bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 345: Immunomodulatory Biomaterials (IMB) For The Disruption Of Chronic Inflammation In Diabetic Foot Ulcers
Authors: ADUMBUMKULATH APARNA , Shin Crystal , Acharya Ghanashyam , Chung Jayer , Nelson Adam , Lee Richard
Published: October 2023

Background:
Diabetic foot ulcerations (DFUs) become non-healing wounds once they stall in the chronic inflammatory phase (CIP), making it vulnerable to infection. Dysregulated secretion of pro-inflammatory cytokines and matrix metalloproteinase (MMPs) are responsible for DFUs that remain in the CIP. Limitations for the development of therapeutics and biologics are the focus upon a single target biomarker, or cellular process that ignores the multiple molecules responsible. Herein, we aim to evaluate a new class of immunomodulatory biomaterials (IMBs) based on phosphate crosslinked (Px) polyvinyl alcohol (PVA) biopolymers (PxPVA) to modulate multiple proinflammatory cytokines and shift the wound out of the CIP.

Methods:
We fabricated immunomodulatory biomaterials (IMB) by the chemical synthesis of PxPVA. Subsequently, we modified the physicochemical properties (hydrophilicity, surface pH, and surface charge; zeta potential) to selectively sequester pro-inflammatory cytokines IL-1β, IL-6, TNF-α. We measured the ability of IMBs to sequester these cytokines from in vitro cytokine solutions, in vivo mouse excisional skin wound model, and ex vivo human DFU samples by Luminex assays and quantitative polymerase chain reaction (q-PCR). Histology was performed on the mouse samples to analyze for re-epithelialization and collagen deposition.

Results:
Surface analysis on the IMBs demonstrated that the material is highly porous and hydrophilic with a negative surface charge. Our IMBs were implanted in mouse excisional skin wounds followed by q-PCR analysis on the 5
th
day revealed that IMBs were able to capture significantly higher concentrations of IL-1β, IL-6, and TNF-α (*
P
≤ 0.05, ***
P
≤ 0.001) compared to polypropylene and PVA controls. Histological analysis of the IMB treated wounds demonstrated a thin and continuous epithelium formation with basket weave arrangement of collagen fibrils with a decreased type-1/type-3 collagen ratio.

Conclusions:
We demonstrated that the physicochemical properties of IMBs can be calibrated to reduce the inflammatory cytokine burden in the wound site and stimulate regenerative wound healing in the mouse model. This is mirrored by similar reductions in cytokine levels in ex vivo human DFU samples.
Abstract 349: Ume (Prunus Mume) Attenuates Angiotensin II-related Hypertension and Vascular Inflammation
Authors: Kuroda Ryohei , Okuno Keisuke , Torimoto Keiichi , Cicalese Stephanie M , Okuno Yoshiharu , Kono Ryohei , Marumoto Shinsuke , Hashimoto Tomoki N , Utsunomiya Hirotoshi , Eguchi Satoru
Published: October 2023

Ume (Prunus mume), is a traditional and familiar food in Asian countries, and is most often pickled (ume-boshi) or used to make juice concentrate (bainiku-ekisu) and wine. Recently, these products are attracting attention as a health food. Angiotensin II (AngII) plays a pivotal role in development of hypertension. However, how Ume works on hypertension still remains unknown. For in vivo experiment, 8- to 10-week-old male C57BL/6 mice were infused with AngII for 2 weeks. Mice are concurrently fed with 10 g/L bainiku-ekisu or normal water. Cardiac function and blood pressure was measured using echocardiography and tail cuff plethysmography. Heart weight and body weight were measured to assess cardiac hypertrophy. For in vitro experiment, primary cultured fibroblasts and vascular smooth muscle cells (VSMC) of 12-week-old Sprague-Dawley rat thoracic aorta was explanted. Bainiku-ekisu or ume-boshi extracts were pretreated to fibroblast and VSMC respectively, prior to AngII stimulation. In in vivo analysis, AngII-induced development of hypertension was significantly attenuated with the bainiku-ekisu treatment. AngII infusion increased heart weight body weight ratio, which was not altered by the bainiku-ekisu treatment. Body weight difference was not observed. Echocardiography data indicated that the protective effect of bainiku-ekisu. In vascular fibroblasts, bainiku-ekisu attenuated induction of vascular cell adhesion molecule-1 (VCAM1) by AngII. In VSMC, hexane fraction extracted from ume-boshi also attenuated VCAM1 induction. Our findings indicate Ume products prevent AngII-induced hypertension and VCAM1 induction. VCAM1 is a key modulator of ER stress and vascular inflammation, that leads to cellular aging. Ume might be a good candidate for a supplement or drug for cardiovascular disease. Further research to elucidate the active ingredient or pursue more efficient processing methods is desired.
Abstract 351: Developing A Human Bone Marrow Organoid Culture System To Study The Diabetic Immune System
Authors: Nguyen Tammy T , Yang Loureiro Zinger , Desai Anand , DeSouza Tiffany , Khair Lyne , Samant Amruta , Cirka Haley , Messina Louis M , Corvera Silvia
Published: October 2023

Objectives:
There is limited understanding on how diabetes negatively impacts wound healing after ischemic tissue damage. Murine models for limb ischemia and myocardial infarction have demonstrated that recruitment of bone marrow hematopoietic stem cell (HSC)-derived immune cells at all stages of ischemic tissue remodeling and healing. By leveraging our lab’s three-dimensional (3D) bone marrow organoid culture system used to study human HSC, we aim to study the effect of diabetes on immune cell development.

Methods:
Surgically discarded bone marrow tissue samples from patients undergoing lower extremity amputation for non-healing ulcer or tissue loss was used to develop our bone marrow 3D organoid culture system. Human HSCs are identified using fluorescence-tagged antibodies against cell surface markers. Hematopoietic potential of bone marrow-derived HSCs was tested using colony forming unit assay.
In vitro
immune cell differentiation into monocyte and macrophage was performed. Bulk RNA libraries were sequenced on the NextSeq 500 system (Illumina) and analysis done on DeBrowser.

