Circulating Sex Hormone Levels and Colon Cancer Risk in Men: A Nested Case–Control Study and Meta-Analysis

S. Harlid received grants from the Cancer foundation in Northern Sweden (grant nos. AMP 17-856, AMP 18-915, and AMP 19-967), the Lions Cancer Research Fund in Northern Sweden (grant no. LP 20-2227), and internal funds from the Department of Radiation Sciences, Umea~ University. The coordination of EPIC is financially supported by IARC and also by the Department of Epidemiology and Biostatistics, School of Public Health, Imperial College London, which has additional infrastructure support provided by the NIHR Imperial Biomedical Research Centre (BRC). The national cohorts are supported by Danish Cancer Society (Denmark); Ligue Contre le Cancer, Institut Gustave Roussy, MutuelleG~en~erale de l'Education Nationale, Institut National de la Sant~e et de la RechercheM~edicale (INSERM; France); German Cancer Aid, German Cancer Research Center (DKFZ), German Institute of Human Nutrition Potsdam-Rehbruecke (DIfE), Federal Ministry of Education and Research (BMBF; Germany); Associazione Italiana per la Ricerca sul Cancro-AIRC-Italy, Compagnia di SanPaolo and National Research Council (Italy); Dutch Ministry of Public Health, Welfare and Sports (VWS), Netherlands Cancer Registry (NKR), LK Research Funds, Dutch Prevention Funds, Dutch ZON (Zorg Onderzoek Nederland), World Cancer Research Fund (WCRF), Statistics Netherlands (the Netherlands); Health Research Fund (FIS) -Instituto de Salud Carlos III (ISCIII), Regional Governments of Andalucia, Asturias, Basque Country, Murcia and Navarra, and the Catalan Institute of Oncology -ICO (Spain); Swedish Cancer Society, Swedish Research Council and Regions of Ska~ ne and V_asterbotten (Sweden); Cancer Research UK (14136 to EPICNorfolk; C8221/A29017 to EPIC-Oxford), Medical Research Council (1000143 to EPIC-Norfolk; MR/M012190/1 to EPIC-Oxford). We would like to thank the participants and staff of the EPIC and NSHDS cohorts for their valuable contribution to this research. We thank the Biobank Research Unit at Umea~ University, VIP, the Northern Sweden MONICA study, and the County Council of V_asterbotten for providing data and samples and acknowledge the contribution from Biobank Sweden, supported by the Swedish Research Council (VR 2017-00650). Special thanks also to Emmanouil Bouras for assistance on the meta-analysis. Furthermore, we acknowledge the use of data and biological samples from the following cohorts: EPIC-Florence [principal investigator (PI) Dr Domenico Palli], EPIC-Asturias (PI Dr J. Ram~on Quir~os) and EPIC-Cambridge (PI Professor Nick Wareham). Finally, on behalf of the EPIC-Norfolk study, we are grateful to all the participants who have been part of the project and to the many members of the study teams at the University of Cambridge (Cambridge, United Kingdom) who have enabled this research.Background: Endogenous sex hormonesmay contribute to higher colorectal cancer incidence rates in men compared with women, but despite an increased number of studies, clear evidence is lacking. Methods: We conducted a comprehensive nested case–control study of circulating concentrations of sex hormones, sex hormone precursors, and sex hormone binding globulin (SHBG) in relation to subsequent colon cancer risk in European men. Concentrations were measured using liquid LC/MS-MS in prospectively collected plasma samples from 690 cases and 690 matched controls from the European Prospective Investigation into Cancer and Nutrition (EPIC) andtheNorthern SwedenHealth and Disease Study (NSHDS) cohorts. Multivariable conditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI). In addition, we conducted a meta-analysis of previous studies on men. Results: Circulating levels of testosterone (OR, 0.68; 95% CI, 0.51–0.89) and SHBG (OR, 0.77; 95% CI, 0.62–0.96) were inversely associated with colon cancer risk. For free testosterone, there was a nonsignificant inverse association (OR, 0.83; 95% CI, 0.58–1.18). In a dose–response meta-analysis of endogenous sex hormone levels, inverse associations with colorectal/colon cancer risk were found for testosterone [relative risks (RR) per 100 ng/dL ¼ 0.98; 95% CI, 0.96–1.00; I2 ¼ 22%] and free testosterone (RR per 1 ng/dL ¼ 0.98; 95% CI, 0.95–1.00; I2 ¼ 0%). Conclusions: Our results provide suggestive evidence for the association between testosterone, SHBG, and male colon cancer development. Impact: Additional support for the involvement of sex hormones in male colon cancer.Cancer foundation in Northern Sweden AMP 17-856 AMP 18-915 AMP 19-967Lions Cancer Research Fund in Northern Sweden LP 20-2227Department of Radiation Sciences, Umea UniversityNIHR Imperial Biomedical Research Centre (BRC)Danish Cancer SocietyLigue nationale contre le cancerIARCDepartment of Epidemiology and Biostatistics, School of Public Health, Imperial College LondonInstitut Gustave Roussy, MutuelleGenerale de l'Education NationaleInstitut National de la Sante et de la Recherche Medicale (Inserm)Deutsche KrebshilfeHelmholtz AssociationGerman Institute of Human Nutrition Potsdam-Rehbruecke (DIfE)Federal Ministry of Education & Research (BMBF)Fondazione AIRC per la ricerca sul cancroCompagnia di San PaoloConsiglio Nazionale delle Ricerche (CNR)Netherlands Government Netherlands GovernmentWorld Cancer Research Fund International (WCRF)Netherlands GovernmentHealth Research Fund (FIS) -Instituto de Salud Carlos III (ISCIII) (Spain)Junta de AndaluciaPrincipality of AsturiasRegional Government of Basque Country (Spain) Regional Government of Murcia (Spain) Regional Government of Navarra (Spain) Catalan Institute of Oncology -ICO (Spain)Swedish Cancer Society Swedish Research Council Region of Skane (Sweden) Region of Vasterbotten (Sweden)Cancer Research UK 14136 C8221/A29017 UK Research & Innovation (UKRI)Medical Research Council UK (MRC)European Commission 1000143 MR/M012190/1Swedish Research CouncilEuropean Commission VR 2017-0065

