Exploiting DNA Ligase III addiction of multiple myeloma by flavonoid Rhamnetin.
DNA Ligase III
DNA repair
Flavonoid
Genomic Instability
LIG3
Multiple myeloma
Natural compounds
Polyphenols
Rhamnetin
Journal
Journal of translational medicine
ISSN: 1479-5876
Titre abrégé: J Transl Med
Pays: England
ID NLM: 101190741
Informations de publication
Date de publication:
22 10 2022
22 10 2022
Historique:
received:
16
09
2022
accepted:
11
10
2022
entrez:
23
10
2022
pubmed:
24
10
2022
medline:
26
10
2022
Statut:
epublish
Résumé
DNA ligases are crucial for DNA repair and cell replication since they catalyze the final steps in which DNA breaks are joined. DNA Ligase III (LIG3) exerts a pivotal role in Alternative-Non-Homologous End Joining Repair (Alt-NHEJ), an error-prone DNA repair pathway often up-regulated in genomically unstable cancer, such as Multiple Myeloma (MM). Based on the three-dimensional (3D) LIG3 structure, we performed a computational screening to identify LIG3-targeting natural compounds as potential candidates to counteract Alt-NHEJ activity in MM. Virtual screening was conducted by interrogating the Phenol Explorer database. Validation of binding to LIG3 recombinant protein was performed by Saturation Transfer Difference (STD)-nuclear magnetic resonance (NMR) experiments. Cell viability was analyzed by Cell Titer-Glo assay; apoptosis was evaluated by flow cytometric analysis following Annexin V-7AAD staining. Alt-NHEJ repair modulation was evaluated using plasmid re-joining assay and Cytoscan HD. DNA Damage Response protein levels were analyzed by Western blot of whole and fractionated protein extracts and immunofluorescence analysis. The mitochondrial DNA (mtDNA) copy number was determined by qPCR. In vivo activity was evaluated in NOD-SCID mice subcutaneously engrafted with MM cells. Here, we provide evidence that a natural flavonoid Rhamnetin (RHM), selected by a computational approach, counteracts LIG3 activity and killed Alt-NHEJ-dependent MM cells. Indeed, Nuclear Magnetic Resonance (NMR) showed binding of RHM to LIG3 protein and functional experiments revealed that RHM interferes with LIG3-driven nuclear and mitochondrial DNA repair, leading to significant anti-MM activity in vitro and in vivo. Taken together, our findings provide proof of concept that RHM targets LIG3 addiction in MM and may represent therefore a novel promising anti-tumor natural agent to be investigated in an early clinical setting.
Sections du résumé
BACKGROUND
DNA ligases are crucial for DNA repair and cell replication since they catalyze the final steps in which DNA breaks are joined. DNA Ligase III (LIG3) exerts a pivotal role in Alternative-Non-Homologous End Joining Repair (Alt-NHEJ), an error-prone DNA repair pathway often up-regulated in genomically unstable cancer, such as Multiple Myeloma (MM). Based on the three-dimensional (3D) LIG3 structure, we performed a computational screening to identify LIG3-targeting natural compounds as potential candidates to counteract Alt-NHEJ activity in MM.
METHODS
Virtual screening was conducted by interrogating the Phenol Explorer database. Validation of binding to LIG3 recombinant protein was performed by Saturation Transfer Difference (STD)-nuclear magnetic resonance (NMR) experiments. Cell viability was analyzed by Cell Titer-Glo assay; apoptosis was evaluated by flow cytometric analysis following Annexin V-7AAD staining. Alt-NHEJ repair modulation was evaluated using plasmid re-joining assay and Cytoscan HD. DNA Damage Response protein levels were analyzed by Western blot of whole and fractionated protein extracts and immunofluorescence analysis. The mitochondrial DNA (mtDNA) copy number was determined by qPCR. In vivo activity was evaluated in NOD-SCID mice subcutaneously engrafted with MM cells.
