Pharmacological Inhibition of Cyclin-Dependent Kinases Triggers Anti-Fibrotic Effects in Hepatic Stellate Cells In Vitro

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2020 May 9

PMID 32380742

Citations 4

Authors

Anna Hubbers

Julia Hennings

Daniela Lambertz

Ute Haas

Christian Trautwein

Yulia A Nevzorova

Roland Sonntag

Christian Liedtke

Affiliations

Soon will be listed here.

Abstract

Liver fibrosis is a wound healing process in response to chronic liver injury, which is characterized by the accumulation of extracellular collagen produced by Hepatic Stellate Cells (HSCs). This process involves cell cycle re-entry and proliferation of normally quiescent HSCs controlled by cyclins and associated cyclin-dependent kinases (Cdks). Cdk2 mediates the entry and progression through S-phase in complex with E-and A-type cyclins. We have demonstrated that cyclin E1 is essential for liver fibrogenesis in mice, but it is not known if this is dependent on Cdk2 or related Cdks. Here, we aimed to evaluate the benefit of the pan-Cdk inhibitor CR8 for treatment of liver fibrosis in vitro. CR8-treatment reduced proliferation and survival in immortalized HSC lines and in addition attenuated pro-fibrotic properties in primary murine HSCs. Importantly, primary murine hepatocytes were much more tolerant against the cytotoxic and anti-proliferative effects of CR8. We identified CR8 dosages mediating anti-fibrotic effects in primary HSCs without affecting cell cycle activity and survival in primary hepatocytes. In conclusion, the pharmacological pan-Cdk inhibitor CR8 restricts the pro-fibrotic properties of HSCs, while preserving proliferation and viability of hepatocytes at least in vitro. Therefore, CR8 and related drugs might be beneficial for the treatment of liver fibrosis.

Citing Articles

Mitochondrial stress induces hepatic stellate cell activation in response to the ATF4/TRIB3 pathway stimulation.

Ye M, Lu W, Rao Q, Li M, Hong H, Yang X J Gastroenterol. 2023; 58(7):668-681.

PMID: 37150773 DOI: 10.1007/s00535-023-01996-7.

SNS-032 attenuates liver fibrosis by anti-active hepatic stellate cells inhibition of cyclin dependent kinase 9.

He X, Hu Y, Chen J, Zhang D, Yang H, Zhang L Front Pharmacol. 2022; 13:1016552.

PMID: 36313366 PMC: 9597511. DOI: 10.3389/fphar.2022.1016552.

Liver Fibrosis-From Mechanisms of Injury to Modulation of Disease.

Liedtke C, Nevzorova Y, Luedde T, Zimmermann H, Kroy D, Strnad P Front Med (Lausanne). 2022; 8:814496.

PMID: 35087852 PMC: 8787129. DOI: 10.3389/fmed.2021.814496.

Synthesis of Novel 2-Thiouracil-5-Sulfonamide Derivatives as Potent Inducers of Cell Cycle Arrest and CDK2A Inhibition Supported by Molecular Docking.

Fatahala S, Sayed A, Mahgoub S, Taha H, El-Sayed M, El-Shehry M Int J Mol Sci. 2021; 22(21).

PMID: 34769385 PMC: 8584424. DOI: 10.3390/ijms222111957.

References

Akiyama T, Ohuchi T, Sumida S, Matsumoto K, Toyoshima K . Phosphorylation of the retinoblastoma protein by cdk2. Proc Natl Acad Sci U S A. 1992; 89(17):7900-4. PMC: 49822. DOI: 10.1073/pnas.89.17.7900. View

Hernandez-Gea V, Friedman S . Pathogenesis of liver fibrosis. Annu Rev Pathol. 2010; 6:425-56. DOI: 10.1146/annurev-pathol-011110-130246. View

Benson C, White J, de Bono J, ODonnell A, Raynaud F, Cruickshank C . A phase I trial of the selective oral cyclin-dependent kinase inhibitor seliciclib (CYC202; R-Roscovitine), administered twice daily for 7 days every 21 days. Br J Cancer. 2006; 96(1):29-37. PMC: 2360206. DOI: 10.1038/sj.bjc.6603509. View

Cosimo E, McCaig A, Carter-Brzezinski L, Wheadon H, Leach M, Le Ster K . Inhibition of NF-κB-mediated signaling by the cyclin-dependent kinase inhibitor CR8 overcomes prosurvival stimuli to induce apoptosis in chronic lymphocytic leukemia cells. Clin Cancer Res. 2013; 19(9):2393-405. DOI: 10.1158/1078-0432.CCR-12-2170. View

Nevzorova Y, Tschaharganeh D, Gassler N, Geng Y, Weiskirchen R, Sicinski P . Aberrant cell cycle progression and endoreplication in regenerating livers of mice that lack a single E-type cyclin. Gastroenterology. 2009; 137(2):691-703, 703.e1-6. PMC: 2730664. DOI: 10.1053/j.gastro.2009.05.003. View

