bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of Village Digitization through the Smart Village Concept in Permis and Rajik Villages, South Bangka Regency
Authors: Darman Darman Saputra , Julia Julia , Herdiyanti Herdiyanti
Published: November 2022

The rapid use of information technology in all aspects of life from the government to the community has encouraged several cities in Indonesia to implement the connectedness of the use of information technology in the form of smart cities. The development of this smart city apart from integrating all elements of the city in an information technology system that is easily accessible to the public is also for the provision of transparent, efficient and fair public services for all communities. In line with the smart city with a smaller context, namely the smart village, which is said to be smart if it innovatively uses information technology to achieve increased quality of life, efficiency and competitiveness in economic, social and environmental aspects. Permis Village, Simpang Rimba Sub-district, South Bangka Regency is a village directly adjacent to Rajik Village and Sebagin Village which is a potential village supported by a strategic location. When viewed in the context of a smart village, there is no understanding as to what ideally the concept of "smart" is when it is attached to the village. A smart village concept that is not only able to apply the use of information technology, but is also able to develop village potential, improve the economy and create. On the basis of these problems, this article constructs a smart village concept that can be applied to villages in Indonesia. This is also supported by the lack of studies that address the development of smart villages in Indonesia. It is hoped that this article will become an alternative for developing smart villages for villages in Indonesia so that in practice it will give rise to a conceptual picture of the realization and elements of smart village development.
Pendampingan Hukum Pemberdayaan Usaha Mikro Dan Pengembangan Bumdes Di Desa Blumbungan, Pamekasan
Authors: Prihatiningtyas Wilda
Published: November 2022

Mitra sasaran dalam pengabdian masyarakat ini yaitu masyarakat Desa Blumbungan, Kecamatan Larangan, Kabupaten Pamekasan, Jawa Timur. Kondisi sosial-ekonomi dan budaya masyarakat serta aktivitas masyarakat desa Blumbungan banyak dipengaruhi oleh kegiatan sosial keagamaan, diantaranya yaitu adanya kegiatan karang taruna, remaja masjid, PKK desa, kelompok pengajian, kelompok tani, serta pengembangan industri kecil/rumah tangga (kripik singkong, pembuatan rokok, permeubelan, pembuatan pilar, produksi tahu). Adanya industri kecil/rumah tangga ini tentu saja sangat membantu peningkatan pendapatan ekonomi masyarakat Blumbungan, walaupun masih ada kendala dalam praktiknya. Beberapa permasalahan mitra yang telah diinventarisasi, khususnya terkait pemberdayaan usaha mikro dan pengembangan BUMDes, diantaranya yaitu: (1) Minimnya pemahaman tentang aspek hukum dalam pengelolaan usaha, (2) Minimnya pengetahuan tentang metode pemasaran dan pengemasan produk, (3) Minimnya akses terkait modal usaha bagi WUB (Wira Usaha Baru), serta (4) Belum ditentukannya arah dan pengembangan BUMDES. Metode pelaksanaan kegiatan yakni meliputi persiapan, penyuluhan, pendampingan terhadap masyarakat desa sesuai kebutuhan yang telah dirumuskan pada tahap sebelumnya, dan terakhir evaluasi. Dengan adanya kegiatan pengabdian masyarakat ini diharapkan membawa perubahan positif bagi kemajuan Desa Blumbungan, sebagaimana slogannya yakni terwujudnya Blumbungan Sejahtera.
Abstract 104: R952Q Mutation In ApoE Receptor-2 Exacerbates Hypercholesterolemia And Accelerates Atherosclerosis Progression In Ldl Receptor-knockout Mice
Authors: Turkson Vanessa M , Haller April , Jaeschke Anja , Hui David
Published: November 2022

Background and objective:
Apolipoprotein E Receptor-2 (apoER2) is a type I transmembrane receptor in the LDL receptor family and is expressed in vascular smooth muscle cells (VSMC), endothelium, and macrophages. An apoER2 mutation referred to as ApoER2-R952Q (R952Q) is associated with elevated plasma triglyceride levels and increased risk of premature coronary artery disease and myocardial infarction. Disease phenotypes emerging from the loss of apoER2 have been established, with results showing that apoER2 inactivation accelerates atherosclerosis progression through macrophage-mediated lipid accumulation and VSMC senescence to accelerate necrosis. This study investigated the effects of the R952Q mutation and its influence on atherosclerosis.

