Aryl hydrocarbon receptor nuclear translocator-like (ARNTL/BMAL1) is associated with bevacizumab resistance in colorectal cancer via regulation of vascular endothelial growth factor AResearch in context
Elke Burgermeister,
Francesca Battaglin,
Fagr Eladly,
Wen Wu,
Frank Herweck,
Nadine Schulte,
Johannes Betge,
Nicolai Härtel,
Jakob N. Kather,
Cleo-Aron Weis,
Timo Gaiser,
Alexander Marx,
Christel Weiss,
Ralf Hofheinz,
Ian S. Miller,
Fotios Loupakis,
Heinz-Josef Lenz,
Annette T. Byrne,
Matthias P. Ebert
Affiliations
Elke Burgermeister
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany; Corresponding author at: Dept. of Medicine II, Universitätsklinikum Mannheim, Medical Faculty Mannheim, University Heidelberg, Theodor-Kutzer Ufer 1-3, D-68167 Mannheim, Germany.
Francesca Battaglin
Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, CA, United States; Unit of Medical Oncology 1, Clinical and Experimental Oncology Department, Veneto Institute of Oncology IOV - IRCCS, Padua, Italy
Fagr Eladly
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Wen Wu
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Frank Herweck
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Nadine Schulte
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Johannes Betge
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Nicolai Härtel
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Jakob N. Kather
Division of Gastroenterology, Hepatology and Hepatobiliary Oncology, University Hospital RWTH Aachen, Aachen, Germany
Cleo-Aron Weis
Institute of Pathology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Timo Gaiser
Institute of Pathology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Alexander Marx
Institute of Pathology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Christel Weiss
Department of Medical Statistics, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Ralf Hofheinz
Department of Medicine III, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Ian S. Miller
Department of Physiology & Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland
Fotios Loupakis
Unit of Medical Oncology 1, Clinical and Experimental Oncology Department, Veneto Institute of Oncology IOV - IRCCS, Padua, Italy
Heinz-Josef Lenz
Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, CA, United States
Annette T. Byrne
Department of Physiology & Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland; UCD School of Biomolecular and Biomedical Science, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland
Matthias P. Ebert
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
Background: The identification of new biomarkers and the development of novel, targetable contexts of vulnerability are of urgent clinical need in drug-resistant metastatic colorectal cancer (mCRC). Aryl-Hydrocarbon-Receptor-Nuclear-Translocator-Like (ARNTL/BMAL1) is a circadian clock-regulated transcription factor promoting expression of genes involved in angiogenesis and tumour progression. We hypothesised that BMAL1 increases expression of the vascular endothelial growth factor A VEGFA gene and, thereby, confers resistance to anti-angiogenic therapy with bevacizumab (Beva), a clinically used antibody for neutralization of VEGFA. Methods: PCR and immunohistochemistry were employed to assess BMAL1 expression in mice (C57BL/6 J Apcmin/+; BALB/c nu/nu xenografts) and CRC patients under combination chemotherapy with Beva. BMAL1 single nucleotide gene polymorphisms (SNPs) were analysed by DNA-microarray in clinical samples. BMAL1 functions were studied in human CRC cell lines using colorimetric growth, DNA-binding and reporter assays. Findings: In murine CRCs, high BMAL1 expression correlated with poor preclinical response to Beva treatment. In CRC patients' tumours (n = 74), high BMAL1 expression was associated with clinical non-response to combination chemotherapy with Beva (*p = .0061) and reduced progression-free survival (PFS) [*p = .0223, Hazard Ratio (HR) = 1.69]. BMAL1 SNPs also correlated with shorter PFS (rs7396943, rs7938307, rs2279287) and overall survival (OS) [rs11022780, *p = .014, HR = 1.61]. Mechanistically, Nuclear-Receptor-Subfamily-1-Group-D-Member-1 (NR1D1/REVERBA) bound a − 672 bp Retinoic-Acid-Receptor-Related-Orphan-Receptor-Alpha-responsive-element (RORE) adjacent to a BMAL1 DNA-binding motif (E-box) in the VEGFA gene promoter, resulting in increased VEGFA synthesis and proliferation of human CRC cell lines. Interpretation: BMAL1 was associated with Beva resistance in CRC. Inhibition of REVERBA-BMAL1 signalling may prevent resistance to anti-angiogenic therapy. Fund: This work was in part supported by the European Commission Seventh Framework Programme (Contract No. 278981 [ANGIOPREDICT]). Keywords: BMAL1, ARNTL, REVERBA, Bevacizumab, Colorectal cancer, VEGFA
