Yan Gong

Background/Objectives: Carfilzomib (CFZ) and bortezomib (BTZ) are proteasome inhibitors used as the first-line therapy for relapsed or refractory multiple myeloma (MM) but are associated with cardiovascular adverse events (CVAEs). This study aims to identify differentially methylated positions (DMPs) and regions (DMRs), and enriched pathways associated with CVAEs related to CFZ or BTZ-based treatment. Methods: Baseline germline DNA methylation profiles from 79 MM patients (49 on CFZ and 30 on BTZ) in the Prospective Study of Cardiac Events During Proteasome Inhibitor Therapy (PROTECT) were analyzed. Epigenome-wide analyses were performed within each group, followed by meta-analyses to identify signals common to CVAEs associated with both medicines. Results: Four DMPs were significantly associated with CFZ-CVAEs, including cg15144237 within ENSG00000224400 (p = 9.45 × 10⁻¹⁰), cg00927646 within TBX3 (p = 9.78 × 10⁻⁸), and cg10965131 within WDR86 (p = 1.00 × 10⁻⁷). One DMR was identified in the FAM166B region (p = 5.46 × 10⁻⁷). There was no evidence of any DMPs in BTZ-treated patients, however two DMPs and one DMR reached a suggestive level of significance (p < 1.00 × 10⁻⁵): cg09666417 in DNAJC18 (p = 3.41 × 10⁻⁷) and cg12987761 in USP18 (p = 5.00 × 10⁻⁷), and a DMR mapped to the WDR86/WDR86-AS1 region (p = 8.11 × 10⁻⁸). Meta-analysis did not find any significant DMPs, with the top CpG being cg17933807 in GNL2 (p = 7.38 × 10⁻⁵). Pathway enrichment analyses identified peroxisome, MAPK, Rap1, adherens junction, phospholipase D, autophagy, and aldosterone-related pathways to be implicated in CVAEs. Conclusions: Our study identified distinct DMPs, DMRs, and pathways enrichment associated with CVAE, suggesting epigenetic contributors to CVAEs and supporting the need for larger validation studies.

Carfilzomib is highly effective in the treatment of multiple myeloma, but it has been associated with cardiovascular adverse events that impact patient outcomes. Our prior global metabolomic analyses indicated an association between hydrophilic bile acids and carfilzomib‐cardiotoxicity risk, although a causal relationship remained to be determined. Here, our objective was to validate the previously identified bile acids in an independent cohort and investigate whether these hydrophilic bile acids play a causal role in carfilzomib‐cardiotoxicity using Mendelian randomization. Using targeted metabolomics, we validated the association between glycoursodeoxycholic acid and carfilzomib‐cardiotoxicity in an independent cohort ( n = 61). We then performed two‐sample Mendelian randomization analyses, using metabolome‐wide association study results to provide the genetic instruments for bile acid levels as exposure and genome‐wide association study summary statistics from the UK Biobank ( n = 484,598) as the outcome data for cardiovascular adverse events. Causal inference was assessed with the inverse‐variance weighted method, followed by multiple sensitivity analyses. Higher glycoursodeoxycholic acid concentration (odds ratio = 0.34, P = 0.032) was associated with lower cardiotoxicity risk after adjusting for hypertension and high levels of brain natriuretic peptides. Mendelian randomization analysis demonstrated a robust causal effect of glycoursodeoxycholic acid on cardiotoxicity risk (β = −0.00065, P = 6.2 × 10 ⁻⁵ ). Gene enrichment analysis indicated pathways involving potassium channel regulation and thromboxane signaling to be implicated. This integrative metabolomic and genetic investigation supports a potential protective role of glycoursodeoxycholic acid in cardiovascular vulnerability and motivates larger, carfilzomib‐specific studies to evaluate its utility for risk stratification.

