bluuebunny/crossref_metadata_2025 huggingface dataset:
Data from Primary Prevention and Interception Studies in RAS-Mutated Tumor Models Employing Small Molecules or Vaccines
Authors: Dragnev Konstantin H. , Lubet Ronald A. , Miller Mark Steven , Sei Shizuko , Fox Jennifer T. , You Ming
Published: October 2023

Abstract
Therapeutic targeting of RAS-mutated cancers is difficult, whereas prevention or interception (treatment before or in the presence of preinvasive lesions) preclinically has proven easier. In the A/J mouse lung model, where different carcinogens induce tumors with different KRAS mutations, glucocorticoids and retinoid X receptor (RXR) agonists are effective agents in prevention and interception studies, irrespective of specific KRAS mutations. In rat azoxymethane-induced colon tumors (45% KRAS mutations), cyclooxygenase 1/2 inhibitors and difluoromethylornithine are effective in preventing or intercepting KRAS-mutated or wild-type tumors. In two KRAS-mutant pancreatic models multiple COX 1/2 inhibitors are effective. Furthermore, combining a COX and an EGFR inhibitor prevented the development of virtually all pancreatic tumors in transgenic mice. In the N-nitroso-N-methylurea-induced estrogen receptor–positive rat breast model (50% HRAS mutations) various selective estrogen receptor modulators, aromatase inhibitors, EGFR inhibitors, and RXR agonists are profoundly effective in prevention and interception of tumors with wild-type or mutant HRAS, while the farnesyltransferase inhibitor tipifarnib preferentially inhibits HRAS-mutant breast tumors. Thus, many agents not known to specifically inhibit the RAS pathway, are effective in an organ specific manner in preventing or intercepting RAS-mutated tumors. Finally, we discuss an alternative prevention and interception approach, employing vaccines to target KRAS.
Alvin J. Siteman Cancer Center: Cancer Prevention Perspective
Authors: Colditz Graham A. , Drake Bettina F. , Eberlein Timothy J.
Published: October 2023

We summarize Siteman Cancer Center catchment that covers 82 counties in southern Illinois and eastern Missouri. We note both the high poverty and cancer rates in many rural counties. Siteman Community Outreach and Engagement has developed a number of strategies to move towards achieving health equity. These include NCI-funded research projects in rural clinics and outreach to improve access to cancer prevention services. To increase capacity for community-engaged research, we have developed and refined a Community Research Fellows Training Program.
Data from Change in Neighborhood Socioeconomic Status and Adherence to the Cancer Prevention Lifestyle Guidelines in Hispanic/Latino Adults: Results from the HCHS/SOL Study
Authors: Pichardo Margaret S. , Pichardo Catherine M. , Talavera Gregory A. , Gallo Linda C. , Kuo Charlene C. , Castañeda Sheila F. , Chambers Earle C. , Daviglus Martha L. , Pirzada Amber , Perreira Krista M. , Sotres-Alvarez Daniela , Peña Ortiz Tania Yadhira , Plascak Jesse J.
Published: October 2023

Neighborhood conditions are dynamic; the association of changing neighborhood socioeconomic factors with cancer preventive behaviors remains unclear. We examined associations of neighborhood socioeconomic deprivation, gentrification, and change in income inequality with adherence to the American Cancer Society Guidelines on Nutrition and Physical Activity for Cancer Prevention in The Hispanic Community Health Study/Study of Latinos (HCHS/SOL). The HCHS/SOL enrolled 16,415 adults, ages 18–74 years, at baseline (2008–2011), from communities in the Bronx, NY, Chicago, IL, Miami, FL, and San Diego, CA. Geocoded baseline addresses were linked to the 2000 decennial Census and 5-year American Community Survey (2005–2009 and 2012–2016) tracts to operationalize neighborhood deprivation index (NDI), gentrification, and income inequality. Complex survey multinominal logistic regression models estimated the relative risk ratio (RRR) with overall guideline adherence level (low, moderate, high) and by components—diet, physical activity, body mass index (BMI), and alcohol intake. Overall, 14%, 60%, and 26% of the population had low, moderate, and high ACS guideline adherence, respectively. NDI was negatively associated with risk of high (vs. low) guideline adherence [RRR = 0.87, 95% confidence interval (CI) = 0.78–0.98], although attenuated after controlling for individual socioeconomic status (SES; RRR = 0.89, 95% CI = 0.80–1.00), and associated with lower adherence to BMI recommendations (low vs. moderate RRR = 0.90, 95% CI = 0.84–0.97; high RRR = 0.86, 95% CI = 0.77–0.97). Gentrification was associated with higher likelihood of meeting the dietary recommendations (low vs. moderate RRR = 1.04, 95% CI = 1.01–1.07), but not with overall adherence or individual components. Change in income inequality was not associated with outcomes. Neighborhood deprivation may be negatively associated with ACS guideline adherence among Hispanic/Latino adults.
Significance:
This study provides new evidence on the link between neighborhood gentrification, changing income inequality and adoption and maintenance of cancer preventive behaviors in an understudied population in cancer research. We observed that while neighborhood deprivation may deter from healthy lifestyle behaviors, positive changes in neighborhood SES via the process of gentrification, may not influence lifestyle guideline adherence among Hispanic/Latino adults.
Cx43-Delivered miR-181b Negatively Regulates Sirt1/FOXO3a Signalling Pathway-Mediated Apoptosis on Intestinal Injury in Sepsis
Authors: Zou Zhaowei , Yu Jianyang , Huang Renli , Yu Jinlong
Published: October 2023

