Prediagnostic Plasma Samples Research Articles

Validation of a Blood-Based Protein Biomarker Panel for a Risk Assessment of Lethal Lung Cancer in the Physicians' Health Study.

This study aimed to assess a four-marker protein panel (4MP)'s performance, including the precursor form of surfactant protein B, cancer antigen 125, carcinoembryonic antigen, and cytokeratin-19, for predicting lung cancer in a cohort enriched with never- and ever-smokers. Blinded pre-diagnostic plasma samples collected within 2 years prior to a lung cancer diagnosis from 25 cases and 100 sex-, age-, and smoking-matched controls were obtained from the Physicians' Health Study (PHS). The 4MP yielded AUC performance estimates of 0.76 (95% CI: 0.61-0.92) and 0.69 (95% CI: 0.56-0.82) for predicting lung cancer within one year and within two years of diagnosis, respectively. When stratifying into ever-smokers and never-smokers, the 4MP had respective AUCs of 0.77 (95% CI: 0.63-0.92) and 0.72 (95% CI: 0.17-1.00) for a 1-year risk of lung cancer. The AUCs of the 4MP for predicting metastatic lung cancer within one year and two years of the blood draw were 0.95 (95% CI: 0.87-1.00) and 0.78 (95% CI: 0.62-0.94), respectively. Our findings indicate that a blood-based biomarker panel may be useful in identifying ever- and never-smokers at high risk of a diagnosis of lung cancer within one-to-two years.

Abstract 468: Associations of circulating miRNAs with pancreatic cancer risk differ by years between blood collection & and cancer diagnosis

Abstract Pancreatic adenocarcinoma (PDAC) is one of the most fatal cancers and currently used biomarkers for PDAC diagnosis are not adequate for early detection. Therefore, new biomarkers for PDAC early detection are urgently needed. We conducted a large-scale study to prospectively evaluate the association of circulating miRNAs with PDAC risk. Specially, we aimed to examine whether the associations of miRNAs expression levels and changes of those levels from blood collection to cancer diagnosis (BCCD) with PDAC risk. We used pre-diagnostic plasma samples of 1307 PDAC cases from 5 cohort studies (290 from the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial, 395 from the Shanghai Women’s and Men’s Health Studies, 154 from the Southern Community Cohort Study, and 468 from the Multiethnic Cohort). Cancer free controls (n=1307) were selected and individually matched to index cases in each cohort. All miRNAs were measured by the NanoString nCounter Analysis System using the Human v3 miRNA Expression panel (798 miRNAs total). Data were pooled and quantile normalized prior to analysis. Associations of circulating miRNAs with PDAC risk were assessed by odds ratio per decile change in miRNA levels using conditional logistic regression. Fold-change of miRNA for cases versus controls by BCCD was analyzed via linear regression. Median time of BCCD was 7.13 years (range: 0.03-19.61 years). We first conducted the association analysis stratified by BCCD: < 5 years, 5-10 years, and >10 years. We found 20 miRNAs were significantly associated with PDAC risk in the <5 years after FDR correction. In the analysis of regressing fold-change of these 20 miRNAs on BCCD, we found the fold change of miR-155-5p was inversely associated with BCCD (β=-0.04, P<0.01), i.e., the case-control difference was larger when blood was drawn closer to PDAC diagnosis, suggesting its potential utility as disease biomarkers for early detection. Conversely, the fold-change of miR-93-5p (β=0.04, P=0.01), miR-223-3p (β=0.03, P=0.02), let-7i-5p (β=0.03, P<0.01), and miR-191-5p (β=0.05, P<0.01) were positively associated with BCCD, i.e., the case-control difference increased with an increasing the time interval of BCCD, indicating they are more likely biomarkers for PDAC risk. Ingenuity pathway analyses of these five significant miRNAs revealed that they were significantly enriched in ribonucleotide reductase signaling pathways at p<0.05. Notably, the silencing of ribonucleotide reductase M2 subunit suppresses tumorigenesis in pancreatic cancer by deactivating the PI3K/AKT/mTOR pathway, further supporting our findings. Findings of this large-scale multiethnic study suggest that changes in circulating levels of let-7i-5p, miR-155-5p, miR-93-5p, miR-191-5p, and miR-223-3p may help identify individuals at an elevated risk of developing PDAC, allowing for screening tests and close surveillance. Citation Format: Hui Cai, Veronica Wendy Setiawan, Xingyi Guo, Jie Wu, Rachael Z Stolzenberg-Solomon, Claire Zhu, Yu-Tang Gao, Jordan Berlin, Fei Ye, Qiuyin Cai, Wei Zheng, Xiao-ou Shu. Associations of circulating miRNAs with pancreatic cancer risk differ by years between blood collection & and cancer diagnosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 468.

Plasma microRNA ratios associated with breast cancer detection in a nested case–control study from a mammography screening cohort