Cirkulerande markörer för risk och etiologi för kolorektal cancer

Background: Colorectal cancer is the third most commonly diagnosed cancer in men and women. Worldwide around 2 million individuals are diagnosed each year – a number expected to increase as colorectal cancer risk factors become more prevalent. In men and women there is a difference in incidence, which possibly could be explained by inherent differences, including sex hormone profiles. The prognosis of colorectal cancer is highly dependent on the stage at diagnosis, with individuals diagnosed at early stages having the best long-term survival. However, as onset of symptoms can be diffuse, many individuals are diagnosed at later stages when survival rates are significantly poorer. Therefore, screening and prevention strategies to detect colorectal cancer at earlier stages or remove cancer precursors such as polyps may be key to increasing survival. Commonly used screening tools today include fecal blood tests and colonoscopy, but they have modest accuracy or may not be cost-effective. Being able to identify markers in blood, either for early detection, as a complementary or alternative screening method, or for risk stratification, could aid in solving this problem.  Aim: The overall of aim of the thesis was to improve our understanding of underlying factors contributing to CRC etiology and to find biomarkers associated with CRC that could aid in the future development of effective risk prediction models.  Methods: All studies included in this thesis were based on a case-control cohort nested within the Northern Sweden Health and Disease Study (NSHDS). Additionally in paper I, we also used data from the European Prospective Investigation into Cancer and Nutrition (EPIC), a large multi-center cohort study. In this paper we examined associations between sex hormones, sex hormone binding globulin (SHBG), and colon cancer in men. The study included 690 colon cancer cases and 690 matched controls. Paper II was a longitudinal study, using repeated samples from 80 men, on circulating sex hormones, SHBG, and DNA methylation in white blood cells. Papers III and IV were nested case-control studies on proteins and colorectal cancer risk with Paper III divided into a discovery and a validation phase. In the first phase, which included 69 colorectal cancer case-control pairs with repeated samples, 160 unique proteins related to inflammation and oncology were analyzed. In the second phase, 13 proteins that were significantly associated with colorectal cancer risk, together with 8 proteins identified from the literature, were measured on a custom panel, and validated in a larger material consisting of 1000 case-control pairs. In paper IV, which included 195 colorectal cancer case-control pairs, the protein analysis was extended to include 1536 proteins linked to oncology, inflammation, neurology, and metabolism. In papers using a matched case-control design, conditional i logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CI) for the associations. For longitudinal analyses, mixed effects models were used to estimate associations.  Results: In paper I, we observed a statistically significant inverse association between circulating levels of testosterone and colon cancer. For SHBG there was a statistically significant inverse association prior to adjustment of testosterone and estradiol levels. In paper II, we found one novel genome-wide significant association between circulating levels of dehydroepiandrosterone and DNA methylation at the cg14319657 CpG site. In addition, we also identified more than 40 differentially methylated regions associated with levels of sex hormones and SHBG. In paper III, we first identified 13 proteins associated with CRC risk in the discovery phase. In the validation phase, however, none of the proteins remained significantly associated with colorectal cancer. When stratifying by tumor site, FGF-21 and PPY, were statically significant in colon and rectal cancer respectively, and showed some modest increase in predictive performance. In paper IV, we identified 20 proteins surpassing a significance threshold of 0.005. One protein, TFF3 (Trefoil Factor 3), which was positively associated with colorectal, also withstood strict Bonferroni correction. In addition, we validated several proteins, including AREG, CEA, and LGALS4, which were identified as biomarker candidates in previous studies.  Conclusions: Our results support the hypothesis that circulating sex hormones play a role in male colon cancer etiology and that this may partly explain the difference in colorectal cancer incidence between men and women. Furthermore, our findings suggest a possible link between circulating sex hormones, SHBG and DNA methylation, which could be of interest in the etiology of colorectal cancer as well as other hormone-dependent diseases. Finally, we also identified several proteins associated with colorectal cancer, some of which have shown potential as screening markers.