Introduction: Gap junctions can transmit signals between cells, including miRNAs, leading to the amplification of adjacent cell damage. No previous study has addressed gap junctions and miRNAs in sepsis because the internal mechanism of sepsis-induced intestinal injury is complex. Therefore, we studied the relationship between connexin43 (Cx43) and miR-181b and provided a research direction for further study of sepsis. Methods: A mouse caecal ligation and puncture method was used to construct a mouse sepsis model. Firstly, damage to intestinal tissues at different time points was analysed. The levels of Cx43, miR-181b, Sirt1, and FOXO3a in intestinal tissues and the transcription and translation of the apoptosis-related genes Bim and puma, which are downstream of FOXO3a were analysed. Secondly, the effect of Cx43 levels on miR-181b and Sirt1/FOXO3a signalling pathway activity was explored by using the Cx43 inhibitor heptanol. Finally, luciferase assays were used to determine miR-181b binding to the predicted target sequence. Results: The results show that during sepsis, intestinal injury becomes increasingly worse with time, and the expression of Cx43 and miR-181b increase. In addition, we found that heptanol could significantly reduce intestinal injury. This finding indicates that inhibiting Cx43 regulates the transfer of miR-181b between adjacent cells, thereby reducing the activity of the Sirt1/FOXO3a signalling pathway and reducing the degree of intestinal injury during sepsis. Conclusions: In sepsis, the enhancement of Cx43 gap junctions leads to an increase in miR-181b intercellular transfer, affects the downstream SIRT1/FOXO3a signalling pathway and causes cell and tissue damage.
Long-Term Outcomes of Endoscopic Submucosal Dissection for Early Remnant Gastric Cancer: A Retrospective Multicenter Study
Authors: Tsuda Kazunori , Tanaka Shinwa , Yoshizaki Tetsuya , Yamamoto Yoshinobu , Ose Takayuki , Ishida Tsukasa , Kitamura Yasuaki , Obata Daisuke , Iwatate Mineo , Ikeda Atsushi , Ariyoshi Ryusuke , Kawara Fumiaki , Takihara Hiroshi , Abe Hirofumi , Takao Toshitatsu , Morita Yoshinori , Sano Yasushi , Umegaki Eiji , Nishisaki Hogara , Toyonaga Takashi , Kodama Yuzo
Published: October 2023

Introduction: Favorable long-term outcomes of endoscopic submucosal dissection (ESD) for early remnant gastric cancer (ERGC) have been reported in single-center studies from advanced institutions. However, no studies have examined the long-term outcomes using a multicenter database. This study aimed to investigate the long-term outcomes of the aforementioned approach using a large multicenter database. Methods: This retrospective multicenter cohort study included 242 cases with 256 lesions that underwent ESD for ERGC between April 2009 and March 2019 across 12 centers. We investigated the long-term outcomes of these patients with the Kaplan-Meier method, and the relationship between curability, additional treatment, or hospital category, and the survival time was evaluated using the log-rank test. Results: During the median follow-up period of 48.4 months, the 5-year overall survival rate was 81.3%, and the 5-year gastric cancer-specific survival rate was 98.1%. The survival time of patients of endoscopic curability (eCura) C-2 without additional surgery was significantly shorter than the corresponding of patients of eCura A/B/C-1 and eCura C-2 with additional surgery. There was no significant difference in either overall survival or gastric cancer-specific survival rate between the high-volume and non-high-volume hospitals. Conclusion: The gastric cancer-specific survival of ESD for ERGC using a multicenter database was favorable. ESD for ERGC is widely applicable regardless of the hospital case volume. Management in accordance with the latest guidelines will lead to long-term survival.
Custom Laser-Cut Silastic® Implants for Type I Thyroplasty in a Preclinical Model
Authors: Wilson Azure , Sayce Lea , Li Zheng , Gartling Gary , Luo Haoxiang , Rousseau Bernard
Published: October 2023

This study aimed to produce customized silicone elastomer implants of varied size and shape for optimization of type I thyroplasty procedures in a rabbit model. Computer-aided design models of different implant designs were designed and used to program laser cutting of a medical-grade Silastic® sheet. Laser-cut implants were produced rapidly and cost-efficiently. Surgical implantation demonstrated vocal fold medialization and phonation in 5 test subjects. This technique may provide a low-cost alternative or adjunct method to hand-carving or commercial implants.
Study on the Efficacy of Sacubitril/Valsartan in Patients with Heart Failure with Preserved Ejection Fraction Undergoing Peritoneal Dialysis
Authors: Sheng Yuping , Ma Xiaoying , Liu Ye , Yang Xingmeng , Sun Fuyun
Published: October 2023

Introduction: Cardiovascular disease is the most common cause of death and morbidity in patients with end-stage renal disease. Sacubitril/valsartan (SAC/VAL) can reduce the risk of cardiovascular mortality among patients with heart failure (HF). The present study set out to evaluate the efficacy of SAC/VAL in the treatment of patients with HF with preserved ejection fraction (HFpEF) undergoing peritoneal dialysis (PD) (HFpEF&PD). Methods: A total of 160 patients with HFpEF&PD were enrolled and randomly divided into the control group (N = 80) and SAC/VAL group (N = 80). The cardiac function efficacy, HF scoring efficacy, echocardiographic parameters, serological indicators, and 6-minute walking test were compared before and after treatment. Results: After 6 months of treatment, the total number of patients who responded to treatment in the SAC/VAL group was higher than that of the control group in terms of cardiac function and HF scoring efficacy. After treatment, levels of early diastolic/late diastolic filling velocity and left ventricular ejection fraction were increased in both groups, while the levels of left atrial diameter, left ventricular end-diastolic diameter, left ventricular end-systolic diameter, inter-ventricular septal diameter, and left ventricular posterior wall diameter were decreased; the NT-proBNP levels were diminished in both groups, while hemoglobin levels and the 6-minute walk distance were increased; the systolic blood pressure, diastolic blood pressure, and 24-h ultrafiltration volume were lowered in all patients. The changes in these indexes in the SAC/VAL group were more obvious than those in the controls. Conclusion: SAC/VAL can significantly improve cardiac function in patients with HFpEF&PD.
School Organizational Climate and Leadership Styles among School Admin-istrators: Effect on the Teacher’s Level of Performance
Authors: Pulumbarit Jaime
Published: October 2023