Mammographic breast cancer screening is effective in reducing breast cancer mortality. Nevertheless, several limitations are known. Therefore, developing an alternative or complementary non-invasive tool capable of increasing the accuracy of the screening process is highly desirable. The objective of this study was to identify circulating microRNA (miRs) ratios associated with BC in women attending mammography screening. A nested case–control study was conducted within the ANDROMEDA cohort (women of age 46–67 attending BC screening). Pre-diagnostic plasma samples, information on life-styles and common BC risk factors were collected. Small-RNA sequencing was carried out on plasma samples from 65 cases and 66 controls. miR ratios associated with BC were selected by two-sample Wilcoxon test and lasso logistic regression. Subsequent assessment by RT-qPCR of the miRs contained in the selected miR ratios was carried out as a platform validation. To identify the most promising biomarkers, penalised logistic regression was further applied to candidate miR ratios alone, or in combination with non-molecular factors. Small-RNA sequencing yielded 20 candidate miR ratios associated with BC, which were further assessed by RT-qPCR. In the resulting model, penalised logistic regression selected seven miR ratios (miR-199a-3p_let-7a-5p, miR-26b-5p_miR-142-5p, let-7b-5p_miR-19b-3p, miR-101-3p_miR-19b-3p, miR-93-5p_miR-19b-3p, let-7a-5p_miR-22-3p and miR-21-5p_miR-23a-3p), together with body mass index (BMI), menopausal status (MS), the interaction term BMI * MS, life-style score and breast density. The ROC AUC of the model was 0.79 with a sensitivity and specificity of 71.9% and 76.6%, respectively. We identified biomarkers potentially useful for BC screening measured through a widespread and low-cost technique. This is the first study reporting circulating miRs for BC detection in a screening setting. Validation in a wider sample is warranted.Trial registration: The Andromeda prospective cohort study protocol was retrospectively registered on 27-11-2015 (NCT02618538).

Abstract 6491: Temporal changes in circulating sex hormones and aromatase activity and associations with lung cancer survival in postmenopausal women

Abstract Background: Exogenous sex hormone supplements had been associated with increased lung cancer mortality in a large clinical trial. No study to date has evaluated temporal trajectories of circulating sex hormones and aromatase activity and their associations with lung cancer survival. Objective: To characterize temporal changes in prediagnostic levels of circulating sex hormones and aromatase activity and to evaluate their associations with overall survival in lung cancer patients. Methods: Included in the analysis were 385 incident lung cancer patients identified in a large prospective cohort among postmenopausal women who had never used cigarette products nor exogenous sex hormone supplements. Concentrations of sex hormones were quantitated using LC-MS/MS assays in prediagnostic plasma samples. The product-substrate molar ratio of estrone to androstenedione was used as an index of aromatase activity (IAA). A multivariable Cox model with restricted cubic spline functions was used to calculate hazard ratio (HR) for overall survival. Results: Of 385 patients with lung cancer, 308 died during a median follow-up of 18.1 years. Initial analyses in all patients showed that higher levels of circulating estrone and IAA and lower levels of androstenedione and testosterone were associated with poorer overall survival after adjusting for nonclinical covariates. In analyses restricted to those with clinical data (n=281), stronger associations were found after further adjustment for tumor stage and treatment regimens. Compared with patients at the median level of IAA, adjusted HRs (95% CI) for total mortality in those at the 30th, 70th, 90th and 95th percentiles were 0.98 (0.84-1.13), 1.05 (0.94-1.16), 1.28 (1.06-1.54), and 1.64 (1.27-2.12), respectively, with P-overall < 0.001. A similar positive association was observed for estrone. In contrast, inverse associations were seen for androgens. For example, compared with the median level of testosterone, HRs (95% CI) for those at the 5th, 10th, 30th, 70th percentiles were 2.10 (1.43-3.09), 1.69 (1.29-2.23), 1.45 (1.19-1.76), and 0.86 (0.78-0.94), respectively, with P-overall = 0.001. Further, we found that circulating levels of sex hormones changed during disease progression. The closer the hormone measurement to cancer diagnosis the higher the IAA and estrone levels (P-overall < 0.001 for both), and the temporal change plateaued 10 years before cancer diagnosis (P-nonlinearity < 0.001 for both). An opposite temporal pattern was found for testosterone. Moreover, the significant association between sex hormones and lung cancer survival was only observed when sex hormones were measured within 10 years before diagnosis. Conclusions: Findings from our study, for the first time, demonstrate temporal changes in circulating sex hormones and their associations with lung cancer survival in postmenopausal women. Citation Format: Yingya Zhao, Xiao-Ou Shu, Yu-Tang Gao, Mark M. Kushnir, Qiuyin Cai, Hui Cai, Qing Lan, Nathaniel Rothman, Wei Zheng, Gong Yang. Temporal changes in circulating sex hormones and aromatase activity and associations with lung cancer survival in postmenopausal women. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6491.

Helicobacter pylori Seropositivity, ABO Blood Type, and Pancreatic Cancer Risk from 5 Prospective Cohorts.

Helicobacter pylori infection may be a risk factor for pancreatic cancer, particularly infection by strains without the cytotoxin-associated gene A (CagA) virulence factor. Non-O blood type is a known risk factor for pancreatic cancer, and H. pylori gastric colonization occurs largely from bacterial adhesins binding to blood group antigens on gastric mucosa. We included 485 pancreatic cancer cases and 1,122 matched controls from 5 U.S. prospective cohorts. Prediagnostic plasma samples were assessed for H. pylori and CagA antibody titers. Conditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) for pancreatic cancer. ABO blood type was assessed using genetic polymorphisms at the ABO gene locus or self-report. Compared to H. pylori seronegative participants, those who were seropositive did not demonstrate an increased risk for pancreatic cancer (OR 0.83, 95% CI 0.65-1.06). This lack of association was similar among CagA seropositive (OR 0.75, 95% CI 0.53-1.04) and seronegative (OR 0.89, 95% CI 0.65-1.20) participants. The association was also similar when stratified by time between blood collection and cancer diagnosis (P-interaction=0.80). Consistent with prior studies, non-O blood type was associated with increased pancreatic cancer risk, but this increase in risk was similar regardless of H. pylori seropositivity (P-interaction=0.51). In this nested case-control study, history of H. pylori infection as determined by H. pylori antibody serology was not associated with pancreatic cancer risk, regardless of CagA virulence factor status. The elevated risk associated with non-O blood type was consistent in those with or without H. pylori seropositivity.