Approach and results:
We generated R952Q-expressing mice using a CRISPR-directed approach. This mouse line was crossed with the LDL-receptor knockout mouse line (

Ldlr
-/-

) and the progenies were fed a high cholesterol diet for 24 weeks. Results showed that R952Q mice were similar to apoER2 knockout mice in displaying increased lesion necrosis compared to

Ldlr
-/-

mice with functional apoER2. However, R952Q mutant mice differed from apoER2 knockout mice, exhibiting increased lipid accumulation in the lesion area. Plasma cholesterol levels were also elevated in the R952Q mutant mice compared to their apoER2 knockout counterparts. Interestingly, unlike the apoER2 deficient VSMCs, the R952Q mutation did not affect VSMC proliferation and showed no signs of accelerated senescence. Thus, the accelerated atherosclerosis observed in the R952Q mutant mice is not due to smooth muscle cell defects, and possibly due to increased plasma cholesterol levels. Additional experiments performed via bone marrow transplant of control and R952Q cells into

Ldlr
-/-

mice showed that the R952Q bone marrow recipients recapitulated the elevated plasma cholesterol levels phenotype of the R952Q mice, suggesting a novel mechanism by which the R952Q mutation in macrophages may accelerate atherosclerosis.

Conclusion:
The R952Q mutation in apoER2 accelerates atherosclerosis via a gain-of-function mechanism in macrophages with a unique phenotype that exacerbates hypercholesterolemia.
Abstract 130: Thymidine Phosphorylase Enhances Abdominal Aortic Aneurysm Formation In Mice
Authors: Hong Liang , Zhuang Jian , Yue Hong , Zhou Xianwu , DeHart Autumn , Toloza-Alvarez Gretel , Du Lili , Li Wei
Published: November 2022

Abdominal aortic aneurysm (AAA) is affected by multiple factors, including vessel wall cells, extracellular matrix, genetic disorders, etc.; however, the pathophysiology of AAA remains unclear. We recently found that thymidine phosphorylase (TYMP) is a signaling protein, and an increase of TYMP inhibits the proliferation of VSMCs, the major cell population that affects AAA progression. TYMP enhances platelet activity, thrombosis, and inflammation. These data suggest that TYMP may play a role in AAA formation. By using human AAA samples, we found that TYMP expression was significantly increased in AAA vessel walls than in healthy aortas. We thus further examined the role of TYMP in AAA formation using a mouse model. WT C57BL/6J and

Tymp
-/-

mice were fed a western diet (TD.88137), started at 4 weeks (wks) old for 8 wks, and then received implantation of Ang II mini pump, which delivers Ang II to mice at a dose of 1 μg/kg/min for 4 wks. By echography, we found that Ang II perfusion dramatically increased inner aorta diameter at the diastolic phase in WT, but not

Tymp
-/-

mice. Mice were sacrificed 4 wks after Ang II perfusion. AAA was confirmed in 3 of the 12 WT mice, but no AAA was found in all 11

Tymp
-/-

mice used. The AAA was mainly formed at the suprarenal artery level. Histological examination confirmed the presence of freshly formed or fibrotic hematoma. By using TYMP overexpressing rat VSMC cell line, C2, and its control cells, PC, we found that TYMP increased MMP2 production, secretion, and activation, which was diminished by tipiracil, a selective TYMP inhibitor. Ang II did not affect extracellular MMP2 activity in both PC and C2. TYMP facilitated TNF-α induced MMP2 activation. TYMP induced constitutive AKT phosphorylation, which was further enhanced by TNF-α. TYMP increased expression of active TGFβ1. Consequently, TYMP significantly enhanced thrombospondin-1 type 1 repeat domain (TSR) stimulated TGFβ1 signaling activation and expression of CTGF, which is correlated with severe fibrotic disorders and AAA. In summary, our study, for the first time, demonstrated that TYMP, a pyrimidine salvage pathway enzyme, plays an important role in regulating vascular biology, especially, VSMC function. TYMP-mediated microenvironment changes enhance the development of AAA.
Abstract 139: Precision Nanomedicine Targeting Novel Endothelial Mechano-sensing Mechanisms To Treat Vascular Diseases
Authors: Fang Yun
Published: November 2022

