Chronic psychosocial stressors contribute to cardiovascular disease (CVD) and related risk factors, particularly in under-resourced communities. Psychosocial stressors activate the hypothalamic-pituitary-adrenal (HPA) axis, leading to sustained cortisol action on immune cells. Natural killer (NK) cells are altered in distribution and function in cardio-metabolic diseases; however, pathways linking cortisol as a marker of chronic stress to NK cell dysfunction remain understudied. We recruited 186 African American women from the Step It Up Community-Engaged, Digital Health Physical Activity Intervention study and collected cortisol, NK cell measures, and psychosocial stress measures using validated questionnaires, including chronic stress and loneliness. Plasma cortisol levels were negatively associated with NK cell proportions, and self-reported levels of loneliness modified this relationship (p = 0.011); among participants with higher loneliness, higher plasma cortisol levels were associated with lower proliferative NK cell proportions (β=-0.30, p = 0.04). In a subset of study participants (n = 24), plasma cortisol levels were directly associated with a loss of NK cell degranulation (β=-0.38, p = 0.03). Using an RNA sequencing dataset, we found that higher plasma cortisol levels were associated with suppressed gene expression related to NK cell cytotoxicity. Finally, in vitro experiments on freshly isolated primary NK cells revealed that cortisol reduces NK cell degranulation (p = 0.006) via FABP-4-induced upregulation of PD-1 expression (p = 0.01). In summary, our study provides evidence that in individuals with higher loneliness, cortisol may alter NK cell distribution and function. Furthermore, cortisol-mediated NK cell dysfunction may be facilitated in a FABP-4/PD-1 dependent manner, leading to impaired innate immune function and, potentially, contributing to worsening CVD risk.

Abstract African American (AA) women face disproportionate cardiovascular disease (CVD) risk due to adverse social determinants of health (SDoH), including lower socioeconomic status (SES). Physical activity (PA) and plasma extracellular vesicles (EVs) modulate CVD risk, but their relationships with SDoH remain unclear. This study examines associations between SDoH, PA, and plasma EVs in a Washington, DC-based cohort of at-risk AA women. Participants (N = 24, Age: 57 ± 12, BMI: 35 ± 6, ASCVD: 9 ± 5) joined the Step It Up Community-Engaged, Digital Health Physical Activity Intervention pilot study. EVs were isolated from fasting plasma samples. Multivariable regression, adjusted for BMI and ASCVD 10-year risk, showed that neighborhood socioeconomic deprivation (NSD) was associated with decreased EV size (β=-0.49, p = 0.01), while higher daily step count was associated with increased EV size (β = 0.48, p = 0.02). EV miRNA cargo, including miR-1246, miR-28-5p, and miR-765, showed distinct expression patterns with EV size, NSD, and PA. In vitro, endothelial cell (EC) barrier integrity directly correlated with miR-28-5p (r = 0.73, p = 0.007) and miR-765 (r=-0.58, p = 0.049). EC migration correlated negatively with miR-28-5p (r=-0.67, p = 0.02). Findings highlight smaller EV size associated with high NSD and low PA, suggesting miR-28-5p may be a biomarker or mediator for CVD pathogenesis. PA interventions may mitigate adverse SDoH effects and establish EV miRNA cargo as biomarkers and/or intervention mediators.

