Background: Receptor activator of nuclear factor-κB (RANK) and its ligand (RANKL) regulate bone metabolism, immune tolerance, immunity, and mammary gland biology. RANKL, induced by progesterone, prolactin, vitamin D3, PTHrP, TNF-α, prostaglandin E2, and interleukins, impacts osteoclast differentiation, lymph node formation, breast cell proliferation, and immune regulation. RANKL/RANK is essential for mammary stem and progenitor cell populations linked to progesterone and BRCA1 mutation-driven breast cancers. This pilot investigated RANKL inhibition with denosumab, a therapeutic anti-RANKL antibody, to assess effects on breast density in postmenopausal women. Methods: This single-arm, open-label pilot administered 120 mg of denosumab subcutaneously monthly for 6 doses to postmenopausal early-stage breast cancer patients. Patients may have received aromatase inhibitors. Those on SERMs were ineligible. Breast density was measured by MR imaging of the breast at baseline and after 6 months. Analysis was performed on 25 evaluable patients. The primary endpoint was the MRI-measured breast density (MRD) change from baseline to the end of study, approximately one month after last injection. A ≥5% relative decrease in breast density in ≥ 30% of participants was predefined to warrant a larger study. Results: Thirty postmenopausal women with a history of breast cancer received the intervention, 25 evaluable with 5 excluded (withdrew consent, image distortion, or ineligibility). All evaluable patients completed baseline and 6-month MRI measurements and received at least 5 doses of monthly denosumab. Median age was 60 years, median BMI was 28.5 kg/m2, and 44% were on aromatase inhibitors. Median baseline MRD was 16.2% (range 4.91-45.34%). Fourteen (56%) were characterized as heterogeneously dense (BI-RADS c), one (4%) extremely dense (BI-RADS d), and ten (40%) as scattered fibroglandular by mammography. Excluding two high density outliers, baseline RANKL correlated with breast density (Spearman’s rho = 0.51). All patients had undetectable RANKL levels after 6 months of denosumab. Relative MRD decreased by -8.19% (mean) and -6.34% (median) for the primary endpoint after 6 months. Fifteen subjects (60%, 95% CI= 42-75%) had a relative MRD decrease of ≥5%, with only two (8%) increasing. There was no significant correlation between changes in MRD and baseline MRD, age, BMI, or baseline RANKL levels. As a reference, a similar population of postmenopausal women on aromatase inhibitors (N=39) followed for 6 months, 21% had a decrease in MRD of ≥5%, 46% had no change, and 33% had an increase of ≥5%. The difference between DMAB and the ref group remained significant after multivariate adjustment (p=0.01). Denosumab was well tolerated, with no significant or unexpected toxicities. Seventeen doses were administered at home by an RN. Exploratory analyses of a subset of patients with available breast biopsies revealed potential correlations between change in MRD following denosumab treatment and baseline immune cell phenotypes including for example higher baseline RANKL+/CD8+ T cells in participants with greater decreases in MRD at 6 months (rho= -0.49, p= 0.10). Conclusions: In this pilot, therapeutic suppression of RANKL for putative breast cancer prevention was well-tolerated and could be administered at home. RANKL levels correlated with baseline breast density, and treatment for 6 months was associated with a reduction in breast density that favors effects of RANKL inhibition on breast tissue. Our results support further studies to define denosumab and RANKL's effects on breast density, including effects that may be mediated through a RANKL-positive breast immune microenvironment in postmenopausal women. Citation Format: Patricia Thompson, Krisha Mehta, Jie Yang, Chuan Huang, Yunhan Liao, Eduardo Scandinari Manzolli, Christina Preece, Caterina Vacchi-Suzzi, Alison Stopeck. Phase II, Single Arm Trial of Denosumab’s Effect on Breast Density and Breast Tissue Biomarkers [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P3-02-04.