Endothelial mechano-transduction mechanisms are instrumental to vascular health and disease but targeting disease-causing mechano-sensing pathways remains extremely challenging. Atherosclerosis preferentially develops at arterial curvatures and bifurcations where disturbed blood flow activates endothelium, leading to peripheral artery disease, carotid artery disease and ischemic stroke. Nevertheless; current atherosclerosis therapies mainly target systematic risk factors but not the vasculature
per se
. This underscores the significance and unique opportunity to identify and target novel mechanosensitive mechanisms in activated endothelium subjected to disturbed flow. Our objectives are to (1) delineate novel endothelial mechano-sensing mechanisms and (2) devise innovative precision nanomedicine approaches targeting these disease-causing mechano-sensitive pathways. Using multiple new transgenic mouse lines, our studies demonstrated that disturbed flow (DF) significantly increases the endothelial expression of TXNDC5 (thioredoxin domain containing 5) and microRNA-92a (miR-92a), two atherogenic molecules driving atherogenesis. Mechanistically, DF-induced TXNDC5 increases proteasome-mediated degradation of heat shock factor 1 and consequently promotes eNOS/NOS3 protein degradation. Meanwhile, DF induces endothelial miR-92a to suppress key anti-inflammatory molecules such as KLF2, KLF4, and PLPP3. We successfully formulated nanoparticles to deliver TXNDC5-targeting CRISPR/Cas9 plasmids driven by an endothelium-specific promoter (CDH5), which effectively deleted endothelial TXNDC5 expression and reduced atherosclerosis in Apoe
-/-
mice. Moreover, VCAM1-tageting polyelectrolyte complex micelles were engineered to selectively deliver miR-92a inhibitors to inflamed endothelium, which markedly decreased arterial stenosis and atherosclerosis in Apoe
-/-
mice. These results elucidate novel endothelial mechano-sensing mechanisms and provide a proof of concept of innovative targeted nanomedicine approaches, addressing an unmet medical need in vascular therapies.
Abstract 147: Endothelial Regulation By Enhancer Associated Long Non-coding RNA
Authors: Tang Xiaofang , Luo Yingjun , Yuan Dongqiang , Malhi Naseeb , Sriram Kiran , Chen Zhen Bouman
Published: November 2022

Lining the critical interface between circulating blood and vascular wall, endothelial cells (ECs) play vital functions in health and disease. The optimal gene expression in ECs is essential to maintain endothelial homeostasis, and its dysregulation can lead to EC dysfunction, a common mechanism underlying many metabolic and cardiovascular diseases, e.g. diabetes and diabetes-associated vasculopathy. We identified an enhancer-associated long non-coding RNA (lncRNA) that enhances endothelial nitric oxide synthase (eNOS) expression, aka LEENE in ECs. LEENE is suppressed in diabetes conditions in ECs in vitro, in vivo, and in human arteries. Knockout of
leene
homologue in mouse resulted in impaired microvascular function, evident in a diabetic hindlimb ischemia model. Overexpression of human LEENE RNA in the knockout mice rescued the ischemic recovery, resembling that of wildtype animals at tissue function and transcriptome, and single cell gene expression levels. Mechanistically, LEENE binds to the promoters of a set of pro-angiogenic genes to induce their transcription, as revealed by chromatin isolation with RNA pulldown combined with sequencing. Taken together, our work demonstrates an essential role for LEENE in the regulation of angiogenesis and tissue perfusion. Functional enhancement of LEENE to restore angiogenesis and blood flow perfusion may provide a novel strategy to tackle ischemic diseases such as peripheral arterial disease.
Abstract 148: A Tissue-specific Crispr/cas9 Vector System For Conditional Gene Disruption In Zebrafish Reveal The Role Of Neuropilin-1 In Heart Regeneration
Authors: Mukhopadhyay Debabrata , Angom Ramcharan Singh , Wang Ying
Published: November 2022

