Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 May 5:13:1145001.
doi: 10.3389/fonc.2023.1145001. eCollection 2023.

Association of RANKL and EGFR gene expression with bone metastases in patients with metastatic non-small cell lung cancer

Anita J W M Brouns  1   2   3 , Lizza E L Hendriks  2   3 , Iris J Robbesom-van den Berge  4 , Annemariek J H M Driessen  5 , Guido M J M Roemen  6 , Britt L J van Herpen  6 , Zoë Dekkers  6 , Bas Heitzer  6 , Daphne J G Leunissen  6 , Laura Moonen  6 , Ragnar Lunde  7 , Marcel Westenend  8 , Marjolein van Driel  4 , Ernst-Jan M Speel  6 , Anne-Marie C Dingemans  2   3   9
Affiliations

Association of RANKL and EGFR gene expression with bone metastases in patients with metastatic non-small cell lung cancer

Anita J W M Brouns et al. Front Oncol. .

Abstract

Introduction: Bone metastases are frequent in patients with non-small cell lung cancer (NSCLC). The receptor activator of Nuclear Factor κB (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) pathway is important in bone metastases development. Furthermore, epidermal growth factor receptor (EGFR) signaling promotes osteoclast formation and stimulation. The understanding of the biological mechanism of bone metastases development might have implications for treatment strategies. Therefore, we studied whether there is an association between EGFR, RANKL, RANK and OPG gene expression in the tumor and presence of bone metastases in patients with NSCLC.

Methods: From an updated multicenter study, including patients with EGFR mutated (EGFR+), Kirsten rat sarcoma (KRAS+) and EGFR/KRAS wildtype metastatic NSCLC, all patients with available formalin-fixed paraffin-embedded (FFPE) tumor samples were selected. Ribonucleic Acid (RNA) was isolated from these samples and gene expressions of EGFR, RANKL, OPG and RANKL were determined via quantitative Polymerase Chain Reaction (qPCR). Data on demographics, histology and molecular subtyping, sample origin, presence of bone metastasis, SREs and bone progression were collected. Primary endpoint was relation between EGFR, RANK, RANKL, OPG gene expression, RANKL: OPG ratio and bone metastases.

Results: In 73/335 (32% EGFR+, 49% KRAS+, 19% EGFR/KRAS wildtype) samples from unique patients, gene expression analysis could be performed. Of these 73 patients, 46 (63%) had bone metastases at diagnosis or developed bone metastases during the disease course. No association was found between EGFR expression and presence of bone metastases. Patients with bone metastases had a significantly higher RANKL expression and RANKL: OPG ratio compared to those without. An increased RANKL: OPG ratio resulted in a 1.65x increased risk to develop bone metastases, especially in the first 450 days after diagnosis of metastatic NSCLC.

Conclusion: Increased RANKL gene expression and RANKL: OPG ratio, but not EGFR expression, was associated with presence of bone metastases. Additionally, an increased RANKL: OPG gene ratio was associated with a higher incidence of bone metastases development.

Keywords: bone metastases; epidermal growth factor expression; epidermal growth factor mutation; lung adenocarcinoma; osteoprotegerin; receptor activator of nuclear factor κb ligand.

