Novel Insights into RAD52’s Structure, Function, and Druggability for Synthetic Lethality and Innovative Anticancer Therapies
Abstract
:Simple Summary
Abstract
1. Introduction
2. RAD52 Functions
2.1. Homologous Recombination (HR)
2.2. Single-Strand Annealing (SSA)
2.3. Stalled Replication Fork
2.4. RNA-Dependent DNA Repair
2.5. Regulatory Role
3. RAD52’s Structure
3.1. DNA/RNA Binding
3.2. Post-Translational Modification
4. RAD52 in Cancer
5. RAD52 Inhibition in Synthetic Lethality Therapies
5.1. F79 Peptide Aptamer
5.2. 6-OH-DOPA
5.3. A5MP/ZMP
5.4. D-I03
5.5. Epigallocatechin; Epigallocatechin-3-Monogallate; NP-004255
5.6. F779-0434
5.7. Curcumin
5.8. C791-0064
5.9. Quinacrine, Mitoxantrone, Doxorubicine
6. Conclusions and Future Perspectives
Author Contributions
Funding
Conflicts of Interest
References
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Inhibitor | Initial Screening (HTS) | In Vitro Assays | In Cellulo Assays | In Vivo Assays | Group [Ref] |
---|---|---|---|---|---|
F79 aptamer | Rational design after mutagenic assay (ssDNA groove and protomer-protomer interaction) | RAD52-ssDNA (EMSA) | BRCA1/2± cell survival (viability and clonogenic: BRCA1/2− cells over BRCA1/2+ and combination with PARPi and leukemia standard treatments); foci (γH2AX: increase at 5 μM); apoptosis (flow cytometry: increase at 5 μM) | PARPi + F79 (2.5 mg/kg/day) synergic against BRCA1-deficient primary AML xenograft in NSG Mice | T. Skorski [84,96] |
6-OH-DOPA | RAD52-ssDNA (FP: 18,304 cpds – Sigma Lopac) | RAD52-ssDNA (FP: IC50 = 1.6 μM, EMSA); DSB repair (ssDNA annealing); RAD52 binding (ITC: Kd = 17.8 μM); oligomer dissociation (DLS; native gel analysis) | BRCA1/2± cell survival (viability and clonogenic: BRCA1/2− selective over BRCA1/2+); DSB repair (SSA selective over HR, NHEJ); foci (RAD52: decrease at 10 μM, γH2AX: increase at 10 μM); apoptosis (flow cytometry: increase at 20–40 μM); siRNA-RAD52 (western blot) | none | R. T. Pomerantz [65] |
ZMP/A5MP | Virtual screening (docking – PDB: 1KN0 ssDNA groove: 1217 FDA and 139,735 NCI drug-like cpds – ZINC library, DOCK6.6 software) | RAD52-ssDNA (EMSA) | BRCA1/2± cell survival (growth rate: BRCA1/2− selective over BRCA1/2+); DSB repair (SSA: decrease at 20 μM); foci (RAD52: decrease at 20 μM) | none | T. Skorski [104] |
D-I03 | DSB repair (ssDNA: 93,672 cpds from Diversity Oriented Synthesis (DOS) library and 279,231 cpds from Molecular Libraries Probe Center Network (MPLCN) - Broad Institute) | DSB repair (ssDNA annealing: IC50 = 5 μM, D-loop: IC50 = 8 μM); RAD52 binding (SPR: Kd = 25.8 μM) | BRCA1/2± cell survival (viability and clonogenic: IC50 = 14.5 μM for BRCA1/2− cells over BRCA1/2+ and combination with PARPi); foci (2.5 μM ≈ 2.0-fold reduction of RAD52 and not of RAD51); DSB repair (SSA, 30 μM ≈ 3.4-fold reduction and selective over HR) | PARPi (talazoparib)+D-I03 (50 mg/kg/day) synergic against BRCA1-deficient solid tumor growth in nude mice | A. V. Mazin [96,106] |
Epigallocatechines (NP-004255) | RAD52-ssDNA (FRET: 2320 cpds from MicroSource SPECTRUM collection); Virtual screening (docking – PDB: 1KN0 ssDNA groove: AnalytiCon Discovery MEGx Natural Products Screen Library) | RAD52-ssDNA (FRET: IC50 = 1.5 μM); DSB repair (ssDNA annealing: IC50 = 7 μM); RAD52 binding (WaterLOGSY); | BRCA2± cell survival (viability); DSB repair (comet assay); siRNA-RAD52 (western blot) | none | M. A. Spies; M. Spies [108] |
F779-0434 | Virtual screening (docking – PDB: 5JRB ssDNA groove: 47,737 cpds Targeted Diversity Library (TDL) – Chemdiv database, DOCK6.5 software) | RAD52-ssDNA (pull-down assay: 50% disruption at 5 μM); | BRCA2± cell survival (viability: BRCA2− selective over BRCA2+, 50% growth inhibition at 10 μM) | none | R. Sun; Q. Zhao [107] |
C791-0064 | Virtual screening (docking – PDB: 5JRB self-association pocket: 66,608 cpds - Chemdiv database, DOCK6.5 software) | RAD52-ssDNA (EMSA); DSB repair (ssDNA annealing); RAD52 binding (MST); | BRCA2± cell survival (viability and clonogenic: IC50 = 29 μM for BRCA2− cells over BRCA+); siRNA-RAD52 and shRNA-BRCA (western blot); foci (γH2AX: increase at 40 μM); apoptosis (flow cytometry: increase at 40 μM, western blot) | none | J. Li [105] |
mitoxantrone | RPA:RAD51 PPIs (FluorIA: 335 cpds – SellekChem, 1200 cpds - Prestwick, 100,000 cpds – Chembridge: EC50 = 30 μM) | - | BRCA1/2± cell survival (viability: EC50 = 0.1–2.5 μM for BRCA2− cells over BRCA+); DSB repair (SSA reduction and selective over HR up to 6 nM); apoptosis (western blot: increase at 0.2 μM); foci (RAD52: decrease at 3 nM over RAD51); RPA:RAD51 PPIs (co-immunoprecipitation) | none | T. Bessho; G. E. O. Borgstahl [109] |
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Balboni, B.; Rinaldi, F.; Previtali, V.; Ciamarone, A.; Girotto, S.; Cavalli, A. Novel Insights into RAD52’s Structure, Function, and Druggability for Synthetic Lethality and Innovative Anticancer Therapies. Cancers 2023, 15, 1817. https://doi.org/10.3390/cancers15061817
Balboni B, Rinaldi F, Previtali V, Ciamarone A, Girotto S, Cavalli A. Novel Insights into RAD52’s Structure, Function, and Druggability for Synthetic Lethality and Innovative Anticancer Therapies. Cancers. 2023; 15(6):1817. https://doi.org/10.3390/cancers15061817
Chicago/Turabian StyleBalboni, Beatrice, Francesco Rinaldi, Viola Previtali, Andrea Ciamarone, Stefania Girotto, and Andrea Cavalli. 2023. "Novel Insights into RAD52’s Structure, Function, and Druggability for Synthetic Lethality and Innovative Anticancer Therapies" Cancers 15, no. 6: 1817. https://doi.org/10.3390/cancers15061817