CRISPR/Cas9 technology-mediated genome editing has significantly improved inactivation of genes both
in vitro
and
in vivo
in many organisms in a targeted manner. This study reports a novel CRISPR-based vector system for tissue-specific gene inactivation in zebrafish. In this vector, the cardiac-specific,
cardiac myosin light chain 2
(
cmlc2)
promoter drives the Cas9 expression to silence the
neuropilin1(nrp1)
gene in cardiomyocytes in a heat-shock inducible manner. This vector system establishes an inimitable tool to regulate the gene knockout in both the developmental and adult stages. Hence, it widens the possibility of loss-of-function studies in zebrafish at different stages of development. In contrast to mammals, adult zebrafish hearts can regenerate after an injury. Neuropilins (nrp) are co-receptors that have been reported earlier to play crucial roles in zebrafish heart regeneration. Using this novel tool, we investigated the role of neuropilin isoforms,
nrp1a
and
nrp1b,
in response to cardiac injury and regeneration in adult zebrafish hearts. We observed that both the isoforms (
nrp1a
and
nrp1b
) are upregulated in the injured heart. Interestingly, the
nrp1b
-knockout significantly altered heart regeneration and reduced heart function, including heart rate, ejection fraction, and fractional shortening in the adult zebrafish. Moreover, the knockdown of
nrp1b
, but not
nrp1a,
further induces the cardiac remodeling genes activation in response to cryoinjury. These results identify a crucial role for
nrp1b
isoform in zebrafish heart function. To our knowledge, this is the first study where we have reported a heat shock-mediated conditional knockdown of nrp1a and nrp1b isoforms in the heart by using CRISPR/Cas9 technology and reveal a new molecular role of neuropilins in cardiac regeneration.
Abstract 152: Myeloid Cell ADAM17 Impairs Hepatic Triglyceride-Rich Lipoprotein Clearance in an LDL Receptor-Deficient Mouse Model of Prediabetes
Authors: Tang Jingjing , Kothari Vishal , Vaisar Tomas , Shimizu-Albergine Masami , Kramer Farah , Frey Jeremy M , Kanter Jenny E , Bornfeldt Karin E
Published: November 2022

ADAM17 (A Disintegrin-like And Metalloproteinase 17) is a transmembrane protease that cleaves its substrates at the cell surface. Cell type-specific substrates cleaved by ADAM17, such as TNFα, can function locally and systemically. When fed a diet enriched in saturated fat, sucrose, and added cholesterol (DDC),

Ldlr
-/-

mice develop prediabetes characterized by elevated triglyceride-rich lipoproteins (TRLs), obesity, glucose intolerance and atherosclerosis. Our results demonstrate that when ADAM17 is deleted from hematopoietic cells, DDC-fed

Adam17
H-/-
Ldlr
-/-

bone marrow transplant chimeras (n=15-30) are less obese and more insulin sensitive than wildtype

Ldlr
-/-

chimeras.

Adam17
H-/-

chimeras show decreased plasma cholesterol and triglycerides, primarily due to a reduction in plasma VLDL assayed by FPLC fractionation. Atherosclerotic lesions are significantly (p<0.05) decreased in the brachiocephalic artery, aortic sinus and the aorta. When ADAM17 is deleted from myeloid cells, DDC-fed