A prospective case–cohort analysis of plasma metabolites and breast cancer risk

BackgroundBreast cancer incidence rates have not declined despite an improvement in risk prediction and the identification of modifiable risk factors, suggesting the need to identify novel risk factors and etiological pathways involved in this cancer. Metabolomics has emerged as a promising tool to find circulating metabolites associated with breast cancer risk.MethodsUntargeted metabolomic analysis was done on prediagnostic plasma samples from a case–cohort study of 1695 incident breast cancer cases and a 1983 women subcohort drawn from Cancer Prevention Study 3. The associations of 868 named metabolites (per one standard deviation increase) with breast cancer were determined using Prentice-weighted Cox proportional hazards regression modeling.ResultsA total of 11 metabolites were associated with breast cancer at false discovery rate (FDR) < 0.05 with the majority having inverse association [ranging from RR = 0.85 (95% CI 0.80–0.92) to RR = 0.88 (95% CI 0.82–0.94)] and one having a positive association [RR = 1.14 (95% CI 1.06–1.23)]. An additional 50 metabolites were associated at FDR < 0.20 with inverse associations ranging from RR = 0.88 (95% CI 0.81–0.94) to RR = 0.91 (95% CI 0.85–0.98) and positive associations ranging from RR = 1.13 (95% CI 1.05–1.22) to RR = 1.11 (95% CI 1.02–1.20). Several of these associations validated the findings of previous metabolomic studies. These included findings that several progestogen and androgen steroids were associated with increased risk of breast cancer in postmenopausal women and four phospholipids, and the amino acids glutamine and asparagine were associated with decreased risk of this cancer in pre- and postmenopausal women. Several novel associations were also identified, including a positive association for syringol sulfate, a biomarker for smoked meat, and 3-methylcatechol sulfate and 3-hydroxypyridine glucuronide, which are metabolites of xenobiotics used for the production of pesticides and other products.ConclusionsOur study validated previous metabolite findings and identified novel metabolites associated with breast cancer risk, demonstrating the utility of large metabolomic studies to provide new leads for understanding breast cancer etiology. Our novel findings suggest that consumption of smoked meats and exposure to catechol and pyridine should be investigated as potential risk factors for breast cancer.

Abstract C081: Associations of plasma tocopherol levels with lung cancer risk among low-income Americans: Results from the Southern Community Cohort Study

Abstract Background: Previous studies reported inverse associations of plasma tocopherol levels with lung cancer risk. No study has been conducted among African Americans (AAs) or the low-income population. Objectives: We conducted a nested case-control study to investigate the associations of plasma tocopherol levels with lung cancer risk among predominantly low-income AAs and European Americans (EAs) living in the Southeastern United States. Methods: This study included 209 incident lung cancer cases and 406 matched controls nested within the Southern Community Cohort Study. Using pre-diagnostic plasma samples collected at the baseline survey, levels of α-, β/γ-, δ-, and total tocopherol were measured by High-Performance Liquid Chromatography with photo-diode array detection. Levels for tocopherol biomarkers―α-, β/γ-, δ-, and total tocopherol―were categorized on the tertile distribution of controls. Conditional logistic regression was used to estimate odds ratios (ORs) and 95% CIs of lung cancer risk after adjusting for potential confounders, including enrollment age, smoking status, pack-years, alcohol consumption, education, household income, self-report of ever having been diagnosed with chronic obstructive pulmonary disease, body mass index, and intake of vitamin supplements. Results: Mean levels of tocopherol biomarkers (ng/ml) were significantly higher among EAs than AAs (12,446 vs. 9,166 for α-tocopherol, 3,325 vs. 2,860 for β/γ-tocopherol, 373 vs. 353 for δ-tocopherol, and 16,145 vs. 12,378 for total tocopherol, respectively; all p&amp;lt;0.05). Overall, higher plasma levels of total tocopherol were significantly inversely associated with lung cancer risk (ORT3 vs. T1 = 0.53, 95% CI: 0.30-0.92; p-trend=0.023 among all participants). The inverse assocaiton was only significant among EAs (ORT3 vs. T1 = 0.19, 95% CI: 0.05-0.64), not AAs (ORT3 vs. T1 = 1.32, 95% CI: 0.64-2.69; p-interaction=0.18). We also found a significant association between total tocopherol and lung cancer risk among men (ORT3 vs. T1 = 0.44, 95% CI: 0.22-0.92), current smokers (ORT3 vs. T1 = 0.51, 95% CI: 0.27-0.95), and cases who were diagnosed within two years following sample collection (ORT3 vs. T1 = 0.36, 95% CI: 0.15-0.87). We also evaluated the associations of tocopherols structural isomers with lung cancer risk. α-tocopherol was significantly associated with decreased risk of lung cancer among EAs (ORT3 vs. T1 = 0.16, 95% CI: 0.04-0.64), but a non-significantly elevated risk among AAs (ORT3 vs. T1 = 1.32, 95% CI: 0.64-2.69; p-interaction=0.01). No significant associations were found for β/γ- and δ-tocopherols after adjustment for all potential confounders. Conclusion: Higher total and α-tocopherol were significantly associated with reduced lung cancer risk, particularly among EAs, men, current smokers, and shorter duration of a lung cancer diagnosis. Further studies to investigate the racial differences in the biological mechanisms between tocopherol biomarkers and lung cancer are warranted to confirm our findings. Citation Format: Hyung-Suk Yoon, Chris Shidal, Wei Zheng, Yan Sun, Jie Wu, Adrian A. Franke, William J. Blot, Xiao-Ou Shu, Qiuyin Cai. Associations of plasma tocopherol levels with lung cancer risk among low-income Americans: Results from the Southern Community Cohort Study [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr C081.