Results:
Our novel human bone marrow co-culture system provides a method for bone marrow cell expansion from primary human bone marrow tissue explants.
De novo
sprouting cells can be observed growing from bone marrow tissue explants harvested from non-diabetic and diabetic donors. Using fluorescence-activated cell sorting analysis, we identified a trend towards reduced HSC numbers in diabetic when compared to non-diabetic donors. HSC derived from both diabetic and non-diabetic bone marrow tissue explants were able to undergo hematopoiesis and immune cell differentiation. Bulk RNA sequencing demonstrates distinct gene expression profiles of the cells expanded from non-diabetic and diabetic bone marrow cells.

Conclusions:
Our 3D bone marrow organoid culture system will allow for future comparative studies between the non-diabetic and diabetic immune system. The preliminary data suggest that there are gene expression profile differences between bone marrow progenitor cells from diabetic and non-diabetic donors. These gene expression differences may attribute to alterations in the HSC number and immune cell development observed in diabetes.
Abstract 352: Neonatal Mesenchymal Cells Promote Recovery Of Infracted Myocardium Through Immune Modulation Of CD44 Mediated Foxp3 Tregulatory Cells After Vascular Infusion
Authors: Saha Progyaparamita , Guru Sameer A , Bilewska Agata , Stefanowicz Artur , Chen Ling , Ge Zhi-Dong , Bolli Roberto , Kaushal Sunjay
Published: October 2023

Intravenous infusion has been used as the method of cell delivery in many preclinical studies as well as in some early clinical trials. Among its advantages are broad distribution, ability to handle a large-volume infusion, and ease of access. Progenitor cells used in cell-based therapy act through their secretomes, rather than their ability to differentiate into lineage-specific cell type. Since not all progenitor cells have similar secretome potency, the innate abilities of the secretome of cells used in clinical trials will obviously dictate their efficacy. We previously showed intramyocardial injection of cardiac neonatal mesenchymal stromal cells (nMSCs) are more effective in repairing the infarcted myocardium compared to adult mesenchymal stromal cells (aMSCs) due to their robust secretome. In this study, we explored the efficacy of intravenous (IV) delivery of nMSCs for myocardial recovery. Six-week-old male Brown Norway rats underwent acute MI by ligation of the left anterior descending artery, followed by IV infusion of cell dose 5 х 10
6
nMSCs/rat body weight (kg) or saline on days 0 and 5. We found cardiac parameters in the rodent ischemia model improved 1 month after nMSCs infusion, and the result is comparable with the intramyocardial injection of nMSCs. Tracking the infused cells in target organs revealed that their movement after IV delivery was mediated by the cell surface receptor CD44. Systemic injection of nMSCs stimulated immunomodulatory responses specifically by increasing FoxP3
+
T-regulatory cell influenced anti-inflammatory macrophages (M2) in heart. These data demonstrate that nMSCs promote immunogenic tolerance via CD44-driven T-reg/M2 stimulation for functional recovery of injured heart. Our data also provide a rationale for a clinical trial of IV infusion of nMSCs to promote functional myocardial improvement in patients with myocardial infarction.
Abstract 353: Management Of Retroperitoneal Fibrosis With Endovascular Aneurysm Repair In Patients Refractory To Medical Management
Authors: Sultan Sherif , Acharya Yogesh , Hynes Niamh
Published: October 2023

Background:
Early diagnosis and treatment of under-recognized retroperitoneal fibrosis (RPF) are essential before reaching the poorly responsive fibrotic stage. Although most patients respond to medical therapy, relapses are common. Open surgery in medically resistant patients is associated with major adverse clinical events. Therefore, we aim to compare the outcomes of endovascular repair in RPF patients who were refractory to the optimal medical management.

Methods:
This is a single-centre longitudinal study of optimal medical therapy (OMT) vs endovascular aneurysm repair (EVAR) in patients with RPF to our tertiary referral vascular centre.

Results:
Seventeen patients were referred to us with peri-aortic RPF over the study period. Eleven received OMT only, while six underwent EVAR after the failure of OMT. Eighty-two percentages (n=14) were male, and the mean follow-up was 74.40 ± 58.40 months. Nine (52%) had immunoglobulin G4-related disease (4 OMT vs 5 EVAR). EVAR patients had 100% technical success without perioperative mortality. Furthermore, all the EVAR patients were symptom-free following the intervention. Pre-operative aortic RPF index (maximum peri-aortic soft tissue diameter/maximum aortic diameter) was higher in the EVAR than in OMT. However, there was a significant decrement in the aortic RPF index following EVAR (p=0.04).

Conclusion:
EVAR in RPF medical-resistant patients is a valuable option in the armamentarium of nephrologists and rheumatologists. Our study shows that an endovascular intervention provides a promising outcome in RPF/IgG4-RD periaortitis, which is refractory to medical therapy. We recommend further studies to investigate and establish the long-term effectiveness of endovascular repair in RPF.
Abstract 355: Cross-omics Identified That Early Hyperlipidemia Increased S-adenosylhomocysteine-hypomethylation And 24 Proinflammatory Metabolites May Synergize In Trained Immunity
Authors: Xu Keman , Saaoud Fatma , Shao Ying , Lu Yifan , Zhao Huaqing , Chen Kaifu , Jiang Xiaohua , Wang Hong , Yang Xiaofeng
Published: October 2023

Introduction:
Metabolomics is the profiling of all small molecules participating in the metabolism of a cell, tissue, or organism. However, multiple-tissue metabolomics has never been reported in early hyperlipidemia. This study aims to investigate the tissue metabolomic mechanisms of how hyperlipidemia impacts the early stage of atherogenesis.

Hypothesis:
We hypothesize that early hyperlipidemia may induce atherogenesis by upregulating trained immunity (innate immune memory)-enhancing proinflammatory tissue metabolomes.

