Final Report Summary - RANKL_OPG (The RANK-Axis in Breast Cancer: A comprehensive prospective evaluation of promising novel biomarkers)
Description of the Project Objectives
Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer mortality in women worldwide. The recently identified Receptor Activator of Nuclear Factor (NF)-κB (RANK)-axis has stimulated novel research in the field of breast cancer etiology, treatment, and prevention. Experimental data implicate RANK signalling in mammary tumor development. Preliminary human data show increased RANK expression correlated with poor outcome, and circulating concentrations of osteoprotegerin, the decoy RANK receptor inhibiting RANK-axis signalling, are increased in several conditions associated with decreased risk of breast cancer (e.g. pregnancy).
The research project object was to provide the first evidence on the role of pre-diagnosis RANK-axis members in the etiology of breast cancer. Specifically, the project aims were to:
• Aim 1: Examine the association of prediagnostic concentrations of sRANKL, and the sRANKL/OPG ratio with risk of breast cancer overall in premenopausal and postmenopausal women in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort
• Aim 2: Evaluate associations between sRANKL, OPG, and sRANKL/OPG and breast cancer risk factors and endogenous hormone concentrations in the EPIC cohort
• Aim 3: Evaluate sRANKL and OPG as mediators of the associations between established risk factors and endogenous hormones and breast cancer in the EPIC cohort
• Aim 4: Examine concentrations of sRANKL, OPG and the sRANKL/OPG ratio across trimesters of pregnancy in a Finnish Maternity Cohort (FMC) methodological study
• Aim 5: Initiate and lead submission of a grant application to fund sRANKL and OPG study in the FMC
Research Project Progress and Main Results
Aims 1-3: EPIC cohort
Since the initiation of the project in January 2014, the research project aims have been achieved. The first manuscript from Aims 1 and 2, “Osteoprotegerin and breast cancer risk by hormone receptor subtype: a nested case-control study in the EPIC cohort,” was submitted in September 2016 and published in BMC Medicine in January 2017 (Fortner et al). In this first, large-scale investigation of OPG and breast cancer risk by hormone receptor subtype, we observed significantly higher risk of estrogen receptor (ER)-negative breast cancer among women with relatively high OPG concentrations. This finding is particularly important as relatively little is known about risk factors for ER-negative disease. The results on sRANKL and sRANKL/OPG will be presented in oral presentation at the American Association for Cancer Research Annual Meeting, a leading international conference, in April 2017. Due to conference regulations limiting publication of study results before the meeting presentation this manuscript is fully prepared but will be submitted directly after the meeting. Basic cross-sectional associations between risk factors and endogenous hormones and OPG and sRANKL were described in the manuscripts on risk, a manuscript more fully describing these associations is in preparation. In Aim 3, we proposed to evaluate whether sRANKL and OPG served as mediators between breast cancer risk factors and breast cancer risk. We completed this analysis, and found no mediation effect. While outside of the scope of the aims proposed here, we additionally evaluated pre-diagnosis circulating OPG and sRANKL, and the ratio, and breast cancer survival using the data generated for this project. An abstract on this study has been submitted to the European Society of Medical Oncology Impakt 2017 Breast Cancer conference.
Aims 4-5: Finnish Maternity Cohort
Aim 4 included a unique analysis of sRANKL and OPG concentrations both across trimesters of a single pregnancy (i.e. participants with three samples collected across one pregnancy). This study demonstrated that concentrations of sRANKL were stable throughout pregnancy; therefore, the single, first-trimester sample available in the FMC is likely a good estimate of exposure throughout pregnancy. OPG is less stable throughout pregnancy; this is an important consideration for the interpretation of results from the FMC. The manuscript describing this study, “Hormone concentrations throughout uncomplicated pregnancies: a longitudinal study,” was published in July 2016 (Schock et al.).
In Aim 5, we proposed a grant application to fund a study on sRANKL and OPG in the FMC. This application was submitted in May 2016. Despite receiving a score in the “Excellent” category, unfortunately, the project was not funded. Internal funding was subsequently acquired. The study samples were requested from the FMC in October 2016 and the sRANKL and OPG assays will be completed at the German Cancer Research Center in the 2nd quarter of 2017.
Transfer of Knowledge Objectives
The fellow`s knowledge transfer objectives capitalized on almost a decade of research experience and established networks in U.S. pregnancy cohorts and Harvard University`s Nurses` Health Study. The fellow: (1) Promoted scientific exchange with U.S.-based cohorts and provide subject area expertise; (2) Broadened data analysis expertise at DKFZ to foster long-term collaborations among colleagues; and, (3) Led and initiated cancer research projects and form strategic professional networks with European colleagues to develop ongoing collaborations.