Diril M, Ratnacaram C, Padmakumar V, Du T, Wasser M, Coppola V . Cyclin-dependent kinase 1 (Cdk1) is essential for cell division and suppression of DNA re-replication but not for liver regeneration. Proc Natl Acad Sci U S A. 2012; 109(10):3826-31. PMC: 3309725. DOI: 10.1073/pnas.1115201109. View

Weiskirchen S, Tag C, Sauer-Lehnen S, Tacke F, Weiskirchen R . Isolation and Culture of Primary Murine Hepatic Stellate Cells. Methods Mol Biol. 2017; 1627:165-191. DOI: 10.1007/978-1-4939-7113-8_11. View

Nevzorova Y, Bangen J, Hu W, Haas U, Weiskirchen R, Gassler N . Cyclin E1 controls proliferation of hepatic stellate cells and is essential for liver fibrogenesis in mice. Hepatology. 2012; 56(3):1140-9. PMC: 3396430. DOI: 10.1002/hep.25736. View

Pietrangelo A, Dierssen U, Valli L, Garuti C, Rump A, Corradini E . STAT3 is required for IL-6-gp130-dependent activation of hepcidin in vivo. Gastroenterology. 2007; 132(1):294-300. DOI: 10.1053/j.gastro.2006.10.018. View

10.

Kabadi S, Stoica B, Loane D, Luo T, Faden A . CR8, a novel inhibitor of CDK, limits microglial activation, astrocytosis, neuronal loss, and neurologic dysfunction after experimental traumatic brain injury. J Cereb Blood Flow Metab. 2014; 34(3):502-13. PMC: 3948132. DOI: 10.1038/jcbfm.2013.228. View

11.

Puche J, Saiman Y, Friedman S . Hepatic stellate cells and liver fibrosis. Compr Physiol. 2013; 3(4):1473-92. DOI: 10.1002/cphy.c120035. View

12.

Bressenot A, Marchal S, Bezdetnaya L, Garrier J, Guillemin F, Plenat F . Assessment of apoptosis by immunohistochemistry to active caspase-3, active caspase-7, or cleaved PARP in monolayer cells and spheroid and subcutaneous xenografts of human carcinoma. J Histochem Cytochem. 2008; 57(4):289-300. PMC: 2664986. DOI: 10.1369/jhc.2008.952044. View

13.

Aziz K, Limzerwala J, Sturmlechner I, Hurley E, Zhang C, Jeganathan K . Ccne1 Overexpression Causes Chromosome Instability in Liver Cells and Liver Tumor Development in Mice. Gastroenterology. 2019; 157(1):210-226.e12. PMC: 6800187. DOI: 10.1053/j.gastro.2019.03.016. View

14.

Liedtke C, Groger N, Manns M, Trautwein C . Interferon-alpha enhances TRAIL-mediated apoptosis by up-regulating caspase-8 transcription in human hepatoma cells. J Hepatol. 2005; 44(2):342-9. DOI: 10.1016/j.jhep.2005.07.020. View

15.

Kim S, BRANDSEN J, de Azevedo Junior W . Structural basis for chemical inhibition of CDK2. Prog Cell Cycle Res. 1996; 2:137-45. DOI: 10.1007/978-1-4615-5873-6_14. View

16.

Delehouze C, Godl K, Loaec N, Bruyere C, Desban N, Oumata N . CDK/CK1 inhibitors roscovitine and CR8 downregulate amplified MYCN in neuroblastoma cells. Oncogene. 2013; 33(50):5675-87. PMC: 4087096. DOI: 10.1038/onc.2013.513. View

17.

Bettayeb K, Baunbaek D, Delehouze C, Loaec N, Hole A, Baumli S . CDK Inhibitors Roscovitine and CR8 Trigger Mcl-1 Down-Regulation and Apoptotic Cell Death in Neuroblastoma Cells. Genes Cancer. 2011; 1(4):369-80. PMC: 3092200. DOI: 10.1177/1947601910369817. View

18.

Vassilev L . Cell cycle synchronization at the G2/M phase border by reversible inhibition of CDK1. Cell Cycle. 2006; 5(22):2555-6. DOI: 10.4161/cc.5.22.3463. View

19.

Wu J, Kharebava G, Piao C, Stoica B, Dinizo M, Sabirzhanov B . Inhibition of E2F1/CDK1 pathway attenuates neuronal apoptosis in vitro and confers neuroprotection after spinal cord injury in vivo. PLoS One. 2012; 7(7):e42129. PMC: 3405037. DOI: 10.1371/journal.pone.0042129. View

20.

de Wind N, Dekker M, Berns A, Radman M, Te Riele H . Inactivation of the mouse Msh2 gene results in mismatch repair deficiency, methylation tolerance, hyperrecombination, and predisposition to cancer. Cell. 1995; 82(2):321-30. DOI: 10.1016/0092-8674(95)90319-4. View