Understanding the Radiobiological Mechanisms Induced by 177Lu-DOTATATE in Comparison to External Beam Radiation Therapy
Abstract
:1. Introduction
2. Results
2.1. Intrinsic Radiosensitivity to 177Lu-DOTATATE and EBRT
2.2. Effect of 177Lu-DOTATATE and EBRT on Cell Cycle
2.3. Effect of 177Lu-DOTATATE and EBRT on Apoptosis
2.4. Effect of 177Lu-DOTATATE and EBRT on Autophagy
2.5. Effect of 177Lu-DOTATATE and EBRT on Reactive Oxygen Species (ROS)
2.6. Effect of 177Lu-DOTATATE and EBRT on DNA Damage
2.7. Effect of Olaparib in Combination with 177Lu-DOTATATE or EBRT on Cell Survival
2.8. Absorbed Dose Calculation of 177Lu-DOTATATE Treatment
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Cell Culture
4.2. The 177Lu-DOTATATE Production
4.3. The 177Lu-DOTATATE, EBRT and Olaparib Treatments
- 177Lu-DOTATATE: 5 MBq was added in each well, in four replicates. After 4 h of incubation at 37 °C, the medium containing 177Lu-DOTATATE was removed and replaced with fresh medium.
- EBRT: cells were irradiated at a dose of 2 Gy with a 6 MV beam from a Clinac 600 linear accelerator (Varian Medical Systems, Palo Alto, CA, USA). The collimator opening was set to 40 × 40 cm2, which gave the possibility of irradiating several plates in one batch. In order to achieve a good dose homogeneity and electronic equilibrium, a 6 mm thick polystyrene build-up was put on top of the plates. Plates were placed on a 5 cm thick polystyrene phantom for adequate backscattering conditions. The dose rate was set to 4 Gy per minute. In order to be consistent with 177Lu-DOTATATE treatment conditions, medium was replaced right after EBRT.
4.4. Cell Dosimetry
4.4.1. Geometry Set-Up
4.4.2. Radioactive Source
4.4.3. Absorbed Dose Calculation
4.5. Crystal Violet Assay
4.6. Cell Cycle Analysis
4.7. Apoptosis Analysis
4.8. Autophagy Analysis
4.9. ROS Measurement
4.10. DNA Damage Detection
4.11. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A
Day 0 | Day 3 | Day 7 and 10 | |
---|---|---|---|
HBL | 80,000 | 40,000 | 1000 |
MM162 | 50,000 | 25,000 | 2000 |
COLO-677 | 160,000 | 80,000 | 1000 |
EJM | 160,000 | 80,000 | 4000 |
MIA-PACA-2 | 80,000 | 40,000 | 200 |
HT-29 | 80,000 | 40,000 | 200 |
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CDI | AF (%) | |||
---|---|---|---|---|
177Lu-DOTATATE | EBRT | 177Lu-DOTATATE | EBRT | |
HBL | 0.84 | 0.72 | 20 | 34 |
MM162 | 0.95 | 0.80 | 12 | 26 |
COLO-677 | NA | NA | 0 | 5 |
EJM | NA | NA | 7 | 5 |
MIA-PACA-2 | NA | 0.58 | 0 | 43 |
HT-29 | NA | 0.63 | 4 | 34 |
% of Branching Ratios Used for the Simulations [44] | Energies (keV) | |
---|---|---|
Gamma | 17.3 | 71, 113, 136, 208, 250 and 321 |
X-rays | 8.5 | 9, 55, 56 and 64 |
β particles | 100.0 | 0 to 497 (Emax) |
Auger electrons | 22.1 | 8, 48, 103 and 111 |
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Delbart, W.; Karabet, J.; Marin, G.; Penninckx, S.; Derrien, J.; Ghanem, G.E.; Flamen, P.; Wimana, Z. Understanding the Radiobiological Mechanisms Induced by 177Lu-DOTATATE in Comparison to External Beam Radiation Therapy. Int. J. Mol. Sci. 2022, 23, 12369. https://doi.org/10.3390/ijms232012369
Delbart W, Karabet J, Marin G, Penninckx S, Derrien J, Ghanem GE, Flamen P, Wimana Z. Understanding the Radiobiological Mechanisms Induced by 177Lu-DOTATATE in Comparison to External Beam Radiation Therapy. International Journal of Molecular Sciences. 2022; 23(20):12369. https://doi.org/10.3390/ijms232012369
Chicago/Turabian StyleDelbart, Wendy, Jirair Karabet, Gwennaëlle Marin, Sébastien Penninckx, Jonathan Derrien, Ghanem E. Ghanem, Patrick Flamen, and Zéna Wimana. 2022. "Understanding the Radiobiological Mechanisms Induced by 177Lu-DOTATATE in Comparison to External Beam Radiation Therapy" International Journal of Molecular Sciences 23, no. 20: 12369. https://doi.org/10.3390/ijms232012369