This study's main goal was to determine the effect of the school's organizational climate and administrator's leadership styles on the teacher's level of performance in the selected Apostolic Vicariate of Bulacan (AVB) Parochial Schools. The teachers at six chosen AVB parochial schools served as the study's primary respondents. The tool utilized was a four-part questionnaire about the culture of the organization, the leadership style, and the effectiveness of the teachers. Frequency distribution, standard deviation, descriptive measures, and standard linear regression analysis were the statistical techniques utilized to quantify and infer the relationship between the relevant variables. The findings showed that gender and the highest educational degree are important predictors of instructors' performance levels at the 0.05 significance level. It was also determined that leadership style and organizational climate had no significant impacts on the teachers' performance. Suggestions on a collaborative effort by the administration, teachers, and students should be looked into and evaluated for the further enhancement of this study.
Relationship of Personality Traits and Organizational Climate on Work Engagement Among Higher Education Personnel in the Philippines
Authors: Cainday Marie Rosette L. , Chan Francis Alvin O. , Bestuer Gaspar M.
Published: October 2023

This study investigated the relationship of personality traits and organizational climate on the work engagement of the teaching and non-teaching personnel of West Visayas State University-Janiuay Campus in 2021. This descriptive research was conducted among 81 personnel of the higher education institution. The statistical tools used were frequency count, ranking, mean, and Spearman rho to analyze the data from the survey. Manchester Personality Test (MPQ) version 14.2 was utilized to measure the personality traits of the participants; the Organizational Climate Questionnaire (OCQ) was employed to determine the organizational climate; and the Utrecht Work Engagement Scale (UWES) was utilized to determine their work engagement. The dominant personality traits among teaching and non-teaching personnel are conscientiousness, rationality, and empathy. None of the personality traits have a significant relationship with work engagement.  Their work engagement is high, and similarly, they perceived the organizational climate as high as well. Additionally, the dimensions of organizational climate, namely respect, planning and decision-making, innovations, relationships, commitment and morale, role clarity, communication, quality service, and training and learning, have a significant relationship with work engagement. Personnel highly engage themselves in their work given a favorable campus organizational climate, resulting in an enhanced degree of enthusiasm and dedication. Points derived from the findings were identified for consideration for inclusion in the continuous enhancement of the faculty and staff development program on campus, the human resource development plan, and policy-making pertaining to the development of human resources at the university.
Bullying in the SUC's Classroom: A Hidden Epidemic?
Authors: Clores Novy R.
Published: October 2023

Several literatures have already pointed to the effects of bullying to the learners’ journey in educational institution.  Acts of bullying have even evolved from the actual or physical acts to virtual acts brought by the ever increasing ICT saturation around the world.  The study explored the prevalence of bullying in SUCs by looking into the profile, the personal interpretation of bullying, the prevalence of bullying in the classroom, the effects of bullying, and the awareness on policies of bullying among tertiary students.

Using cross-sectional survey method with a questionnaire as the main instrument, 95 respondents were tapped. For the findings, majority of the respondents were females aged 18 – 24 years old, Varied definitions of bullying were presented indicating that these acts are done in multiple and in combination of ways by the perpetrators.  The most dominant effects of bullying were low self-esteem, depression, shyness, perceived negative school climate and wanting to be left alone.  These effects somehow diminished the motivation or desire of the learner to further seeking knowledge in school. Furthermore, learners expressed their frustration and interest in reaching their dreams and aspirations. One of the academic heads of the institution expressed during the interview that while there are existing policies in the institution against bullying, reporting cases of bullying is often problematic. This implies that responsible office could not immediately act on it unless victims come forward. Some said that bullying is part of the growing process and taking it seriously is a sign of immaturity thus it should not be considered as big deal. With respect of the EQ of the victims and non – victims there were significant differences in Self awareness, self motivation and empathy and no significant differences on managing emotions and social skills implying the victims are effective in keeping it among themselves. In conclusion, bullying is prevalent in SUCs and it is the method of reporting of cases that causes a seemingly lack of action on the part of the authorities.  It is therefore recommended that inclusion of reporting mechanism and vigorous campaign against bullying must be included in the school academic manual. Furthermore, everyone in the institution should work hand in hand to halt this hidden epidemic.
Assessment of the No Plastic Policy Implementation: A Basis to Plastic Waste Management Action Plan
Authors: Valenzuela Victoria P. , Sultan Lianne Mar DC , Calatrava Ma. Cecilia A. , Esguerra Pauline G. , Montero Diane Grace B.
Published: October 2023

This paper assessed the extent of the implementation of the Provincial Ordinance 2012-09 or the No Plastic Policy in the Municipality of Balagtas, Bulacan. Since plastic waste is one of the evident issues in the municipality, the study aimed to assess the extent and determine the challenges encountered in the implementation of the No Plastic Policy which was signed in order to address the issues related to plastic wastes. The extent of the implementation of the policy was gathered through survey questionnaires distributed to business sectors and households while semi-structured interviews were conducted among the key implementers to determine the challenges encountered. The findings of the study showed that the extent of the implementation of the No Plastic Policy was somewhat implemented and the challenges encountered include costly and limited alternatives; lack of clarity in the operational guidelines; non-compliance of the residents; and lack of strict implementation efforts. Thus, the findings of the study showed that there is a need to strengthen the existing ordinance in the Municipality of Balagtas, Bulacan.
Abstract 109: A Spatial Transcriptomic Approach to Understanding Coronary Atherosclerotic Plaque Stability
Authors: Gastanadui Maria Gabriela G , Litovsky Silvio , Margaroli Camilla , Richter Robert , Wang Dezhi , Xing Dongqi , Wells James , Gaggar Amit , Nanda Vivek , Patel Rakesh P , Payne Gregory A
Published: October 2023