Introduction: Adverse psychosocial factors worsen cardiovascular risk through chronic inflammation and dysregulation of the sympathetic nervous system and hypothalamic-pituitary-adrenal axis. Social determinants of health are closely related to psychosocial risk in underserved populations within the United States, though few studies have examined relationships between psychosocial factors and lipoproteins in communities with elevated social risks. We hypothesized that higher psychosocial risk would be associated with less favorable lipid profiles among African American women living in resource-limited Washington, DC neighborhoods. Methods: African American women with overweight or obesity aged 21-75 years residing in Wards 5, 7, and 8 of Washington, D.C. and Prince George’s County in Maryland were recruited for the Step It Up study. Principal component analysis of baseline psychosocial survey data was performed and identified high loadings for depression, social isolation, and loneliness. Participants were assigned psychosocial risk based on sums of these survey scores, with higher scores indicating greater psychosocial adversity. Fasting lipid profiles were measured using Nuclear Magnetic Resonance spectroscopy, quantifying lipoprotein particle concentrations, numbers, and sizes as well as apolipoprotein (Apo) A/B levels. Associations between psychosocial risk and lipid profiles were assessed using linear regression models, adjusting for demographic, socioeconomic, and comorbidity covariates. Results: Participants with psychosocial survey data were included in analyses (n=165, age 57.4±12.6 years, BMI 36.2±6.7). Women in the high psychosocial risk tertile had higher BMI and systolic blood pressure and greater proportions of lipid-lowering therapy compared to those in the low risk tertile. High-risk individuals also had lower Healthy Eating Index scores and lower socioeconomic status. Increased psychosocial risk was associated with low density lipoprotein (LDL) concentration (β= 4.75, p=0.01), LDL particle numbers (β= 68.61, p=0.01) and ApoB concentrations (β= 3.50, p=0.01) (Table). Psychosocial factor scores were not associated with lipid size. Conclusions: Psychosocial adversity was associated with more atherogenic lipid profiles, including higher LDL concentration and particle numbers and ApoB concentrations. These findings suggest that psychosocial risk factors among at-risk populations may contribute to cardiovascular risk through lipid dysregulation.

Introduction:The authors explored the relationship between lifestyle behavior, self-efficacy, and self-reported physical activity among individuals with overweight/obesity residing in resource-limited neighborhoods. Methods:Using data from the Washington, DC Cardiovascular Health and Needs Assessment and the Step It Up digital health intervention study, the authors examined the relationships between self-efficacy and self-reported physical activity participation. Individuals completed the Physical Activity and Nutrition Self-Efficacy scale and the Global Physical Activity Questionnaire, which assesses self-reported physical activity participation. Logistic and gamma regression models were performed to assess the associations between Physical Activity and Nutrition Self-Efficacy quintiles and (1) the odds of any physical activity participation and (2) the duration of physical activity participation. Results:The sample (N=227) had a mean age of 57.71 (±12.60) years and BMI of 34.55 (±7.19) and were predominantly female (90.75%). The majority had college education (57.55%) and less than $60,000 annual income (44.15%). Individuals who scored in the third quintile were 3.26 (95% CI=1.18, 9.02) and 3.97 (95% CI=1.46, 10.82) times more likely to report moderate physical activity and muscle-strengthening activity participation, respectively, than those who scored in the first quintile, after adjusting for sociodemographic factors. Those who scored in the third and fifth quintiles were 3.91 (95% CI=1.28, 11.97) and 5.92 (95% CI=1.90,18.47) times more likely to meet the guideline-recommended 150 minutes weekly moderate-to-vigorous physical activity. Conclusions:Higher physical activity self-efficacy is associated with higher odds of participation, physical activity duration, and the odds of meeting the guideline-recommended weekly moderate-to-vigorous physical activity. Interventions that augment self-efficacy may be particularly important in improving cardiovascular health by enhancing physical activity in individuals with obesity.

Chronic stress is associated with cardiovascular disease (CVD) risk and elevated amygdala activity. Previous research suggests a plausible connection between amygdala activity, hematopoietic tissue activity, and cardiovascular events; however, the underlying biological mechanisms linking these relationships are incompletely understood. Chronic stress is thought to modulate epigenomic modifications. Our investigation focused on associations between amygdala activity (left (L), right (R), maximum (M), and average (Av) AmygA), and splenic (SpleenA), and bone marrow activity (BMA) as determined by 18Fluorodeoxyglucose (FDG) on Positron Emission Tomography/Computed Tomography (PET/CT) scans. Subsequently, we assessed how these markers of chronic stress and hematopoietic activity might relate to the DNA methylation of stress-associated genes in a community-based cohort of African American individuals from Washington D.C. at risk for CVD. To assess the relationships between AmgyA, SpleenA, BMA, and DNA methylation, linear regression models were run and adjusted for body mass index and 10-year predicted atherosclerotic CVD risk. Among 60 participants (93.3% female, mean age 60.8), M-AmygA positively associated with SpleenA (β = 0.29; p = 0.001), but not BMA (β = 0.01; p = 0.89). M-AmygA (β = 0.37; p = 0.01 and β = 0.31; p = 0.02, respectively) and SpleenA (β = 0.73; p < 0.01 and β = 0.59; p = 0.005, respectively) were associated with both IL-1β and TNFα. Decreased M-AmygA, SpleenA, IL-1β, and TNFα were associated with methylation of NFκB1 at cg07955720 and STAT3 at cg19438966. Our findings suggest a potential association between AmygA, SpleenA, and pro-inflammatory cytokines in the setting of chronic stress, suggesting an adverse hematopoietic effect. Furthermore, findings reveal associations with epigenetic markers of NFκB and JAK/STAT pathways linked to chronic stress.