A Cross-Sectional and Longitudinal Analysis of Pre-Diagnostic Blood Plasma Biomarkers for Early Detection of Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) is a major cause of cancer death that typically presents at an advanced stage. No reliable markers for early detection presently exist. The prominent tumor stroma represents a source of circulating biomarkers for use together with cancer cell-derived biomarkers for earlier PDAC diagnosis. CA19-9 and CEA (cancer cell-derived biomarkers), together with endostatin and collagen IV (stroma-derived) were examined alone, or together, by multivariable modelling, using pre-diagnostic plasma samples (n = 259 samples) from the Northern Sweden Health and Disease Study biobank. Serial samples were available for a subgroup of future patients. Marker efficacy for future PDAC case prediction (n = 154 future cases) was examined by both cross-sectional (ROC analysis) and longitudinal analyses. CA19-9 performed well at, and within, six months to diagnosis and multivariable modelling was not superior to CA19-9 alone in cross-sectional analysis. Within six months to diagnosis, CA19-9 (AUC = 0.92) outperformed the multivariable model (AUC = 0.81) at a cross-sectional level. At diagnosis, CA19-9 (AUC = 0.995) and the model (AUC = 0.977) performed similarly. Longitudinal analysis revealed increases in CA19-9 up to two years to diagnosis which indicates a window of opportunity for early detection of PDAC.

Identification of prediagnostic metabolites associated with prostate cancer risk by untargeted mass spectrometry-based metabolomics: A case-control study nested in the Northern Sweden Health and Disease Study.

Prostate cancer (PCa) is the most common cancer form in males in many European and American countries, but there are still open questions regarding its etiology. Untargeted metabolomics can produce an unbiased global metabolic profile, with the opportunity for uncovering new plasma metabolites prospectively associated with risk of PCa, providing insights into disease etiology. We conducted a prospective untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics analysis using prediagnostic fasting plasma samples from 752 PCa case-control pairs nested within the Northern Sweden Health and Disease Study (NSHDS). The pairs were matched by age, BMI,and sample storage time. Discriminating features were identified by a combination of orthogonal projection to latent structures-effect projections (OPLS-EP) and Wilcoxon signed-rank tests. Their prospective associations with PCa risk were investigated by conditional logistic regression. Subgroup analyses based on stratification by disease aggressiveness and baseline age were also conducted. Various free fatty acids and phospholipids were positively associated with overall risk of PCa and in various stratification subgroups. Aromatic amino acids were positively associated with overall risk of PCa. Uric acid was positively, and glucose negatively, associated with risk of PCa in the older subgroup. This is the largest untargeted LC-MS based metabolomics study to date on plasma metabolites prospectively associated with risk of developing PCa. Different subgroups of disease aggressiveness and baseline age showed different associations with metabolites. The findings suggest that shifts in plasma concentrations of metabolites in lipid, aromatic amino acid, and glucose metabolism are associated with risk of developing PCa during the following two decades.

Abstract LB109: Circulating miRNA as biomarkers for pancreatic cancer early detection

Abstract Pancreatic cancer is one of the most fatal human cancers, with an overall 5-year survival rate of 10.8%. Early detection is critical for improving pancreatic cancer prognosis, but biomarkers for early detection are lacking. We conducted a two-stage study to identify circulating miRNAs as biomarkers for pancreatic cancer early detection using pre-diagnostic plasma samples, collected within 5 years prior to cancer diagnosis, from case-control studies nested in five prospective studies. The discovery stage included 185 case-control pairs from the Prostate, Lung, Colorectal and Ovarian Cancer (PLCO) Screening Trial. Replication stage samples comprised 277 case-control pairs from diverse cohorts: Shanghai Women’s and Men’s Health Studies, Southern Community Cohort Study (SCCS), and Multiethnic Cohort. Controls were individually matched on age at enrollment, sex, recruitment site (SCCS), race/ethnicity, and date of blood draw in each cohort. Cell-free small RNAs were extracted from plasma samples and miRNAs were measured by the NanoString nCounter Analysis System using the Human v3 miRNA Expression panel (a total of 798 miRNAs). Normalized miRNAs were categorized by decile. For miRNAs that have ≥10% samples with an undetectable level (0), the non-zero level was categorized by approximately 10% increment. Associations of circulating miRNAs with pancreatic cancer risk, measured in odds ratios (ORs) and 95% confidence intervals (CIs) per decile change, were calculated using conditional logistic regression analyses in discovery and replication studies, separately within each cohort, and results meta-analyzed. We identified three miRNAs, hsa-miR-199a-3p+/hsa-miR-199b-3p, hsa-miR-191-5p, hsa-miR-767-5p, being consistently associated with pancreatic risk in both discovery and replication sets, with combined ORs (95% CIs) of 0.89 (0.84-0.95), 0.90 (0.85-0.95), and 1.08 (1.02-1.13), and P of 9.09E-05, 6.95E-05 and 4.03E-03, respectively. Adjustment for age, BMI, smoking, diabetes and family history of pancreatic cancer did not change the associations. Stratified analyses by age at diagnosis found five additional replicated miRNAs: hsa-miR-640, hsa-miR-1299, hsa-miR-22-3p, hsa-miR-874-5p, and hsa-miR-449b-5p among those 65 years or older, with combined ORs (95% CIs) of 1.33 (1.16-1.52), 1.28 (1.12-1.46), 0.76 (0.65-0.89), 1.25 (1.09-1.43), and 1.22 (1.07-1.39), and P-value ranging from 4.75E-05 to 0.003. These results suggest that circulating miRNA biomarkers may be useful in identifying individuals with high risk of developing pancreatic cancer for close surveillance and/or a screening test. Citation Format: Cong Wang, Hui Cai, Qiuyin Cai, Jie Wu, Rachael Stolzenberg-Solomon, Clair Zhu, Yu-Tang Gao, Jordan Berlin, Fei Ye, Wei Zheng, Veronica W. Setiawan, Xiao-Ou Shu. Circulating miRNA as biomarkers for pancreatic cancer early detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB109.

