Cyclin E1 in Murine and Human Liver Cancer: A Promising Target for Therapeutic Intervention during Tumour Progression

Roland Sonntag,Christian Liedtke,Fabio Ticconi,Daniela Lambertz,Marlene Kohlhepp,Nikolaus Gassler,Ute Haas,Ivan G Costa,Frank Tacke,Christian Trautwein,Thorsten Cramer,Christian Penners,Andreas Kroh

doi:10.3390/cancers13225680

Abstract

Simple SummaryThe cell cycle regulator Cyclin E1 is a key mediator and biomarker of liver cancer progression in mice and man independent of its canonical interacting partner Cyclin-dependent kinase 2. Over-expression of Cyclin E1 during hepatocarcinogenesis modulates several distinct biological processes such as proliferation, DNA damage response, stemness, invasion and the tumour microenvironment. Interventional depletion of Cyclin E1 in the course of liver cancer progression significantly reduces tumour burden. In contrast, the expression of Cyclin-dependent kinase 2 is dispensable for the progression of liver cancer in mice and lacked diagnostic or prognostic value in patients. Thus, specific inhibition of Cyclin E1 expression represents a promising strategy for the treatment of liver cancer.Cyclin E1 (CCNE1) is a regulatory subunit of Cyclin-dependent kinase 2 (CDK2) and is thought to control the transition of quiescent cells into cell cycle progression. Recently, we identified CCNE1 and CDK2 as key factors for the initiation of hepatocellular carcinoma (HCC). In the present study, we dissected the contributions of CCNE1 and CDK2 for HCC progression in mice and patients. Therefore, we generated genetically modified mice allowing inducible deletion of Ccne1 or Cdk2. After initiation of HCC, using the hepatocarcinogen diethylnitrosamine (DEN), we deleted Ccne1 or Cdk2 and subsequently analysed HCC progression. The relevance of CCNE1 or CDK2 for human HCC progression was investigated by in silico database analysis. Interventional deletion of Ccne1, but not of Cdk2, substantially reduced the HCC burden in mice. Ccne1-deficient HCCs were characterised by attenuated proliferation, impaired DNA damage response and downregulation of markers for stemness and microinvasion. Additionally, the tumour microenvironment of Ccne1-deficient mice showed a reduction in immune mediators, myeloid cells and cancer-associated fibroblasts. In sharp contrast, Cdk2 was dispensable for HCC progression in mice. In agreement with our mouse data, CCNE1 was overexpressed in HCC patients independent of risk factors, and associated with reduced disease-free survival, a common signature for enhanced chromosomal instability, proliferation, dedifferentiation and invasion. However, CDK2 lacked diagnostic or prognostic value in HCC patients. In summary, CCNE1 drives HCC progression in a CDK2-independent manner in mice and man. Therefore, interventional inactivation of CCNE1 represents a promising strategy the treatment of liver cancer.

Highlights

Hepatocellular carcinoma (HCC) is the fourth most common cause of cancer-related death and classically develops in a multi-step process based on chronic hepatitis, caused, e.g., by viral hepatitis, alcohol consumption or non-alcoholic metabolic injury [1]
To assess the specific contributions of CCNE1 or Cyclin-dependent kinase 2 (CDK2) for HCC progression, DEN was injected into two-week-old conditional Ccne1 or Cdk2 knockout mice with inducible Crerecombinase under control of the Mx1-gene promoter [14]
After HCC initiation (22 weeks), interventional inactivation of Ccne1 (Ccne1−/−) or Cdk2 (Cdk2−/−) in liver, lymphocytes and hematopoietic cells was induced by pIpC injection

Summary

Introduction

Hepatocellular carcinoma (HCC) is the fourth most common cause of cancer-related death and classically develops in a multi-step process based on chronic hepatitis, caused, e.g., by viral hepatitis, alcohol consumption or non-alcoholic metabolic injury [1]. During these processes, hepatocytes accumulate oncogenic mutations due to genetic instability or viral insertions in cell cycle-related pathways, such as MYC, TP53, Retinoblastoma protein (RB) and cyclins [2]. In the last decade, improved drug therapy agents, such as tyrosine kinase inhibitors (e.g., Regorafenib), selective Cyclin-dependent kinase (CDK) inhibitors (e.g., Palbociclib) or immunotherapeutic strategies (e.g., Atezolizumab, Nivolumab) have shown partial benefits for patients with advanced HCC, but the overall prognosis remains limited [5]. Recent data on Regorafenib or Nivolumab indicated that pharmacokinetics, toxicity and clinical responses highly depend on gene polymorphisms (reviewed in [5])

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