Methods:
Metabolome analyses were performed in four tissues (heart, aorta, plasma, and liver) from three-week high-fat diet-fed APOE
-/-
mice and chow diet-fed WT mice. The microarray data of ApoE
-/-
and WT mouse aortas at identical conditions were used for statistical and cross-omics analysis.

Results:
Significant results were found:
a.
Multi-organ metabolome analysis identified upregulation of 30, 122, 67, and 97 metabolites in the aorta, heart, liver, and plasma, respectively, indicating metabolome reprogramming in tissues of early atherogenic ApoE KO mice.
b.
92% (70 out of 76) of the upregulated metabolites are differentially expressed among four tissues, suggesting a tissue specificity of upregulated metabolites in early hyperlipidemia.
c.S
-adenosylhomocysteine (SAH) significantly increased in early atherosclerosis without hyperhomocysteinemia, promoting hypomethylation-driven inflammation and trained immunity.
d.
Early hyperlipidemia induces short-term hyperglycemic fluctuations with increased 1,5-AG and low glycogenolysis, implying that trained immunity induced in early hyperlipidemia is a new type of glycolysis-independent trained immunity.
e.
The novel cross-omic approach identified that the mRNA transcripts of 11 out of 105 synthetases are upregulated during the early stage of Atherosclerosis development.

Conclusion:
Our results have provided novel insights into early hyperlipidemia-induced tissue metabolomic reprogramming and trained immunity, enhancing vascular inflammation and atherogenesis, as well as new therapeutic targets for treating cardiovascular diseases (CVD), inflammations, and immune responses.
Abstract 360: Role Of Mitochondrial Dynamics In Vascular Smooth Muscle Cell Death
Authors: Jadli Anshul S , Gomes Karina , Meechem Megan , Edalat Pariya , Ballasy Noura , Patel Vaibhav B
Published: October 2023

Background:
Vascular smooth muscle cell (VSMC) death has been linked to the pathogenesis of various vascular diseases. Excessive mitochondrial fission in response to pathological stimuli contributes to cell death, thus, inhibition of mitochondrial fission can be a promising therapeutic target in vascular diseases. Mitochondrial division inhibitor 1 (mdivi-1) inhibits Drp1-dependent mitochondrial fission by attenuation of its GTPase activity. As association between fission inhibition & VSMCs cell death has been implicated in vascular diseases, the underlying mechanism warrants further investigation. We assessed the effects of mdivi-1 (an inhibitor of Drp1-mediated mitochondrial fission) on staurosporine (STS)-induced mitochondrial dysfunction, impaired autophagy, & cell death in aortic VSMC.

Methods & Results:
The flow cytometry analysis demonstrated reduced cell death in VSMCs-treated with mdivi-1. The mitochondrial fission was evaluated using Mitotracker Red staining. mdivi-1-treated VSMCs showed significantly higher mitochondrial networks, mean branch length, & mitochondrial footprint compared to STS-treated cells indicating reduced mitochondrial fragmentation. Inhibitory effects of mdivi-1 on fission were exerted by reduced Drp1 phosphorylation at 616. Mitochondrial function via evaluation of mitochondrial permeability transition pore opening, mitochondrial membrane potential & bioenergetic profile showed restored mitochondrial function & metabolic potential in the mdivi-1-treated cells compared to the STS group. The mdivi-1-treated VSMCs showed significantly reduced ROS generation indicating suppressed oxidative stress. The improved mitochondrial structural integrity due to inhibition of fission was associated with reduced mitophagy & cell death-related proteins.

Conclusion:
In response to apoptotic stimuli, excessive Drp1-dependent mitochondrial fission leads to mitochondrial damage, playing a key role in VSMC death. Inhibition of Drp1-dependent mitochondrial fission by mdivi-1 attenuated mitochondrial fission & prevented cell death in VSMCs. Pharmacological inhibition of mitochondrial fission provides a potential therapeutic target in vascular diseases associated with loss of VSMC.
Abstract 362: Pharmacologic Blockade Of Sodium-glucose Cotransporter-2 Inhibits Vascular Smooth Muscle Cell Calcification
Authors: Ji Yan , Vadali Manisha , Schoenherr Alex , Bryson Alaina , Dang Ho , Hotop Austin , Khoukaz Hekmat , Fay William P
Published: October 2023

Background:
Empagliflozin (Empa) is a highly selective sodium glucose cotransporter-2 (SGLT2) inhibitor used to reduce the risk of cardiovascular death. Empa reduces arterial stiffness and blood pressure in humans, as well as atherosclerosis and intimal hyperplasia in preclinical models, though the mechanisms underlying these effects are poorly defined. We investigated effects of Empa on the calcification of human coronary artery smooth muscle cells (SMCs).

Methods:
To determine the effect of Empa on SMC calcification triggered by high phosphate (HP) conditions, SMCs were cultured in standard (
i.e.
, normal phosphate [NP]) medium, HP medium, or HP medium containing Empa (2.5 μM). After 12-days, SMC calcification was quantified by the alizarin red staining method with results quantified by comparison to a standard curve and normalized to NP control. Similar methods were used to study effects of Empa on SMC calcification induced by platelet-derived growth factor-BB (50 ng/mL). Effects of Empa on angiotensin II-induced SMC senescence, a precursor of cell calcification, were studied by quantifying senescence-associated β galactosidase activity. Real-time reverse transcriptase PCR was used to study gene expression of Runx2, a master regulator of SMC osteogenic transition.

Results:
Empa significantly decreased HP-induced SMC calcification (1.14±0.11
vs
. 1.32±0.11 AU/mL, n=5, p<0.05) and PDGF-BB-induced calcification (1.20±0.17
vs
. 1.94±0.22 AU/mL, n=5, p<0.05). Empa significantly inhibited SMC senescence (1.03±0.06
vs
. 1.39±0.04, n=5, p<0.05), while decreasing Runx2 gene expression by 77±35% (n=6, p <0.01).