To evaluate the feasibility of obtaining a 21-gene Oncotype DX Breast Recurrence Score® (RS) result from metastatic biopsies in newly diagnosed hormone receptor positive, HER2− metastatic breast cancer patients and correlate RS results with matched primary samples. Retrospective analysis of metastatic biopsies from HR+, HER2− breast cancer patients. Slides were sent to Genomic Health, Inc. for RNA isolation and RS determination. Success rates were evaluated across metastatic sites, and RS results were compared between matched primary and metastatic samples. RS result was obtained in 46

Sphingomyelin Synthase 1 (gene name: SGMS1 ) participates in regulation of sphingolipid levels by synthesizing sphingomyelin from ceramide and phosphatidylcholine. Evidence have supported SGMS1's functions in regulating proliferation, cell cycle, cell death and migration. While its functions have begun to be explored, very little is known about upstream regulators. Here, we demonstrate that SGMS1 is a direct gene target of the GATA1-TAL1 transcriptional complex in K562 erythroleukemic cells. A predicted GATA1 consensus DNA binding sequence was identified with in a newly characterized alternative SGMS1 promoter (TSS-7 promoter) and ChIP analysis confirmed GATA1 occupancy on the promoter. Down-regulation of GATA1 in K562 cells significantly decreased SGMS1 expression and enzymatic activity, and ChIP-Seq analysis from ENCODE showed colocalized peaks for GATA1 and TAL1 ( a well-established GATA1 DNA binding partner) on the SGMS1 gene. Analysis of publicly available datasets shows that elevated GATA1, TAL1 and SGMS1 expression not only clusters GATA1 positive chronic myelogenous leukemia cells (like K562), but also selectively identifies acute erythrocytic and megakaryocytic leukemias (M6 and M7 AML, respectively). Microarray gene expression analysis after down-regulation of SGMS1 in M6 AML Hel cells revealed alteration of genes regulating G2/M check point and mitotic spindle formation. This phenotype was functionally confirmed by the significant delay in G2/M cell cycle progression of cells with SGMS1 downregulation and sensitization to the clinically relevant anti-mitotic agent, Taxol. Altogether, these results identify SGMS1 as a novel target of GATA1/TAL1 transcriptional complex and they support a role for the GATA1/TAL1/SGMS1 axis in regulating transit through G2/M. Importantly, results also point to combination of anti-mitotic agents and inhibition of SGMS1 as a potential novel therapeutic approach against the aggressive and resilient M6 AMLs. KEY POINTS:The Sphingomyelin Synthase 1 gene ( SGMS1 ) is a novel direct target of GATA1 and TAL1. High SGMS1 levels are associated with high GATA1/TAL1 expression and regulate cell cycle progression through the G2/M checkpoint in GATA1 + erythroleukemic Acute Myeloid Leukemia Hel cells. High SGMS1 is associated with lower probability of survival of patients with Acute Myeloid Leukemia and down-regulation of SGMS1 co-operates with microtubule targeting agents to induce cytotoxicity in GATA1 positive AML Hel cells.