Knowledge transfer achievements include completion of collaborative investigations leading to the following published (or under review) manuscripts:
• “Pre-diagnosis insulin-like growth factor-I and risk of epithelial invasive ovarian cancer by histological subtypes: A collaborative re-analysis from the Ovarian Cancer Cohort Consortium” was accepted at Cancer, Causes and Control (Ose et al. 2017) and “Androgens and ovarian cancer risk by subtype: Results from the Ovarian Cancer Cohort Consortium” is under revision at Cancer Research (Ose et al, under review). Dr. Jennifer Ose included these world-wide pooling projects as part of her doctoral thesis.
• A book chapter titled “Obesity and Breast Cancer”, co-authored with German Cancer Research Center Division of Cancer Epidemiology scientists (Fortner et al, 2016)
• Two manuscripts investigating the potential gain in discrimination between cases and controls realized by integrating circulating hormone concentrations into existing risk prediction models. These manuscripts were co-first authored by Dr. Fortner and statistician Dr. Anika Hüsing. “Endometrial cancer risk prediction including serum-based biomarkers: results from the EPIC cohort” was published in the International Journal of Cancer (Fortner et al, 2017). The revision of “Added value of serum hormone measurements in risk prediction models for breast cancer for women not using menopausal hormone therapy” is in press at Clinical Cancer Research (Hüsing et al. 2017).
• “Human Chorionic Gonadotropin Does Not Correlate with Risk for Maternal Breast Cancer: Results from the Finnish Maternity Cohort”, a cooperation with the Finnish Maternity Cohort, published in Cancer Research (Fortner et al. 2017)
Expected Final Results and Impact
The research project provided the first large-scale prospective study on the RANK-axis in breast cancer risk. We identified OPG as a novel risk marker for ER- breast cancer, the breast cancer subtype few known risk factors, and no targeted treatments. Our research has a translational component, as the RANK axis is targetable pharmacologically, with denosumab, a US-FDA approved drug for osteoporosis which acts as a decoy receptor for RANKL. Our group is expanding its work on the RANK-axis. Using the data generated through this project, we are currently investigating the association between pre-diagnosis OPG and sRANKL and breast cancer survival. To our knowledge, this is the first investigation of its kind, and may lead to new strategies to improve breast cancer survival. An extension of this investigation on breast cancer survival is being initiated within a patient cohort. Our completed project on RANK-axis members throughout pregnancy is critical to inform the interpretation of our ongoing work in the FMC. Our ongoing study on breast cancer risk in the FMC will provide the first data on the RANK-axis in pregnancy and subsequent breast cancer risk in the mother. A breast cancer diagnosis in the years following pregnancy has a disproportionately negative impact on a young family. This research will improve our knowledge on the etiology of this disease, thus improving our ability to predict risk, develop targeted screening programs, and inform prophylactic and treatment strategies.
Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer mortality in women worldwide. The recently identified Receptor Activator of Nuclear Factor (NF)-κB (RANK)-axis has stimulated novel research in the field of breast cancer etiology, treatment, and prevention. Experimental data implicate RANK signalling in mammary tumor development. Preliminary human data show increased RANK expression correlated with poor outcome, and circulating concentrations of osteoprotegerin, the decoy RANK receptor inhibiting RANK-axis signalling, are increased in several conditions associated with decreased risk of breast cancer (e.g. pregnancy).
The research project object was to provide the first evidence on the role of pre-diagnosis RANK-axis members in the etiology of breast cancer. Specifically, the project aims were to:
• Aim 1: Examine the association of prediagnostic concentrations of sRANKL, and the sRANKL/OPG ratio with risk of breast cancer overall in premenopausal and postmenopausal women in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort
• Aim 2: Evaluate associations between sRANKL, OPG, and sRANKL/OPG and breast cancer risk factors and endogenous hormone concentrations in the EPIC cohort
• Aim 3: Evaluate sRANKL and OPG as mediators of the associations between established risk factors and endogenous hormones and breast cancer in the EPIC cohort
• Aim 4: Examine concentrations of sRANKL, OPG and the sRANKL/OPG ratio across trimesters of pregnancy in a Finnish Maternity Cohort (FMC) methodological study
• Aim 5: Initiate and lead submission of a grant application to fund sRANKL and OPG study in the FMC
Research Project Progress and Main Results
Aims 1-3: EPIC cohort
Since the initiation of the project in January 2014, the research project aims have been achieved. The first manuscript from Aims 1 and 2, “Osteoprotegerin and breast cancer risk by hormone receptor subtype: a nested case-control study in the EPIC cohort,” was submitted in September 2016 and published in BMC Medicine in January 2017 (Fortner et al). In this first, large-scale investigation of OPG and breast cancer risk by hormone receptor subtype, we observed significantly higher risk of estrogen receptor (ER)-negative breast cancer among women with relatively high OPG concentrations. This finding is particularly important as relatively little is known about risk factors for ER-negative disease. The results on sRANKL and sRANKL/OPG will be presented in oral presentation at the American Association for Cancer Research Annual Meeting, a leading international conference, in April 2017. Due to conference regulations limiting publication of study results before the meeting presentation this manuscript is fully prepared but will be submitted directly after the meeting. Basic cross-sectional associations between risk factors and endogenous hormones and OPG and sRANKL were described in the manuscripts on risk, a manuscript more fully describing these associations is in preparation. In Aim 3, we proposed to evaluate whether sRANKL and OPG served as mediators between breast cancer risk factors and breast cancer risk. We completed this analysis, and found no mediation effect. While outside of the scope of the aims proposed here, we additionally evaluated pre-diagnosis circulating OPG and sRANKL, and the ratio, and breast cancer survival using the data generated for this project. An abstract on this study has been submitted to the European Society of Medical Oncology Impakt 2017 Breast Cancer conference.