Conclusion:
Empa significantly inhibits SMC calcification triggered by clinically relevant pathophysiologic stimuli. Empa significantly inhibits SMC senescence and Runx2 gene expression, identifying molecular pathways underpinning Empa’s anti-calcific effect. These findings identify an additional mechanism by which Empa may decrease vascular stiffness and provide insights into how SGLT2 inhibitors exert beneficial effects on vascular diseases dependent on SMC calcification and senescence, including hypertension and atherosclerosis.
Abstract 365: Altered Gene Expression And Impaired Repair Functions Of Circulating Endothelial Colony-Forming Cells From Diabetic Patients
Authors: Marei Isra , Thomas Binitha , Ahmetaj-Shala Blerina , Chidiac Omar , Habib Tanwir , Ahmed El-Naas , Ehtesham Anam , Dilawar Azwa , Aboidris Leena E , Jameesh Moidy Muhammed , Jayyousi Amin , Al suwaidi Jassim M , Abi Khalil Charbel A , Mitchell Jane A , Triggle Chris R
Published: October 2023

Introduction:
Vascular repair functions are impaired in type 2 diabetes (T2D), leading to increased cardiovascular disease risk. Endothelial progenitor cells (EPCs) play a key role in re-endothelialization and neovascularization processes. This study aims to compare the transcriptomic profile and repair functions of the EPC subtype endothelial colony forming cells (ECFCs) in healthy and T2D patients.

Hypothesis:
T2D affects the expression profile of ECFCs, which impacts vascular repair functions.

Methods:
ECFCs were isolated from the peripheral blood of 4 healthy and 3 T2D patients (71% males, mean age 48.9 ± 6.9) using selective plating. Isolated cells showed cobblestone morphology and expressed endothelial and progenitor markers. RNAseq was performed using Illumina NextSeq sequencer. Differential expression analysis was performed using R and Ingenuity Pathway Analysis (IPA). Functional assays were done to assess angiogenesis (Matrigel assay), migration (scratch assay), and adhesion molecules expression (ELISA).

Results:
RNAseq analysis revealed 22 differentially expressed genes (adjusted p-value <0.1, Fold Change ±1.5), with 15 up-regulated and 7 down-regulated genes in T2D (Figure 1). Top molecular and cellular functions identified by IPA included: cellular movement, cell-cell signaling, cellular assembly, organization, development, growth, and proliferation. Enrichment analysis also showed an impact on extracellular matrix structure and organization, and cell adhesion, among other functions. Repair functions were impaired in T2D ECFCs as indicated by impaired tube formation and reduced wound closure rate. The expression of VCAM-1 and P-selectin was significantly increased in T2D ECFCs.

Conclusion:
ECFCs in T2D have impaired angiogenesis and migration functions and show a variable transcriptomic profile when compared to healthy cells. Impaired repair functions in ECFCs could contribute to the cardiovascular complications of T2D.
Abstract 370: Corticosterone-mediated Associations Between Neighborhood Environment Perceptions and Lysosomal Acid Lipase Levels - Data From The Washington, D.C. Cardiovascular Health and Needs Assessment
Authors: Baumer Yvonne , Ortiz-Whittingham Lola , Cintron Manuel A , Tarfa Hannatu A , Dave Ayushi , Saurabh Abhinav , Collins Billy S , Mitchell Valerie , Powell-Wiley Tiffany M
Published: October 2023

Experiences of adverse social determinants of health (SDoH), like less favorable neighborhood environment perceptions, are associated with cardiovascular disease (CVD). Lysosomal acid lipase (LAL) plays an important role in lipid metabolism, and genetic LAL deficiency has been linked to CVD risk. We aimed to determine the potential relationship between neighborhood perceptions and LAL and to evaluate stress-related biomarkers as mediators of this relationship in the D.C. Cardiovascular Health and Needs Assessment cohort (60 African American participants, mean age 61yr, 93% female), biomarkers were measured using ELISA, and perceptions of neighborhood violence, physical and social environment, social cohesion, and safety were determined using validated questionnaires with higher scores indicating more favorable perceptions. We explored neighborhood perceptions-LAL relationships with regression models adjusted for BMI and ASCVD risk score and stratified by hyperlipidemia due to the potential confounding role of LDL. Mediation analysis was used to evaluate cortisol and corticosterone as potential mediators of the neighborhood perceptions-LAL relationship. Of the neighborhood perceptions measured, more favorable perceptions of physical and social environment associated with greater LAL (β=0.23, p=0.09); this association persisted only among participants without hyperlipidemia (β=0.42, p=0.04). Only corticosterone associated with both perceived physical and social environment and LAL (Figure); corticosterone mediated 73.1% of this relationship in all participants and 89.9% of this relationship in participants without hyperlipidemia. Our data highlight the relationship between perceived physical and social environment and LAL mediated by corticosterone. Results also emphasize the importance of future interdisciplinary work to better understand mechanisms by which SDoH and associated biomarkers contribute to CVD disparities.
Abstract 371: Functional Characterization Of A Ccm2 Mutation Associated With Coronary Artery Disease
Authors: Fang Shi , Lee-Kim Vivian , Schnitzler Gavin R , Barry Aurelie , Gupta Rajat
Published: October 2023

Introduction:
Coronary Artery Disease (CAD) remains the leading cause of death despite effective therapies to lower serum cholesterol. To identify new mechanisms of disease for prevention and therapy, several recent genome-wide association studies (GWAS) have identified hundreds of loci with statistically significant association with CAD (5x10^-8). In a recent large meta-analysis of 181,522 cases and over 1 million controls subjects, the CARDIOGRAM consortium identified a locus with genome-wide significant association with CAD in the chromosome 7p14.