Background: Acute myeloid leukemia (AML) is an aggressive hematologic malignancy characterized by the clonal expansion of immature myeloblasts from leukemic stem cells (LSCs) and the disruption of normal hematopoiesis by hematopoietic stem cells (HSCs). Currently, treatment response is evaluated at the end of induction therapy (Days 28 ± 7). Despite an overall remission rate of about 70% for newly diagnosed AML and 30-50% for relapsed/refractory AML, many patients endure toxic treatments without clinical benefit. There is a need for biomarkers that enable timely evaluation of treatment response and facilitate real-time therapy adjustments that optimize therapeutic benefit while minimizing unnecessary toxicity. Our prior retrospective study at Stony Brook University Hospital (SBUH) evaluated the dynamic changes of LSC versus HSC in AML patients using flow cytometric markers. Among the five LSC markers studied, changes in LSC/HSC subsets identified by CLL1 and CD45RA showed the strongest correlation with treatment response. Moreover, the presence of CLL1+ or CD45RA+ LSC/HSC subsets in peripheral blood correlated highly with bone marrow aspirate findings, suggesting that peripheral blood monitoring of LSC/HSC subsets could be a timely, non-invasive method to track LSC/HSC dynamics during AML treatment. Aims: The primary aims of this study are to assess: 1) the feasibility of tracking changes in LSC/HSC subsets using peripheral blood CLL1 and CD45RA as biomarkers on Days 3, 5, and 7 of induction therapy in AML patients, and 2) the feasibility of using LSC/HSC subsets to predict therapeutic response as evaluated by bone marrow examination at the end of induction cycle (Days 28 ± 7). Methods: This pilot prospective study plans to recruit 20 AML patients from SBUH who are 18 years or older, with newly diagnosed or relapsed/refractory AML and pre-treatment LSC marker (CLL1 and/or CD45RA) expression ≥50% of total HSCs. Following informed consent, peripheral blood samples are collected and processed for multicolor flow cytometry (MFC) on Days 3, 5, and 7 of induction therapy. The HSC population is defined as CD45dim/SSC and CD34+CD38low/-. CLL1 or CD45RA positive HSCs are identified as LSCs. LSC/HSC subsets are calculated as the percentage of CLL1+ or CD45RA+ cells within the HSC population. Changes in LSC/HSC subsets from pre-treatment will be evaluated and compared between patients with persistent disease and those achieving complete remission after induction chemotherapy. This study is registered at ClinicalTrials.gov (NCT06297551). Results: To date, 7 patients with newly diagnosed or relapsed AML have been enrolled. In the 4 patients who achieved complete remission, LSC/HSC subsets changed by an average of -14.22 ± 18.70% on Day 3, -65.19 ± 29.65% on Day 5, and -78.26 ± 20.76% on Day 7 compared to pre-treatment levels (normalized as 0% change). In the 3 patients with persistent disease after induction therapy, LSC/HSC subsets changed by an average of +19.30 ± 23.55% on Day 3, +11.60 ± 18.34% on Day 5, and -2.57 ± 5.11% on Day 7. Conclusions: Our preliminary results indicate that measuring the change in peripheral blood CLL1+ or CD45RA+ LSC/HSC subsets on Days 5 to 7 of induction therapy is feasible and may predict therapeutic outcome in AML. A significant decrease in CLL1+ or CD45RA+ LSC/HSC subsets in peripheral blood was observed as early as Days 5 to 7 of induction therapy, and this decrease in LSC/HSC subset biomarkers appears to be associated with complete remission by the end of the treatment cycle. Conversely, a lack of reduction in these subsets was seen in patients with persistent disease post induction therapy. Recruitment is ongoing, and we hope that additional data will further refine our findings for this study. In future studies, we hope to determine the optimal threshold of CLL1+ and CD45RA+ LSC/HSC subset change on Days 5 to 7 of induction therapy that is associated with remission versus disease persistence.

Nanoliposomal irinotecan (nal-IRI) is a promising novel hyperthermic intraperitoneal chemotherapy (HIPEC) agent given its enhanced efficacy against gastrointestinal tumors, safety profile, thermo-synergy, and heat stability. This report describes the first in-human phase 1 clinical trial of nal-IRI during cytoreductive surgery (CRS) and HIPEC. Patients with peritoneal surface disease (PSD) from appendiceal and colorectal neoplasms were enrolled in a 3 + 3 dose-escalation trial using nal-IRI (70–280 mg/m2) during HIPEC for 30 min at 41 ± 1 °C. The primary outcome was safety. The secondary outcomes were pharmacokinetics (PK) and disease-free survival. Adverse events (AEs) categorized as grade 2 or higher were recorded. The serious AEs (SAEs) were mortality, grade ≥ 3 AEs, and dose-limiting toxicity (DLT). Irinotecan and active metabolite SN38 were measured in plasma and peritoneal washings. The study enrolled 18 patients, who received nal-IRI during HIPEC at 70 mg/m2 (n = 3), 140 mg/m2 (n = 6), 210 mg/m2 (n = 3), and 280 mg/m2 (n = 6). No DLT or mortality occurred. The overall morbidity for CRS/HIPEC was 39% (n = 7). Although one patient experienced neutropenia, no AE (n = 131) or SAE (n = 3) was definitively attributable to nal-IRI. At 280 mg/m2, plasma irinotecan and SN38 measurements showed maximum concentrations of 0.4 ± 0.6 µg/mL and 3.0 ± 2.4 ng/mL, a median time to maximum concentration of 24.5 and 26 h, and areas under the curve of 22.6 h*µg/mL and 168 h*ng/mL, respectively. At the 6-month follow-up visit, 83% (n = 15) of the patients remained disease-free. In this phase 1 HIPEC trial (NCT04088786), nal-IRI was observed to be safe, and PK profiling showed low systemic absorption overall. These data support future studies testing the efficacy of nal-IRI in CRS/HIPEC.