Background: Lp(a) has gained increased attention as a significant risk factor for cardiovascular disease (CVD), and its plasma levels are up to 3-fold higher in African Americans (AAs) likely contributing to the heightened CVD risk. While it is widely acknowledged that Lp(a) levels are largely determined by genetics, the potential impact of epigenetic alterations and a connection to adverse social determinants of health (SDoH) is unknown. Adverse SDoH are known to accelerate CVD risk, partially through epigenetic changes of the genome. In this pilot study, we aim to investigate a potential connection between socioeconomic status (SES) measures of SDoH, methylation levels within the Lp(a) gene, and circulating plasma Lp(a) levels in AAs. Methods: Sixty AAs (93.3% female, mean age: 61 years, mean BMI 33.7kg/m^2) at moderate CVD risk and living in under-resourced neighborhoods within the Washington DC area, were recruited to the NIH. Baseline sociodemographics were collected, including individual-level SES and census tract-based neighborhood socioeconomic deprivation (NSD). Methylating patterns determined by DNA methylation analysis on the Lp(a) gene ( LPA ) were detected utilizing the Illumina technique. Lp(a) plasma levels were measured using ELISA. We conducted a multivariable regression analysis with adjustment for ASCVD 10-year risk score and BMI to explore associations between NSD, methylation levels on LPA , and Lp(a) plasma levels. Results: We examined 20 LPA methylation sites. Four sites were significantly associated with circulating Lp(a) levels. None of these four methylation sites was significantly associated with SES. However, two methylation sites associated negatively with NSD (cg17028067 b= -0.27, p=0.04; cg22888279 b= -0.38, p=0.003). Neither SES nor NSD were directly associated with Lp(a) plasma levels. In a final step, we determined if either cg17028067 or cg22888279 would associate with 18 FDG-PET/CT-measured amygdala activity, a pathophysiological measure of chronic stress. Amygdala activity was negatively associated with LPA -cg22888279 (b= -0.28, p = 0.03) (Figure). Conclusion: Our data highlight the importance of chronic stress-related epigenetic modification of the LPA gene, especially in the cg22888279 locus, to increasing plasma Lp(a) levels, potentially further accelerating CVD development and progression in AAs.

Loneliness is known to be an important contributor to cardiovascular disease (CVD). However, little is known about the impact of loneliness on endothelial barrier integrity, a crucial hallmark of CVD development and progression. In this study, we aimed to investigate how loneliness might impact the endothelium. We found greater perceived loneliness associated with the product of circulating epinephrine and TNFα (E/T), which, in turn, associated with lower VE-cadherin on endothelial cells in an ex vivo experiment. Additionally, circulating plasma levels of soluble VE-cadherin were associated significantly with subclinical CVD. To explore the mechanistic aspects of these associations, we measured the effects of E/T on endothelial barrier function in vitro. E/T treatment decreased endothelial VE-cadherin expression and dampened endothelial barrier integrity, involving at least partial JAK/Stat signaling, highlighting a potential additive effect of epinephrine and TNFα on endothelial dysfunction, potentially accelerating CVD development and progression in individuals experiencing loneliness-related chronic stress.

