TP5: A Novel Therapeutic Approach Targeting Aberrant and Hyperactive CDK5/p25 for the Treatment of Colorectal Carcinoma
Abstract
:1. Introduction
2. Results
2.1. TP5 Inhibits CDK5 Activity and Decreases Cell Viability and Migration
2.2. TP5 Alone Induces Cell Apoptosis by Impairing the Cell Cycle and DNA Damage Repair
2.3. TP5 Acts Synergistically with Chemotherapy by Increasing DNA Damage
2.4. TP5 Suppresses CRC Tumor Growth In Vivo
3. Discussion
4. Materials and Methods
4.1. Reagents and Compounds
4.2. Cell Line Preparation
4.3. Cell Viability Assay
4.4. Colony Formation Assay
4.5. CDK5 Activity Assay
4.6. Western Blotting
4.7. Immunofluorescence Staining
4.8. Wound Healing Assay
4.9. Apoptosis Assay
4.10. Cell Cycle Analysis
4.11. MitoSOX Staining
4.12. Subcutaneous CRC Model and Treatment
4.13. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Ferlay, J.; Colombet, M.; Soerjomataram, I.; Mathers, C.; Parkin, D.M.; Pineros, M.; Znaor, A.; Bray, F. Estimating the global cancer incidence and mortality in 2018: GLOBOCAN sources and methods. Int. J. Cancer 2019, 144, 1941–1953. [Google Scholar] [CrossRef] [Green Version]
- Van Cutsem, E.; Cervantes, A.; Adam, R.; Sobrero, A.; Van Krieken, J.H.; Aderka, D.; Aranda Aguilar, E.; Bardelli, A.; Benson, A.; Bodoky, G.; et al. ESMO consensus guidelines for the management of patients with metastatic colorectal cancer. Ann. Oncol. 2016, 27, 1386–1422. [Google Scholar] [CrossRef] [PubMed]
- Salvatore, L.; Aprile, G.; Arnoldi, E.; Aschele, C.; Carnaghi, C.; Cosimelli, M.; Maiello, E.; Normanno, N.; Sciallero, S.; Valvo, F.; et al. Management of metastatic colorectal cancer patients: Guidelines of the Italian Medical Oncology Association (AIOM). ESMO Open 2017, 2, e000147. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Das, S.; Ciombor, K.K.; Haraldsdottir, S.; Goldberg, R.M. Promising New Agents for Colorectal Cancer. Curr. Treat. Options Oncol. 2018, 19, 29. [Google Scholar] [CrossRef] [PubMed]
- Dhavan, R.; Tsai, L.H. A decade of CDK5. Nat. Rev. Mol. Cell Biol. 2001, 2, 749–759. [Google Scholar] [CrossRef]
- Hisanaga, S.; Saito, T. The regulation of cyclin-dependent kinase 5 activity through the metabolism of p35 or p39 Cdk5 activator. Neurosignals 2003, 12, 221–229. [Google Scholar] [CrossRef]
- Lopes, J.P.; Agostinho, P. Cdk5: Multitasking between physiological and pathological conditions. Prog. Neurobiol. 2011, 94, 49–63. [Google Scholar] [CrossRef] [Green Version]
- Pozo, K.; Castro-Rivera, E.; Tan, C.; Plattner, F.; Schwach, G.; Siegl, V.; Meyer, D.; Guo, A.; Gundara, J.; Mettlach, G.; et al. The role of Cdk5 in neuroendocrine thyroid cancer. Cancer Cell 2013, 24, 499–511. [Google Scholar] [CrossRef] [Green Version]
- Pozo, K.; Bibb, J.A. The Emerging Role of Cdk5 in Cancer. Trends Cancer 2016, 2, 606–618. [Google Scholar] [CrossRef] [Green Version]
- Yu, H.P.; Xie, J.M.; Li, B.; Sun, Y.H.; Gao, Q.G.; Ding, Z.H.; Wu, H.R.; Qin, Z.H. TIGAR regulates DNA damage and repair through pentosephosphate pathway and Cdk5-ATM pathway. Sci. Rep. 2015, 5, 9853. [Google Scholar] [CrossRef] [Green Version]