Background: Identifying when to start CDK4/6 inhibitors or use chemotherapy in hormone receptor positive (HR+) metastatic breast cancer (MBC) remains challenging. The 21-gene Oncotype DX Breast Recurrence Score® test is validated to predict chemotherapy benefit in early stage HR+ breast cancer but has not been studied for use in the MBC setting.Objective: To assess the feasibility of obtaining Recurrence Scores from metastatic sites after standard of care biopsy in HR+, HER2 negative patients and correlate Recurrence Score results from matched primary breast cancer when available. Methods: A total of 48 metastatic biopsies were retrieved retrospectively from the residual tissue of patients with primary HR+, HER2 negative breast cancer. This included 36 from bone and 12 from other sites [liver (7), lung (1), rectum (1), brain (1), skin (2)]. Slides were sent to Genomic Health Inc. for RNA isolation and Recurrence Score result determination using standardized protocols. Recurrence Score results were available for 18 matched primary and metastatic biopsy samples. The percent success rate for Recurrence Score result was determined for the various metastatic sites and results compared between matched primary and metastatic site. Results: Recurrence Score results were obtained in 48% of metastatic biopsies (23 of 48 samples) including bone (17), liver (4), lung (1), and skin (1). Reasons for Recurrence Score failure included insufficient RNA (17), poor quality RNA (1), failed QC (4), and other (3). The mean Recurrence Score from the 23 metastatic sites was 35 (range: 1–66). Notably, 70% (16/23) of successful metastatic biopsies yielded Recurrence Scores in the high-risk range (>25). None of the 23 metastatic biopsies gained HER2 by RT-PCR. Among the 18 paired samples, higher recurrence score results were observed in all but three of the metastatic biopsy samples with mean Recurrence Score results of 20 (range 7 to 41) for the primary and 35 (range 1-66) for the metastatic site. For paired samples, 72% of metastatic biopsies yielded Recurrence Scores >25 compared to 17% of primary sample. Primary Recurrence Scores were not predictive of metastatic scores (r2=0.052). Estrogen receptor (ER) expression status was conserved in 87% whereas progesterone receptor (PR) was lost in 69% of the metastatic lesion. Among the pairs, 5 had de novo metastatic disease. In these, the Recurrence Score was higher in the metastatic biopsy in each case compared to the matched primary (mean 36 versus 23, respectively). Among de novo cases, there was 100% concordance in ER positive and HER2 negative expression and only 60% concordance in PR expression between primary and metastatic sites. Conclusion: Using standard of care metastatic biopsy samples, a Recurrence Score result was successfully generated in 48% of samples including bone. This small series demonstrates wide variability in Recurrence Score results in metastatic disease with overall higher scores, common loss of PR, and minimal correlation to matched primary disease. Further examination of the potential significance of the Recurrence Score for treatment decisions in the metastatic setting requires additional tissue sampling during biopsy as insufficient RNA was the primary reason for failure. Citation Format: Julie Anne L Gemmill, Patricia Thompson, Rebecca Batiste, Caterina Vacchi-Suzzi, Christina Preece, Jules Cohen, Lea Baer, Carolyn Mies, Michelle Turner, Christy A Russell, Alison Stopeck. The feasibility of obtaining oncotype DX breast recurrence score® results from metastatic sites of patients with hormone receptor positive metastatic breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS5-37.