RESULTS
Here, we provide evidence that a natural flavonoid Rhamnetin (RHM), selected by a computational approach, counteracts LIG3 activity and killed Alt-NHEJ-dependent MM cells. Indeed, Nuclear Magnetic Resonance (NMR) showed binding of RHM to LIG3 protein and functional experiments revealed that RHM interferes with LIG3-driven nuclear and mitochondrial DNA repair, leading to significant anti-MM activity in vitro and in vivo.
CONCLUSION
Taken together, our findings provide proof of concept that RHM targets LIG3 addiction in MM and may represent therefore a novel promising anti-tumor natural agent to be investigated in an early clinical setting.
Identifiants
pubmed: 36273153
doi: 10.1186/s12967-022-03705-z
pii: 10.1186/s12967-022-03705-z
pmc: PMC9588242
doi:
Substances chimiques
Annexin A5
0
DNA Ligase ATP
EC 6.5.1.1
DNA Ligases
EC 6.5.1.-
DNA, Mitochondrial
0
Flavonoids
0
Phenols
0
Recombinant Proteins
0
rhamnetin
71803L5F4S
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
482Informations de copyright
© 2022. The Author(s).
Références
Nat Med. 2015 Jun;21(6):572-80
pubmed: 26005854
J Hematol Oncol. 2019 Mar 21;12(1):32
pubmed: 30898137
Cell Death Dis. 2017 Dec 13;8(12):3208
pubmed: 29238067
Int J Cancer. 2020 Nov 15;147(10):2658-2668
pubmed: 32383203
Cell Death Dis. 2012 Nov 29;3:e436
pubmed: 23190608
PLoS Genet. 2008 Jun 27;4(6):e1000110
pubmed: 18584027
Mutat Res Genet Toxicol Environ Mutagen. 2015 Nov;793:166-75
pubmed: 26520387
Biomed Res Int. 2016;2016:2346585
pubmed: 27642590
Mol Pharm. 2014 Jan 6;11(1):283-93
pubmed: 24256025
Cell. 2011 Mar 4;144(5):646-74
pubmed: 21376230
J Transl Med. 2017 Dec 21;15(1):261
pubmed: 29268770
Mol Cancer Res. 2012 Jan;10(1):96-107
pubmed: 22112941
Cancers (Basel). 2021 May 29;13(11):
pubmed: 34072312
Nat Rev Mol Cell Biol. 2010 Mar;11(3):220-8
pubmed: 20177397
N Engl J Med. 2022 Aug 11;387(6):558-561
pubmed: 35947712
Transl Cancer Res. 2013 Jun;2(3):
pubmed: 24224145
Eur J Med Chem. 2019 Oct 15;180:465-485
pubmed: 31330448
Clin Cancer Res. 2012 Apr 1;18(7):1888-900
pubmed: 22351691
N Engl J Med. 2021 Feb 25;384(8):705-716
pubmed: 33626253
Haematologica. 2020 Feb 20;106(1):185-195
pubmed: 32079692
J Transl Med. 2018 Mar 20;16(1):75
pubmed: 29558948
Clin Pharmacol Ther. 2019 Aug;106(2):422-431
pubmed: 30739312
J Transl Med. 2020 May 12;18(1):195
pubmed: 32398139
Clin Transl Sci. 2021 Jan;14(1):113-119
pubmed: 33089968
Mol Cancer Res. 2016 Apr;14(4):363-73
pubmed: 26850462
J Transl Med. 2016 Jan 14;14:13
pubmed: 26762586
Eur J Med Chem. 2019 Dec 1;183:111715
pubmed: 31550663
J Biochem. 2003 Jun;133(6):757-65
pubmed: 12869532
Genome Med. 2017 Apr 19;9(1):34
pubmed: 28420421
J Biol Chem. 2013 Sep 20;288(38):27343-27357