The mechanisms underlying carfilzomib (CFZ)-induced cardiotoxicity remain incompletely elucidated. In this study, we used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to characterize the transcriptional impact of CFZ and to evaluate whether atorvastatin could prevent these deleterious transcriptional changes. hiPSC-CMs were treated with 1 µM CFZ, CFZ + atorvastatin, atorvastatin, or vehicle control, followed by RNA sequencing, differential expression analyses, and pathway analyses. Transcriptomic profiling revealed a marked upregulation of genes in multiple proteasome subunits, including ATPase components (PSMC1, PSMC4, PSMC5, PSMC6) and non-ATPase regulatory subunits (PSMD1, PSMD2, PSMD12), suggesting a strong compensatory activation of proteostasis and protein quality-control pathways in response to CFZ exposure. In addition, several of the most significantly altered genes were those implicated in cardiomyopathy and heart failure, such as BAG3 and FLNC, and many heat-shock proteins, indicating the activation of cardiac stress–response pathways relevant to CFZ-associated cardiotoxicity. Atorvastatin co-treatment partially reversed a subset of CFZ-induced transcriptional changes, particularly within cholesterol biosynthesis and lipid-regulatory pathways (e.g., ACAT2 and ACTA1) but did not restore the CFZ-mediated downregulation of sarcomeric genes. Together, these findings define a multifactorial signature of deleterious CFZ-induced transcriptional changes and suggest that atorvastatin may provide partial metabolic, but not structural, cardio protection.

Background/Objectives: Carfilzomib (CFZ) and bortezomib (BTZ) are proteasome inhibitors used as the first-line therapy for relapsed or refractory multiple myeloma (MM) but are associated with cardiovascular adverse events (CVAEs). This study aims to identify differentially methylated positions (DMPs) and regions (DMRs), and enriched pathways in patients who developed CFZ- and BTZ- related CVAEs. Methods: Baseline germline DNA methylation profiles from 79 MM patients (49 on CFZ and 30 on BTZ) in the Prospective Study of Cardiac Events During Proteasome Inhibitor Therapy (PROTECT) were analyzed. Epigenome-wide analyses within each group identified DMPs, DMRs, and enriched pathways associated with CVAEs compared with individuals without CVAEs. Results: Four DMPs were significantly associated with CFZ-CVAE: cg15144237 within ENSG00000224400 (p = 9.45x10⁻¹⁰), cg00927646 within TBX3 (p = 9.78x10⁻⁸), and cg10965131 within WDR86 (p = 1.00x10⁻⁷). One DMR was identified in the FAM166B region (p = 5.46x10⁻⁷). There was no evidence of any DMPs in BTZ-CVAE patients, however two DMPs and one DMR reached a suggestive level of significance (p < 1.00x10⁻⁵): cg09666417 in DNAJC18 (p = 3.41x10⁻⁷) and cg12987761 in USP18 (p = 5.00x10⁻⁷), and a DMR mapped to the WDR86/WDR86-AS1 region (p = 8.11x10⁻⁸). Meta-analysis did not find any significant DMPs, with the top CpG being cg17933807 in GNL2 (p = 7.38 x10⁻⁵). Pathway enrichment analyses identified peroxisome, MAPK, Rap1, adherens junction, phospholipase D, autophagy, and aldosterone-related pathways to be implicated in CVAEs. Conclusions: Our study identified distinct DMP, DMR, and pathway enrichment associated with CVAE, suggesting epigenetic contributors to CVAEs and supporting the need for larger validation studies.