Automated alarm and root-cause analysis based on real time high-dimensional process data : Part of a joint research project between UmU, Volvo AB & Volvo Cars

Today, a large amount of raw data are available within manufacturing industries. Unfortunately, most of it is not further analyzed in search of valuable information regarding the optimization of processes. In the painting process at the Volvo plant in Umeå, adjusted settings on the process equipments (e.g. robots, machines etc.) are mostly based on the experience of the personnel rather than actual facts (i.e. analyzed data). Consequently, time- and cost waste caused by defects is obtained when painting the commercial heavy-duty truck bodies (cabs). Hence, the aim of this masters thesis is to model the quality as a function of available background- and process data. This should be presented in an automated alarm and root-cause system. A variety of supervised learning algorithms were trained in order to estimate the probability of having at least one defect per cab. Even with a small amount of data, results have shown that such algorithms can provide valuable information. Later in this thesis work, one of the algorithms was chosen and used as the underlying model in the prototype of an automated alarm system. When this probability was considered as too high, an intuitive root-cause analysis was presented. Ultimately, this research has demonstrated the importance and possibility of analyzing data with statistical tools in the search of limiting costs- and time waste

Automated alarm and root-cause analysis based on real time high-dimensional process data : Part of a joint research project between UmU, Volvo AB & Volvo Cars

Today, a large amount of raw data are available within manufacturing industries. Unfortunately, most of it is not further analyzed in search of valuable information regarding the optimization of processes. In the painting process at the Volvo plant in Umeå, adjusted settings on the process equipments (e.g. robots, machines etc.) are mostly based on the experience of the personnel rather than actual facts (i.e. analyzed data). Consequently, time- and cost waste caused by defects is obtained when painting the commercial heavy-duty truck bodies (cabs). Hence, the aim of this masters thesis is to model the quality as a function of available background- and process data. This should be presented in an automated alarm and root-cause system. A variety of supervised learning algorithms were trained in order to estimate the probability of having at least one defect per cab. Even with a small amount of data, results have shown that such algorithms can provide valuable information. Later in this thesis work, one of the algorithms was chosen and used as the underlying model in the prototype of an automated alarm system. When this probability was considered as too high, an intuitive root-cause analysis was presented. Ultimately, this research has demonstrated the importance and possibility of analyzing data with statistical tools in the search of limiting costs- and time waste

Automated alarm and root-cause analysis based on real time high-dimensional process data : Part of a joint research project between UmU, Volvo AB & Volvo Cars

Today, a large amount of raw data are available within manufacturing industries. Unfortunately, most of it is not further analyzed in search of valuable information regarding the optimization of processes. In the painting process at the Volvo plant in Umeå, adjusted settings on the process equipments (e.g. robots, machines etc.) are mostly based on the experience of the personnel rather than actual facts (i.e. analyzed data). Consequently, time- and cost waste caused by defects is obtained when painting the commercial heavy-duty truck bodies (cabs). Hence, the aim of this masters thesis is to model the quality as a function of available background- and process data. This should be presented in an automated alarm and root-cause system. A variety of supervised learning algorithms were trained in order to estimate the probability of having at least one defect per cab. Even with a small amount of data, results have shown that such algorithms can provide valuable information. Later in this thesis work, one of the algorithms was chosen and used as the underlying model in the prototype of an automated alarm system. When this probability was considered as too high, an intuitive root-cause analysis was presented. Ultimately, this research has demonstrated the importance and possibility of analyzing data with statistical tools in the search of limiting costs- and time waste

A two-tiered targeted proteomics approach to identify pre-diagnostic biomarkers of colorectal cancer risk