Introduction:
Classically, vulnerable plaques at risk for acute coronary syndrome (ACS) have been defined by cellular architecture. However, contemporary imaging studies have shed light into new mechanisms of ACS (e.g., plaque erosion), challenging plaque rupture as the sole mechanism of disease. Furthermore, mechanisms within the vascular microenvironment that may modulate these disease phenotypes are poorly understood. Our objective was to examine differential programming of smooth muscle and macrophage cells amongst patients with stable and unstable plaques using spatial transcriptomics.

Methods:
Autopsy-derived coronary arteries were reviewed by an independent pathologist and segregated into stable (N=8) and unstable (defined by the presence of CD68+ cells and a large lipid core, N=8) plaques for spatial profiling. In combination with immunofluorescent histology, our approach enabled cell-specific, spatially resolved transcriptional profiling of plaques with mapping of cell populations via gene expression patterns.

Results:
We observed unique spatial and cell-specific transcriptional signatures for stable and unstable plaques including regional differences in the tunica intima and media. These regions differentially expressed pro-inflammatory (e.g., IFN-gamma, MHC Class II, pro-inflammatory cytokines) and pro-thrombotic signaling pathways. Cell-type-specific expression analysis using spatial deconvolution revealed heterogeneous CD68+ cell populations which shared gene signatures with endothelial, smooth muscle, and myeloid cells.

Conclusion:
Our results highlight cell-specific and regional pro-inflammatory and coagulation transcriptional changes within unstable coronary arterial plaques and supports the idea of cellular dedifferentiation and intra-plaque plasticity as a contributor to plaque instability. These findings may help to identify novel mediators of ACS and potential therapeutic targets.
Abstract 121: TMEM16E And TMEM16F Are Required For The Formation Of Prothrombotic Extracellular Vesicles From Endothelial Cells
Authors: Anderson Papa Freduah A , Schmaier Alec A
Published: October 2023

Procoagulant extracellular vehicles (EVs) have been implicated in thrombotic cardiovascular disease. Externalization of phosphatidylserine (PS) supports the assembly of coagulation enzyme complexes and the formation of EVs. Prior work has demonstrated that the endothelial cell (EC) membrane is a significant source of procoagulant PS and that the transmembrane proteins TMEM16E and TMEM16F are required for procoagulant PS externalization. We sought to determine whether formation of procoagulant EC EVs is regulated by TMEM16E or TMEM16F. TMEM16E or TMEM16F were silenced in HUVECS using siRNA and cells were treated with TNF-α (16 h, 10 ng/mL) to stimulate tissue factor expression and PS externalization. EC EVs were isolated from conditioned media via ultracentrifugation. EV charge, particle number, and size were analyzed using zeta potential, nanoparticle tracking analysis, and transmission electron microscopy (TEM). Procoagulant activity was measured via factor VIIa-catalyzed factor Xa generation, prothrombinase complex assay, and by thrombin generation in plasma. Silencing of TMEM16E or TMEM16F reduced factor Xa generation on EC EVs by 90% (P < 0.0001, 2 independent siRNA). TMEM16E and TMEM16F were required for thrombin generation, a process also inhibited by lactadherin, which blocks the accessibility of PS to coagulation enzymes. TNF-α resulted in a ~400-fold increase in total EC EV generation compared to vehicle control (P < 0.0001). TNF-α-induced EC EVs were smaller than that induced by vehicle control (88 nM vs 110 nM, P < 0.02). Silencing of TMEM16E or TMEM16F reduced TNF-α-induced EV production by 100-fold (P < 0.0001) and EVs from TMEM16E or TMEM16F silenced cells trended larger in size. We confirmed the exosome identity of EC EVs via CD63 immunogold staining and TEM. Zeta potential showed that EC EVs were negatively charged and EC EVs from TNF-α-stimulated cells showed the most negative charge. Silencing of TMEM16F reduced the negative charge of EC EVs (-38.57mV vs -20.51mV, P < 0.004) suggesting less abundance of negatively charged phospholipids. This study identifies TMEM16E and TMEM16F as essential for the production of PS-positive, procoagulant EC EVs. TMEM16 proteins may therefore be therapeutic targets to protect against thrombosis.
Abstract 125: Sex-dependent Effects of Sortilin Antisense Oligonucleotide on Vascular Calcification
Authors: Lupieri Adrien , Becker-Greene Dakota , Le Thanh-Dat , Roschel Marina M , Jha prabhash , Sonawane Abhijeet , Tsimikas Sotirios , Kuraoka Shiori , Singh Sasha A , Aikawa Masanori , Aikawa Elena
Published: October 2023

Background:
The development of atherosclerotic calcification is a complex process associated with an increased risk of cardiovascular events. Despite decades of extensive studies, this disorder has no effective therapeutic strategies. Previous studies demonstrated that genetic deletion of sortilin (Sort1) decreased vascular smooth muscle cell (SMC) calcification in experimental atherosclerosis. We hypothesized that silencing sortilin expression
in vivo
using antisense oligonucleotide (ASO-Sort1) will offer a promising strategy to prevent vascular calcification.