The mechanisms underlying carfilzomib (CFZ)-induced cardiotoxicity remain incompletely elucidated. In this study, we used human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) to char-acterize the transcriptional impact of CFZ and to evaluate whether atorvastatin could prevent these dele-terious transcriptional changes. hiPSC-CMs were treated with 1 µM CFZ, CFZ + atorvastatin, atorvastatin, or vehicle control, followed by RNA sequencing and differential expression analysis and pathway analyses. Transcriptomic profiling revealed a marked upregulation of genes in multiple proteasome subunits, in-cluding ATPase components (PSMC1, PSMC4, PSMC5, PSMC6) and non-ATPase regulatory subunits (PSMD1, PSMD2, PSMD12), suggesting a strong compensatory activation of proteostasis and protein quality-control pathways in response to CFZ exposure. In addition, several of the most significantly altered genes were those implicated in cardiovascular disease and heart failure, such as BAG3 and FLNC and many heat-shock proteins, indicating activation of cardiac stress-response pathways relevant to CFZ-associated cardiotoxicity. Atorvastatin co-treatment partially reversed a subset of CFZ-induced transcriptional changes, particularly within cholesterol biosynthesis and lipid regulatory pathways (e.g. ACAT2 and ACTA1), but did not restore CFZ-mediated downregulation of sarcomeric genes. Together, these findings define a mul-tifactorial signature of CFZ-induced deleterious transcriptional changes and suggest that atorvastatin may provide partial metabolic, but not structural, cardioprotection.

Hypertension is a known modifiable risk factor for Alzheimer’s disease and related dementia (ADRD). However, it is unknown how variance in hypertension control, antihypertensive medications, and social determinants of health, such as social deprivation index (SDI), influence the risk of developing ADRD. Validated hypertension computable phenotype algorithms were applied to electronic health record data from the OneFlorida Data Trust (1/1/2013–12/31/2016), to identify apparent treatment‐resistant hypertension (aTRH), and hypertension‐control levels (well‐controlled hypertension, intermediate‐controlled hypertension, uncontrolled hypertension). The primary outcome was a new ADRD diagnosis using validated ICD‐9/10 codes. Multiple adjusted stepwise logistic regression models were used to identify factors associated with ADRD development. ADRD cumulative hazard incidence per hypertension control levels was assessed using the Nelson–Aalen estimator and log‐rank test. A total of 57,273 hypertension patients with 6401 (11%) incident ADRD cases were included in the analysis. The average age was 67 years, with 57% females and 32% identifying as Black or African American. aTRH was a significant ADRD predictor (OR: 1.327, 95% CI: 1.234–1.427), compared to other hypertension phenotypes. aTRH was also significantly associated with a higher incidence of ADRD over time ( P < 0.0001). Patients prescribed thiazide diuretics (OR: 0.894, 95% CI: 0.837–0.956) and fixed‐dose combination medications (OR: 0.804, 95% CI: 0.732–0.882) had a lower risk of ADRD. A linear relationship between SDI quartiles and ADRD risk was found. aTRH was significantly associated with the development of ADRD. Our study also highlights the importance of comprehensive hypertension control and socioeconomic interventions in preventing or reducing ADRD risk in hypertension patients.

Medication-related osteonecrosis of the jaw (MRONJ) is a severe adverse effect of antiresorptive agents, including bisphosphonates (BPs) and denosumab (DMB). We conducted a case–control epigenome-wide association study (EWAS) of 24 cancer patients treated with BPs or BPs + DMB using the Infinium^® MethylationEPIC v2.0 to explore epigenetic differences associated with MRONJ. Differentially methylated positions (DMPs) and regions (DMRs) were assessed across three analyses: MRONJ vs. controls (main), BPs-MRONJ vs. BPs-controls, and BPs/DMB-MRONJ vs. BPs/DMB-controls. Eight plasma bone biomarkers were quantified by Luminex and correlated with top methylation sites. We identified 10 DMPs and 4 DMRs at suggestive significance (p < 1 × 10⁻⁵). cg1913766 in the NOP56 promoter was hypomethylated in the main analysis (p = 2.19 × 10⁻⁷) and in BPs-MRONJ (p = 4.80 × 10⁻⁶), correlating with osteocalcin (p = 0.02 and 0.03, respectively). TNXB cg21289669 was hypermethylated in the main analyses (p = 6.31 × 10⁻⁶), and TNXB locus formed a DMR (p = 3.30 × 10⁻¹⁰) in the main and BPs-MRONJ analyses (p = 2.76 × 10⁻⁷). cg11392877 in PDE8A was hypomethylated in BPs/DMB-MRONJ (p = 5.35 × 10⁻⁷). TRIM15 was a significant DMR in BPs-MRONJ and the main analysis (p = 3.30 × 10⁻¹⁰). TRIM15, TNXB, and PDE8A regulate collagen I, while NOP56 supports ribosome biogenesis, potentially contributing to MRONJ. Given the small sample size, these findings are preliminary and validation in larger studies is warranted.

