Osteoporosis and related fractures are major health concerns in aging populations. We aimed to develop and validate a polygenic score (PGS) for osteoporosis in a Japanese population using heel quantitative ultrasound (QUS)-derived T-scores. Using genome-wide association study data from 12,371 participants in the Tohoku Medical Megabank Community-Based Cohort, we identified genome-wide significant loci, including MBL2, TMEM135, and WNT16. PGS models were constructed and evaluated using independent datasets for model selection (n = 1,419) and validation (n = 8,711). Adding the PGS to age and sex resulted in only a modest increase in discrimination (AUC = 0.664 to 0.728) but supported genetic risk stratification. Individuals in the lowest PGS quintile (bottom 20%, genetically high-risk) had an increased risk of osteoporosis (odds ratio (OR) = 1.22, 95% confidence interval (CI): 1.07-1.40), whereas those in the highest PGS quintile (top 20%, genetically low-risk) had a reduced risk (OR = 0.85, 95% CI: 0.71-0.98). Prospective analysis over a mean 3.4-year follow-up confirmed the gradient: the lowest-quintile (high-risk) group had a higher incidence rate ratio (IRR) than the intermediate group (IRR = 1.42, 95% CI: 1.17-1.73), whereas the highest-quintile (low-risk) group had a lower IRR (IRR = 0.70, 95% CI: 0.54-0.89). Age-stratified analyses revealed no statistically significant interaction between age and PGS in predicting osteoporosis, and no differences in the slope of age-related T-score declines across genetic risk groups. Observed T-scores in young adults (20-44 years) and extrapolation to age 20 suggested lower bone status around peak bone mass among genetically high-risk individuals. These findings indicate that a Japanese-specific PGS can stratify osteoporosis risk and may help identify individuals at elevated genetic risk earlier in adulthood. ### Competing Interest Statement T.H. is a board member of Genome Analytics Japan, Inc. The other authors declare no conflicts of interest associated with this manuscript. ### Funding Statement This study was supported by the Tohoku Medical Megabank Project of AMED (JP23tm0124006; JP21tm0424601) and JSPS KAKENHI (JP23K09696, YOY and AS). ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Institutional Review Board of Iwate Medical University gave ethical approval for this work (approval ID: HG H25-2). I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes The QUS-derived T-score GWAS summary statistics are available at jMorp (https://jmorp.megabank.tohoku.ac.jp/gwas-studies/TGA000015). Individual-level data are not publicly available; access may be granted upon request and approval by the Information Distribution Review Committee of the TMM Project.

Hydrocephalus in mice is frequently associated with impaired motile cilia function. We previously reported that the loss of Hoatz , a motile cilia and flagella-associated gene, causes ventriculomegaly of variable severity. Here, we characterize the ventricular phenotype of Hoatz −/− mice using in vivo magnetic resonance imaging (MRI), histology, and transcriptomics. Severe hydrocephalus occurred in only 4.9% (4/82) of homozygotes. High-resolution MRI revealed a reproducible ~ 4.7-fold enlargement of the lateral and third ventricles, while overall brain size was preserved. Despite the modest degree of ventriculomegaly, detailed volumetric analyses demonstrated deformation of the surrounding brain parenchyma, most prominently affecting the hippocampus. Ventricular enlargement emerged rapidly during early postnatal development and was occasionally associated with periventricular edema. Morphological and transcriptomic analyses further revealed changes in hippocampal microglia consistent with an altered activation state. Collectively, these findings define Hoatz deficiency as a cause of early-onset ventriculomegaly of the lateral and third ventricles, characterized by preserved overall brain size and regional hippocampal deformation, and establish a complementary mouse model for investigating the consequences of altered cerebrospinal fluid dynamics.

AIM:Cerebral small vessel disease (CSVD)-related MRI findings, including white matter hyperintensities (WMHs), are not rare in general elderly populations. The aim of this study was to elucidate the contribution of hereditary CSVD-related genes to CSVD-related MRI findings in a general Japanese population. METHODS:We analyzed datasets from 324 individuals aged ≥ 50 years in Tohoku Medical Megabank (TMM), focusing on MRI markers and variants of NOTCH3, ABCC6, COL4A1, COL4A2, GLA, HTRA1, and TREX1 genes. Background factors included age, sex, hypertension, diabetes, dyslipidemia, hyperuricemia, alcohol drinking, and smoking. RESULTS:Pathogenic variant carriers were identified within ABCC6 (n = 20), but not other genes. To compare with previous studies including rare NOTCH3 variants regardless of pathogenicity, we included 24 rare functional variants of NOTCH3. We performed a gene-based analysis using the burden test and sequence kernel association test (SKAT) adjusted for background factors, between WMH/lacune and ABCC6/NOTCH3. The only significant finding was the correlation between WMH volume and rare NOTCH3 variants by SKAT, both with the basic model, adjusted for age, sex, and hypertension (p = 0.045), and full model, adjusted for all background factors (p = 0.027). We also analyzed the association between intracranial major artery stenosis/occlusion (ICASO) and RNF213 p.Arg4810Lys, the East Asian-specific variant susceptible to ICASO; however, we failed to identify a significant correlation. CONCLUSIONS:This study suggests that NOTCH3 may contribute to WMH volume in a general Japanese population.

The association between body mass index (BMI) and pancreatic cancer remains elusive in Asian populations. Clarifying the causal relationship between type 2 diabetes (T2D) and pancreatic cancer warrants triangulation of evidence, with Mendelian Randomization (MR) providing a robust approach. A two-sample, multivariable MR analysis was performed to investigate the associations between BMI, T2D, and pancreatic cancer risk in the Japanese population. Single nucleotide polymorphisms (SNPs) associated with BMI and T2D were selected from the Genome-Wide Association Study (GWAS) Catalog, and summary statistics for pancreatic cancer were derived from our previous GWAS in the Japanese population. MR analyses were performed using a random-effects inverse variance-weighted (IVW) method as well as sensitivity analyses. Genetic predisposition to T2D, assessed using 891 SNPs, was associated with increased pancreatic cancer risk (IVW odds ratio [OR]: 1.16; 95% confidence interval [CI]: 1.08-1.25 per 1-unit increase in the ln odds of T2D). MR-Egger and weighted median MR analyses yielded similar results. By contrast, genetically indexed BMI showed no association with pancreatic cancer risk (IVW OR: 1.00; 95% CI: 0.95-1.04 per 1 kg/m2 increase). The association between T2D and pancreatic cancer remained significant after adjusting for BMI (OR: 1.15; 95% CI: 1.07-1.24), while BMI was not significant in univariable or multivariable analyses. Our MR study supports a modest causal association of T2D, independent of BMI, with pancreatic cancer risk.

Social isolation, characterized by a lack of social connections with family, friends, and others, is associated with adverse health outcomes. However, the genetic contribution to the susceptibility to social isolation remains unclear. This study aimed to identify genetic loci associated with social isolation using the Lubben Social Network Scale (LSNS-6) in a Japanese population. The Tohoku Medical Megabank Community-Based Cohort Study was conducted between 2013 and 2016. The participants were genotyped using the Affymetrix Axiom Japonica Array. The LSNS-6 was used to assess familial and friend ties through six questions and social isolation statuses were defined using the total scale, family subscale, and friend subscale. Genome-wide association studies (GWASs) were conducted using a generalized linear mixed model, adjusting for age, sex, 10 genetic principal components and batch effects. In total, 63,497 participants who completed genotyping and the LSNS-6 were included. The mean age was 59.4 ± 11.9 years, and 41,126 (64.8%) were female. Significant genetic loci were identified in GWASs for the total scale (rs10736933 near ACADSB and HMX3) and friend subscale of LSNS-6 (rs1778366 near LINC02315 and LRFN5). This study provides the first genome-wide evidence of social isolation in the Japanese population, suggesting associations with ACADSB, HMX3, LINC02315, and LRFN5. These findings could enable personalized prevention and intervention for social isolation and related psychiatric disorders.

Abstract Hydrocephalus in mice is frequently associated with impaired motile cilia function. We previously reported that the loss of Hoatz, a motile cilia and flagella-associated gene, causes ventriculomegaly of variable severity. Here, we characterize the ventricular phenotype of Hoatz −/− mice using in vivo magnetic resonance imaging (MRI), histology, and transcriptomics. Severe hydrocephalus occurred in only 4.9% (4/82) of homozygotes. High-resolution MRI revealed a reproducible ~4.7-fold enlargement of the lateral and third ventricles, while overall brain size was preserved. Despite the modest degree of ventriculomegaly, detailed volumetric analyses demonstrated deformation of the surrounding brain parenchyma, most prominently affecting the hippocampus. Ventricular enlargement emerged rapidly during early postnatal development and was occasionally associated with periventricular edema. Morphological and transcriptomic analyses further revealed changes in hippocampal microglia consistent with an altered activation state. Collectively, these findings define Hoatz deficiency as a cause of early-onset ventriculomegaly of the lateral and third ventricles, characterized by preserved overall brain size and regional hippocampal deformation, and establish a complementary mouse model for investigating the consequences of altered cerebrospinal fluid dynamics.

AIMS:Adverse childhood experiences (ACEs), especially in early life, can affect psychosocial development and increase lifelong risk for mental disorders. ACEs are also known to induce persistent epigenetic changes. This study aimed to explore ACE-associated DNA methylation signatures using an epigenome-wide association study (EWAS) targeting inter-individual differentially methylated regions (DMRs). METHODS:We developed a targeted capture probe system covering ~1.3 million CpG sites within inter-individual DMRs. This system was applied to salivary DNA from drug-naïve children aged 6-12 years with exposure to multiple early-life ACEs (n = 23) or who had no ACEs (n = 21). RESULTS:We identified 15 novel DMRs significantly associated with ACEs. A cluster of six CpG sites within an exon of the EIF4G2 gene showed consistently increased methylation in children with ACEs, with strong inter-site correlations. Enrichment analysis indicated that genes near these DMRs are involved in neurodevelopmental disorders, suggesting that early adversity may influence brain development through epigenetic mechanisms. CONCLUSION:Our findings suggest that early adversity may contribute to lasting epigenetic modifications in children. The identified DMRs may serve as noninvasive biomarkers for retrospective ACE assessment and provide insights into the biological embedding of early-life stress.

Background: Integrating polygenic scores (PGS) into population-based stroke prevention strategies may help reduce the stroke burden at both individual and population levels. However, prospective evidence regarding the interaction between genetic and modifiable risk factors remains limited, particularly in East Asian populations. Methods: We evaluated the influence of modifiable risk factors across PGS levels in the prospective population-based Tohoku Medical Megabank Community-Based Cohort Study. Stroke PGS was calculated using a model developed by the GIGASTROKE project. Results: During a median follow-up of 4.84 years (121,843.2 person-years), 246 incident stroke events were identified. Compared with participants in the intermediate PGS group (third quintile), those in the highest PGS group (fifth quintile) had a 60% greater risk of incident stroke (hazard ratio [HR], 1.60; 95% confidence interval [CI], 1.10–2.34). When participants with intermediate genetic risk and no modifiable risk factors were used as the reference group, hypertension (HR, 2.17; 95% CI, 1.11–4.25) and diabetes (HR, 2.10; 95% CI, 1.04–4.27) were significantly associated with an increased stroke risk in the intermediate PGS group. In contrast, multiple modifiable risk factors were significantly associated with stroke risk in the highest PGS group, including hypertension (HR, 2.90; 95% CI, 1.53–5.48), diabetes (HR, 2.07; 95% CI, 1.02–4.20), dyslipidemia (HR, 2.20; 95% CI, 1.28–3.79), obesity (HR, 1.98; 95% CI, 1.19–3.30), and smoking (HR, 2.57; 95% CI, 1.34–4.89). Participants with both the highest PGS and ≥ 4 modifiable risk factors had a substantially elevated stroke risk (HR, 2.76; 95% CI, 1.28–5.92). The combination of genetic and modifiable risk factors significantly influenced the cumulative incidence of stroke (P<0.001). Conclusions: Modifiable risk factors substantially influenced stroke risk even among participants with high genetic susceptibility. The relative importance of modifiable risk factors for stroke prevention may differ according to an individual's genetic risk level.

The use of medical databases, known as real-world data (RWD), enables accurate and efficient estimation of disease prevalence in population-based studies, making it a potential game-changer in epidemiology. However, the lack of standardized data formats across hospitals complicates integration across institutions. In Japan, health insurance claims data are standardized under a nationally unified format, providing a reliable source of structured RWD. We evaluated the utility of insurance claims data in epidemiological research. Incorporating both diagnosis and prescription information into case definitions resulted in four- and six-fold increases in the estimated prevalence of Alzheimer’s disease (AD) and Parkinson’s disease (PD), respectively, compared with conventional self-reported definitions. Subsequent genome-wide association studies (GWAS) for AD showed increased model log-likelihood and identified a characteristic APOE signal, findings observed only with extended case definitions. The APOE effect size was consistent with large case–control studies, while standard errors remained comparable to smaller studies. These results indicate that claims-based phenotyping improves case identification without loss of accuracy and supports scalable approaches for genomic epidemiology and public health surveillance. All participants provided written informed consent and the study protocol was approved by the Institutional Review Board of Iwate Medical University (Approval number HG2021-009) No materials requiring permission from other sources have been used in this manuscript. This study does not involve interventional components and therefore was not registered as a clinical trial.

BACKGROUND:Lysinuric protein intolerance (LPI) is a rare genetic disorder characterized by an inherited defect in cationic amino acid transport caused by pathogenic variants in the SLC7A7 gene. While LPI causes systemic complications, the underlying cellular mechanisms remain poorly understood. This study investigated the cellular characteristics of LPI, focusing on intracellular metabolite profiles and astrocyte response to hyperammonemia. OBJECTIVES:To examine intracellular metabolite changes in LPI patients and to evaluate the response of patient-derived astrocytes to ammonia exposure. METHODS:Peripheral blood mononuclear cells (PBMCs) from three LPI patients and three healthy controls were analyzed for intracellular metabolite profiles using capillary electrophoresis-fourier transform mass spectrometry. Induced pluripotent stem cells were generated from a patient's PBMCs and differentiated into astrocytes. We evaluated LPI-astrocytes and their response to ammonia treatment by RNA sequencing, gene expression profiling, and cell viability assays. RESULTS:Metabolite analysis revealed significant intracellular metabolite imbalances in LPI patients, with increases of 21 metabolites including 11 amino acids. LPI-astrocytes exhibited distinct cellular characteristics regarding altered gene expression and enhanced cell cycle progression. When exposed to ammonia, the astrocytes demonstrated markedly lower cell viability and increased reactive oxygen species (ROS) production compared to control astrocytes. N-acetylcysteine supplementation significantly ameliorated ammonia-induced cytotoxicity. CONCLUSIONS:SLC7A7 dysfunction leads to intracellular metabolite disturbances and an increase in vulnerability to ammonia toxicity through ROS production of astrocyte, suggesting hyperammonemia and amino acid deficiencies as potential therapeutic targets in LPI patient care.

Plasma amino acids (AAs) have emerged as promising biomarkers for metabolic disorders, yet their causality remains unclear. We aimed to investigate the genetic determinants of AA levels in a cohort of 10,333 individuals and their causal effects on cardiometabolic traits using Mendelian randomization (MR). Plasma levels of 20 AAs were quantified using capillary electrophoresis mass spectrometry. Genome-wide association studies were conducted using BOLT-LMM and heritability estimation via LDSC analysis. Causal effects of AAs on 11 cardiometabolic traits were examined using two-sample MR analyses. We identified 85 locus-metabolite associations across 43 genes for 18 AAs, including 44 novel loci linked to metabolic genes. Heritability for AAs was estimated at 16%. MR analysis demonstrated cystine to positively associate with systolic blood pressure (SBP) (beta = 0.056, SE = 0.010), while serine indicated protective effects on SBP (beta = - 0.040, SE = 0.011), diastolic BP (beta = - 0.044, SE = 0.010), and coronary artery disease (odds ratio 0.888, SE = 0.028). We identified potentially novel genetic loci associated with AA levels and demonstrated robust causal associations between several AAs and cardiometabolic traits. These findings reinforce the importance of AAs as potential biomarkers and therapeutic targets in cardiometabolic health.

Jawless vertebrates, which diverged from jawed vertebrates approximately 550 million years ago, possess an adaptive immune system characterized by two distinct lineages of lymphocytes bearing unique antigen receptors. However, despite decades of research, the mechanism of antigen presentation in these organisms remains unclear. In this study, we report that chromosomes 22, 50, and 59 in the sea lamprey ( Petromyzon marinus ) exhibit synteny with the human major histocompatibility complex (MHC) region. The syntenic region spans from DHX16 to RING1 (chromosome 6: 30,517,301-33,034,069) in the human genome (T2T-CHM13v2.0). Similar to the human MHC region, P. marinus chromosome 59 harbors homologs of DDX39B, NOTCH, PBX, TNF, and BRD genes, yet notably lacks the core MHC class I, II, and III (complement) genes. Genotyping of three independent lampreys showed no signatures of balancing selection, a characteristic feature of the human MHC. Additionally, these lamprey chromosomes exhibit synteny with chromosome 9 in amphioxus ( Branchiostoma lanceolatum ), which contains conserved homologs that form its "proto-MHC." These findings suggest that chromosomes 22, 50, and 59 in P. marinus are a proto-MHC or its paralog. ### Competing Interest Statement The authors have declared no competing interest.

Hydrocephalus is often associated with motile ciliary dysfunction in mice. It is believed that ependymal ciliary abnormalities impede cerebrospinal fluid (CSF) circulation, resulting in increased intracranial pressure. We previously identified Hoatz as a cilia-related gene and demonstrated that knockout mice develop ventriculomegaly. In this study, we combined in vivo magnetic resonance imaging (MRI), transcriptomics, and histology to elucidate the potential mechanisms underlying ventricular enlargement. High-resolution T2-weighted MRI revealed a ~ 4.5-fold increase in ventricular volume, occasionally accompanied by edematous fluid accumulation in the corpus callosum. However, the overall brain size was preserved, indicating parenchymal atrophy rather than simple intracranial hypertension. Unexpectedly, transcriptome profiling of ependymal cultures revealed robust upregulation of microglia-associated genes. Immunostaining confirmed activation of coexisting microglia, regardless of intracranial pressure. Similarly, RNA sequencing (RNA-seq) of hippocampal tissues identified multiple upregulated genes, enriched in immune-related pathways, including Trem2, Tyrobp, C1qa/b, Lyz2, and Ctss. Furthermore, confocal imaging revealed an amoeboid, activated morphology of hippocampal microglia. These findings indicate that ventriculomegaly in Hoatz-null mice may result not only from impaired CSF flow but also from neuroinflammation-driven neurodegeneration. This dual mechanism suggests potential relevance to normal pressure hydrocephalus and other neurodegenerative disorders.

Childhood obesity is a growing global health concern, contributing to numerous non-communicable diseases and long-term health complications. The prevalence of obesity in children and adolescents continues to rise, driven by complex interactions among various factors. The key risk factors include both environmental and genetic influences. Environmental factors include family elements like household conditions and lifestyle, while genetic factors refer to inherited predispositions. More recently, epigenetic factors have gained attention, focusing on chemical modifications such as DNA methylation that are influenced by the prenatal and early-life environment and may contribute to obesity risk. Unlike obesity in adults, the risk factors for obesity in children are largely dependent on their family environments rather than individual behaviors. For effective intervention, it is important to identify at-risk children and their families as early as possible after birth. Despite advances in machine learning, polygenic risk scores, and epigenomic markers-which show promise as being more accurate and comprehensive prediction methods-no risk prediction models are currently in clinical use. Achieving predictions with higher accuracy, external validation, and consideration of population-specific factors (e.g., ethnic variability) while avoiding bias or stigma in targeted interventions is needed for effective childhood obesity prevention. Herein, we summarize environmental, genetic, and epigenetic risk factors for childhood obesity and review the unique situations and regional factors in Japan, which are the focus of our study. Furthermore, we introduce the major advances in risk prediction models for childhood obesity.

BackgroundDry eye disease (DED) is a common ocular condition with diverse and heterogeneous symptoms. Current treatment standards of DED include the post facto management of associated symptoms through topical eye drops. However, there is a need for predictive, preventive, personalized, and participatory medicine. The DryEyeRhythm mobile health app enables real-time data collection on environmental, lifestyle, host, and digital factors in a patient’s daily environment. Combining these data with genetic information from biobanks could enhance our understanding of individual variations and facilitate the development of personalized treatment strategies for DED. ObjectiveThis study aims to integrate digital data from the DryEyeRhythm smartphone app with the Tohoku Medical Megabank database to create a comprehensive database that elucidates the interplay between multifactorial factors and the onset and progression of DED. MethodsThis prospective observational cohort study will include 1200 participants for the discovery stage and 1000 participants for the replication stage, all of whom have data available in the Tohoku Medical Megabank database. Participants will be recruited from the Community Support Center of Sendai, Miyagi Prefecture, Japan. Participant enrollment for the discovery stage was conducted from August 1, 2021, to June 30, 2022, and the replication stage will be conducted from August 31, 2024, to March 31, 2026. Participants will provide demographic data, medical history, lifestyle information, DED symptoms, and maximum blink interval measurements at baseline and after 30 days using the DryEyeRhythm smartphone app. Upon scanning a registration code, each participant’s cohort ID from the Tohoku Medical Megabank database will be linked to their smartphone app, enabling data integration between the Tohoku Medical Megabank and DryEyeRhythm database. The primary outcome will assess the association between genetic polymorphisms and DED using a genome-wide association study. Secondary outcomes will explore associations between DED and various factors, including sociodemographic characteristics, lifestyle habits, medical history, biospecimen analyses (eg, blood and urine), and physiological measurements (eg, height, weight, and eye examination results). Associations will be evaluated using logistic regression analysis, adjusting for potential confounding factors. ResultsThe discovery stage of participant enrollment was conducted from August 1, 2021, to June 30, 2022. The replication stage will take place from August 31, 2024, to March 31, 2026. Data analysis is expected to be completed by September 2026, with results reported by March 2027. ConclusionsThis study highlights the potential of smartphone apps in advancing biobank research and deepening the understanding of multifactorial DED, paving the way for personalized treatment strategies in the future. International Registered Report Identifier (IRRID)DERR1-10.2196/67862

Dementia, particularly Alzheimer’s disease (AD), continues to be a major public health concern due to population aging, yet minimally invasive biomarkers for early diagnosis have not been established. DNA methylation (DNAm) has recently attracted considerable attention as a promising biomarker. This study aimed to identify blood-based DNAm biomarkers for early detection of AD. We analysed blood-derived DNA from 48 patients with AD (from Biobank Japan) and 48 age- and sex-matched controls (from the Tohoku Medical Megabank Biobank) using Apolipoprotein ε type 4 (APOE)-associated genotype analysis and targeted-bisulfite sequencing. High-risk APOE genotypes were more frequent in AD patients (23/48, [47.9

Although previous studies reported that BP PRS is associated with CVD, it is less explored whether BP PRS and BP are jointly associated with CVD, especially among non-European populations. Therefore, we examined joint associations of BP control and BP polygenic risk score (PRS) with CVD mortality in a Japanese population. Data were obtained from the Japan Multi-Institutional Collaborative Cohort (J-MICC) Study, the multi-centered cohort study with 14 study areas throughout Japan. Of which, we analyzed 35,000 Japanese individuals (Mean age: 55 years old, Men: 44

Background: The potential impacts of polygenic scores (PGS) on health-behavior changes are not fully understood. The Iwate PGS Assessment and Risk Communication Study aims to investigate the effects of reporting PGS-based risk for ischemic stroke on health behaviors. Methods: Participants wishing to know their PGS-based ischemic stroke risk were recruited from health checkup venues for workers in Iwate Prefecture in 2023. Health checkup data, biospecimens, and questionnaire responses were collected for biochemical testing, genotyping, and storage in the Tohoku Medical Megabank integrated biobank. The risk was calculated using an integrative PGS model for East Asians. Participants were randomly assigned to two groups, and one group received their risk report as the intervention group. The impacts of the risk notification will be investigated in follow-up surveys. Results: Of 3,599 workers, 2,088 participated in the study (consent rate, 58.0%). The demographic profile of the eligible 2,083 participants was as follows: 80.7% males, and dominance of participants aged 18-29 years (25.2%), in their 30's (25.3%), and in their 40's (24.7%). Two hundred participants (9.7%) had a risk of 1.0 as the reference; 57 (2.7%), 927 (44.7%), and 888 (42.9%) participants had 2.1-3.4-, 1.4-1.9-, and <1.0-fold that risk, respectively. Conclusion: We collected health information and biospecimens from over 2,000 workers, and disclosed the PGS-based ischemic stroke risk. Behavioral effects will be evaluated 1 year after disclosure, with follow-up until 2030. As Japan's first large-scale PGS risk communication study, it will provide initial insights for implementing PGS in personalized preventive medicine.

Sex differences shape human physiology and disease risk, yet their autosomal epigenetic basis remains incompletely understood. Using whole genome bisulfite sequencing data from iMETHYL database (∼100 adults) across three purified blood cell types (CD4⁺ T cells, monocytes, neutrophils), we performed comprehensive mapping of cell-type specific sex-associated DNA methylation. Using genome-wide Z -test and confidence-interval method, we identified thousands of autosomal differentially methylated sites (DMSs), the majority of which were cell type-specific. We found that neutrophils exhibited pronounced female-biased hypermethylation, whereas CD4⁺ T cells and monocytes showed more balanced patterns. DMSs were enriched within gene bodies and annotated to neuronal and adhesion functions, with T-cell activation uniquely associated in CD4⁺ T cells. Transcription factor binding site enrichment indicated hematopoietic regulators, suggesting that sex-associated methylation is tightly linked to early developmental processes within blood lineages. Differentially methylated regions overlapped genome-wide association study loci for lifetime smoking, multiple sclerosis, and psoriasis, indicating convergence between sex-associated epigenetic states and genetic susceptibility. Evolutionary analysis revealed limited conservation but identified a conserved intronic CpG within FIGN . This study provides the first genome-wide, cell type-resolved map of autosomal sex-associated DNA methylation in human blood and establishes a foundation for mechanistic and translational studies in sex-informed biology and medicine. ![Figure][1] ### Competing Interest Statement The authors have declared no competing interest. Japan Society for the Promotion of Science, JP22K07583, JP23K27531, JP25H01314, JP25H01309, JP23H03838, JP24K22098, and JP25K10839 Japan Agency for Medical Research and Development, JP19dm0207074 and JP24wm0625302 [1]: pending:yes