Circulating Insulin-Like Growth Factor 1-Related Biomarkers and Risk of Lethal Prostate Cancer.

BackgroundExperimental and epidemiologic evidence supports the role of circulating insulin-like growth factor-1 (IGF-1) levels with the risk of prostate cancer. Most circulating IGF-1 is bound to specific binding proteins, and only about 5% circulates in a free form. We explored the relation of free IGF-1 and other components of the IGF system with lethal prostate cancer.MethodsUsing prospectively collected samples, we undertook a nested case-only analysis among 434 men with lethal prostate cancer and 524 men with indolent, nonlethal prostate cancer in the Physicians’ Health Study and the Health Professionals Follow-up Study. Prediagnostic plasma samples were assayed for free IGF-1 and total IGF-1, acid labile subunit, pregnancy-associated plasma protein A (PAPP-A), and intact and total IGF binding protein 4. We estimated odds ratios (ORs) and corresponding 95% confidence intervals (CIs) for the associations between IGF-1–related biomarkers and lethal prostate cancer using unconditional logistic regression models adjusted for age, height, and body mass index.ResultsMen in the highest quartile of PAPP-A levels had 42% higher odds of lethal prostate cancer (pooled adjusted OR = 1.42, 95% CI = 1.04 to 1.92) compared with men in the lowest 3 quartiles. There were no statistically significant differences in the other plasma analytes. The positive association between PAPP-A and lethal prostate cancer was present among men with intact PTEN but not among those with tumor PTEN loss (2-sided Pinteraction = .001).ConclusionsOur study provides suggestive evidence that among men who later develop prostate cancer, higher plasma PAPP-A levels measured prior to diagnosis are associated with increased risk of lethal compared with indolent disease.

Circulating Tissue Polypeptide-Specific Antigen in Pre-Diagnostic Pancreatic Cancer Samples.

Simple SummaryDetecting cancer early significantly increases the chances of successful (surgical) treatment. Pancreatic cancer is one of the deadliest cancer forms, since it is usually discovered at a late and already spread stage. Finding biomarkers showing pancreatic cancer at an early stage is a possible approach to early detection and improved treatment. The aim of our study was to assess the potential of tissue polypeptide specific antigen (TPS) as a biomarker for early pancreatic cancer detection. We studied TPS levels in blood plasma samples from a population-based biobank in Västerbotten, Sweden that were collected before individuals were diagnosed with pancreatic cancer. Although TPS levels are raised at diagnosis, this occurs late, and thus TPS does not seem to hold promise as an early detection marker for pancreatic cancer.Early detection of pancreatic ductal adenocarcinoma (PDAC) is challenging, and late diagnosis partly explains the low 5-year survival. Novel and sensitive biomarkers are needed to enable early PDAC detection and improve patient outcomes. Tissue polypeptide specific antigen (TPS) has been studied as a biomarker in PDAC diagnostics, and it has previously been shown to reflect clinical status better than the ‘golden standard’ biomarker carbohydrate antigen 19-9 (CA 19-9) that is most widely used in the clinical setting. In this cross-sectional case-control study using pre-diagnostic plasma samples, we aim to evaluate the potential of TPS as a biomarker for early PDAC detection. Furthermore, in a subset of individuals with multiple samples available at different time points before diagnosis, a longitudinal analysis was used. We assessed plasma TPS levels using enzyme-linked immunosorbent assay (ELISA) in 267 pre-diagnostic PDAC plasma samples taken up to 18.8 years before clinical PDAC diagnosis and in 320 matched healthy controls. TPS levels were also assessed in 25 samples at PDAC diagnosis. Circulating TPS levels were low both in pre-diagnostic samples of future PDAC patients and in healthy controls, whereas TPS levels at PDAC diagnosis were significantly increased (odds ratio 1.03; 95% confidence interval: 1.01–1.05) in a logistic regression model adjusted for age. In conclusion, TPS levels increase late in PDAC progression and hold no potential as a biomarker for early detection.

Abstract 749: Plasma insulin-like growth factor 1-related biomarkers and risk of lethal prostate cancer

Abstract Background Experimental and epidemiologic evidence supports the role of plasma IGF-1 and risk of prostate cancer. About 5% of IGF-1 circulates in a free or bioavailable form, and is only weakly correlated with total IGF-1; we hypothesized that higher levels of free IGF-1 would be associated with risk of lethal prostate cancer. Methods Lethal prostate cancer was defined as fatal prostate cancer plus metastatic prostate cancer. Non-lethal prostate cancer was defined as cases in which the men remained free of known metastases for at least eight years. Using prospectively collected samples in a nested design, we identified 434 lethal cases and 524 men with non-lethal prostate cancer in two prospective cohorts: the Physicians' Health Study (mean years of follow-up 33.2) and the Health Professionals Follow-up Study (mean years of follow-up 18.5). Circulating levels in prediagnostic plasma samples were assayed for IGF-1-related biomarkers, including free and total IGF-1, acid labile subunit (ALS), pregnancy-associated plasma protein A (PAPP-A, a protease that cleaves the IGF complex), intact IGF binding protein 4 (IGFBP-4), and total IGFBP-4, with risk of lethal prostate cancer. We estimated odds ratios (ORs) and corresponding 95% confidence intervals (CI) for the associations between IGF-1-related biomarkers (in quartiles) and lethal prostate cancer using unconditional logistic regression models adjusted for age, height, weight, and body mass index. Subgroup analyses were conducted by time from blood draw to diagnosis, and tumor biomarkers, ERG as a marker of the TMPRSS2:ERG fusion, phosphatase and tensin homolog (PTEN) loss, and IGF-1 receptor (IGF1R) protein expression. Results We observed no significant association between free IGF-1 and lethal prostate cancer (pooled adjusted OR for the highest versus lowest group 0.93, 95% CI 0.64 to 1.35) after adjusting for potential covariates. However, men in the highest quartile of PAPP-A levels had 43% higher odds of developing lethal prostate cancer (pooled adjusted OR 1.43, 95% CI 1.05 to 1.95) compared to men in the lowest three quartiles. The positive association between PAPP-A and lethal prostate cancer was present among men without PTEN loss, but not among those with (P for interaction = 0.002). There were no significant differences across the two cohorts (P for heterogeneity &amp;gt; 0.05 for all) and no significant associations were observed between other plasma biomarkers and lethal prostate cancer. Conclusions We found no significant association between free IGF-1 and lethal prostate cancer, but provide suggestive evidence that higher PAPP-A levels are associated with an increased risk of developing lethal prostate cancer. This observation merits testing in other cohorts. Citation Format: Chaoran Ma, Ye Wang, Lorelei A. Mucci, Meir J. Stampfer, Michael Pollak, Kathryn L. Penney. Plasma insulin-like growth factor 1-related biomarkers and risk of lethal prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 749.