Background: This study evaluated the impact of diabetes mellitus (DM) and its treatments on cardiovascular outcomes in prostate cancer (PC) patients aged 66 years and older, with or without androgen deprivation therapy (ADT) exposure. Methods: Using the SEER-Medicare database (2009–2017), two cohorts were created: Cohort 1 included all PC patients enrolled in Medicare Parts A and B; Cohort 2 was a subset of Cohort 1 receiving ADT and enrolled in Medicare Part D. Exposures were DM and DM medications. Outcomes included cardiovascular events (CVEs), cardiovascular mortality (CVm), PC-specific mortality (PCsm), and all-cause mortality, analyzed using multivariable Fine-Gray and Cox models. Results: Cohort 1 included 150,647 PC patients (32% with DM, median age 72). DM was associated with higher risk of CVE (subdistribution hazard ratio [sHR] 1.20, 95% CI 1.17–1.22), CVm (sHR 1.35, 1.28–1.43), and all-cause mortality (adjusted HR [aHR] 1.22, 1.19–1.26) (all p < 0.001). Non-Hispanic Blacks (NHBs) and patients from lower socioeconomic (SES) and education areas experienced comparable or worse outcomes. In Cohort 2 (n = 14,938), DM patients on non-metformin therapies had higher all-cause mortality (aHR 1.33, 1.11–1.25; p = 0.002) than those on metformin, particularly in NHB and low education groups. Sensitivity analyses with follow-up limited to two years showed consistent results as overall. Conclusions: Diabetic PC patients, especially NHB, lower SES and lower education subgroups, were associated with worse cardiovascular and all-cause mortality outcomes. Metformin may be associated with better outcomes in these populations, warranting further research on the disparities in PC and diabetes, and cardioprotective effects of DM medications across different subpopulations.

CYP2C9 metabolizes approximately 20% of clinically administered drugs. Several single-nucleotide polymorphisms (SNPs) of CYP2C9 (e.g., *2, *3, *8, and rs12777823) are used as biomarkers to predict CYP2C9 activity. However, a large proportion of variability in CYP2C9 expression remains unexplained. Background/Objectives: We previously identified a variable number tandem repeat (pVNTR) polymorphism in the CYP2C9 promoter. The short repeat (pVNTR-S) showed reduced transcriptional activity in reporter gene assays and was associated with decreased CYP2C9 mRNA expression. However, because pVNTR-S is in high linkage disequilibrium (LD) with CYP2C9*3 in the European population, whether pVNTR-S directly impacts CYP2C9 expression remains unclear. The objective of this study was to clarify the association between the pVNTR-S and CYP2C9 mRNA expression in human liver samples and to assess its impact on CYP2C9 expression independently of known CYP2C9 biomarkers. Methods: Gene expression was measured by real-time qPCR. SNPs and pVNTRs were genotyped using SNapShot assays and fragment analysis, respectively. Associations between CYP2C9 and the pVNTR-S or SNPs were analyzed using multiple linear regression. Results: Our results showed that pVNTR-S was associated with lower CYP2C9 expression (34% reduction, p-value = 0.032) in human liver samples (n = 247), while the known CYP2C9 biomarkers (CYP2C9*2, *3, *8, or rs12777823) were not. These results suggest that pVNTR-S reduces CYP2C9 expression independently of known biomarkers. Therefore, pVNTR-S may explain additional variability in CYP2C9 expression when present alone or in conjunction with other CYP2C9 alleles.