Objectives: There is growing evidence that urine cadmium is a temporally stable biomarker indicative of long-term cadmium exposure; however questions remain with regard to generalizability to older persons, the impact of changes in smoking behavior, and the degree of temporal stability when repeat sample collection spans years instead of weeks or months. Methods: Using archived samples from cohorts of older men (Osteoporotic Fractures in Men (MrOS-US)) and women (Study of Osteoporotic Fractures (SOF)) (mean age = 80 at study visit 2), we analyzed two morning urine samples each from 39 men and 18 women with a diverse self-reported smoking history. For MrOS, samples were collected approximately 6 years apart, and 4 years apart for SOF. Intra-class correlations were computed to assess temporal stability, and adjusted for age and body mass index. Results: The median creatinine-adjusted urinary cadmium levels (0.39 mu g/g for men, 0.89 mu g/g for women) were similar to levels expected for these age/sex groups in the US according to the National Health and Nutrition Examination Survey. The overall intra-class correlation was high (ICC = 0.85; 95% CI: 0.76-0.91) and similar between cohorts (MrOS: ICC = 0.74; 95% CI: 0.58-0.86; SOF: ICC = 0.81; 95% CI: 0.59-0.93), but slightly lower among those who stopped smoking between visits of sample collection (ICC = 0.64; 95% CI: 0.31-0.87) or among former smokers who quit prior to the first sample collection (ICC = 0.68; 95% CI: 0.25-0.93). Conclusions: We report good-to-excellent reproducibility of urine cadmium using morning urine samples collected 4-6 years apart from older men and women, but slightly lower correlations among those with a history of smoking. Single measures of urine cadmium are a reliable biomarker in older men and women.

Exposure to lead (Pb) is implicated in a plethora of health threats in both adults and children. Increased exposure levels are associated with oxidative stress in the blood of workers exposed at occupational levels. However, it is not known whether lower Pb exposure levels are related to a shift toward a more oxidized state. To assess the association between blood lead level (BLL) and glutathione (GSH) redox biomarkers in a population of healthy adults, BLL and four GSH markers (GSH, GSSG, GSH/GSSG ratio and redox potential E h ) were measured in the blood of a cross-sectional cohort of 282 avid seafood-eating healthy adults living on Long Island (NY). Additionally, blood levels of two other metals known to affect GSH redox status, selenium (Se) and mercury (Hg), and omega-3 index were tested for effect modification. Regression models were further adjusted for demographic and smoking status. Increasing exposure to Pb, measured in blood, was not associated with GSSG, but was associated with lower levels of GSH/GSSG ratio and more positive GSH redox potential E h , driven by its association with GSH. No effect modification was observed in analyses stratified by Hg, Se, omega-3 index, sex, age, or smoking. Blood Pb is associated with lower levels of GSH and the GSH/GSSG ratio in this cross-sectional study of healthy adults.

There is growing evidence of immunotoxicity related to exposure to toxic trace metals, and an examination of gene expression patterns in peripheral blood samples may provide insights into the potential development of these outcomes. This pilot study aimed to correlate the blood levels of three heavy metals (mercury, cadmium, and lead) with differences in gene expression in 24 participants from the Long Island Study of Seafood Consumption. We measured the peripheral blood mRNA expression of 98 genes that are implicated in stress, toxicity, inflammation, and autoimmunity. We fit multiple linear regression models with multiple testing correction to correlate exposure biomarkers with mRNA abundance. The mean blood Hg in this cohort was 16.1 µg/L, which was nearly three times the Environmental Protection Agency (EPA) reference dose (5.8 µg/L). The levels of the other metals were consistent with those in the general population: the mean Pb was 26.8 µg/L, and the mean Cd was 0.43 µg/L. The expression of three genes was associated with mercury, four were associated with cadmium, and five were associated with lead, although none were significant after multiple testing correction. Little evidence was found to associate metal exposure with mRNA abundance for the tested genes that were associated with stress, toxicity, inflammation, or autoimmunity. Future work should provide a more complete picture of physiological reactions to heavy metal exposure.

Our aim was to explore the association between blood cadmium (BCd) and depressive symptoms, adjusting for pack years and blood cotinine, and also stratifying by smoking status. Using data from the US National Health and Nutrition Examination Survey (NHANES) 2005-2012, we categorized depressive symptoms using the PHQ-9 (Patient Health Questionnaire-9) survey and modeled depressive symptoms in relation to BCd adjusted for blood cotinine, pack years of smoking, and other covariates. We also stratified by self-reported smoking status (current, former, never). There were 11,209 subjects from 2005 to 2012, age ≥ 18 with PHQ-9, smoking, and blood cadmium data available. 876 (7.8%) met criteria for depressive symptoms. Depressive symptoms were associated with BCd levels in a crude model and with adjustment for pack years and cotinine. The association disappeared when analyzed among current, former, or never smokers. Consistent with the literature, we found an association between BCd and depressive symptoms; however, that association disappears in analyses stratified by smoking status. This suggests residual confounding may be present. It is important to stratify by smoking status when investigating health outcomes associated with BCd.