Intra-Individual Changes in Immune Marker Concentrations and Multiple Myeloma Risk: A Longitudinal Prospective Study Using Repeated Pre-Diagnostic Plasma Samples

Background: We studied immune marker trajectories (i.e. intra-individual changes in immune marker concentrations over time) of ten immune markers in relation to multiple myeloma (MM) risk. Low blood concentrations of MCP-3, MIP-1α, VEGF, FGF-2, Fractalkine, and TGF-α have been associated with myeloma risk in a previous study of single blood samples per participant collected around six years before diagnosis (Vermeulen et al., submitted). Longitudinal studies regarding these immune markers may provide substantial input to understanding their natural history in myeloma development and how these markers could be used in clinical routine.Methods: We identified 66 MM cases who had donated two blood samples before myeloma diagnosis, as well as matched cancer-free controls from the Northern Sweden Health and Disease Study. Blood levels of MCP-3, MIP-1α, MIP-1β, VEGF, FGF-2, Fractalkine, TGF-α, IL-13, TNF-α, and IL-10 were assessed in duplicate using Luminex bead-based multiplex assays (Millipore, Billerca, MA). Samples from matched cases and controls, as well as repeated samples from the same individuals, were included in random order within the same analytical batch. Linear mixed effects modeling was applied to investigate differences in immune marker trajectories between cases and controls.Findings: Samples were donated on median 12.3 ± 4.7 and 4.7 ± 3.8 years (± SD) prior to MM diagnosis for the baseline and the repeated sample, respectively. Median age at myeloma diagnosis was 62.9 ± 8.6 years (± SD). MM cases had significantly lower plasma levels of MCP-3, VEGF, FGF-2, Fractalkine, and TGF-α compared to controls. Notably, these immune markers also decreased significantly over time, while remaining temporally stable among control individuals. Decreasing marker trajectories were especially pronounced for VEGF (β = -0.012, P= 3.450 x 10-4), FGF-2 (β = -0.010, P= 5.700 x 10-5), and TGF-α (β = -0.019, P= 3.640 x 10-4) among future myeloma patients the closer to diagnosis the sample was donated. Subgrouping of samples by time between blood draw and diagnosis showed that plasma levels of MCP-3 were inversely associated with myeloma risk in samples collected as long as 12 to 24 years before diagnosis (OR = 0.23, Ptrend = 0.036) (OR = 4th vs. 1st quartile).Interpretation: Our data provide further support of inverse associations between MM risk and plasma levels of MCP-3, VEGF, FGF-2, Fractalkine, and TGF-α. Low MCP-3 levels might be involved in early myeloma development. Considering that monoclonal gammopathy of unclear significance (MGUS) is detectable in the majority of myeloma patients in samples collected up to 15 years before diagnosis (Landgren et. al. Blood 2009, Weiss et. al. Blood 2009), decreasing immune marker levels of VEGF, FGF-2, and TGF-α might be indicative of progression to MM among MGUS patients. Therefore, these markers could be of potential use as biomarkers to improve prediction of myeloma progression in patients followed for MGUS. However, larger studies including samples from both, myeloma and MGUS patients together with matched controls are needed to further evaluate this hypothesis. DisclosuresNo relevant conflicts of interest to declare.

Prospective analysis of circulating metabolites and endometrial cancer risk

BackgroundEndometrial cancer is strongly associated with obesity and dysregulation of metabolic factors such as estrogen and insulin signaling are causal risk factors for this malignancy. To identify additional novel metabolic pathways associated with endometrial cancer we performed metabolomic analyses on pre-diagnostic plasma samples from 853 case-control pairs from the European Prospective Investigation into Cancer and Nutrition (EPIC). MethodsA total of 129 metabolites (acylcarnitines, amino acids, biogenic amines, glycerophospholipids, hexoses, and sphingolipids) were measured by liquid chromatography-mass spectrometry. Conditional logistic regression estimated the associations of metabolites with endometrial cancer risk. An analysis focusing on clusters of metabolites using the bootstrap lasso method was also employed. ResultsAfter adjustment for body mass index, sphingomyelin [SM] C18:0 was positively (OR1SD: 1.18, 95% CI: 1.05–1.33), and glycine, serine, and free carnitine (C0) were inversely (OR1SD: 0.89, 95% CI: 0.80–0.99; OR1SD: 0.89, 95% CI: 0.79–1.00 and OR1SD: 0.91, 95% CI: 0.81–1.00, respectively) associated with endometrial cancer risk. Serine, C0 and two sphingomyelins were selected by the lasso method in >90% of the bootstrap samples. The ratio of esterified to free carnitine (OR1SD: 1.14, 95% CI: 1.02–1.28) and that of short chain to free acylcarnitines (OR1SD: 1.12, 95% CI: 1.00–1.25) were positively associated with endometrial cancer risk. Further adjustment for C-peptide or other endometrial cancer risk factors only minimally altered the results. ConclusionThese findings suggest that variation in levels of glycine, serine, SM C18:0 and free carnitine may represent specific pathways linked to endometrial cancer development. If causal, these pathways may offer novel targets for endometrial cancer prevention.