Methods and Results:
We functionally interrogated the chromosome 7p14 GWAS association with a combination of population genetics and in vitro experimental methods. Statistical fine-mapping identified a lead variant in the locus, rs2107732, with a posterior probability of association 63.5%. This variant has a significant association with CAD of
P
= 1.53 x 10^-8). The minor allele is present in 9.2% of subjects, reduced risk of CAD (OR=0.92), and encodes a valine to isoleucine substitution in CCM2 at amino acid position 74 (V74I). To understand the importance of CCM2 and the coding mutation V74I in CAD, we first knocked-down CCM2 in immortalized human aortic endothelial cells (teloHAECs). KLF2, a critical athero-protective gene, was upregulated upon CCM2 knockdown. However, overexpressing wildtype CCM2 decreased the level of KLF2, and overexpressing CCM2 with the V74I mutation downregulated KLF2 to a comparable level. To circumvent the potential interference from the endogenous wildtype CCM2, we used CRISPR-Cas9-mediated homology-directed repair in teloHAECs and generated multiple clones carrying homozygous CCM2 V74I mutation. These clones will serve as useful tools to understand the mechanism by which CCM2 V74I mutation decreases the risk of CAD.

Conclusions:
We identified a missense mutation in CCM2 which is associated with a decreased risk of coronary artery disease. Functional analysis of CCM2 in arterial endothelial cells suggests that the protective effect is through upregulation of KLF2 expression.
Abstract 372: Association Between Vasomotor Symptoms And Arterial Stiffness Among Women In Menopausal Transition
Authors: Choi Hye Rin , Chang Yoosoo , Kwon Ria , Lim Ga-young , Park Eunkyo , Ryu Seungho
Published: October 2023

Obejectives:
We examined the association between vasomotor symptoms (VMSs) and arterial stiffness, using brachial-ankle pulse wave velocity (baPWV), among Korean women in menopausal transition.

Study design:
This cross-sectional study comprised 945 women aged 42-52 years.

Methods:
VMSs, including hot flashes and night sweats, were assessed using the Menopause-Specific Quality of Life questionnaire (Korean version). Moderate/severe VMSs were defined as having a score of equal to or greater than 3 points (range: 0 to 6, 6 being most bothersome). Arterial stiffness was measured by baPWV. The high-risk group was defined as baPWV over the 90th percentile. Multivariable logistic regression models were performed to determine whether prevalent VMSs were significantly associated with high level of baPWV in menopausal transitioning women.

Results:
Compared to women without VMSs, those with VMSs had a prevalence ratio (PR) of 2.12 (95% confidence interval (CI), 1.17-3.84) for high baPWV after adjusted for age, menopausal status, education levels, body mass index, total and low-density lipoprotein cholesterol, triglycerides, diabetes, hypertension, physical activity, amount of alcohol intakes, and smoking status. The multivariable-adjusted PRs for high baPWV were 1.54 (95% CI: 0.73-3.23) and 2.72 (95% CI: 1.36-5.44) in women with mild and moderate/severe VMSs, respectively (P for trend = 0.005).

Conclusion:
Our findings suggest that VMSs, including moderate/severe VMSs, were significantly associated with arterial stiffness among Korean menopausal transitioning women.
Abstract 398: Impaired Macrophage Interleukin-1β-Dependent Wound Healing In The Context Of Diabetes
Authors: Sharma Sheila , Mantsounga Chris S , Lee Cadence , Neverson Jade , Pierce Julia , Amelotte Elizabeth , Carley Rachel T , Butler Celia , Choudhary Gaurav , Morrison Alan R
Published: October 2023

Peripheral Artery Disease (PAD), associated with diabetes, is an atherosclerotic vascular disease with impaired angiogenesis and poor wound healing. Increasing data support early inflammation as playing a key role in setting the state for adequate recovery following acute vascular injury. Here, we sought to understand the impact of diabetes upon IL-1β-dependent angiogenesis during wound healing. We identified an important role for inflammatory macrophage IL-1β-driven vascular endothelial growth factor-A (VEGF-A) production during angiogenesis required for wound healing. To determine the role of IL-1β signaling in the context of diabetes, we used a macrophage-specific
IL-1
β
-deletion model (
mIL-1
β
), a macrophage-specific
IL-1R
-deletion model (
mIL-1R
), and a Leptin (

Lepr
db/db
)

model. Using a punch biopsy model, wound closure was quantified over ten days. Mice with diabetes (

Lepr
db/db
)

demonstrate initial wound expansion, followed by delayed wound closure. Comparable results were found using
mIL-1
β
mice, suggesting that mice with diabetes may phenocopy disrupted IL-1β signaling. Analysis from all three models confirmed decreased macrophage expression of pro-angiogenic VEGF-A in response to IL-1β signaling
in vitro
and reduced VEGF-A expression in wounds
in vivo
. We examined the impact of upregulating IL-1β-dependent transcription factors, STAT3 and NF-κB, to bypass the impairment conferred by diabetes and found that constitutively active STAT3 and IKK-2 (NF-κB activator) appear to reconstitute VEGF-A expression in macrophages. In summary, angiogenesis dependent healing requires the contribution of macrophage IL-1β-dependent VEGF-A expression in the early inflammatory state, and diabetes is associated with an impairment in this pathway. Therapeutic angiogenesis strategies in the context of diabetes may require upregulation of transcription factors downstream of IL-1β to restore macrophage VEGF-A expression that is required for angiogenesis-dependent wound healing.
Abstract 405: Short Stapled ApoC-II Mimetic Peptides Activate Lipoprotein Lipase And Decrease Plasma Triglycerides In Mouse Models
Authors: Sviridov Denis , Dasseux Amaury , Wolska Anna T , Reimund Mart , Pryor Milton , Drake Steve , Remaley Alan
Published: October 2023