Adam17
M-/-
Ldlr
-/-

bone marrow transplant chimeras recapitulate the phenotypes of

Adam17
H-/-

mice, suggesting that the phenotypes are mediated by myeloid cells. Mechanistic studies indicate that the reduced plasma VLDL levels in ADAM17-deficient chimeras are not due to reduced VLDL production or reduced intestinal uptake of triglycerides. Furthermore, analysis of pre- and post-heparin plasma demonstrates that lipoprotein lipase activity is not increased in the ADAM17-/- chimeras. Instead, characterization of the TRLs by proteomics and differential ion mobility analysis revealed that ADAM17-deficient chimeras have reduced levels of 25-30 nm TRL remnant particles (e.g., p=0.027 for 25 nm particles; n=8-10) and reduced levels of APOC1, APOC2 and APOC3 per particle. Together, these results are consistent with the proposal that deletion of ADAM17 in myeloid cells results in increased hepatic clearance of TRLs and their remnants. Our work suggests that myeloid cells play important roles in regulating hepatic TRL remnant clearance and associated atherosclerosis in a model of prediabetes.
Abstract 158: Bioactive Lipids As Biomarkers Of Severe Adverse Reaction Associated With Apheresis Platelet Concentrate Transfusion
Authors: Duchez Anne-Claire , FAUTEUX-DANIEL Sebastien , SUT Caroline , EBERMEYER Theo , ARTHAUD Charles-Antoine , EYRAUD Marie-Ange , PRIER Amelie , AUDOUX Estelle , BERTRAND-MICHEL Justine , PAYRASTRE Bernard , GARRAUD Olivier , Boilard Eric , HAMZEH-COGNASSE2 Hind , COGNASSE Fabrice
Published: November 2022

Blood transfusion is a life-saving procedure in which whole blood, or blood components are provided to a patient. Platelet concentrates (PC) may sometimes induce adverse reactions, that are occasionally severe. PC comprise active biomolecules such as cytokines and lipid mediators. Platelet storage lesions are structural and biochemical changes in PC and vary in collection and processing conditions. We investigated lipid mediators as bioactive molecules along storage conditions and in adverse reactions after transfusion. The PC left over were analysed after transfusion, classified with/out severe reaction. Our data presented a decrease of lysophosphatidylcholine species to produce lysophosphatidic acid species. This balance could lead and be correlated with complication symptoms, as other lipids. Lysophosphatidic acid increased with primarily platelet-inhibitor-lipids when single donor apheresis platelet upon storage. Anti-inflammatory and platelet-induced-inhibition lipids were weakly expressed in case of severe adverse reaction. Thus, a decrease in lysophosphatidylcholine and an increase in lysophosphatidic acid could be reliable predictors of serious adverse transfusion reactions transfusion. These data highlighted the importance of PC manufacturing and storage processing, not only to maximize clinical efficacy but also to minimize adverse reactions.
Abstract 197: Proteomic Characterization Of Human Brain Lipoproteins Speciated By Size Exclusion Chromatography
Authors: Merrill Nathaniel , He Yi , McDermott Jason , Swertfeger Debi , Heinecke Jay , Davidson Sean , Melchior John T
Published: November 2022

Like plasma, the cerebrospinal fluid (CSF) contains lipid-protein complexes called lipoproteins tasked with transporting lipids and protecting against cerebrovascular dysfunction. These brain lipoproteins (BLps) are generated
de novo
in the central nervous system (CNS), but little is known about them, because they exist in CSF at <0.5% the concentration of lipoproteins in plasma making them difficult to study. BLps are thought to resemble plasma high-density lipoproteins (HDL), a heterogenous population of compositionally distinct particles important for transporting lipid, protecting against oxidative stress, and modulating immune and inflammatory responses in the periphery. Mass spectrometry (LC-MS/MS) studies have shown that 260 different proteins can interact with different HDLs in the periphery and specific combinations of these proteins cluster on the lipid surface to govern particle function. In the current study, we developed a lipoprotein profiling technology that overcomes the BLp abundance problem in CSF to capture BLp subspecies with unprecedented sensitivity and resolution. Using high-resolution size exclusion chromatography, we identified at least 10 distinctly sized BLp populations in human CSF with a majority of the particles eluting in size ranges between plasma low-density lipoproteins (LDL) and HDL. BLps were isolated from fractionated CSF using a lipid-binding resin. Proteomic analysis of the particles using LC-MS/MS identified 299 different lipid-associated proteins across fractions with 189 of those proteins not currently reported on the human HDL Proteome Watch. Co-migration analysis further revealed at least 27 unique protein communities across the fractions indicating that multiple distinct subspecies exist within the same sized subfractions. DAVID functional analysis shows that the communities are highly enriched in proteins important in the immune response and neurometabolic functions such as neuron generation and development. In summary, our study shows that similar to plasma HDL, BLps are a highly speciated population of compositionally distinct particles but are enriched in CNS-derived proteins that are essential for maintaining cognitive health.
Abstract 206: Into The Wormhole: Understanding SR-B1’s Cholesterol Transport Functions Through Structural Studies
Authors: Powers Hayley R , Jenjak Shawn , Volkman Brian , Sahoo Daisy
Published: November 2022