PubMed Disclaimer

Conflict of interest statement

There are no conflicts of interest related to this manuscript. Outside this manuscript: Author AB: participation on advisory board of Jansen. Author LH: research funding Roche Genentech. AstraZeneca, Boehringer Ingelheim, Takeda, Beigene under negotiation all payments were paid to the institution. Speaker’s fee from MSD, Lilly. Fees for educational webinars: Benecke, Medtalks, VJOncology self, high5oncology institution. Support for attending meetings and/or travel: Roche Genentech. Advisory board BMS, Eli Lilly, Roche Genentech, Pfizer, Takeda, MSD, Merck, Novartis, Boehringer Ingelheim, Amgen, Janssen. Mentorship program with key opinion leaders: funded by AstraZeneca. Interview sessions funded by Roche Genentech, Bayer, Lilly. Local PI of clinical trials: AstraZeneca, Novartis, BMS, MSD/Merck, GSK, Takeda, Blueprint Medicines, Roche Genentech, Janssen Pharmaceuticals, Mirati, Abbvie, Gilead. Author A-MD: Roche: Advisory Board, Steering Committee Eli Lilly: Honorarium Boehringer Ingelheim: Advisory Board Astra Zeneca: Honorarium, Advisory Board Jansen: Honorarium industry sponsored symposium Chiesi: Honorarium Amgen: Advisory Board, research support Pfizer: Honorarium Bayer: Advisory Board Takeda: Honorarium Pharmamar: Advisory Board Sanofi: Advisory Boar. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Flowchart of pathology sample selection. The flowchart showed the process of sample selection and reasons for exclusion of samples. n, number; FFPE, formalin-fixed paraffin-embedded; RNA, Ribonucleic acid.
Figure 2
Figure 2
EGFR, RANKL, OPG, RANK gene expression in all patients. This figure shows the relative EGFR, RANKL, OPG and RANK gene expression measured on pathology samples of all patients. EGFR, Epidermal Growth Factor Receptor; RANKL, Receptor Activator of Nuclear Factor κB ligand; OPG, osteoprotegerin; RANK, Receptor Activator of Nuclear Factor κB.
Figure 3
Figure 3
(A–E) EGFR, OPG, RANKL gene expression, RANKL: OPG ratio and RANK gene expression in relation to presence of bone metastases. Patients were subdivided in groups by EGFR expression. The first quartile is the lowest and the fourth quartile is the highest EGFR gene expression. (A) EGFR gene expression, (B) OPG gene expression, (C) RANKL gene expression, (D) RANKL: OPG ratio, (E) RANK gene expression. An asterisk denotes a significant difference between groups. EGFR, Epidermal Growth Factor Receptor; OPG, osteoprotegerin; RANKL, Receptor Activator of Nuclear Factor κB ligand; RANK, Receptor Activator of Nuclear Factor κB.
Figure 4
Figure 4
(A–E) EGFR, RANKL, OPG gene expression, RANKL: OPG ratio and RANK gene expression in relation to site of tumor biopsy. (A) EGFR expression, (B) OPG expression, (C) RANKL expression, (D) RANKL: OPG ratio, (E) RANK expression, all expressions are shown in primary tumor, non-bone metastases and bone metastases. An asterisk denotes a significant difference between groups. EGFR, Epidermal Growth Factor Receptor; OPG, osteoprotegerin; RANKL, Receptor Activator of Nuclear Factor κB ligand; RANK, Receptor Activator of Nuclear Factor κB.
See this image and copyright information in PMC

References

    1. Hendriks LE, Smit EF, Vosse BA, Mellema WW, Heideman DA, Bootsma GP, et al. . EGFR mutated non-small cell lung cancer patients: more prone to development of bone and brain metastases? Lung Cancer (2014) 84(1):86–91. doi: 10.1016/j.lungcan.2014.01.006 - DOI - PubMed
    1. Peters S, Danson S, Hasan B, Dafni U, Reinmuth N, Majem M, et al. . A randomized open-label phase III trial evaluating the addition of denosumab to standard first-line treatment in advanced NSCLC: the European thoracic oncology platform (ETOP) and European organisation for research and treatment of cancer (EORTC) SPLENDOUR trial. J Thorac Oncol (2020) 10:1647–56. doi: 10.1016/j.jtho.2020.06.011 - DOI - PubMed
    1. Weilbaecher KN, Guise TA, McCauley LK. Cancer to bone: a fatal attraction. Nat Rev Cancer (2011) 11(6):411–25. doi: 10.1038/nrc3055 - DOI - PMC - PubMed
    1. Pantano F, Clezardin P, Santini D. RANK/RANKL pathway in cancer: biological activity beyond bone? J Bone Oncol (2012) 1(3):67–8. doi: 10.1016/j.jbo.2012.11.001 - DOI - PMC - PubMed
    1. Schneider MR, Sibilia M, Erben RG. The EGFR network in bone biology and pathology. Trends Endocrinol Metab (2009) 20(10):517–24. doi: 10.1016/j.tem.2009.06.008 - DOI - PubMed