Background: Dyslipidemia disproportionately affects African American (AA) women and contributes to disparate cardiovascular disease (CVD) outcomes. We previously showed that Natural Killer (NK) cells are functionally impaired by hyperlipidemia, particularly LDL, in a DUSP1 -dependent mechanism. However, the impact of dyslipidemia on NK cell activity-regulating receptor expression (e.g., NKp46) is not completely understood. Therefore, we sought to elucidate the potential impact of LDL on NK cell NKp46 expression in AA women. Methods: NK cell-NKp46 expression was measured by flow cytometry in fresh blood samples from the Step It Up: Physical Activity Intervention participants (n=154). We used multivariable regression to examine associations between plasma lipid profile (LDL, HDL, triglycerides, and total cholesterol) and NK-NKp46 expression. Subsequently, we performed in vitro experiments with freshly isolated naïve NK cells with and without overnight LDL treatment. We used flow cytometry to determine a potential causal impact of LDL on NKp46 expression on NK cells. Results: All Step It Up study participants were AA women (mean age 57 years, mean BMI 36.1 kg/m^2, at intermediate risk for CVD). In the unadjusted model, LDL associated negatively with NKp46 expression on all NK cells (b=-0.18, p=0.03), while no significant associations were found with HDL, triglycerides, or total cholesterol levels. These observed associations persisted after adjustment for BMI and ASCVD 10-year risk (b= -0.17, p=0.04). When focusing on NK cell subsets, LDL trended to significance with NKp46 on cytotoxic NK cells (p=0.08), while no associations were seen with NKp46 expression on proliferative NK cells (p=0.47). For NK cells treated overnight with LDL, NKp46 expression on all NK cells was reduced by 6.78% (p=0.008), mainly driven by a 6.37% reduction on the cytotoxic NK cell subset (p<0.001). The proliferative NK cell subset showed no significant decrease in NKp46 expression levels (p=0.56) compared to vehicle-treated controls (Figure). Conclusion: Our data demonstrate that an LDL-induced reduction in NKp46 expression on NK cells could present an additional pathway by which hyperlipidemia impairs NK cell function, potentially accelerating CVD development and progression in AA women with hyperlipidemia and CVD.

Introduction: Chronic neighborhood stressors contribute to disparate CVD outcomes, with neighborhood socioeconomic deprivation (NSD) linked to inflammation. Separately, relationships have been seen with CVD and specific monocyte phenotypes. However, the connection between neighborhood exposures and monocyte subsets is less clear. Thus, we examined NSD with monocyte phenotypes, hypothesizing that chronic NSD cross-sectionally associates with monocyte subsets. Methods: This study utilized data from the Multi-Ethnic Study of Atherosclerosis (MESA), a population-based prospective cohort of adults aged 45-84 years (N=6814). NSD was scored from principal factor analyses using U.S. Census data (2000), with higher values indicating higher deprivation. Monocyte phenotypes were measured from cryopreserved peripheral blood mononuclear cells by flow cytometry at MESA Exam 1 (2000-02). Subsets were characterized as classical monocytes (CMs, CD14++CD16-), intermediate monocytes (IMs, CD14+CD16+), and non-classical monocytes (NCMs, CD14+CD16++). Linear regression models were used to examine associations between NSD and monocyte phenotypes, adjusting for individual-level covariates. Results: Of the MESA cohort, participants with monocyte phenotypes (n=1527) were included in analyses (age 62.9±10.5 years, 50.5% male, 37.4% White, 28.6% Black, 20.6% Hispanic). Higher NSD was associated with lower CMs but higher IMs and NCMs (Table). When gender-stratified, relationships remained significant for CMs but not for IMs. In men, higher NSD was associated with higher NCMs but not when adjusted for covariates. Conclusion: Neighborhood deprivation as a marker of chronic stress was associated with shifts in monocyte subsets in a partially sex-dependent manner, with differential relationships with CMs, IMs, and NCMs. With IMs and NCMs associated with accelerated CVD, these findings may help illuminate the role of monocytes in how neighborhood exposures lead to CVD. Future analyses will examine interactions with race/ethnicity and inflammatory biomarkers.