pubmed: 23902763
Nat Rev Clin Oncol. 2018 Sep;15(9):564-576
pubmed: 29955114
Lancet Oncol. 2019 Jan;20(1):e15-e28
pubmed: 30614472
J Agric Food Chem. 2016 Apr 20;64(15):3048-53
pubmed: 27045836
Curr Cancer Drug Targets. 2012 Sep;12(7):757-67
pubmed: 22671925
J Transl Med. 2016 Jun 23;14(1):186
pubmed: 27338244
J Transl Med. 2022 Feb 10;20(1):82
pubmed: 35144648
J Chem Theory Comput. 2019 Apr 9;15(4):2734-2742
pubmed: 30807148
Biochemistry. 2010 Jul 27;49(29):6165-76
pubmed: 20518483
Cancer Res. 2008 May 1;68(9):3169-77
pubmed: 18451142
Lancet Haematol. 2021 Feb;8(2):e122-e134
pubmed: 33347814
High Throughput. 2018 Oct 26;7(4):
pubmed: 30373182
Front Oncol. 2014 Feb 28;4:42
pubmed: 24616882
Nat Cancer. 2021 Jun;2(6):598-610
pubmed: 34179826
J Transl Med. 2014 Aug 03;12:219
pubmed: 25086598
Oxid Med Cell Longev. 2009 Nov-Dec;2(5):270-8
pubmed: 20716914
Cancers (Basel). 2021 Mar 30;13(7):
pubmed: 33808190
Dev Cell. 2021 Sep 13;56(17):2427-2439.e4
pubmed: 34352222
J Exp Clin Cancer Res. 2020 Jun 20;39(1):117
pubmed: 32563270
Blood. 2014 Apr 10;123(15):2355-66
pubmed: 24505083
Nutrients. 2016 Aug 22;8(8):
pubmed: 27556486
Clin Cancer Res. 2013 Apr 15;19(8):2096-106
pubmed: 23446999
Br J Haematol. 2008 Nov;143(4):520-31
pubmed: 18986388
Leukemia. 2019 Feb;33(2):487-498
pubmed: 30120376
J Transl Med. 2015 Jul 08;13:218
pubmed: 26152229
Mol Cancer Res. 2015 Mar;13(3):470-82
pubmed: 25563294
Curr Pharm Des. 2004;10(9):1011-33
pubmed: 15078130
Genome Biol. 2002;3(4):REVIEWS3005
pubmed: 11983065
Chem Rev. 2006 Feb;106(2):687-99
pubmed: 16464020
Nutrients. 2010 Nov;2(11):1106-31
pubmed: 22254000
Clin Cancer Res. 2018 Feb 1;24(3):696-707
pubmed: 29138344
Expert Opin Biol Ther. 2013 Jun;13 Suppl 1:S125-37
pubmed: 23692413
J Nutr Sci. 2016 Dec 29;5:e47
pubmed: 28620474
High Throughput. 2018 Dec 18;7(4):
pubmed: 30567415
J Transl Med. 2021 Nov 4;19(1):457
pubmed: 34736494
Mol Cancer Ther. 2009 Jan;8(1):26-35
pubmed: 19139110
Cancers (Basel). 2021 Mar 19;13(6):
pubmed: 33808562
Cancers (Basel). 2021 Aug 29;13(17):
pubmed: 34503175
Cancer Res. 2016 Sep 15;76(18):5431-41
pubmed: 27503931
Glycobiology. 2018 Dec 1;28(1):42-49
pubmed: 29087468
PLoS Genet. 2011 Jun;7(6):e1002080
pubmed: 21655080
Mol Cancer Res. 2015 Apr;13(4):699-712
pubmed: 25828893
Nat Genet. 2013 Sep;45(9):977-83
pubmed: 23852168
Nucleic Acids Res. 2000 Sep 15;28(18):3558-63
pubmed: 10982876
Molecules. 2021 Feb 19;26(4):
pubmed: 33669817
Expert Rev Hematol. 2018 Oct;11(10):793-803
pubmed: 30148649
Nat Rev Clin Oncol. 2019 Feb;16(2):81-104
pubmed: 30356138
Oncol Lett. 2019 Jan;17(1):676-682
pubmed: 30655816
Blood. 2008 Nov 1;112(9):3835-46
pubmed: 18633129
Haematologica. 2022 Aug 01;107(8):1891-1901
pubmed: 35045690