Longitudinal change of metabolite profile and its relation to multiple risk factors for the risk of developing hepatitis B-related hepatocellular carcinoma.

Despite considerable knowledge of viral pathogenesis, the pathophysiological changes related to the multifactorial, multistep process of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) development remains unclear. Longitudinal metabolomics study can reveal biological process for disease progression. We performed metabolite profiling with longitudinal prediagnostic plasma samples from two nested case-control studies of hepatitis B surface antigen carriers participating in ultrasound screening for HCC, one within a government employee cohort (870 samples from 109 HCC cases and 107 controls) and the other within a hospital-based cohort (266 samples from 63 HCC cases and 114 controls). Of the 34 measured metabolites, tyrosine, isoleucine, and glutamine were consistently associated with HCC. In analyses combining longitudinal data, a high metabolic risk score based on the three amino acids was robustly associated with increased risk of HCC (OR = 3.71, 95% confidence interval: 2.53-5.42), even after adjustment for clinical factors, or when assessed for different times up to ≥8 years before diagnosis. Similar association was observed in an independent, prospective analysis comprising 633 randomly selected individuals of the government employee cohort. More importantly, this metabolite signature was longitudinally influenced by HBV-infection phase and involved in gradual progression to liver fibrosis and cirrhosis. Furthermore, mediation analysis showed that the score mediated substantial proportions of the associations of key viral factors, insulin resistance, and diabetes status with HCC risk. Our results suggest that an amino-acid dysregulation metabotype may play a role in HBV-related HCC development, and may also be linked to common pathways that mediate increased HCC risks.

Abstract A06: Associations of prediagnostic metabolomic profiles with colorectal cancer risk—a collaborative reanalysis within the COnsortium of METabolomic Studies (COMETS)

Abstract Background: An improved understanding of the etiology of colorectal cancer is needed to allow more accurate disease prediction. This study will use a state-of-the-art metabolomics strategy leveraging existing data from 8 prospective cohort studies to discover and validate novel blood-based biomarkers that are associated with colorectal cancer risk. This project is in its early phase and the abstract is focusing on describing the study design. Methods: We will perform a meta-analysis using data from 8 large-scale prospective cohort studies that are part of the Consortium of Metabolomics Studies (COMETS): European Prospective Investigation into Cancer and Nutrition, Estonian Biobank, Women Health Study – Observational Study, Prostate, Lung, Colorectal and Ovarian Cancer Observational Study, Nurses’ Health Study, Health Professional Follow-up Study, Shanghai Women’s and Men’s Health Studies. All studies used a nested case-control study design (1:1 matching at a minimum by age and sex). Prediagnostic plasma and serum samples were analyzed using state-of-the art metabolomics platforms: Metabolon, Inc., Broad Institute and Biocrates. We expect n~150 overlapping identified metabolites across platforms. Data will be split into discovery and validation set. We will investigate associations of metabolites with colorectal cancer risk using multivariable conditional logistic regression models and random effects meta-analysis. Analyses will be stratified by age (&amp;lt;45, 45-55, ≥55 years) and site (colon versus rectal). Heterogeneity between studies will be assessed using Q-value and I2 statistics. Metabolites at the 0.05 false discovery rate adjusted significance levels will be carried forward to the validation set. Results: The present study includes n=2,093 case-control pairs in the discovery dataset and n=1,082 case-control pairs in the validation dataset. Demographic data such as age at blood draw and age at diagnosis, race, body mass index, smoking, and dietary intake at baseline (red meat, vegetables, fruits, fat, and fiber) are collected. Data on tumor site are &amp;gt;95% complete. Tumor stage is available for &amp;gt;85% of patients and is expected to increase substantially since medical data abstraction is under way in some cohorts. Conclusions: This is the largest study, to our knowledge, investigating the associations of prediagnosis metabolomic profiles with colorectal cancer risk. The proposed analyses have the potential to identify biomarkers that may enhance population-wide and tailored screening of colorectal cancer risk. Citation Format: Jennifer Ose, Cornelia M. Ulrich, Mary Playdon, Ken Boucher, A. Heather Elisassen, Xia-Ou Shu, Wei Zheng, Andres Metspalu, Tonu Esku, Jessica Lasky-Su, Steve Moore, Marc Gunter. Associations of prediagnostic metabolomic profiles with colorectal cancer risk—a collaborative reanalysis within the COnsortium of METabolomic Studies (COMETS) [abstract]. In: Proceedings of the AACR Special Conference on Modernizing Population Sciences in the Digital Age; 2019 Feb 19-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(9 Suppl):Abstract nr A06.