Introduction: Previous studies reported associations between blood cadmium (BCd) and both depression and obstructive lung disease (OLD) after controlling for cigarette smoking, which is an important source of cadmium exposure and a potential confounder. We hypothesize that residual confounding may still be present even after controlling for pack-years and cotinine, and that stratifying by cigarette smoking may be necessary to remove the confounding influence of cigarette smoking. Methods: We used available data from the US National Health and Nutrition Examination Survey- 2005-2012 for PHQ-9 scores as a measure of depression and 2007-2012 for FEV1/FVC ratios as a measure of OLD. Depression, defined as PHQ-9 ≥10, was modeled in a logistic regression adjusted for age, race, alcohol use, poverty income ratio (PIR) and blood cotinine as well as pack years, and was also stratified by self-reported smoking status (current, former, never). FEV1/FVC ratios were modeled in a linear regression with the same adjustment as well as adjustment for medical conditions, and similar smoking stratification. Results: Depression was associated with BCd levels in a crude model and with adjustment for pack years and cotinine (OR= 1.21 95% CI: 1.09-1.35), but the association disappeared when stratifying by smoking status (e.g., never smokers: OR= 0.70 95% CI: 0.33-1.47). OLD was associated with BCd levels in a crude continuous model and with adjustment for pack years and cotinine (β= -0.02; p<0.01); when stratified by smoking status, never-smokers showed no association (β=-0.0001; p=0.98). Conclusion: Consistent with the literature, we found an association between BCd and both depression and OLD; however, that association disappears in analyses stratified by never smokers. This suggests residual confounding may be present. It is important to stratify by smoking status when exploring health outcomes associated with BCd.

Red blood cell (RBC) traits are important heritable clinical biomarkers and modifiers of disease severity. To identify coding genetic variants associated with these traits, we conducted meta-analyses of seven RBC phenotypes in 130,273 multi-ethnic individuals from studies genotyped on an exome array. After conditional analyses and replication in 27,480 independent individuals, we identified 16 new RBC variants. We found low-frequency missense variants in MAP1A (rs55707100, minor allele frequency [MAF] = 3.3%, p = 2 x 10(-10) for hemoglobin [HGB]) and HNF4A (rs1800961, MAF = 2.4%, p < 3 x 10(-8) for hematocrit [HCT] and HGB). In African Americans, we identified a nonsense variant in CD36 associated with higher RBC distribution width (rs3211938, MAF = 8.7%, p = 7 x 10(-11)) and showed that it is associated with lower CD36 expression and strong allelic imbalance in ex vivo differentiated human erythroblasts. We also identified a rare missense variant in ALAS2 (rs201062903, MAF = 0.2%) associated with lower mean corpuscular volume and mean corpuscular hemoglobin (p < 8 x 10(-9)). Mendelian mutations in ALAS2 are a cause of sideroblastic anemia and erythropoietic protoporphyria. Gene-based testing highlighted three rare missense variants in PKLR, a gene mutated in Mendelian non-spherocytic hemolytic anemia, associated with HGB and HCT (SKAT p < 8 x 10(-7)). These rare, low-frequency, and common RBC variants showed pleiotropy, being also associated with platelet, white blood cell, and lipid traits. Our association results and functional annotation suggest the involvement of new genes in human erythropoiesis. We also confirm that rare and low-frequency variants play a role in the architecture of complex human traits, although their phenotypic effect is generally smaller than originally anticipated.