The metabolism of triglyceride-rich lipoproteins plays an important role in cardiovascular diseases and dyslipidemias. Lowering plasma triglycerides (TGs) by activating lipoprotein lipase (LPL), the main plasma enzyme that hydrolyses TGs, with ApoC-II mimetic peptides is a potential strategy for treatment of ApoC-II deficiency, potentially other forms of hypertriglyceridemia.We previously described a dual ApoC-II mimetic/ApoC-III antagonist peptide called D6-PV. It’s a bihelical, 41 amino acid long peptide with the first helix designed to bind to lipoproteins and the second helix to activate LPL. D6-PV showed a marked reduction in plasma TGs in ApoC-II Knock-Out and ApoC-III transgenic mouse models.We employed a hydrocarbon stapling technique to make a new single helix SP1 peptide based on the last 21 C-terminal amino acids (A59-E79) of ApoC-II. The staple increased the peptide’s ability to bind to lipoproteins and improved amphipathicity without interfering with the lipoprotein lipase activating motif located on the hydrophilic face of the peptide.The SP1 peptide was more potent than D6-PV in its ability to increase LPL activity
in vitro
. Intraperitoneal injection of the SP1 peptide into ApoC-II Knock-Out or ApoC-III transgenic mice (1-5 umole/kg) showed similar to the D6-PV a 70-90 percent reduction in plasma triglycerides 1 hour after injection.We also used two consecutive staples or “stitching” to make the SP2 peptide. These modifications along with Glycine77 to
N
-methyl-glycine substitution and replacing two C-terminal Glutamic acids with their D-isomer analogs made the SP2 peptide resistant to proteolysis by trypsin, pepsin and Proteinase K.In summary, the new SP1 and SP2 peptides could be a treatment for chronic or acute hypertriglyceridemia, acute pancreatitis and a potential oral therapy for ApoC-II deficiency.
Abstract 406: Normalization Of Erythrocyte Morphology In Scarb1
-/-
Mice By Bacterial Serum Opacity Factor Is Dependent On HDL-free Cholesterol
Authors: Rosales Corina , Wang Ziyi , Yelamanchili Dedipya , Gotto Antonio M , Gillard Baiba K , Pownall Henry J
Published: October 2023

Compared to wild-type mice, HDL-receptor-deficient (Scarb1
-/-
) mice have higher plasma levels of free cholesterol (FC)-rich HDL and exhibit multiple pathologies—female infertility and deranged platelet and erythrocyte morphology and function. These pathologies are associated with a low mol% FC in ovaries, platelets, and erythrocytes, effects that are reversed by the HDL-lowering drug probucol. Some strains of Streptococcus pyogenes secrete a protein, serum opacity factor (SOF), which catalyzes the clouding i.e., opacification, of plasma. SOF specifically targets and quantitatively converts HDL into three products, that include a cholesteryl ester-rich microemulsion containing all the cholesterol content of >100,000 HDL particles as well as APOE and its heterodimer with APOA2 as its sole apolipoproteins. Delivery of SOF with an adeno-associated virus (AAV
SOF
) constitutively lowers plasma HDL-FC and reverses female infertility in Scarb1
-/-
mice. Thus, we tested the hypothesis—AAV
SOF
delivery to Scarb1
-/-
mice will normalize erythrocyte morphology in an HDL-FC dependent way. The erythrocyte morphology and FC content expressed as mol% FC of three groups of mice—WT, untreated Scarb1
-/-
mice (control) and Scarb1
-/-
mice receiving AAV
SOF
—were compared and correlated with their respective HDL-mol% FC. Among Scarb1
-/-
mice, AAV
SOF
treatment normalized reticulocyte number, erythrocyte morphology and erythrocyte mol% FC. Plasma- and HDL-mol% FC positively correlated (p < 0.0001) across all three groups of mice. HDL-mol% FC also positively correlated with erythrocyte mol% FC (p
<
0.0001). Finally, the erythrocyte mol% FC positively correlated with both the number of reticulocytes (p = 0.006) and abnormal erythrocytes (p < 0.0001). Although less profound, AAV
SOF
treatment also reduced the FC contents of some extravascular tissues. HDL-FC spontaneously transfers from plasma lipoproteins to cell membranes in multiple tissues sites on a time scale of minutes to a few hours. FC-enrichment of erythrocytes, which are in contact with plasma HDL, was more profound than FC enrichment in tissues. AAV
SOF
treatment lowers both plasma HDL-FC and erythrocyte-FC and normalizes erythrocyte morphology and lipid composition in an HDL-FC dependent way.
Abstract 407: The Role Of Peroxisomes And Bile Acid-initiated Signaling In Vascular Smooth Muscle Cell Phenotype
Authors: Holden Claire M , Bogan Bethany , Paredes Felipe , San Martin Alejandra
Published: October 2023

Background:
Atherosclerosis is a chronic inflammatory process characterized by the accumulation of foam cells (lipid-laden macrophage-like cells). Vascular smooth muscle cells (VSMCs) are not terminally differentiated, and their phenotype is modulated in response to various stimuli, such as cholesterol (CHO), which increase susceptibility to vascular diseases. Modulated SMCs contribute to foam cell formation within atherosclerotic plaques. The intracellular metabolism of CHO via peroxisomes in hepatocytes produces bile acids like chenodeoxycholic acid (CDCA), which agonizes farnesoid X nuclear receptor (FXR) and liver X receptor (LXR). FXR regulates bile acid, glucose, and lipid homeostasis. LXR facilitates the first step of reverse CHO transport. It is unknown if VSMCs synthesize bile acids and whether peroxisomes and the bile acids they produce play a role in VSMC-derived foam cell formation.

Results:
Using human aortic VSMC (HASMCs), we found that CHO treatment decreased expression of several peroxisomal proteins. Peroxisomal loss-of-function experiments demonstrated increased intracellular lipid accumulation, as visualized with Oil Red O (ORO) staining. Peroxisomal biogenesis associated with fenofibrate treatment completely eradicated intracellular ORO. We have shown that HASMCs synthesize bile acids after exposure to CHO, which led us to hypothesize that bile acid-initiated signaling participates in the phenotypic modulation initiated by CHO in VSMCs. Treatment of HASMCs with LXR agonists increased expression of the efflux CHO transporter ABCA1 and differentiation markers SMA and CNN1, and decreased expression of macrophage marker LGALS3.