Psoriasis, a systemic autoimmune disease, is associated with metabolic dysfunction, vascular inflammation, and atherosclerotic cardiovascular disease (CVD). The IL-23/IL-17 pathway is critical for psoriatic inflammation; targeting these reduces skin, joint, and eye involvement but does not affect CVD outcomes. Hence, the drivers of psoriatic CVD are still to be elucidated. IL-9-producing Th9 cells are found in the circulation of psoriatic patients. We found a significant association of Th9 cells with early radiographic atherosclerotic CVD in a cohort of patients with psoriasis. We observed increased aortic plaque size, and skin/aortic Th9 cells in a psoriatic atherogenesis model (ApoE KO mice treated with IMQ). To understand the involvement of IL-9 in disease development, we inhibited IL-9 in this model. Mice treated with anti-IL-9 ab had decreased skin inflammation compared to isotype-treated mice, accompanied by reduced aortic plaque formation. A novel spontaneous psoriatic-atherogenesis model also corroborated this finding. In vitro, IL-9 induced endothelial cell (HAoEC) dysfunction through STAT3. Together, these results suggest that Th9/IL-9 may be a therapeutic target to prevent atherosclerotic CVD in psoriatic patients. Our findings were further supported by IL-9RKO/ApoEKO mice model, and conditional IL9R deletion model. Also, RNAseq data of IL9-treated HAoEC provided us with some leads about downstream targets of IL-9/STAT3 involved in psoriatic CVD development. Supported by NIAID intramural research program 1ZIAAI001251; National Psoriasis Foundation Cytokines and Chemokines and Their Receptors (CCR)

OBJECTIVES:Atherosclerotic cardiovascular disease (ASCVD) is a leading cause of death. Systemic autoimmune and inflammatory diseases are associated with increased ASCVD risk, severity, and mortality. The inflammatory cytokine interleukin 9 (IL-9) has been linked to murine atherogenesis, raising fundamental questions about the populations in which and the mechanisms by which IL-9 drives ASCVD. METHODS:Circulating T helper subsets and coronary computed tomography angiography data were analysed in patients with psoriasis. Murine models of psoriatic atherogenesis (imiquimod-ApoE-/-, IL-23-ApoE-/-, and Card14ΔE138-ApoE-/-) were investigated using IL-9 blockade or global and endothelial-specific Il9r deletion. Primary human aortic endothelial cells were used to assess IL-9/STAT3-dependent endothelial responses. RESULTS:Here, we found that expansion of IL-9-producing T helper cells (Th9) was significantly associated with high-risk radiographic ASCVD in patients with the autoimmune disease psoriasis. Th9 cells were poised to migrate to coronary vessels and were identified in human atherosclerotic plaque from individuals with psoriasis. In vivo, murine inflammatory atherogenesis was prevented by IL-9 blockade and by IL-9 receptor (IL-9R) deletion in endothelial cells. In human arterial endothelial cells, IL-9R/STAT3 signalling promoted endothelial dysfunction via diverse mechanisms including adhesion, activation, angiogenesis, and release of leukocyte chemoattractants. CONCLUSIONS:These findings suggest the Th9high state may represent a novel psoriatic ASCVD endotype that could be targeted using precision approaches to prevent ASCVD in at-risk individuals.

Atherosclerotic cardiovascular disease (ASCVD) is a leading cause of death, and understanding its pathogenic drivers is critical for effective prevention and treatment. Inflammation has a critical role in ASCVD, and patients with inflammatory diseases are at increased risk. However, the key inflammatory mediator promoting ASCVD are incompletely understood, a major barrier when targeting inflammation to prevent ASCVD. Here, we found that interleukin-9 (IL-9) producing T helper cells (Th9) were significantly associated with ASCVD in patients with the autoimmune disease psoriasis. Th9 cells were poised to migrate to coronary vessels and were identified in atherosclerotic plaque. In vivo, murine inflammatory atherogenesis was prevented by IL-9 blockade and by IL-9 receptor (IL-9R) deletion in endothelial cells. In human arterial endothelial cells, IL-9R/STAT3 signaling promoted endothelial dysfunction, angiogenesis, and release of leukocyte chemoattractants. These findings suggest that in autoimmune diseases like psoriasis, Th9/IL-9 promote atherosclerosis by directly targeting endothelial cells, and that IL-9R/STAT3 signaling could be a promising therapeutic target for ASCVD.