- Tian, B.; Yang, Q.; Mao, Z. Phosphorylation of ATM by Cdk5 mediates DNA damage signalling and regulates neuronal death. Nat. Cell Biol. 2009, 11, 211–218. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ruiz de Porras, V.; Bystrup, S.; Cabrero-de Las Heras, S.; Musulen, E.; Palomero, L.; Alonso, M.H.; Nieto, R.; Arango, D.; Moreno, V.; Queralt, C.; et al. Tumor Expression of Cyclin-Dependent Kinase 5 (Cdk5) Is a Prognostic Biomarker and Predicts Outcome of Oxaliplatin-Treated Metastatic Colorectal Cancer Patients. Cancers 2019, 11, 1540. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Robb, C.M.; Kour, S.; Contreras, J.I.; Agarwal, E.; Barger, C.J.; Rana, S.; Sonawane, Y.; Neilsen, B.K.; Taylor, M.; Kizhake, S.; et al. Characterization of CDK(5) inhibitor, 20-223 (aka CP668863) for colorectal cancer therapy. Oncotarget 2018, 9, 5216–5232. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhuang, K.; Zhang, J.; Xiong, M.; Wang, X.; Luo, X.; Han, L.; Meng, Y.; Zhang, Y.; Liao, W.; Liu, S. CDK5 functions as a tumor promoter in human colorectal cancer via modulating the ERK5-AP-1 axis. Cell Death Dis. 2016, 7, e2415. [Google Scholar] [CrossRef]
- Binukumar, B.K.; Zheng, Y.L.; Shukla, V.; Amin, N.D.; Grant, P.; Pant, H.C. TFP5, a peptide derived from p35, a Cdk5 neuronal activator, rescues cortical neurons from glucose toxicity. J. Alzheimer’s Dis. 2014, 39, 899–909. [Google Scholar]
- Binukumar, B.K.; Shukla, V.; Amin, N.D.; Grant, P.; Bhaskar, M.; Skuntz, S.; Steiner, J.; Pant, H.C. Peptide TFP5/TP5 derived from Cdk5 activator P35 provides neuroprotection in the MPTP model of Parkinson’s disease. Mol. Biol. Cell 2015, 26, 4478–4491. [Google Scholar] [CrossRef] [Green Version]
- Tabouret, E.; Wang, H.; Amin, N.; Jung, J.; Appay, R.; Cui, J.; Song, Q.; Cardone, A.; Park, D.M.; Gilbert, M.R.; et al. TP5, a Peptide Inhibitor of Aberrant and Hyperactive CDK5/p25: A Novel Therapeutic Approach against Glioblastoma. Cancers 2020, 12, 1935. [Google Scholar] [CrossRef]
- Fitzgerald, J.C.; Camprubi, M.D.; Dunn, L.; Wu, H.C.; Ip, N.Y.; Kruger, R.; Martins, L.M.; Wood, N.W.; Plun-Favreau, H. Phosphorylation of HtrA2 by cyclin-dependent kinase-5 is important for mitochondrial function. Cell Death Differ. 2012, 19, 257–266. [Google Scholar] [CrossRef] [Green Version]
- NavaneethaKrishnan, S.; Rosales, J.L.; Lee, K.Y. Loss of Cdk5 in breast cancer cells promotes ROS-mediated cell death through dysregulation of the mitochondrial permeability transition pore. Oncogene 2018, 37, 1788–1804. [Google Scholar] [CrossRef] [Green Version]
- Yu, I.S.; Cheung, W.Y. Metastatic Colorectal Cancer in the Era of Personalized Medicine: A More Tailored Approach to Systemic Therapy. Can. J. Gastroenterol. Hepatol. 2018, 2018, 9450754. [Google Scholar] [CrossRef]
- Andre, T.; Shiu, K.K.; Kim, T.W.; Jensen, B.V.; Jensen, L.H.; Punt, C.; Smith, D.; Garcia-Carbonero, R.; Benavides, M.; Gibbs, P.; et al. Pembrolizumab in Microsatellite-Instability-High Advanced Colorectal Cancer. N. Engl. J. Med. 2020, 383, 2207–2218. [Google Scholar] [CrossRef] [PubMed]
- Vogel, A.; Kirstein, M.M. First-line molecular therapies in the treatment of metastatic colorectal cancer—A literature-based review of phases II and III trials. Innov. Surg. Sci. 2018, 3, 85–86. [Google Scholar] [CrossRef]