Mechanisms of mercury (Hg) toxicity at low doses from seafood consumption, the most common exposure route, are not well understood. We tested the hypothesis that seafood Hg exposure is related to a shift in redox status, indicated by a decrease in the ratio of reduced to oxidized glutathione (GSH:GSSG) in blood, or increase in redox potential (Eh). We also examined whether key seafood nutrients (selenium (Se), omega-3 fatty acids) confound or modify this shift. We measured blood concentrations of total Hg, Se, GSH, GSSG, and the Omega-3 Index (% omega-3s of total fatty acids in red blood cell membranes) in seafood consumers in Long Island, NY. We examined relationships between Hg, GSH:GSSG ratio and Eh. Elevated blood Hg (>5.8µgL−1) was associated with lower GSH:GSSG (β=−116.73, p=0.01), with no evidence of confounding by Se or Omega-3 Index. However, in models stratified by Omega-3 Index levels, Hg-GSH:GSSG associations were weakened among those with high Omega-3 Index levels (>6% of fatty acids, β=−63.46, p=0.28), and heightened among those with low Omega-3 Index (β=−182.53, p<0.01). We observed comparable patterns for Eh in relation to Hg. These results support the hypothesis that Hg exposure from seafood is linked to a shift in redox status toward oxidative stress, modified by omega-3 fatty acids in this population. Further work should examine the role of different seafood nutrients and Hg-induced shifts in redox status in the diverse health effects associated with elevated Hg exposure.

Introduction: Although lead (Pb) is considered a redox-inert metal, it binds to proteins' sulfhydryl groups and is known to act via oxidative stress. Human studies of Pb in relation to glutathione-related biomarkers have been conducted in occupationally exposed workers; however no studies have been conducted at lower levels of exposure (<10 µg/dL). The objective of this study is to assess the association between blood Pb in a non-occupationally exposed population and reduced and oxidized glutathione (measured singularly) and glutathione-derived biomarkers (i.e., reduced GSH/oxidized GSSG ratio and calculated redox potential Eh), as indicators of oxidative stress. Methods: Blood Pb and four GSH markers (GSH, GSSG, GSH:GSSG ratio and redox potential Eh) were measured in the blood of a cohort of 282 avid seafood-eating healthy adults living on Long Island (NY, USA). Linear regression models were adjusted for demographic characteristics and smoking status. Results: The mean blood Pb level in the overall study population (n = 282) was 1.77 ± 1.03 µg/dL. As the Pb level increases, there is a shift to a more oxidized state, demonstrated by the β coefficient of -233.3 for GSH:GSSH, p <0.01; and +5.6 for Eh, p <0.01 per 1 µg/dL increase in Pb; results are consistent across unadjusted and adjusted models. Effect modification was not present in analyses stratified by smoking, sex, age, or blood selenium or mercury. There was no association between Pb and GSH or GSSG. Conclusion: To our knowledge this is the first study to report on the association between blood glutathione redox markers (including GSH:GSSG ratio and Eh redox potential) and Pb in an adult low-exposure population. Even at low levels of Pb exposure, we observe a significant trend toward a more oxidized state with rising blood lead levels, suggesting Pb may induce a shift toward oxidative stress even at very low levels of exposure.

White blood cells play diverse roles in innate and adaptive immunity. Genetic association analyses of phenotypic variation in circulating white blood cell (WBC) counts from large samples of otherwise healthy individuals can provide insights into genes and biologic pathways involved in production, differentiation, or clearance of particular WBC lineages (myeloid, lymphoid) and also potentially inform the genetic basis of autoimmune, allergic, and blood diseases. We performed an exome array-based meta-analysis of total WBC and subtype counts (neutrophils, monocytes, lymphocytes, basophils, and eosinophils) in a multi-ancestry discovery and replication sample of similar to 157,622 individuals from 25 studies. We identified 16 common variants (8 of which were coding variants) associated with one or more WBC traits, the majority of which are pleiotropically associated with autoimmune diseases. Based on functional annotation, these loci included genes encoding surface markers of myeloid, lymphoid, or hematopoietic stem cell differentiation (CD69, CD33, CD87), transcription factors regulating lineage specification during hematopoiesis (ASXL1, IRF8, IKZF1, JMJD1C, ETS2-PSMG1), and molecules involved in neutrophil clearance/apoptosis (C10orf54, LTA), adhesion (TNXB), or centrosome and microtubule structure/function (KIF9, TUBD1). Together with recent reports of somatic ASXL1 mutations among individuals with idiopathic cytopenias or clonal hematopoiesis of undetermined significance, the identification of a common regulatory 3 ' UTR variant of ASXL1 suggests that both germline and somatic ASXL1 mutations contribute to lower blood counts in otherwise asymptomatic individuals. These association results shed light on genetic mechanisms that regulate circulating WBC counts and suggest a prominent shared genetic architecture with inflammatory and autoimmune diseases.