Conclusions:
These data show that VSMCs have the metabolic capacity to synthesize bile acids, which is impaired if peroxisomes are non-functional or absent. Our data indicates that activation of bile acid receptors upregulates ABCA1, favoring a differentiated VSMC phenotype. Together, these data suggest that bile acid-mediated activation of FXR and LXR have differential roles in CHO-induced VSMC to foam cell transition and that LXR activation may represent a novel atheroprotective target which increases cellular efflux of CHO, promoting VSMC differentiation.
Abstract 410: Effect Of Total Sphingomyelin Synthase Activity On Low Density Lipoprotein Catabolism
Authors: Li Zhiqiang , He Mulin , Jiang Xian C
Published: October 2023

Objective:
Sphingomyelin (SM) and cholesterol are two key lipid partners on cell membrane and on lipoproteins. Many studies have indicated the influence of cholesterol on SM metabolism. This study examined the influence of SM biosynthesis on cholesterol metabolism.

Approach and Results:
Inducible global Sms1 KO/global Sms2 KO mice were prepared to evaluate the effect of whole-body SM biosynthesis deficiency on lipoprotein metabolism. Tissue cholesterol, SM, ceramide, glucosylceramide levels were measured. TG production rate and LDL catabolism were measured. Lipid rafts were isolated and LDL receptor mass and function were evaluated. Also, the effect of exogenous sphingolipids on hepatocytes were investigated. We found that total SMS depletion significantly reduced plasma SM levels. Also, the total deficiency significantly induced plasma cholesterol, apoB, and apoE levels. Importantly, total SMS deficiency, but not SMS2 deficiency, dramatically decreased LDL receptor in the liver and attenuated LDL uptake through the receptor. Further, we found that total SMS deficiency greatly reduced LDL receptor in the rafts which contained significantly lower SM and significantly higher glucosylceramide as well as cholesterol. Furthermore, we treated primary hepatocytes and Huh7 cells (a human hepatoma cell line) with SM or ceramide or glucosylceramide, we found that only SM could up-regulate LDL receptor levels in the dose-dependent fashion.

Conclusions:
Whole body SM biosynthesis plays an important role in LDL-cholesterol catabolism. The total SMS deficiency, but not SMS2 deficiency, reduces LDL uptake and causes LDL-cholesterol accumulation in the circulation. Given the fact that serum SM level is a risk factor for cardiovascular diseases, inhibiting SMS2 but not SMS1 should be the desirable approach.
Abstract 419: Liberation Of Soluble Urokinase Plasminogen Receptor (supar) Occurs In Response To Coronavirus Infection But Not To Influenza Virus Infection
Authors: Sharma Sriganesh B , Kumar Kiran , Wu Weisheng , Hayek Salim S , Moore Bethany B , Obi Andrea T
Published: October 2023

Objectives:
Urokinase (uPA), and its receptor, uPAR, mediate fibrinolysis and matrix remodeling. Liberation of monocyte/macrophage (MO/Mϕ) bound uPAR to yield soluble uPAR (suPAR) occurs after infection with coronaviruses (CoVs) such as SARS-CoV-2, and suPAR levels correlate with systemic inflammation and degree of end organ injury. How suPAR liberation from MO/Mϕs occurs after CoV infection, and whether suPAR induction occurs after pneumotropic viral infections such as influenza A (IAV) is unclear.

Methods:
Murine bone marrow derived Mϕs (BMDMs) and immortalized Mϕs (RAW264.7) were infected with CoV [MHVA59 (murine model for SARS-CoV-2 infection) or MHV1] or with IAV (A/PR/8/34) and levels of mRNA and protein were measured using qRT-PCR and ELISA, respectively. uPA activity of cell supernatants was quantitated using a commercially available assay. uPA knockdown of Mϕs was performed using siRNA transfection. C57BL6/J mice underwent intranasal inoculation of either 2x10
5
pfu
of MHVA59, 25
pfu
of IAV, or with PBS (sham). Plasma levels of suPAR and splenic Mϕ (surrogate for circulating MOs) levels of mRNA and protein were analyzed at post-infection day 3.

Results:
Though both CoV or IAV infection resulted in induction of mRNA and protein levels of cellular uPAR and uPA, and uPA activity, only CoV infection yielded liberation of suPAR from Mϕs (Fig. 1A-C). Indicating a functional role in suPAR liberation, infection of uPA-silenced Mϕs yielded a blunted induction of suPAR release despite not affecting mRNA levels of uPAR (Fig 1D). Finally, inoculation of mice with MHVA59, but not with IAV, resulted in elevated plasma levels of suPAR, despite inducing a robust induction of uPAR in splenic Mϕs (Fig. 1E).

Conclusions:
suPAR induction results after CoV infection, but not IAV infection, and occurs through a uPA-dependent pathway. Antagonism of co-regulators of uPAR and uPA may prove to be a therapeutic strategy to combat post-CoV tissue injury mediated by Mϕs.
Abstract 420: In-hospital Statin Use Is Not Associated With Lower Mortality In Patients Hospitalized With Covid-19
Authors: Yan Crystal L , Rivera Rodriguez Beatriz , Andrade-Bucknor Sharon
Published: October 2023

Background:
Morbidity and mortality from COVID-19 largely stem from a severe systemic inflammatory response. As such, medications with anti-inflammatory properties such as statins may have protective effects in COVID-19 patients. The purpose of our study was to determine whether in-hospital statin use was associated with lower morbidity or mortality in patients hospitalized with COVID-19.

Methods:
We conducted a two-center retrospective study of 692 adult patients hospitalized from March 2020 to October 2021 with COVID-19 confirmed by RT-PCR. Groups were stratified based on in-hospital statin use. Baseline characteristics, biomarkers, and in-hospital outcomes were described. A multivariate regression model was used to estimate the association of the primary outcome of interest with in-hospital statin use. Statin use was defined as any dose of atorvastatin, rosuvastatin, simvastatin, pravastatin, or fluvastatin. Statistical analyses were performed using SPSS 28.

Results:
Statin use was present in 45.7% (315/692) of patients. These patients were older (68 vs. 62 years old, p<0.001) and had higher rates of hyperlipidemia (31.9% vs. 16.3%, p<0.001), hypertension (78.7% vs. 54.9%, p<0.001), type 2 diabetes (50.2% vs. 31.5%, p<0.001), coronary artery disease (21.9% vs. 6.4%, p<0.001), heart failure (19.4% vs. 7.5%, p<0.001), and chronic kidney disease (16.8% vs. 5.1%, p<0.001). There was no difference in levels of low-density lipoprotein or inflammatory markers (D-dimer, CRP, and ferritin) between groups. However, patients with in-hospital statin use were less often treated with remdesivir (53.8% vs 61.3%, p=0.03), corticosteroids (71.8% vs 77.9%, p=0.04), or tocilizumab (10.4% vs 18.9%, p=0.001). Patients with in-hospital statin use had shorter length of stay (18 vs. 21 days, p<0.001) and lower adjusted odds of intubation (0.65 [0.46-0.92], p=0.01). There was no difference in all-cause mortality.

Conclusion:
COVID-19 patients treated with in-hospital statin therapy had more cardiovascular risk factors and received less treatment for COVID-19, yet they had shorter length of stay and lower odds of intubation. In-hospital statin use was not associated with lower risk of mortality.
Abstract 439: Metabolic Syndrome Induced Metabolomic Alterations In A Porcine Model Of Chronic Myocardial Ischemia
Authors: Broadwin Mark , Harris Dwight , Banerjee Debolina , Stanley Madigan , sellke frank , Usheva Anny
Published: October 2023

Objective:
Metabolic syndrome (MetS) has been shown to be a risk factor for cardiovascular disease. Despite this there remains a paucity of data that elucidates the exact mechanisms by which MetS causes cardiac tissue to become more vulnerable to ischemic insult. The purpose of this multi omics study was to establish the role of MetS on chronically ischemic myocardium in a porcine model. We hypothesized that metabolic syndrome altered fatty acid metabolism and predisposes ischemic myocardium to oxidative stress.

Methods:
12 four-week-old Ossabaw Swine were fed an obesogenic diet to induce metabolic syndrome (n=6) or a control normal diet (n=6) at 8 weeks pigs underwent surgical placement of an ameroid constrictor to the left circumflex artery. After an additional 8 weeks pigs were humanly euthanized and underwent tissue procurement. Tissue metabolites were extracted and analyzed via a 5500 QTRAP hybrid triple quadrupole mass spectrometer. For gene expression responses, RNAseq was used.

Results:
Pigs with metabolic syndrome demonstrated a significant alteration in metabolites associated with fatty acid oxidation and ketogenesis. MetS was associated with an increase in acetoacetate (P=0.01), 3-hydroxybuterate (P=0.048), and acetoacetyl CoA (P=0.03). Succinyl-CoA was significantly decreased in MetS swine (P<0.01) (figure 1A). Furthermore, when compared to lean diet counterparts, MetS caused a significant increase in malonyl-CoA (P<0.01), propionyl-CoA (P<0.01), and HMG-CoA (P=0.03). (figure 1A). The expression of the HMG-CoA lyase (HMGCL) required for fatty acid β-oxidation and ketogenesis is significantly higher in MetS than in LD (figure 1B).

Conclusion:
Metabolic syndrome leads to an increase in fatty acid β-oxidation and ketogenesis in response to ischemic insult when compared with lean diet controls. This response may predispose ischemic cardiac tissue in MetS to increased oxidative stress.
Abstract 441: Relationship Between Diabetes, Glucose Control, And Vascular Health: Findings From The American Heart Association Cardiometabolic Health Strategically Focused Research Network
Authors: Garshick Michael , Barrett Tessa A , Jindal Manila , Newman Jonathan D , Fadzan Maja , Bredefeld Cindy , Levy Natalie , Akinlonu Adedoyin , Heguy Adriana , Drenkova Kamelia , Schlamp Florencia , Giannarelli Chiara , Fisher Edward A , Goldberg Ira J , Berger Jeffrey
Published: October 2023

Background:
Diabetes mellitus (DM) increases the risk of vascular endothelial damage and subsequent cardiovascular (CV) events. This study was designed to investigate mechanisms of endothelial cell (EC) damage in DM.

Methods:
CHORD (CHOlesterol lowering and Residual Risk in Diabetes) is an ongoing study evaluating mechanisms of CV disease (CVD) risk in patients with 1) LDL-C >100 mg/dL, 2) no known CVD and 3) with or without DM. In a subset of participants, EC harvesting was performed at baseline by inserting a J-wire through an angiocatheter into the brachial vein. ECs were isolated with magnetic beads directed against CD146 and transcript expression assessed using next generation RNA sequencing.

Results:
We performed EC harvesting on participants with DM (n=11, mean age 47 ± 15 years, 45% female, mean HbA1c 6.4%) and controls (n=11, mean age 50 ± 15 years, 56% female, mean HbA1c 5.2%). Comparison of brachial vein EC RNA sequencing in DM vs controls after adjustment for age, sex, and race/ethnicity demonstrated 1043 upregulated genes and 26 downregulated genes with excellent supervised hierarchical clustering between groups (p<0.05). Using a linear regression model adjusting for age, sex, and race/ethnicity, 899 EC genes were significantly associated with HbA1c (p<0.05). Of the original 1069 EC genes differentially expressed in diabetes (over control), only 257 genes displayed a concordant relationship to HbA1c (HbA1c dependent) while the rest (n=812) were HbA1c independent. (Figure 1). Pathways related to EC genes that were HbA1c dependent included those related to endosomal processing whereas HbA1c independent pathways included those related to energy utilization (e.g. PGC-1 alpha phosphorylation), and cell signaling (RAC1, TGFBR1).

Conclusion:
Our data demonstrate the impact of DM on EC abnormalities and suggest differences in EC gene expression between glucose dependent and independent processes.