Aims 4-5: Finnish Maternity Cohort
Aim 4 included a unique analysis of sRANKL and OPG concentrations both across trimesters of a single pregnancy (i.e. participants with three samples collected across one pregnancy). This study demonstrated that concentrations of sRANKL were stable throughout pregnancy; therefore, the single, first-trimester sample available in the FMC is likely a good estimate of exposure throughout pregnancy. OPG is less stable throughout pregnancy; this is an important consideration for the interpretation of results from the FMC. The manuscript describing this study, “Hormone concentrations throughout uncomplicated pregnancies: a longitudinal study,” was published in July 2016 (Schock et al.).
In Aim 5, we proposed a grant application to fund a study on sRANKL and OPG in the FMC. This application was submitted in May 2016. Despite receiving a score in the “Excellent” category, unfortunately, the project was not funded. Internal funding was subsequently acquired. The study samples were requested from the FMC in October 2016 and the sRANKL and OPG assays will be completed at the German Cancer Research Center in the 2nd quarter of 2017.
Transfer of Knowledge Objectives
The fellow`s knowledge transfer objectives capitalized on almost a decade of research experience and established networks in U.S. pregnancy cohorts and Harvard University`s Nurses` Health Study. The fellow: (1) Promoted scientific exchange with U.S.-based cohorts and provide subject area expertise; (2) Broadened data analysis expertise at DKFZ to foster long-term collaborations among colleagues; and, (3) Led and initiated cancer research projects and form strategic professional networks with European colleagues to develop ongoing collaborations.
Knowledge transfer achievements include completion of collaborative investigations leading to the following published (or under review) manuscripts:
• “Pre-diagnosis insulin-like growth factor-I and risk of epithelial invasive ovarian cancer by histological subtypes: A collaborative re-analysis from the Ovarian Cancer Cohort Consortium” was accepted at Cancer, Causes and Control (Ose et al. 2017) and “Androgens and ovarian cancer risk by subtype: Results from the Ovarian Cancer Cohort Consortium” is under revision at Cancer Research (Ose et al, under review). Dr. Jennifer Ose included these world-wide pooling projects as part of her doctoral thesis.
• A book chapter titled “Obesity and Breast Cancer”, co-authored with German Cancer Research Center Division of Cancer Epidemiology scientists (Fortner et al, 2016)
• Two manuscripts investigating the potential gain in discrimination between cases and controls realized by integrating circulating hormone concentrations into existing risk prediction models. These manuscripts were co-first authored by Dr. Fortner and statistician Dr. Anika Hüsing. “Endometrial cancer risk prediction including serum-based biomarkers: results from the EPIC cohort” was published in the International Journal of Cancer (Fortner et al, 2017). The revision of “Added value of serum hormone measurements in risk prediction models for breast cancer for women not using menopausal hormone therapy” is in press at Clinical Cancer Research (Hüsing et al. 2017).
• “Human Chorionic Gonadotropin Does Not Correlate with Risk for Maternal Breast Cancer: Results from the Finnish Maternity Cohort”, a cooperation with the Finnish Maternity Cohort, published in Cancer Research (Fortner et al. 2017)
Expected Final Results and Impact
The research project provided the first large-scale prospective study on the RANK-axis in breast cancer risk. We identified OPG as a novel risk marker for ER- breast cancer, the breast cancer subtype few known risk factors, and no targeted treatments. Our research has a translational component, as the RANK axis is targetable pharmacologically, with denosumab, a US-FDA approved drug for osteoporosis which acts as a decoy receptor for RANKL. Our group is expanding its work on the RANK-axis. Using the data generated through this project, we are currently investigating the association between pre-diagnosis OPG and sRANKL and breast cancer survival. To our knowledge, this is the first investigation of its kind, and may lead to new strategies to improve breast cancer survival. An extension of this investigation on breast cancer survival is being initiated within a patient cohort. Our completed project on RANK-axis members throughout pregnancy is critical to inform the interpretation of our ongoing work in the FMC. Our ongoing study on breast cancer risk in the FMC will provide the first data on the RANK-axis in pregnancy and subsequent breast cancer risk in the mother. A breast cancer diagnosis in the years following pregnancy has a disproportionately negative impact on a young family. This research will improve our knowledge on the etiology of this disease, thus improving our ability to predict risk, develop targeted screening programs, and inform prophylactic and treatment strategies.