Colorectal cancer prognosis is dependent on stage, and measures to improve early detection are urgently needed. Using prospectively collected plasma samples from the population-based Northern Sweden Health and Disease Study, we evaluated protein biomarkers in relation to colorectal cancer risk. Applying a two-tiered approach, we analyzed 160 proteins in matched sequential samples from 58 incident colorectal cancer case–control pairs. Twenty-one proteins selected from both this discovery phase and the literature were then analyzed in a validation set of 450 case–control pairs. Odds ratios were estimated by conditional logistic regression. LASSO regression and ROC analysis were used for multi-marker analyses. In the main validation analysis, no proteins retained statistical significance. However, exploratory subgroup analyses showed associations between FGF-21 and colon cancer risk (multivariable OR per 1 SD: 1.23 95% CI 1.03–1.47) as well as between PPY and rectal cancer risk (multivariable OR per 1 SD: 1.47 95% CI 1.12–1.92). Adding protein markers to basic risk predictive models increased performance modestly. Our results highlight the challenge of developing biomarkers that are effective in the asymptomatic, prediagnostic window of opportunity for early detection of colorectal cancer. Distinguishing between cancer subtypes may improve prediction accuracy. However, single biomarkers or small panels may not be sufficient for effective precision screening

An epigenome-wide analysis of sex hormone levels and DNA methylation in male blood samples

Endogenous sex hormones and DNA methylation both play important roles in various diseases. However, their interplay is largely unknown. A deeper understanding of their interrelationships could provide new insights into the pathology of disease development. We, therefore, investigated associations between circulating sex hormones, sex hormone binding globulin (SHBG), and DNA methylation in blood, using samples from 77 men (65 with repeated samples), from the population-based Northern Sweden Health and Disease Study (NSHDS). DNA methylation was measured in buffy coat using the Infinium Methylation EPIC BeadChip (Illumina). Sex hormone (oestradiol, oestrone, testosterone, androstenedione, dehydroepiandrosterone, and progesterone) and SHBG concentrations were measured in plasma using a high-performance liquid chromatography tandem mass spectrometry (LC/MS-MS) method and an enzyme-linked immunoassay, respectively. Associations between sex hormones, SHBG, and DNA methylation were estimated using both linear regression and mixed-effects models. Additionally, we used the comb-p method to identify differentially methylated regions based on nearby P values. We identified one novel CpG site (cg14319657), at which DNA methylation was associated with dehydroepiandrosterone, surpassing a genome-wide significance level. In addition, more than 40 differentially methylated regions were associated with levels of sex hormones and SHBG and several of these mapped to genes involved in hormone-related diseases. Our findings support a relationship between circulating sex hormones and DNA methylation and suggest that further investigation is warranted, both for validation, further exploration and to gain a deeper understanding of the mechanisms and potential consequences for health and disease.Originally included in thesis in manuscript form. </p

Plasma concentrations of gut hormones acyl ghrelin and peptide YY and subsequent risk of colorectal cancer and molecular tumor subtypes

Obesity and metabolic dysfunction are implicated in colorectal cancer development. Appetite-regulating gut hormones might have a role in colorectal cancer risk. We investigated whether circulating levels of the gut hormones ghrelin (analyzed as acyl ghrelin) and Peptide YY (PYY) were associated with subsequent colorectal cancer risk, including clinical and molecular tumor subtypes. We also provide descriptive data on these hormones in relation to background participant characteristics and metabolic biomarkers. This population-based study included 1,010 matched case-control pairs with a median of 12.3 years of follow-up. Acyl ghrelin and PYY were measured by multiplex immunoassay. Data on KRAS and BRAF mutations and microsatellite instability (MSI) status were available for 704 and 708 cases, respectively. Conditional logistic regression models estimated association to colorectal cancer risk. Partial correlation and linear regression were used to investigate relationships between background and metabolic variables and variation in plasma gut hormone concentrations. Acyl ghrelin was not clearly associated with colorectal cancer risk (multivariable OR per 1 SD increase: 1.11; 95% CI, 1.00-1.23). Positive associations were observed for specific subtypes, in particular BRAF-mutated colorectal cancer and right-sided colon cancer, although with nonsignificant heterogeneity. PYY was not related to colorectal cancer risk (multivariable OR per 1 SD: 1.04; 95% CI, 0.95-1.14) or any tumor subtype. In the control participants, ghrelin was inversely correlated with BMI, and PYY was positively correlated with C-peptide and insulin levels. These findings provide limited support for a possible role for ghrelin in colorectal cancer development, primarily in specific anatomical and molecular tumor subtypes. PREVENTION RELEVANCE: The findings of this study do not support a major role for the metabolic gut hormones ghrelin and PYY in colorectal cancer development but suggest the possibility of an involvement for ghrelin in specific tumor subtypes. Elucidating subtype-specific risk factors and mechanisms of carcinogenesis may have implications for precision prevention