Methods and Results:
Male and female Ldlr-/- mice (n=10 per group) were fed a high-fat diet and administered with ASO-Sort1 for 15 weeks. While ASO treatment reduced aortic sortilin expression at a similar range of 52% and 57% in female and male mice, respectively, female mice displayed lower basal expression of Sort1 than male. After ASO-Sort1 treatment, we assessed atherosclerotic lesion formation and calcification of the aorta using molecular imaging and histology. While treatment had no effect on body weight, plasma cholesterol, and progression of atherosclerosis in either sex, it prevented the development of aortic calcification exclusively in male mice. To assess the sex-dependent mechanism involved, we analyzed the changes of the aortic proteome in both groups of mice treated with ASO-Sort1. Our results showed a reduction of vesicular trafficking pathways in both male and female mice; however, other biological processes including autophagy showed sex-dependent inhibitions only in male mice. To validate our findings, we explored
in vitro
whether 17β-estradiol (E2) treatment of mouse SMCs promotes calcification independently of sortilin. Results showed that E2 induces calcification in SMC treated with ASO-Sort1.

Conclusion:
Targeting sortilin using antisense technology is efficient to prevent vascular calcification in male mice; however, this effect was lost in female mice due to a sortilin-independent development of calcification mediated by E2. This unexpected outcome highlighted a novel sex-dependent discrepancy of the calcification pathway mediated by sortilin.
Abstract 128: Safety And Efficacy Of Pro-efferocytic Nanoparticles To Treat Atherosclerosis In A Porcine Model
Authors: Bamezai Sharika , Alsaigh Tom , Lofti Mozhgan , Zhang Yapei , Kumar Gayatri S , Wang Fudi , Ye Jianqin , Luo Lingfeng , Maegdefessel Lars , Kojima Yoko , Kumari Manisha , Bell Caitlin , Adkar Shaunak , Smith Bryan R , Leeper Nick J
Published: October 2023

Background and Aims:
Within the atherosclerotic plaque, the formation of a necrotic core is, in part, a consequence of impaired macrophage efferocytosis in which apoptotic vascular cells upregulate “don’t-eat-me,” anti-phagocytic ligand CD47.
In vivo
, translational murine model studies demonstrating off-target effects of anti-CD47 antibody-mediated blockade prompted the development and successful validation of a macrophage-specific nanotherapy loaded with a chemical inhibitor of Src homology 2 domain-containing phosphatase-1 (SHP-1), a small molecule downstream of the CD47-SIRPα signaling axis. We aimed to test the safety and efficacy of this SHP-1 inhibitor nanotherapy in preventing atherosclerosis in a pre-clinical porcine model.

Methods and Results:
Sixteen LDLR
-/-
pigs were fed an atherogenic diet and treated with single-walled carbon nanotubes (SWNT) loaded with SHP1 inhibitor (SWNT-SHP1i) (n=8) or with SWNT alone (n=8), dosed weekly for three months. Blood samples were collected at baseline and at monthly intervals. The lipid panel, CMP, and CBC demonstrated no significant difference between the SWNT-SHP1i and control treatment groups at each timepoint. Survey of major arterial beds by histology revealed early-stage fatty streak disease within the left and right coronary artery (RCA) ostium in addition to the abdominal aorta distal to the renal arteries. Pigs underwent PET/CT in the final week of treatment. The SWNT-SHP1i treated cohort displayed a reduced carotid artery uptake of 18F-FDG compared to the controls (p = 0.017). Bulk RNA sequencing analysis of the carotid artery demonstrated multiple differentially expressed pathways related to hemostasis and the immune response.

Conclusion:
In a porcine model, macrophage-specific SWNT-SHP1i treatment is not significantly associated with anemia, thus circumventing off-target effects of global anti-CD47 blockade. PET/CT reveals evidence of significantly decreased vascular inflammation in the carotid arteries in the SWNT-SHP1i treated cohort. Bulk RNA-seq confirms numerous significant differentially expressed genes. Ongoing blinded, quantitative histological analysis will further compare both plaque burden and plaque M1/M2 macrophage content between the two cohorts.
Abstract 146: Foamy Macrophages Rely On Trem2 For Survival During Atherosclerosis
Authors: Patterson Michael , Firulyova Maria M , Xu Yingzheng , Zhu Alisha , Schrank Patricia , Ronayne Christine , Kennedy Ainsley E , Hillman Hannah , Bold Tyler , Zaitsev Konstantin , Williams Jesse W
Published: October 2023

Atherosclerotic plaque formation is supported by the accumulation of cholesterol loaded foamy macrophages within the artery wall. Molecular regulators of foamy macrophage differentiation and survival remain to be fully understood. We performed trajectory analysis of a murine META scRNAseq dataset and compared it against a genome-wide foamy macrophage CRISPR screen, which identified Triggering Receptor Expressed on Myeloid Cells 2 (Trem2) as a candidate regulator for foamy macrophage lipid uptake and differentiation. Conditional deletion of Trem2 in macrophages led to dramatically reduced atherosclerotic plaque burden in the aorta and aortic root after 8 or 16 weeks of western diet feeding. This was associated with reduced local macrophage proliferation and enhanced cell death within lesions, but independent of circulating cholesterol or inflammatory markers. In addition, Trem2-deficient macrophages showed reduced ability to take up lipids compared to WT macrophages, both in vivo and in vitro. Gene expression analysis of foamy and non-foamy macrophages revealed that Trem2 was required for proliferation gene expression and for the induction of cholesterol efflux genes after lipid loading. Without Trem2 foamy macrophages accumulated excess lipid in the endoplasmic reticulum (ER), leading to an ER-stress response and cholesterol-mediated cytotoxicity. Overall, this study reveals Trem2 as a mediator of foamy macrophage differentiation and atherosclerosis plaque growth, and furthermore identifies the Trem2 pathway as a putative therapeutic target for future intervention studies.
Abstract 150: Targeting The Lipogenic Enzyme ATP Citrate Lyase As A Potential Therapy Against Coronary Artery Disease
Authors: Grobs Yann , Lemay Sarah-Eve S , Romanet Charlotte , Bourgeois Alice , SALEM MABROUKA , Martineau Sandra , Breuils-Bonnet Sandra , Voisine Pierre , Potus Francois , Provencher Steeve , Boucherat Olivier , Bonnet Sebastien
Published: October 2023