Abstract Psoriasis is a systemic autoimmune disease with a global prevalence of 2-3% and is strongly associated with metabolic dysfunction, vascular inflammation, and atherosclerotic cardiovascular disease (CVD). The IL-23/IL-17 pathway is critical for psoriatic inflammation. However, IL23/IL-17 blockade has no effect on CVD outcomes. Hence, the potential drivers of psoriatic CVD are still to be elucidated. IL-9-producing PU.1+/CD3+/CD4+ Th9 cells are found in the skin and circulation of patients with psoriasis. We found a significant association of Th9 cells with early radiographic atherosclerotic coronary artery disease in a psoriatic patient cohort. We observed an increase in aortic plaque size, and skin/aortic Th9 cells in a psoriatic atherogenesis model (IMQ-treated ApoE KO mice). To understand the involvement of IL-9 as a driver of the disease, we pharmacologically inhibited IL-9 in different modified disease models. IL-9 blockade-treated mice showed decreased skin inflammation compared to isotype-treated mice, accompanied by reduced aortic plaque formation. In vitro, IL-9 induced endothelial cell dysfunction through STAT3. Together, these results suggest that Th9/IL-9 may be a therapeutic target to prevent psoriasis-related atherosclerotic CVD. To further support our finding, we developed IL-9RKO/ApoE KO mice model, as well as conditional deletion of IL9R in various cell types. Further, transcriptomic profiling of endothelial cells will elucidate upon IL-9/STAT3 targets.

Addressing the need for more equitable cardio-oncology care requires attention to existing disparities in cardio-oncologic disease prevention and outcomes. This is particularly important among those affected by adverse social determinants of health (SDOH). The intricate relationship of SDOH, cancer diagnosis, and outcomes from cardiotoxicities associated with oncologic therapies is influenced by sociopolitical, economic, and cultural factors. Furthermore, mechanisms in cell signaling and epigenetic effects on gene expression link adverse SDOH to cancer and the CVD-related complications of oncologic therapies. To mitigate these disparities, a multifaceted strategy is needed that includes attention to health care access, policy, and community engagement for improved disease screening and management. Interdisciplinary teams must also promote cultural humility and competency and leverage new health technology to foster collaboration in addressing the impact of adverse SDOH in cardio-oncologic outcomes.

Background: Chronic stress is a significant contributor for cardiovascular disease (CVD) and existing health disparities. Loss of endothelial barrier integrity is a hallmark of CVD, but little is known how chronic stress impacts endothelial barrier integrity. This study aimed to investigate the signaling pathways of how loneliness-related chronic stress could impact endothelial barrier integrity. Methods: We examined the relationship among validated questionnaires of loneliness-related chronic stress and various hormonal and cytokines in a community-based cohort of African American adults at risk for CVD from resource-limited neighborhoods . Then, we conducted ex vivo as well as in vitro experiments to probe these effects and investigate the mechanism of action. Results: We determined that a 1-SD increase in loneliness was associated with a 0.32-increase (p=0.04) in the product of epinephrine (E, a catecholamine increased in individuals experiencing chronic stress) and TNFa (T, a cytokine increased in individuals experiencing loneliness), suggesting a synergism between epinephrine and TNFa. In ex vivo experiments, the product of E*T was associated with a loss in endothelial VE-cadherin expression (p=.03). In subsequent in vitro experiments to investigate this effect we treated human aortic endothelial cells with E, T, or their combination; together, these biomarkers decreased endothelial VE-cadherin expression (E: 0.94 ± 0.11, T: 0.79 ± 0.05 vs E+T: 0.58 ± 0.06-fold change of control, p<0.01) and dampened endothelial barrier integrity by increasing inter-endothelial gap formation (E: 1.59 ± 0.16 and T: 1.65 ± 0.17 vs E+T: 2.25 ± 0.13-fold change of control, p<0.001). Furthermore, we determined that the impact of E+T on endothelial cells is JAK/Stat signaling pathway dependent, establishing a putative mechanism. Conclusions: We found that a combination of epinephrine and TNFa decreased endothelial barrier integrity, suggesting an additive effect potentially accelerating CVD development and progression in individuals experiencing loneliness-related chronic stress. We provide a potential mechanism by which chronic stress could impact endothelial barrier integrity and function - ultimately worsening CVD health disparities.