Abstract LB-152: Associations between circulating sex hormone levels and risk of colon cancer among men from six European countries

Abstract Colorectal cancer (CRC) is the third most common cancer worldwide and its incidence is rising. Identifying etiologic pathways and early detection for CRC are key to prevention and reducing mortality as the prognosis for advanced CRC, despite recent improvements in therapy, remains poor. One area of potentially measurable CRC risk factors, that is still relatively unexplored, are circulating levels of endogenous sex hormones. The incidence of CRC is greater in men than in women and a proposed hypothesis for this sex disparity is that higher levels of endogenous estrogens found in women may be protective against CRC. In addition, androgens, including testosterone, have been hypothesized to confer protective effects for CRC in men. However, few studies have evaluated the association between testosterone and CRC in men, and the limited available data suggest that any observed protective association is likely confined to colon rather than rectal cancer. The aim of this study was therefore to evaluate the relation of colon cancer risk with circulating levels of sex hormones and sex hormone binding globulin (SHBG) in a male population. Preliminary analyses included data from 461 cases of incident colon cancer and 461 matched controls from the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort. Sex hormones (estradiol, estrone, testosterone, androstenedione, dehydroepiandrosterone, and progesterone) were measured in prediagnostic plasma samples using liquid chromatography-tandem /mass spectrometry (LC/-MS/MS), and SHBG was measured using a solid phase “sandwich” immunoradiometric assay. To test potential associations between circulating levels of sex hormones and SHBG with the risk of developing colon cancer, conditional logistic regression models were applied to estimate odds ratios (OR) and 95% confidence intervals (95% CI). Multivariable models were adjusted for colon cancer risk factors, including anthropometric parameters, alcohol consumption, physical activity and smoking status. In addition, models including estrone, estradiol and testosterone were adjusted for SHBG and vice versa. Preliminary results indicated an inverse association for testosterone (multivariable OR per SD: 0.80 95% CI: 0.69 - 0.93) and SHBG (multivariable OR per SD: 0.84 95% CI: 0.72 - 0.99) with CRC risk. However, after additional adjustments to SHBG and sex hormone models respectively these associations were attenuated towards the null. No associations were identified for estradiol, estrone, androstenedione, dehydroepiandrosterone, and progesterone. These preliminary results do not support an association between endogenous estrogen and colon cancer in men, while a potential association between testosterone and SHBG and colon cancer will need to be further evaluated. Citation Format: Justin Harbs, Sabina Rinaldi, Audrey Gicquiau, Xijia Liu, Marc Gunter, Bethany Van Guelpen, Neil Murphy, Sophia Harlid, Epic Network. Associations between circulating sex hormone levels and risk of colon cancer among men from six European countries [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr LB-152.

Early Detection of Post-Transplant Lymphoproliferative Disorder Using Circulating Tumor DNA

Purpose Diffuse large B cell (DLBCL)-like post-transplant lymphoproliferative disorder (PTLD) affects up to 10% of thoracic transplant recipients. Although 50% of PTLDs can be related to Epstein-Barr virus (EBV) infection, and high or increasing EBV viral load may precede EBV+ PTLD, preemptive monitoring of EBV titers does not reliably predict PTLD . Consequently, we assessed the utility of circulating tumor DNA (ctDNA), a promising biomarker for many cancers including DLBCL , for early detection of PTLD. Methods We used Cancer Personalized Profiling by Deep Sequencing (CAPP-Seq) to analyze diagnostic tumor and plasma samples and matched germline DNA in a cohort of 10 heart and lung transplant recipients who developed PTLD to identify somatic alterations. We compared mutational patterns in PTLD to 173 de novo DLBCL cases. We additionally sequenced serial post-transplant plasma samples preceding diagnosis in 5 patients (2 EBV-, 3 EBV+), and during or after treatment in 8 patients, to determine the dynamics of ctDNA and compared these patterns to clinical outcomes and EBV titers. Results There was no significant difference in mutational burden between patients with PTLD and de novo DLBCL (median PTLD variants: 216, IQR = 79 - 321; median de novo DLBCL variants: 143.5, IQR = 45 - 243.8; p = 0.35), although there was a trend toward greater variant count in EBV- compared to EBV+ PTLD (p = 0.064). The prevalence of alterations in known driver genes was similar between PTLD and de novo DLBCL. Prediagnostic plasma samples were available for 5 cases, and we detected ctDNA in all patients prior to clinical diagnosis (mean lead time = 114 days, maximum lead time 1.5 years). Levels of ctDNA during and after treatment were concordant with EBV titers in EBV+ disease and with clinical response in all patients. Conclusion PTLD patients have detectable ctDNA prior to clinical diagnosis. Development of screening tools utilizing both ctDNA and EBV titers would facilitate early detection of PTLD in the transplant population.

Biomarkers of vascular injury in relation to myocardial infarction risk: A population-based study

Little is known about circulating biomarkers of vascular injury in relation to cardiovascular disease risk. Thus, we evaluated associations between six novel markers (E-Selectin, P-Selectin, thrombomodulin, thrombopoietin, intercellular adhesion molecule 3 and GPIIb/IIIa) and established cardiovascular risk factors as well as the risk of myocardial infarction (MI) in a population-based study. Biomarkers were measured in pre-diagnostic plasma samples of a case-cohort subset of EPIC-Heidelberg (incident MI cases: n = 369, random sub-cohort: n = 2,418). Generalized Linear models were used to analyse cross-sectional associations between biomarkers and cardiovascular risk factors. Multivariable Cox Regression analyses were carried out to obtain Hazard Ratios (HRs) of MI across quartiles of biomarkers levels. Cross-sectional analyses showed that sex, smoking, alcohol consumption, diabetes and exogenous hormone use were associated with biomarker levels. However, while fibrinogen was associated with MI risk (HR per standard deviation: 2.97 [95% confidence interval: 1.61, 5.46]), none of the six novel biomarkers was associated with MI risk after multivariable adjustment. In a population-based cohort, biomarkers of vascular injury were associated with established cardiovascular risk factors, but not MI risk. The tested biomarkers may reflect pathophysiological alterations in cardiovascular disease development rather than constituting independent MI risk factors.