Introduction:
Coronary artery disease (CAD) such as coronary stenosis and intra vein graft stenosis are characterized by a cancer-like pro-proliferative and apoptosis-resistant phenotype of smooth muscle cells (SMCs), fueled by a metabolic shift toward glycolysis and interconnected global changes in the epigenetic landscape. The nucleo-cytoplasmic enzyme ATP Citrate Lyase (ACLY) has recently emerged as a key player and therapeutic target in cancer by favoring Warburg effect, lipid synthesis and chromatin remodeling. However, its role in CAD is still unknown. We hypothesized that ACLY is upregulated in CAD and supports the abnormal phenotype of CAD-CoASMCs.

Methods and Results:
ACLY expression positively correlates with vascular remodeling (aSMA labeling) in human distal coronary artery (CoAs, p<0,01) as well as in dog saphenous vein (SV) graft from bypass surgery (p<0,05). Increased expression and activation (phosphorylation) of ACLY were also observed in CoASMCs isolated from CAD patients (immunoblot, p<0,01) with preferential localization in the nucleus (immunofluorescence ACLY labelling (p<0,001)). Pharmacological (BMS303141) or molecular (siRNA) ACLY inhibition resulted in decreased survival, proliferation (immunoblot: PCNA, MCM2 and SURVIVIN (p<0,05) & immunofluorescence Ki67 and Annexin V labelling (p<0,001)), histones acetylation (immunoblot: acH3K27 and acH4 (p<0,05)) and reversed Warburg effect (immunoblot: pPDH, LDHA, PFKBP3 (p<0,01) & seahorse assays (p<0,05)) of CAD-CoASMC. RNA sequencing analysis showed that ACLY molecular inhibition in CAD-CoASMC mostly affectes cell cycle and cell proliferation pathways. Ex-vivo, ACLY inhibition attenuates vascular remodeling (EVG staining) in human CoAs and SVs rings denuded of endothelial cells and exposed to growth factors (p<0,05). In vivo, SMS-targeted loss of Acly in mice model of carotid artery denudation injury prevents vascular remodeling (p<0,05). Same results are observed with pharmacological inhibition of ACLY (BMS303141 & bempedoic acid) in carotid artery wire injury rat model (p<0,05).

Conclusion:
We demonstrated that ACLY is implicated in vascular remodeling in CAD and its Pharmacological inhibition may represent a novel avenue as therapeutic treatment.
Abstract 152: Metabolism And Therapeutic Potential Of Lipidated Amino Acids In Nonalcoholic Steatohepatitis And Atherosclerosis
Authors: Anand Sumit Kumar , Das Sandeep , Finney Alexandra C , Rohilla Sumati , McKinney M. Peyton , Rom Oren
Published: October 2023

Nonalcoholic fatty liver disease (NAFLD) affects nearly one third of the global population. Despite significant advances in our understanding of the underlying causes and considerable efforts in drug development, no pharmacological therapy currently exists for this disease. Accelerated atherosclerosis, independent of traditional risk factors, is the major cause of death in patients with NAFLD, particularly in those with the more severe nonalcoholic steatohepatitis (NASH). Thus, there is an urgent need to identify new pathways for simultaneous targeting of NASH and atherosclerosis. Imbalanced lipid metabolism and dysregulated amino acid metabolism are emerging as common features of both NASH and atherosclerosis, although their crosstalk has not received much attention. Lipidated amino acids or N-acyl amino acids (NAAs) have emerged as endogenous signaling molecules in which an amide bond links an amino acid to the acyl moiety of a long-chain fatty acid. Utilizing unbiased transcriptomics, we uncovered suppression of known NAA biosynthetic genes and upregulation of degradative genes in livers from humans and mice with NASH, concomitant with marked reduction of NAAs, as determined by metabolomics. Similar findings were noted in lipid loaded primary hepatocytes and hepatic cell lines. Hepatic NAAs, particularly N-oleoyl leucine (C18:1-Leu), inversely correlated with NASH and inflammatory indices. Importantly, C18:1-Leu administered to mice with established NASH reduced transaminases, steatohepatitis, hepatic macrophages and fibrosis. RNA-seq revealed induction of hepatic peroxisome proliferator-activated receptor α (PPARα)/fatty acid oxidation (FAO) and suppression of chemokine signaling pathways aligned with reduced liver and plasma C-C motif chemokine ligand 2 (CCL2). Luciferase and qPCR analyses showed that C18:1-Leu induces PPARα transcriptional activity, while stimulating FAO, as determined by Seahorse. In atherosclerotic mice, C18:1-Leu reduced lesional macrophages and atherosclerosis while lowering hepatic steatosis and CCL2. Our findings uncovered a newly identified metabolic pathway linking NASH and atherosclerosis and a novel potential therapy for the simultaneous treatment of these two prominent diseases.
Abstract 156: Role of Platelets in Maintaining Hemostasis and Coagulation Activation in Viral-induced Acute Respiratory Distress Syndrome (ards) in Mice
Authors: Sharma Swati , Williams Willie A , Elzawam Aymen Z , Antoniak Silvio
Published: October 2023

