Studying the DNA Damage Response Pathway in Hematopoietic Canine Cancer Cell Lines, a Necessary Step for Finding Targets to Generate New Therapies to Treat Cancer in Dogs

Overview

Journal Front Vet Sci

Specialty Veterinary Medicine

Date 2023 Sep 1

PMID 37655260

Authors

Beatriz Hernandez-Suarez

David A Gillespie

Ewa Dejnaka

Piotr Kupczyk

Bozena Obminska-Mrukowicz

Aleksandra Pawlak

Affiliations

Soon will be listed here.

Abstract

Background: Dogs present a significant opportunity for studies in comparative oncology. However, the study of cancer biology phenomena in canine cells is currently limited by restricted availability of validated antibody reagents and techniques. Here, we provide an initial characterization of the expression and activity of key components of the DNA Damage Response (DDR) in a panel of hematopoietic canine cancer cell lines, with the use of commercially available antibody reagents.

Materials And Methods: The techniques used for this validation analysis were western blot, qPCR, and DNA combing assay.

Results: Substantial variations in both the basal expression (ATR, Claspin, Chk1, and Rad51) and agonist-induced activation (p-Chk1) of DDR components were observed in canine cancer cell lines. The expression was stronger in the CLBL-1 (B-cell lymphoma) and CLB70 (B-cell chronic lymphocytic leukemia) cell lines than in the GL-1 (B-cell leukemia) cell line, but the biological significance of these differences requires further investigation. We also validated methodologies for quantifying DNA replication dynamics in hematopoietic canine cancer cell lines, and found that the GL-1 cell line presented a higher replication fork speed than the CLBL-1 cell line, but that both showed a tendency to replication fork asymmetry.

Conclusion: These findings will inform future studies on cancer biology, which will facilitate progress in developing novel anticancer therapies for canine patients. They can also provide new knowledge in human oncology.

References

Choi S, Yang H, Lee S, Ki J, Nam D, Yoo H . TopBP1 and Claspin contribute to the radioresistance of lung cancer brain metastases. Mol Cancer. 2014; 13:211. PMC: 4168047. DOI: 10.1186/1476-4598-13-211. View

Xing L, Lin L, Yu T, Li Y, Cho S, Liu J . A novel BCMA PBD-ADC with ATM/ATR/WEE1 inhibitors or bortezomib induce synergistic lethality in multiple myeloma. Leukemia. 2020; 34(8):2150-2162. PMC: 7392808. DOI: 10.1038/s41375-020-0745-9. View

Altschul S, Madden T, Schaffer A, Zhang J, Zhang Z, Miller W . Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997; 25(17):3389-402. PMC: 146917. DOI: 10.1093/nar/25.17.3389. View

Kim H, Xu H, George E, Hallberg D, Kumar S, Jagannathan V . Combining PARP with ATR inhibition overcomes PARP inhibitor and platinum resistance in ovarian cancer models. Nat Commun. 2020; 11(1):3726. PMC: 7381609. DOI: 10.1038/s41467-020-17127-2. View

Daud A, Ashworth M, Strosberg J, Goldman J, Mendelson D, Springett G . Phase I dose-escalation trial of checkpoint kinase 1 inhibitor MK-8776 as monotherapy and in combination with gemcitabine in patients with advanced solid tumors. J Clin Oncol. 2015; 33(9):1060-6. DOI: 10.1200/JCO.2014.57.5027. View

Chong G, Kuo F, Tsai S, Lin C . Validation of reference genes for cryopreservation studies with the gorgonian coral endosymbiont Symbiodinium. Sci Rep. 2017; 7:39396. PMC: 5220285. DOI: 10.1038/srep39396. View

Fournier M, Paulson A, Pavelka N, Mosley A, Gaudenz K, Bradford W . Delayed correlation of mRNA and protein expression in rapamycin-treated cells and a role for Ggc1 in cellular sensitivity to rapamycin. Mol Cell Proteomics. 2009; 9(2):271-84. PMC: 2830839. DOI: 10.1074/mcp.M900415-MCP200. View

Pawlak A, Henklewska M, Hernandez-Suarez B, Siepka M, Gladkowski W, Wawrzenczyk C . Methoxy-Substituted γ-Oxa-ε-Lactones Derived from Flavanones-Comparison of Their Anti-Tumor Activity In Vitro. Molecules. 2021; 26(20). PMC: 8538229. DOI: 10.3390/molecules26206295. View

Pawlak A, Henklewska M, Hernandez Suarez B, Luzny M, Kozlowska E, Obminska-Mrukowicz B . Chalcone Methoxy Derivatives Exhibit Antiproliferative and Proapoptotic Activity on Canine Lymphoma and Leukemia Cells. Molecules. 2020; 25(19). PMC: 7582533. DOI: 10.3390/molecules25194362. View

10.

Ly T, Ahmad Y, Shlien A, Soroka D, Mills A, Emanuele M . A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells. Elife. 2014; 3:e01630. PMC: 3936288. DOI: 10.7554/eLife.01630. View

11.

Yang M, Tian X, Fan Z, Yu W, Li Z, Zhou J . Targeting RAD51 enhances chemosensitivity of adult T‑cell leukemia‑lymphoma cells by reducing DNA double‑strand break repair. Oncol Rep. 2019; 42(6):2426-2434. PMC: 6859462. DOI: 10.3892/or.2019.7384. View

12.

Techer H, Pasero P . The Replication Stress Response on a Narrow Path Between Genomic Instability and Inflammation. Front Cell Dev Biol. 2021; 9:702584. PMC: 8270677. DOI: 10.3389/fcell.2021.702584. View

13.

Brandsma I, Fleuren E, Williamson C, Lord C . Directing the use of DDR kinase inhibitors in cancer treatment. Expert Opin Investig Drugs. 2017; 26(12):1341-1355. PMC: 6157710. DOI: 10.1080/13543784.2017.1389895. View

14.

Broustas C, Lieberman H . DNA damage response genes and the development of cancer metastasis. Radiat Res. 2014; 181(2):111-30. PMC: 4064942. DOI: 10.1667/RR13515.1. View

15.

Grosse N, van Loon B, Rohrer Bley C . DNA damage response and DNA repair - dog as a model?. BMC Cancer. 2014; 14:203. PMC: 3995094. DOI: 10.1186/1471-2407-14-203. View

16.

Sarmento L, Povoa V, Nascimento R, Real G, Antunes I, Martins L . CHK1 overexpression in T-cell acute lymphoblastic leukemia is essential for proliferation and survival by preventing excessive replication stress. Oncogene. 2014; 34(23):2978-90. DOI: 10.1038/onc.2014.248. View

17.

Nakaichi M, Taura Y, Kanki M, Mamba K, Momoi Y, Tsujimoto H . Establishment and characterization of a new canine B-cell leukemia cell line. J Vet Med Sci. 1996; 58(5):469-71. DOI: 10.1292/jvms.58.469. View

18.

Zhu H, Swami U, Preet R, Zhang J . Harnessing DNA Replication Stress for Novel Cancer Therapy. Genes (Basel). 2020; 11(9). PMC: 7564951. DOI: 10.3390/genes11090990. View

19.

Chen G, Gharib T, Huang C, Taylor J, Misek D, Kardia S . Discordant protein and mRNA expression in lung adenocarcinomas. Mol Cell Proteomics. 2002; 1(4):304-13. DOI: 10.1074/mcp.m200008-mcp200. View

20.

Graner A, Hellwinkel J, Lencioni A, Madsen H, Harland T, Marchando P . HSP90 inhibitors in the context of heat shock and the unfolded protein response: effects on a primary canine pulmonary adenocarcinoma cell line. Int J Hyperthermia. 2016; 33(3):303-317. PMC: 5675827. DOI: 10.1080/02656736.2016.1256503. View