- Al-Lazikani, B.; Banerji, U.; Workman, P. Combinatorial drug therapy for cancer in the post-genomic era. Nat. Biotechnol. 2012, 30, 679–692. [Google Scholar] [CrossRef] [PubMed]
- Ehrlich, S.M.; Liebl, J.; Ardelt, M.A.; Lehr, T.; De Toni, E.N.; Mayr, D.; Brandl, L.; Kirchner, T.; Zahler, S.; Gerbes, A.L.; et al. Targeting cyclin dependent kinase 5 in hepatocellular carcinoma—A novel therapeutic approach. J. Hepatol. 2015, 63, 102–113. [Google Scholar] [CrossRef] [PubMed]
- Futatsugi, A.; Utreras, E.; Rudrabhatla, P.; Jaffe, H.; Pant, H.C.; Kulkarni, A.B. Cyclin-dependent kinase 5 regulates E2F transcription factor through phosphorylation of Rb protein in neurons. Cell Cycle 2012, 11, 1603–1610. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Roufayel, R.; Murshid, N. CDK5: Key Regulator of Apoptosis and Cell Survival. Biomedicines 2019, 7, 88. [Google Scholar] [CrossRef] [Green Version]
- Chi, T.F.; Horbach, T.; Gotz, C.; Kietzmann, T.; Dimova, E.Y. Cyclin-Dependent Kinase 5 (CDK5)-Mediated Phosphorylation of Upstream Stimulatory Factor 2 (USF2) Contributes to Carcinogenesis. Cancers 2019, 11, 523. [Google Scholar] [CrossRef] [Green Version]
- Liu, W.; Li, J.; Song, Y.S.; Li, Y.; Jia, Y.H.; Zhao, H.D. Cdk5 links with DNA damage response and cancer. Mol. Cancer 2017, 16, 60. [Google Scholar] [CrossRef] [Green Version]
- Courapied, S.; Sellier, H.; de Carne Trecesson, S.; Vigneron, A.; Bernard, A.C.; Gamelin, E.; Barre, B.; Coqueret, O. The cdk5 kinase regulates the STAT3 transcription factor to prevent DNA damage upon topoisomerase I inhibition. J. Biol. Chem. 2010, 285, 26765–26778. [Google Scholar] [CrossRef] [Green Version]
- Chiker, S.; Pennaneach, V.; Loew, D.; Dingli, F.; Biard, D.; Cordelieres, F.P.; Gemble, S.; Vacher, S.; Bieche, I.; Hall, J.; et al. Cdk5 promotes DNA replication stress checkpoint activation through RPA-32 phosphorylation, and impacts on metastasis free survival in breast cancer patients. Cell Cycle 2015, 14, 3066–3078. [Google Scholar] [CrossRef] [Green Version]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Amin, N.; Wang, H.; Song, Q.; Bhaskar, M.; Yadav, S.P.; Gilbert, M.R.; Pant, H.; Tabouret, E.; Zhuang, Z. TP5: A Novel Therapeutic Approach Targeting Aberrant and Hyperactive CDK5/p25 for the Treatment of Colorectal Carcinoma. Int. J. Mol. Sci. 2023, 24, 11733. https://doi.org/10.3390/ijms241411733
Amin N, Wang H, Song Q, Bhaskar M, Yadav SP, Gilbert MR, Pant H, Tabouret E, Zhuang Z. TP5: A Novel Therapeutic Approach Targeting Aberrant and Hyperactive CDK5/p25 for the Treatment of Colorectal Carcinoma. International Journal of Molecular Sciences. 2023; 24(14):11733. https://doi.org/10.3390/ijms241411733
Chicago/Turabian StyleAmin, Niranjana, Herui Wang, Qi Song, Manju Bhaskar, Sharda Prasad Yadav, Mark R. Gilbert, Harish Pant, Emeline Tabouret, and Zhengping Zhuang. 2023. "TP5: A Novel Therapeutic Approach Targeting Aberrant and Hyperactive CDK5/p25 for the Treatment of Colorectal Carcinoma" International Journal of Molecular Sciences 24, no. 14: 11733. https://doi.org/10.3390/ijms241411733