Platelets are crucial players during viral infections, as bridge hemostasis, inflammation, and immune response. RNA viruses like IAV are associated with pathologic platelet activation and thrombosis. We are currently investigating how platelet depletion before or post initial IAV infection can affect disease outcome. WT mice were infected with IAV (0.02HAU, intranasally) for 7 days to induce viral ARDS. Mice were injected with platelet depletion antibody R300 or control IgG at day 0 (prior) and day 3 (after infection). Mice treated with R300/control IgG were subjected to dsRNA mimetic poly IC intranasally for 3 days to induce virus-like ARDS. Global lung function was recorded, plasma and BALF were collected 7 or 3 dpi or final poly IC treatment, respectively. BALF cellularity and blood cellular composition was analyzed. Markers of lung injury and cytokines were measured. We observed that IAV infection leads to local lung inflammation, coagulation activation and ARDS-like lung dysfunction. Mice treated with R300 showed severe thrombocytopenia in blood. We observed more impaired alveolar hemostasis and increased hemorrhages into the airspace when platelet depletion was initiated 3 dpi compared to mice where platelets were depleted prior to infection. In addition, in dsRNA-induced ARDS model, platelet depletion resulted in increased signs of lung dysfunction, increased lung injury and local inflammation compared to mice with normal platelet levels. In conclusion, depletion of platelets after IAV infection resulted in increased alveolar hemorrhage and was associated with more pronounced lung inflammation, injury, and lung dysfunction. Our study suggests that interfering with platelet before viral infection can be beneficial as it limits the role of platelets as potential multiplicator of an initial pathologic immune reaction subsequently reducing collateral damage and organ dysfunction/failure. However, at later stages of viral infection, platelet activation is beneficial to facilitate tissue hemostasis as their absence later can result in increased hemorrhagic risk and more pronounced lung dysfunction. We aim to address the mechanism by which platelet contribute later to pathologic immune processes in dsRNA-induced or viral ARDS
Abstract 217: Protease Activated Receptor 2 Is Critical For Vascular Smooth Muscle Cell Transition To A Macrophage-like State
Authors: Wadding-Lee Caris A , Coughlin Taylor , Jones Shannon M , Jay Megan , Lusis Jake , Camerer Eric , Civelek Mete , Owens A Phillip
Published: October 2023

Objective:
Recent lineage tracing studies have shown vascular smooth muscle cells (VSMCs) develop macrophage-like characteristics (MLCs) in late-stage atherosclerosis resulting in accelerated inflammation and disease severity. Our objective was to determine if the inflammatory receptor, protease activated receptor 2 (PAR2), accelerates VSMC-MLC in a mouse model of atherosclerosis.

Methods and Results:
Our lab demonstrated

Par2
-/-

mice have decreased atherosclerotic lesion area versus proficient controls via downregulation of VSMC migration and cytokine release. To examine the role of PAR2 in VSMC dedifferentiation,

Par2
+/+

and

Par2
-/-

VSMCs were treated with 10 ug/mL methyl- β-cyclodextrin cholesterol for 72 hours. While

Par2
+/+

VSMCs demonstrated a loss of VSMCs markers (α
-
actin and myosin heavy chain) and upregulated macrophage markers (CD68 and Mac-2),

Par2
-/-

VSMCs remained stable. Looking at macrophage characteristics,

Par2
-/-

VSMCs phagocytized significantly less latex beads than

Par2
+/+

VSMCs. Analysis of a hybrid mouse diversity panel (HMDP), conducted on 101 strains of mice with induced atherosclerosis, determined PAR2 was significantly correlated with the MLC super-gene Krüppel-like factor 4 (KLF4), which was found to have lower genetic expression in
Par2
deficient VSMCs. Previous studies have demonstrated
Par2
mRNA is bound and stabilized by the RNA binding protein human antigen R (HuR). Our studies confirm these findings where

Par2
-/-

VSMCs have less HuR mRNA expression and activity, suggesting a role of HuR in PAR2 mRNA stability.

Par2
flox/flox

mice bred with transgelin (SM22-alpha) Cre and low-density lipoprotein receptor deficient mice l have been establishing in our lab to investigate the effects of cell-specific deletions of PAR2 in atherosclerosis. VSMC-specific
Par2
deficiency blunts the formation of atherosclerosis versus the

Cre
+

littermates.

Conclusions:
These results suggest that VSMC-specific PAR2 activation mediates the dedifferentiation of VSMCs via upregulation of KLF4 and subsequent binding by HuR, which stabilizes PAR2 mRNA and upregulates PAR2. Future studies will continue to study VSMC dedifferentiation utilizing tagged lineage-tracing mouse models and various
in vitro
techniques.
Abstract 245: Endothelin-1 And Procollagen Type-1 Carboxy-terminal Propeptide Are Associated With Functional Tolerance And Cardiac Structure And Function In Heart Failure With Preserved Ejection Fraction
Authors: Imran Tasnim F , Garrett Thomas , Parra Laura , Arcuri Daniel , Nguyen Christopher , JANKOWICH MATTHEW , Avazmohammadi Reza , atalay michael , Wu Wen-Chih H , Choudhary Gaurav
Published: October 2023

Introduction:
The role of emerging biomarkers such as endothelin-1 (ET-1) and collagen peptides in functional tolerance among patients with heart failure with preserved ejection fraction (HFpEF) is not well understood.

Hypothesis:
We hypothesize that plasma ET-1 and procollagen type-1 carboxy-terminal propeptide (PICP) are associated with 6-minute walk distance and parameters of cardiac structure and function.

Methods:
In a cross-sectional cohort study, we evaluated the association of ET-1 and PICP with 6-minute walk distance and parameters of cardiac structure and function on cardiac magnetic resonance (CMR) imaging in patients with HFpEF. In a subset of patients, we used a novel diffusion tensor imaging CMR technique to determine the myocardial fiber orientation in the septum as characterized by the helix angle transmurality (slope of the helix angle versus transmural depth from endocardium to epicardium) (Fig 1C).

