3390/cancers12051180

AMA Style

Kiss A, Ráduly AP, Regdon Z, Polgár Z, Tarapcsák S, Sturniolo I, El‐Hamoly T, Virág L, Hegedűs C. Targeting Nuclear NAD+ Synthesis Inhibits DNA Repair, Impairs Metabolic Adaptation and Increases Chemosensitivity of U-2OS Osteosarcoma Cells. Cancers. 2020; 12(5):1180. https://doi.org/10.3390/cancers12051180

Chicago/Turabian Style

Kiss, Alexandra, Arnold P. Ráduly, Zsolt Regdon, Zsuzsanna Polgár, Szabolcs Tarapcsák, Isotta Sturniolo, Tarek El‐Hamoly, László Virág, and Csaba Hegedűs. 2020. "Targeting Nuclear NAD+ Synthesis Inhibits DNA Repair, Impairs Metabolic Adaptation and Increases Chemosensitivity of U-2OS Osteosarcoma Cells" Cancers 12, no. 5: 1180. https://doi.org/10.3390/cancers12051180

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop