The underlying pathophysiological mechanisms of knee osteoarthritis (KOA) remain incompletely understood. This study aimed to investigate the clinical significance of long non-coding RNA LINC-PINT in KOA and elucidate its molecular mechanism in regulating chondrocyte function via the miR-324-3p/GPX4 axis. 100 KOA patient samples and 40 normal knee cartilage tissue samples were collected, and the relevant gene expression was quantified using RT-qPCR. C28/I2 was cultured in vitro to establish an IL-1β-induced KOA cellular model. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to validate the targeted molecular regulatory interactions. Cell proliferation and apoptosis, as well as inflammatory factors levels, were assessed using cell counting kit-8 (CCK-8), flow cytometry, and enzyme-linked immunosorbent assay (ELISA), respectively. Oxidative stress-related indicators and Fe2+ concentration were evaluated using commercial assay kits. Western blotting was performed to detect protein expression. In KOA tissues, LINC-PINT and glutathione peroxidase 4 (GPX4) were significantly downregulated, whereas miR-324-3p was markedly upregulated. LINC-PINT demonstrated the potential diagnostic discrimination ability and was identified as an independent protective factor in KOA. Overexpression of LINC-PINT effectively counteracted interleukin-1 beta (IL-1β)-induced suppression of chondrocyte proliferation, reduced apoptosis, and attenuated the secretion of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), while significantly alleviating cartilage matrix degradation, as indicated by increased expression of collagen II and aggrecan and decreased matrix metallopeptidase 13 (MMP13) levels. Moreover, LINC-PINT directly bound to miR-324-3p, which in turn directly targeted GPX4. Functional rescue assays further confirmed that transfection with a miR-324-3p mimic reversed the protective effects of LINC-PINT overexpression on chondrocytes, whereas GPX4 overexpression mitigated the detrimental effects induced by the miR-324-3p mimic. IL-1β disrupted the cellular redox balance by increasing the levels of Fe2+, reactive oxygen species (ROS), and (malondialdehyde) MDA and by inhibiting superoxide dismutase (SOD) activity and the GSH/GSSG ratio. Overexpression of LINC-PINT can counteract these effects, and the protective effect was enhanced when combined with Per-1. Transfection with miR-mimic reversed the protective effect of LINC-PINT, suggesting that LINC-PINT may act as a competitive endogenous RNA (ceRNA) to capture this pro-oxidative miR-324-3p. Finally, overexpression of GPX4 could protect cells from miRNA-induced damage, highlighting its crucial role in alleviating iron-dependent oxidative stress and iron-overloaded apoptosis. LINC-PINT demonstrated the potential diagnostic discrimination ability and exerted a protective effect in KOA by modulating the miR-324-3p/GPX4 axis.
Lymphedema is a lymphatic dysfunction leading to an accumulation of fluid and fat in the arm or leg. Here, we performed noncoding RNA profiling of human breast cancer-induced secondary lymphedema. We identified the long intergenic non-protein coding RNA, P53-induced transcript (LINC-PINT), as essential for the lymphedema development. LINC-PINT is the most expressed lncRNA in human lymphatic endothelial cells (LECs) under stress condition. Knocking down LINC-PINT in LECs promotes the expression of inflammation-related genes. Mechanistically, ATAC-seq revealed that LINC-PINT induces the transcription of genes involved in lymphangiogenesis and immune cell adhesion by increasing chromatin accessibility. Notably, LINC-PINT deficiency impairs LEC proliferation, migration, and sprouting. Conditional deletion of Lnc-Pint in mouse lymphatic endothelium (Lnc-Pintlecko) leads to a reduction in dermal lymphatic network density. Lnc-Pintlecko mice exhibit decreased lymphedema, reduced dermal backflow, fibrosis, and inflammation. Our findings unveil a crucial molecular role of LINC-PINT in lymphatic function and hold substantial clinical implications for lncRNA as biomarker of lymphedema.
The identification of predictive biomarkers for nasopharyngeal carcinoma (NPC) treatment response is critical to improving clinical outcomes. Our previous studies demonstrated that the long non-coding RNA (lncRNA) LINC-PINT enhances the radiosensitivity of NPC cells. However, the clinical relevance of functional polymorphisms in LINC-PINT remains unclear. A total of 199 Chinese NPC patients who received concurrent platinum-based chemoradiotherapy were enrolled. The association between the LINC-PINT rs1059698 polymorphism and short-term treatment efficacy (assessed 3 months post-treatment) was evaluated using multivariate logistic regression. LINC-PINT expression in NPC tissues was quantified by real-time polymerase chain reaction (PCR). A luciferase reporter assay was performed to assess the effect of rs1059698 on miR-646 binding. Eight machines were further employed learning algorithms to construct predictive models for treatment efficacy. NPC patients with a complete response (CR) showed significantly higher LINC-PINT expression than those with partial response (PR) or progressive disease (PD) (P < 0.05). The rs1059698 CC genotype was significantly associated with an improved treatment response (CR vs. PR + PD, OR = 0.189, 95% CI: 0.042-0.860, P = 0.031). Individuals carrying the CC genotype also exhibited approximately 1.5-fold higher LINC-PINT expression compared to AA genotype carriers (P < 0.001). Functional analysis showed that the A-to-C substitution at rs1059698 disrupted miR-646 binding to LINC-PINT. Additionally, the Gradient Boosting Machine (GBM) model incorporating rs1059698 and clinical variables achieved moderate predictive accuracy (AUC = 0.765 in training and 0.641 in validation). Our findings suggest that the rs1059698 polymorphism were associated with NPC treatment response possibly by influencing the expression of LINC-PINT or altering miRNA-lncRNA interaction. LINC-PINT rs1059698 may serve as a predictive biomarker for chemoradiotherapy efficacy in NPC.
Salmonella virulence chiefly relies upon two major pathogenicity islands, SPI-1 and SPI-2, which enable host cell invasion and intracellular survival, respectively. Growing evidence suggests post-transcriptional control of SPI gene expression by Hfq-dependent small regulatory RNAs (sRNAs) such as PinT. This 80-nucleotide sRNA is highly expressed after Salmonella enters host cells and modulates the transition from the SPI-1 to SPI-2 program by targeting the mRNAs of different virulence factors. However, it remains unclear how PinT activity can be counteracted when the suppression of virulence genes needs to be relieved. Here, we mapped the RNA interactome of Salmonella recovered from infected macrophages, using an optimized version of RIL-seq. In addition to offering an unprecedented view of Hfq-mediated RNA interactions during Salmonella's intracellular infection stage, RIL-seq uncovered the previously described 3' end-derived sRNA InvS as a direct negative regulator of PinT. Biochemical and genetic experiments suggest a decoy mechanism by which InvS lifts PinT-mediated target repression. Moreover, InvS acts as an mRNA repressor of the adhesion protein MipA and PinT interaction with InvS relieves mipA repression. Together, our work identifies a pair of antagonistic sRNAs in a growing post-transcriptional network of virulence gene regulation.
Signal transduction allows bacterial pathogens to sense the host environment and regulate gene expression accordingly for adaptation and survival. While the success of infection largely depends on the timely induction of virulence genes, the activity of the regulatory pathways controlling their expression must be tightly regulated for pathogens to cause disease. Here, we establish that a small RNA (sRNA) promotes the negative feedback control of a master virulence regulator in Salmonella enterica serovar Typhimurium (S. Typhimurium) by repressing a signaling protein essential for its induction in response to an intracellular cue. We show that the virulence regulatory PhoP/PhoQ pathway is inhibited by the PhoP-activated sRNA PinT in mildly acidic pH, an infection-relevant condition encountered by S. Typhimurium inside macrophages. PinT directly represses the translation of ugtL mRNA, which encodes the PhoP activator UgtL. This negative feedback regulation reduces PhoP activity, thereby decreasing the expression of PhoP-activated virulence genes like pagC. PinT-mediated repression of ugtL is predicted to be conserved in Salmonella enterica, but not in the nonpathogenic species Salmonella bongori, thus suggesting that the regulation is relevant for virulence. Our findings uncover how pathogens achieve proper levels of induction of their virulence programs through the post-transcriptional negative feedback regulation of factors enhancing the signaling activity of virulence pathways. To cause disease, pathogens must express their virulence genes at the right time and in proper levels. Here, we establish that a small RNA (sRNA) restricts the activation of a regulator critical for the virulence of Salmonella enterica serovar Typhimurium (S. Typhimurium). We show that the sRNA PinT inhibits the activity of the master virulence regulator PhoP by repressing its activator UgtL through a direct interaction with ugtL mRNA. This regulation reduces the expression of PhoP-activated genes. Because PhoP activates PinT and UgtL, the three regulators form a negative feedback loop. That the PinT-mediated repression of ugtL is predicted to occur in Salmonella enterica but not in the nonpathogenic species S. bongori suggests it may be a key virulence determinant. Our results unveil a novel layer of fine-tuning of PhoP activity ensuring that S. Typhimurium induces the proper level of its virulence program in response to an infection-relevant stress condition.
Single-nucleotide polymorphisms (SNPs) in the chromosome 17q12-q21 region and, independently, early-life nasal microbiota dominated by Moraxella, Streptococcus, or Haemophilus (MSH) increase risk of chronic wheeze and asthma development. We sought to determine whether 17q12-q21 risk SNPs and nasal microbiota interact to modulate childhood wheeze risk. Nasal wash samples from 12-month-old infants in 2 birth cohorts, COAST (Childhood Origins of Asthma; n = 180) and URECA (Urban Environment and Childhood Asthma; n = 139), underwent 16S ribosomal RNA variable region 4 sequencing. Nasal microbiota dominated by MSH or Corynebacterium, Dolosigranulum, Staphylococcus, or Bacillus (CDSB) were assessed. Paired blood was genotyped for 9 17q12-q21 risk SNPs. Logistic regression tested interactions between 17q12-q21 SNPs and MSH or CDSB on wheeze risk in the first 3 years of life. A549 lung epithelial cells, CRISPR-edited to encode the rs7216389 risk genotype (rs7216389TT) were compared to the heterozygous (rs7216389CT) line using bulk RNA sequencing. SNPs, particularly those in the ORMDL3 (rs8076131; odds ratio [OR]: 1.72; 95% CI: 1.09-2.71; Pint = .031) and GSDMB (rs2305480; OR: 1.72; 95% CI: 1.09-2.71; Pint = 0.042; and rs7216389; OR: 1.73; 95% CI: 1.09-2.70; Pint = .047) genes, interact with MSH microbiota to increase early-life wheeze risk (false discovery rate Pint = .016 for all), while interactions with CDSB reduce risk. A549 airway epithelial cells homozygous for rs7216389TT exhibited decreased expression of genes involved in antimicrobial responses and neutrophil recruitment and evidence increased microbial adherence compared with the heterozygous cell line. Airway microbiota interact with SNPs at the 17q12-q21 locus in genes involved in sphingolipid metabolism and intracellular antimicrobial responses, to modulate wheeze risk.
BACKGROUND: Although the associations between air pollution exposure and thyroid function have been reported, the interactive effects of the genes involved remain unknown. Therefore, we aimed to identify candidate genetic loci involved in thyroid function by interacting with annual average exposure to particulate matter with a diameter of 10 microns or smaller (PM10) in Korean adults. A total of 1,863 and 1,458 adults were included in the discovery and replication steps, respectively. The average annual concentration of PM10 exposure was considered, and the participants were classified into two groups (low-to-moderate exposure and high-exposure groups) for binary analyses. A genome-wide single-nucleotide polymorphism analysis using a PM10 exposure interaction study was performed to determine thyroid-stimulating hormone levels in Korean adults. RESULTS: Although no SNPs surpassed the stringent genome-wide significance threshold of Pint < 5E-08, one SNP (rs11781213) near MSRA reached a suggestive level of significance of Pint < 5E-07. Two genetic susceptibility loci (FAM84B/PCAT1 and STARD13) were replicated at a nominal significance level of Pint < 1E-05 for the discovery cohort, and Pint < 0.05 for the replication cohort. A genetic variant (rs7169081 G > A) between CGNL1 and GCOM1 was of functional interest. CONCLUSIONS: This is the first study reporting genome-wide-air pollution interaction results for thyroid function. The association between long-term PM10 exposure and thyroid hormone levels may be partly explained by identifying several suggestive loci, including MSRA, PCAT1, and GCOM1.
Biomolecular condensates, such as germ granules, organize RNAi pathways critical for fertility and genome regulation. Yet, the protein composition and functional contributions of these condensates remain poorly defined. Here, we applied TurboID proximity labeling to the Caenorhabditis elegans germ granule protein SIMR-1, integrating mass spectrometry with genetic screening, CRISPR-based tagging, and small RNA sequencing. This systematic approach identified several previously uncharacterized germ granule proteins that contribute to fertility, germline immortality, exogenous RNAi, and transgenerational inheritance. Small RNA sequencing of 21 mutants revealed broad and class-specific defects in siRNA and miRNA biogenesis, with distinct factors associated with defects in WAGO-class 22G-RNAs, CSR-class 22G-RNAs, or histone-directed small RNAs. Among these, we identified PINT-1, a highly disordered protein that directly interacts with and is recruited to germ granules by the PIWI Argonaute PRG-1. PINT-1 is required for piRNA-dependent and -independent secondary siRNA biogenesis and germline development. Comparative genomics revealed that PINT-1 has co-evolved with PRG-1 across nematodes, with a conserved structured N-terminus and a rapidly diverging repeat-rich intrinsically disordered region. Together, our findings expand the germ granule proteome and reveal how distinct condensate components contribute to specialized functions within the small RNA pathways, while highlighting an evolutionarily co-adapted PIWI interactor critical for siRNA biogenesis.
Outdoor air pollution, including fine particulate matter (PM2.5), is an established carcinogen, yet few studies evaluate its associations with prostate cancer risk and findings are mixed. We used spatiotemporal prediction models to estimate annual average historical residential PM2.5 (1980 to 2017) and NO2 (1990 to 2017) concentrations for N = 289,299 men aged ≥50 years in the NIH-AARP Diet and Health Study, enrolled from six states and two metro areas in 1995 to 1996, followed through 2018. We used Cox regression to estimate hazard ratios and 95% confidence intervals (HR [CI]) of associations between 5-year average PM2.5 and NO2 and incident prostate cancer, overall and by tumor aggressiveness (ie, advanced [n = 5,791], high-grade [ie, Gleason score ≥8, n = 5,793], fatal [n = 3,057]). We evaluated interactions with hypothesized effect modifiers. Historical PM2.5 and NO2 concentrations were not associated with prostate cancer risk overall (PM2.5 HR10-year lag per 5 µg/m3=1.00 [0.98-1.02], NO2 HR10-year lag per 10 ppb = 1.00 [0.98-1.01]). Some positive trends were observed for aggressive disease (eg, high-grade PM2.5 HR10-year lag=1.03 [0.99-1.09], advanced: NO2 HR10-year lag=1.04 [0.99-1.08]). Stratified analyses also showed stronger associations for aggressive disease among Hispanic men (eg, high-grade: NO2 HR = 1.25 [1.05-1.48], pint=0.01). Heterogeneity was also evident by urbanicity (high-grade: pint=0.01, PM2.5 and NO2) and state (advanced: pint=0.05, NO2), potentially reflecting air pollution mixture heterogeneity. In this large U.S. cohort, historical residential PM2.5 or NO2 concentrations were not associated with prostate cancer risk overall; however, risk was elevated in some subgroups and for aggressive outcomes. Future evaluation of air pollution mixture components may clarify observed subgroup differences.
Although the prevalence of cardiometabolic disease is greater in men vs women, the relative risk of cardiovascular disease (CVD) conferred by cardiometabolic conditions is higher in women than in men. We examined the sex-specific association of cardiometabolic risk factor burden with subclinical echocardiographic cardiac remodeling. In this cross-sectional study, we examined whether sex modifies the association between cardiometabolic disease burden (measured as metabolic syndrome severity [MetSS] score) with echocardiographic markers of subclinical cardiac remodeling (including global longitudinal strain [GLS] and E/e' ratio) using multiplicative interaction terms in multivariable-adjusted linear regression models. Among 6182 Framingham Heart Study participants (mean age 51 ± 15 years; 54% women), we observed that sex modifies the association between the MetSS score and subclinical markers of left ventricular systolic and diastolic function. Specifically, a higher MetSS score was associated with worse GLS and E/e' ratio in women versus men. For example, every 1-point increase in the MetSS score was associated with 0.35% higher (worse) GLS in women compared with 0.23% higher GLS in men (ß 0.35, SE 0.04 in women versus ß 0.23, SE 0.04 in men, pint 0.01). Similarly, the MetSS score was more strongly associated with a higher (worse) E/e' ratio in women versus men (ß 0.29, SE, 0.02 in women versus ß, 0.20, SE, 0.02 in men, pint <0.001). We observed that sex modifies the association of cardiometabolic disease burden with subclinical markers of cardiac dysfunction. Specifically, higher cardiometabolic disease burden was associated with worse GLS and E/e' ratio in women versus men. This study explores if sex modifies with the relationship between cardiometabolic risk and markers of cardiac dysfunction. The relationship between cardiometabolic risk factors and global longitudinal strain and E/e’ ratio varies by sex. Women have a steeper decline in markers of cardiac function than men for any given burden of cardiometabolic risk factors.
Severe aortic stenosis (AS) and mitral regurgitation (MR) are frequently undertreated and characterized by persistent sex, racial and ethnic, socioeconomic, and geographic disparities despite effective valve therapies. Whether automated electronic clinician notification (ECN) alerts improve the evaluation and treatment of AS and MR across health systems is unknown. The purpose of this study was to evaluate whether ECN alerts improve guideline-directed evaluation and treatment of significant AS and MR across multiple health systems. ALERT is a multisystem, cluster-randomized clinical trial including clinicians ordering echocardiograms across 5 U.S. health systems encompassing 35 hospitals between August 2024 and September 2025. Clinicians were randomized 1:1 to receive an ECN alert identifying significant AS or MR with accompanying care recommendations or to no alert with usual care. The primary endpoint was a hierarchical composite of time to surgical or transcatheter valve intervention, followed by time to multidisciplinary heart team clinic evaluation within 90 days, analyzed using the stratified win-ratio method. Secondary outcomes included individual components of the composite. A total of 765 clinicians ordering 2,016 echocardiograms were included. In the win-ratio analysis of the primary endpoint, ECN alert was superior to usual care (win ratio: 1.27; 95% CI: 1.05-1.54; P = 0.007), including higher rates of valve intervention (13.4% vs 9.6%; P = 0.005) and multidisciplinary heart team evaluation (22.7% vs 17.9%; P = 0.005) and shorter times to both endpoint components. Effect sizes were similar in AS (win ratio: 1.29) and MR patients (win ratio: 1.23). No evidence of heterogeneity was noted by valve pathology (Pint = 0.821) or across prespecified subgroups (age, sex, race, social deprivation index, inpatient vs outpatient setting, provider specialty, and rurality; Pint > 0.100 for all) and sensitivity analyses yielded consistent results across modified intention-to-treat, intention-to-treat, and per-protocol populations. In this multisystem cluster randomized trial, automated ECN alerts improved timely guideline-directed evaluation and valve intervention for clinically significant AS and MR. These findings suggest that electronic health record-integrated clinical decision support may represent a scalable strategy to reduce undertreatment and improve access to specialized valve care. (Addressing Under-treatment and Health Equity in AS and MR Using an Integrated EHR Platform; NCT06099665).
Emerging data suggest that, in patients with chronic coronary syndromes (CCS), clopidogrel provides superior antithrombotic protection compared with aspirin for secondary prevention, without an associated increase in bleeding risk. The consistency of the observed benefits across ethnicities remains uncertain. Randomized controlled trials (RCTs) comparing aspirin vs clopidogrel for secondary prevention in patients with CCS were screened. The primary endpoint was trial-defined major adverse cardiovascular events (MACE). Prespecified subgroup interaction by ethnicity (East Asian vs non-East Asian) was performed. Trial sequential analysis was used to assess the statistical power of the results. Seven RCTs were included, encompassing 30 165 patients, of whom 75.1% were East Asians. Median follow-up was 36.0 months. Overall, clopidogrel was associated with significant reductions in MACE (incidence rate ratio [IRR] 0.83, 95% confidence interval [CI] 0.69-0.99) and net adverse clinical events (IRR 0.83, 95% CI 0.69-0.99), compared with aspirin. There were no differences between groups in major bleeding, myocardial infarction, or mortality. The results were consistent in the East Asian population, whereas a significant interaction by ethnicity was observed for MACE (East Asians: IRR 0.76, 95% CI 0.59-0.98; non-East Asians: IRR 1.00, 95% CI 0.74-1.36; Pint = 0.010) and mortality (Pint = 0.014), with no significant benefits observed in non-East Asian patients. The analyses were statistically powered for most outcomes in East Asian populations but for none in non-East Asian populations. In patients with CCS, the overall body of evidence suggests improved outcomes with clopidogrel compared with aspirin, without an apparent effect on mortality. However, a high level of confidence in these findings is currently limited to East Asian populations, and additional evidence is needed to clarify their applicability to non-East Asian patients. PROSPERO (CRD420251145176).
DNA methylation accurately predicts chronological age, including gestational age (GA). Previous studies have used CpGs on the EPIC or 450K arrays to generate epigenetic clocks for estimating GA. Using the Asthma&Allergy (A&A) array, we estimated GA and calculated GA acceleration (GAA) in cord blood DNA from 2,451 ancestrally diverse participants from seven birth cohorts investigating early life risk factors for asthma and allergic diseases and disease onset in childhood. Two gestational epigenetic clocks were constructed: one used GA-associated CpGs in an epigenome-wide association study (EWAS) and a second used CpGs associated with GA in specific cell types. For both, we calculated GAA and tested for associations with six prenatal variables and eight allergy-related childhood outcomes. We then conducted pathway analysis of expressed genes correlated with GAA and validated gene expression signatures in peripheral blood at age 2. Strong correlations between reported GA and estimated GA were observed using the EWAS and the cell-specific clocks (r=0.90 and r=0.83, respectively). Using the cell-specific clock, GAA was associated with two outcomes (higher birthweight, Padj=1.69x10-5; less allergic asthma, Padj=0.025), while the EWAS clock was associated with birthweight (Padj=4.68x10-4). A significant sex by GAA interaction effect on birthweight, with a larger effect size in females, was observed with both clocks (EWAS, Pint=5.77x10-3; cell-specific, Pint=0.021). Cord blood RNA-seq analysis revealed upregulated IL-6 and TNF and downregulated IL-10 signaling pathways associated with GAA, and gene expression in blood at age 2 further revealed associations with asthma at age 7. Positive correlations between GAA and inflammatory gene expression and the negative association with allergic asthma suggest that increased expression of inflammatory genes in cord blood and at age 2 is protective against developing asthma. CpGs on the A&A array are accurate predictors of GA, capturing aging aspects specifically related to inflammatory programs.
Renal dysfunction might affect outcomes in patients with ST-segment elevation myocardial infarction (STEMI) and multivessel coronary artery disease (MVD) undergoing percutaneous coronary intervention (PCI). In MULTISTARS AMI, patients with STEMI and MVD were randomized to immediate or staged PCI of non-culprit lesions. In this pre-specified analysis, patients were stratified according to the presence of renal dysfunction at baseline, defined at an estimated glomerular filtration rate (eGFR) of 60 ml/min/1.73 m2. Patients with an eGFR < 30 ml/min/1.73 m2 were excluded from the trial. The primary endpoint was a composite of death, non-fatal myocardial infarction, stroke, unplanned revascularization, or hospitalization for heart failure at 1 year. In MULTISTARS AMI, 108 (13%) of 832 patients had renal dysfunction. The primary endpoint occurred more frequently in patients with renal dysfunction (19.4% vs. 11.2%, unadjusted HR 1.82, 95% CI 1.13-2.94), primarily driven by higher rates of death. Among patients with renal dysfunction, the rates of the primary end point were 14.5% and 24.5% in the immediate and staged PCI groups (unadjusted HR 0.55, 95% CI 0.23-1.33). There was no interaction between renal dysfunction and the randomized treatment assignment with respect to the primary end point (adjusted HR 1.30, 95% CI 0.8-2.20, pint 0.82). The occurrence of acute renal insufficiency was statistically similar in patients with renal dysfunction who underwent immediate and staged PCI (10.9% vs. 18.9%, unadjusted HR 0.61, 95% CI 0.22-1.72, pint 0.09). Renal dysfunction at baseline emerged as a strong risk factor for the development of acute renal insufficiency (adjusted HR 5.0, 95% CI 2.30-10.70, p < 0.01). Outcomes with immediate compared to staged multivessel PCI did not appear significantly altered by the presence of renal dysfunction at baseline. (Supported by Boston Scientific; MULTISTARS AMI ClinicalTrials.gov number, NCT03135275).
The impact of complex coronary anatomy on outcomes in patients at high bleeding risk (HBR) undergoing percutaneous coronary intervention (PCI) remains unclear. Patients from a large PCI referral center were labeled as HBR in presence of ≥1 major or ≥2 minor Academic Research Consortium HBR criteria and undergoing complex PCI if they had any of the following: ≥3 vessels treated, ≥3 stents implanted, ≥3 lesions treated, stent length >60 mm, bifurcation lesion with ≥2 stents, PCI of chronic total occlusion, left main, saphenous venous graft or requiring the use of atherectomy. Primary outcomes at 1 year were major adverse cardiac event (MACE), a composite of all-cause death, myocardial infarction or target vessel revascularization, and major bleeding. Of 16,966 patients, 7,295 (43.0%) were HBR with 3,611 (49.5%) undergoing complex PCI and 9,671 (57.0%) were non-HBR with 4,103 (42.4%) undergoing complex PCI. At 1 year, patients with complex PCI had a higher risk of MACE in both HBR (adjusted Hazard Ratio [adj. HR] 1.78, 95% Confidence Interval [95% CI] 1.54 to 2.06) and non-HBR (adj. HR 1.82 95% CI 1.55 to 2.12, respectively, pint 0.64) groups and a higher risk of major bleeding (adj. HR 1.55, 95% CI 1.29 to 1.87 and adj. HR 1.36, 95% CI 1.03 to 1.78, respectively; pint = 0.46), as compared to patients undergoing noncomplex PCI. In conclusion, complex PCI was associated with significantly higher risk of ischemic and bleeding adverse outcomes at 1-year in both HBR and non-HBR patients.
Breast cancer characteristics and outcomes vary by tumor subtype, poverty, race, and geography. Persistent poverty (> 20% residents in poverty for 30+ years) has been associated with breast cancer risk, but whether associations differ by subtype is unknown. We examined subtype-specific breast cancer incidence by persistent poverty, stratified by rurality and race. Using county-level Surveillance, Epidemiology, and End Results data from 2017 to 2021 (excluding 2020), we calculated Luminal A, Luminal B, HER2-enriched, and triple-negative breast cancer (TNBC) incidence rates. We estimated rate differences (RDs) by persistent poverty using age-adjusted and multivariable linear regression models, with stratification by rurality and county racial composition. In age-adjusted models, persistent poverty counties had lower incidence of Luminal A (RD - 13.58, 95% CI = - 19.8, - 7.4) and higher TNBC (RD = 3.82, 95% CI = 2.0, 5.6) compared to non-persistently poor counties. Differences were not significant in multivariable models. In stratified analysis, higher TNBC rates were observed in persistently poor rural (multivariable RD =1.70, 95% CI = 0.3, 4.8) but not urban (multivariable RD = - 1.07, 95% CI = - 4.7, 2.5; pint = 0.09) areas. In counties with > 5% non-Hispanic Black population, Luminal B rates were lower in persistently poor vs. non-persistently poor counties (multivariable RD = - 3.96, 95% CI = - 6.7, - 1.2; pint = 0.03). Results from this study suggest that differences in breast cancer subtypes by persistent poverty status are largely explained by other measures of more recent disadvantage including recent poverty, unemployment, uninsurance, and race. Targeted strategies are needed to address breast cancer disparities within socioeconomically disadvantaged communities.
Computational pathology has emerged as an attractive option for improving risk stratification in prostate cancer (PCa), but most approaches either lack interpretability or focus solely on tumor morphology. We aimed to identify an interpretable, immune microenvironment-derived computational pathology biomarker for PCa. We identified two cohorts (Discovery and Validation) with M0 PCa (n=490) who were treated with radical prostatectomy with H&E-stained whole-slide images (WSIs) and data on distant metastasis (DM). We identified a third cohort from TCGA with M0 PCa (n=326) with prostatectomy WSIs and bulk sequencing. Immune cells were identified from WSIs using a deep learning method (CellViT), and spatially dense immune clusters were quantified using DBSCAN. CIBERSORTx was utilized for immune cell deconvolution and TRUST4 for immune receptor repertoire reconstruction. Median follow-up was 12.6 (Discovery) and 8.1 years (Validation). 14% (n=37, Discovery) and 17% (n=38, Validation) had Gleason 8-10 disease. Immune cell abundance was not associated with DM. In Discovery, increased immune cluster was associated with decreased risk of DM for Gleason 8-10 (adjusted hazard ratio 0.42, 95% confidence interval 0.19-0.93) but not Gleason 6-7 (1.26, 0.77-2.05; Pint=0.020), with similar results in Validation (Gleason 8-10 0.60, 0.37-0.98; Gleason 6-7 1.19, 0.74-1.91; Pint=0.043). In Gleason 8-10 but not 6-7, high-cluster samples were enriched for CD8+ T cells, activated memory CD4+ T cells, Tregs (P≤0.037), and clonal T cell populations (P≤0.039). These findings propose immune spatial clustering as a novel, interpretable computational pathology biomarker and provide insight into the unique immune features of high-grade PCa.
Large-scale randomized data comparing clinical outcomes of atrial fibrillation patients of Asian vs non-Asian races are limited. Data from A Collaboration Between Multiple Institutions to Better Investigate Non-Vitamin K Antagonist Oral Anticoagulant Use in Atrial Fibrillation, which pooled patient-level data from the four pivotal randomized trials of direct oral anticoagulants (DOACs) vs warfarin in patients with atrial fibrillation, were analysed. Baseline characteristics and clinical outcomes in patients of Asian race (Asians) vs non-Asian race were compared. The relative efficacy and safety of DOACs compared with warfarin in Asians vs non-Asians, assessing for interactions between race and treatment effect, were investigated. Outcomes across the range of body weight and creatinine clearance in Asians were also explored. A total of 10 212 Asian patients and 61 471 non-Asians were identified. Compared with non-Asians, Asians were on average 3.2 years younger and 20 kg lighter, had worse renal function (mean creatinine clearance 64.9 vs 77.3 mL/min), and had higher rates of prior stroke/transient ischaemic attack (37.2% vs 26.6%) (P < .001 for each). In the warfarin arm (median time in therapeutic range 57.7% for Asians vs 66.2% for non-Asians, P < .001), Asians had a higher adjusted risk of stroke/systemic embolic events (SEEs), major bleeding, intracranial haemorrhage, gastrointestinal bleeding, and primary net clinical outcome (stroke/SEE, major bleeding, or death). Compared with warfarin, standard-dose (SD) DOACs significantly reduced the risks in Asians to a greater degree than non-Asians for stroke/SEE (hazard ratio [HR] .65, 95% confidence interval .53-.80 vs HR .86, 95% confidence interval .78-.95), major bleeding (HR .62 [.52-.75] vs HR .91 [.84-.98]), and primary net clinical outcome (HR .76 [.68-.85] vs HR .94 [.90-.98]; Pint < .02 for each). Standard-dose DOACs increased gastrointestinal bleeding only in non-Asians (Asians HR .92 [.69-1.23] vs non-Asians HR 1.41 [1.25-1.58], Pint = .009). In Asians, SD DOACs reduced the risks of clinical events across the wide range of body weight and creatinine clearance. Compared with SD DOACs, lower-dose DOACs significantly increased the risk of stroke/SEE (HR 1.57 [1.15-2.13]) and secondary net clinical outcome (stroke/SEE, intracranial haemorrhage, or death; HR 1.23 [1.03-1.48]) in Asians. Clinical outcomes with SD DOACs vs warfarin in patients with atrial fibrillation were generally even more favourable in patients of Asian than non-Asian race. Compared with warfarin, SD DOACs did not increase the risk of gastrointestinal bleeding in Asians. Standard-dose DOACs are preferred over warfarin or lower-dose DOACs in patients of Asian race.
Stroke and atrial fibrillation are frequent manifestations of left atrial (LA) myopathy in patients with end-stage kidney disease (ESKD). However, the natural history of LA dysfunction and its reversibility following renal transplantation remain insufficiently defined. We aimed to assess baseline LA function and post-transplantation changes in ESKD using LA emptying fraction (LAEF) and LA function index (LAFI). A total of 111 patients with ESKD listed for renal transplantation and 45 healthy controls were enrolled. All participants underwent two-dimensional echocardiography and treadmill exercise testing at baseline and at 6 months. Patients who did not undergo transplantation during follow-up continued maintenance hemodialysis. At baseline, LA volumes were significantly increased in patients with ESKD regardless of whether they were on hemodialysis; however, no significant difference in LAEF was observed between groups (p = 0.073). Compared to the control group, the hemodialysis cohort exhibited a significantly higher prevalence of elevated LA pressure (p < 0.001) and a reduced LAFI (p = 0.001), and LAFI remained significantly lower in the hemodialysis group following multivariable adjustment. Six months after transplantation, transplant recipients had significantly higher LAEF and LAFI compared to patients remaining on hemodialysis (adjusted Pint = 0.013 and 0.045). Both LAEF and LAFI correlated positively with exercise capacity at baseline, and longitudinal improvement in LAFI correlated with improved exercise duration at follow-up. LA function is impaired in ESKD, particularly among patients on dialysis, and is associated with reduced exercise capacity. Renal transplantation is linked to relative preservation and modest improvement of LA function, paralleling better functional performance.
Iron overload disrupts cellular homeostasis and drives ferroptosis through dysregulated iron metabolism. Non-coding RNAs (ncRNAs) are considered as key regulators of various biological functions and targets for a new generation of RNA therapeutics and biomarkers. However, few studies have investigated the regulatory roles of ncRNAs, particularly competitive endogenous RNAs (ceRNAs) in iron overload. This study performed whole-transcriptome sequencing to characterize the ceRNA network in ferric ammonium citrate (FAC)-induced iron-overloaded HT-1080 fibrosarcoma cells. A total of 208 differentially expressed mRNAs, 83 lncRNAs, and 170 circRNAs (q < 0.05) were identified, with hierarchical clustering revealing distinct expression patterns between control and iron-treated groups. KEGG enrichment implicated vitamin B6 metabolism (q < 0.001) and lysine degradation (q < 0.001) as key disrupted pathways. ceRNA network was conducted and further demonstrated lncRNA/circRNA-mediated regulation of ferroptosis genes via shared miRNA response elements. Notably, LINC-PINT-232 was implicated in the regulation of both ferritin heavy chain (FTH) and sequestosome 1 (SQSTM1), two ferroptosis-associated mRNAs. FTH upregulation mitigates iron toxicity through ferroxidase activity, while SQSTM1 modulates lipid peroxidation in ferroptosis. These findings provide a preliminary transcriptomic landscape for hypothesis generation regarding ncRNA-mediated regulatory mechanisms in iron overload-induced ferroptosis and offer a computational foundation for future functional and therapeutic investigations.