Results:
A total of 36 patients (25 with HFpEF, 11 controls), mean age 70 ± 7.7 yrs, BMI 33 ± 8.5 kg/m
2
, 94% men, 53% had diabetes, and 75% had hypertension. Mean LVEF 58.7 ± 7.7%, RVEF 49.1 ± 10.4%, mean global myocardial mid- LV T1 1285ms, mean LV mass indexed to body surface area 65 ± 12 g/m
2
. Using univariate linear regression, we found that both plasma ET-1 levels and PICP levels were moderately inversely associated with 6-minute walk distance (R
2
-0.56, p<0.001) and (R
2
-0.40, p=0.02) (Fig 1A) respectively, with a non-significant association with age (R
2
= 0.30, p=0.08) and indexed LV mass (R
2
=0.29, p=0.09). ET-1 was moderately associated with septal HAT, a marker of myocardial fiber orientation, (R
2
=0.58, p=0.02), measured in a subset of patients (Fig 1B, C).

Conclusions:
Plasma ET1 and PICP levels are moderately associated with lower 6-minute walk distance; ET-1 is associated with septal myocardial fiber orientation in this exploratory analysis. Future studies are needed to determine the prognostic value of these biomarkers in patients with HFpEF.
Abstract 260: Machine Learning To Identify Heterogeneity Of Treatment Effect Of Intermediate-dose Vs Standard-dose Prophylactic Anticoagulation In The Inspiration Trial
Authors: Bikdeli Behnood , Spicer Alexandra , Bakhshandeh Hooman , Talasaz Azita , Rashidi Farid , Sharif-Kashani Babak , Beigmohammadi Mohammad Taghi , Farrokhpour Mohsen , Sezavar Seyed Hashem , Payandemehr Pooya , Dabbagh Ali , Gohari Moghadam Keivan , Eslami Vahid , Sinha Pratik , Goldhaber Samuel , Piazza Gregory , Krumholz Harlan M , Sadeghipour Parham , Churpek Matthew M
Published: October 2023

Background:
In the Intermediate vs Standard-Dose Prophylactic Anticoagulation in Critically-ill Patients With COVID-19: An Open Label Randomized Controlled Trial (INSPIRATION) trial, intermediate- compared with standard-dose prophylactic anticoagulation did not reduce a composite of venous/arterial thrombosis, treatment with extracorporeal membrane oxygenation or mortality. Whether machine learning (ML) implemented to predict individualized treatment effects (ITE) would identify variations in outcomes using baseline study variables is unknown.

Methods:
We used data from participants in the INSPIRATION trial. Patients were split into separate training (50%) and testing (50%) cohorts. In the training set, 24 different machine learning approaches to predict ITE were fit using baseline variables as predictors. The models were evaluated in the test set using the adjusted Qini index and the c-for-benefit, which illustrate how well the models identify patients deriving benefit or harm by the intervention. A value of 0 for the adjusted Qini index and 0.5 for c-for-benefit suggest the model did not identify any heterogeneous treatment effect.

Results:
A total of 562 patients were included. Baseline characteristics were balanced in the study groups. The single-model (s-learner) gradient boosted machine (GBM) model performed best, followed by the Tian transformation GBM model, and the causal forests model. Only the s-learner GBM model had an adjusted Qini >1, and only the s-learner GBM and Tian transformation GBM had c-for-benefit >0.50 (
Figure
). All other algorithms found no evidence of heterogeneous treatment effect.

Conclusions:
Of 24 machine learning models designed to predict ITE, only 2 found evidence of heterogeneous treatment effect. Considering multiple comparisons, these results should be considered as hypothesis-generating. The utility of ML models such as s-learner GBM for identification of ITE requires validation in additional trials.
Abstract 270: Multi-transcriptomic Profiling Reveals Novel Molecular Mediators During Abdominal Aortic Aneurysm Expansion.
Authors: Chernogubova Ekaterina - , Sachs Nadja , Wu Zhiyuan , Li Zhaolong , Pauli Jessica , Winski Greg , Paloschi Valentina , Busch Albert , Eckstein Hans-Henning , Maegdefessel Lars
Published: October 2023

Abdominal aortic aneurysm (AAA), a life-threatening condition associated with high morbidity and mortality rates, is despite decades of intense research still mostly treated by surgical repair as currently no medical therapy is available. Long non-coding RNAs (lncRNAs) are highly cell type specific and potential molecular disease modulators, posing potential drug targets. We performed single cell (sc) RNA and combined total (bulk) RNA sequencing to study the transcriptome of AAAs from patients undergoing open aortic repair. Further, we implemented hybridization-based RNA in situ sequencing (HybRISS) to localize highly intriguing genes within the tissue.In this current study, we examined three different sequencing datasets (total RNA from dilated vs. non- dilated AAA specimen from an individual patient each, AAA vs. control from individual patients and one scRNA-seq dataset). Six lncRNAs could be cross-referenced between all three datasets. All of these lncRNAs presented a cell-type specific expression pattern, with three lncRNAs being highly enriched within the SMC cluster. We found all newly identified lncRNAs conserved in our bulk and scRNA-seq datasets of experimentally-induced AAA in mice and LDLR-deficient Yucatan mini-pigs. Further, potential novel regulator genes could be visualized in human AAA tissue sections using HybRISS to resolve their distinct location and co-expression pattern within the diseased tissue. In summary, our approach utilizing dilated and non-dilated human tissue specimens, different preclinical models and resulting datasets demonstrate that combining bulk Seq, scRNA-Seq data and spatially resolved sequencing methods are powerful tools to detect and describe novel lncRNAs as disease modulators.