BACKGROUND:Neurodevelopmental effects of omega-3 fatty acids and mercury from fish consumption have been characterized in children. In contrast, neurobehavioral outcomes associated with fish are not well studied in adults.OBJECTIVE:This study of avid seafood consumers on Long Island (NY, USA) sought to define associations between mercury, seafood consumption, omega-3 fatty acids and neurobehavioral outcomes.METHODS:A computer-based test system was used to assess neurobehavioral function. Blood total Hg (Hg) and omega-3 index were measured in 199 adult avid seafood eaters, who also completed the neurobehavioral assessment and an extensive food and fish frequency and demographic questionnaire.RESULTS:For most of the outcomes considered, neither Hg nor omega-3 index was associated with neurobehavioral outcomes after adjustment for key confounding variables. Fish consumption, however, was associated with decreased odds of both self-reported fatigue (OR 0.85; 95 % CI 0.72, 1.01) and a constellation of neurologic symptoms (OR 0.79; 95 % CI 0.66, 0.96).CONCLUSIONS:Results from our study provide little evidence that omega-3 fatty acids or Hg is associated with cognitive function in adult avid seafood consumers. Larger studies are needed to confirm our finding of associations between fish consumption and decreased self-reported fatigue and neurologic impairment.

Cadmium is a naturally-occurring element, and humans are exposed from cigarettes, food, and industrial sources. Following exposure, cadmium accumulates in the kidney and is slowly released into the urine, usually proportionally to the levels found in the kidneys. Cadmium levels in a single spot urine sample have been considered indicative of long-term exposure to cadmium; however, such a potentially exceptional biomarker requires careful scrutiny. In this review, we report good to excellent temporal stability of urinary cadmium (intraclass correlation coefficient 0.66–0.81) regardless of spot urine or first morning void sampling. Factors such as changes in smoking habits and diseases characterized by increased excretion of proteins may produce short-term changes in urinary cadmium levels. We recommend that epidemiologists use this powerful biomarker in prospective studies stratified by smoking status, along with thoughtful consideration of additional factors that can influence renal physiology and cadmium excretion.

Platelet production, maintenance, and clearance are tightly controlled processes indicative of platelets' important roles in hemostasis and thrombosis. Platelets are common targets for primary and secondary prevention of several conditions. They are monitored clinically by complete blood counts, specifically with measurements of platelet count (PLT) and mean platelet volume (MPV). Identifying genetic effects on PLT and MPV can provide mechanistic insights into platelet biology and their role in disease. Therefore, we formed the Blood Cell Consortium (BCX) to perform a large-scale meta-analysis of Exomechip association results for PLT and MPV in 157,293 and 57,617 individuals, respectively. Using the low-frequency/rare coding variant-enriched Exomechip genotyping array, we sought to identify genetic variants associated with PLT and MPV. In addition to confirming 47 known PLT and 20 known MPV associations, we identified 32 PLT and 18 MPV associations not previously observed in the literature across the allele frequency spectrum, including rare large effect (FCER1A), low-frequency (IQGAP2, MAP1A, LY75), and common (ZMIZ2, SMG6, PEAR1, ARFGAP3/PACSIN2) variants. Several variants associated with PLT/MPV (PEAR1, MRVI1, PTGES3) were also associated with platelet reactivity. In concurrent BCX analyses, there was overlap of platelet-associated variants with red (MAP1A, TMPRSS6, ZMIZ2) and white (PEAR1, ZMIZ2, LY75) blood cell traits, suggesting common regulatory pathways with shared genetic architecture among these hematopoietic lineages. Our large-scale Exomechip analyses identified previously undocumented associations with platelet traits and further indicate that several complex quantitative hematological, lipid, and cardiovascular traits share genetic factors.