Scavenger receptor Class B type 1 (SR-B1) is a glycosylated integral membrane protein that serves as the primary receptor for high density lipoprotein (HDL) and plays a central role in the reverse cholesterol transport pathway (RCT). In RCT, HDL removes cholesterol within atherosclerotic plaques and delivers it to the liver for excretion. HDL-cholesterol is delivered into hepatocytes through interaction with SR-B1, highlighting the need to improve our understanding of SR-B1 structure-function relationships to effectively lower plasma cholesterol levels and cardiovascular disease risk. Currently, structural information about SR-B1 remains limited to a structure of short transmembrane-spanning peptide segment, homology models, and transient transfection studies in cultured cells. Towards our long-term goal of resolving a full-length structure of human SR-B1, we have expressed and purified full-length human SR-B1 from an Sf9 insect cell baculoviral infection system. To verify function, human full-length SR-B1 expressed in plated Sf9 cells displayed a significant increase in DiI-HDL binding and DiI-lipid uptake compared to empty vector controls. Using PFO-PAGE, we demonstrated that SR-B1 forms dimers and higher order oligomers in Sf9 cells. After functional verification, we successfully incorporated purified protein into detergent micelles. Thermal shift assays revealed purified SR-B1 remained stable over a 6-week period. Purified SR-B1 also maintained the ability to bind to apolipoprotein A-I (the main protein component of HDL) and holoparticle HDL with high affinity (K
D
= 42.77 ± 7.15 μg/mL) by microscale thermophoresis (MST). Additionally, SR-B1 was able to bind oxidized low density lipoprotein (oxLDL). Further, while glycosylation remains important for SR-B1 expression, we are the first to demonstrate by MST that SR-B1’s glycosylation status did not impact HDL binding affinity (K
D
of 42.77 ± 7.15 μg/mL for glycosylated vs. 41.25 ± 4.92 μg/mL for unglycosylated SR-B1). Availability of functional purified human SR-B1 is a critical step towards a high-resolution structure and these studies lay the foundation for understanding the dynamics of SR-B1-mediated cholesterol transport and novel ways to modulate cardiovascular disease risk.
Abstract 220:
Smad3
Regulates Smooth Muscle Cell Fate And Governs Adverse Remodeling And Calcification Of Atherosclerotic Plaque
Authors: Cheng Paul , Wirka Robert , Kim Juyong B , Nguyen Trieu , Kundu Ramendra K , Zhao Quanyi , Sharma Disha , Pedroza Albert J , Nagao Manabu , Iyer Dharini , Fischbein Michael P , Quertermous Thomas
Published: November 2022

Atherosclerotic plaques consist mostly of smooth muscle cells (SMC), and genes that influence SMC phenotype can modulate coronary artery disease (CAD) risk. Allelic variation at 15q22.33 has been identified by genome-wide association studies to modify the risk of CAD, and is associated with expression of
SMAD3
in SMC
,
however the mechanism by which this gene modifies CAD risk remains poorly understood. SMC-specific deletion of
Smad3
in a murine atherosclerosis model resulted in greater plaque burden, more positive remodeling, and increased vascular calcification. Single-cell transcriptomic analyses revealed that loss of
Smad3
altered SMC transition cell state toward two fates: a novel SMC phenotype that governs both vascular remodeling and recruitment of inflammatory cells, as well as a chondromyocyte fate. The remodeling population was marked by uniquely high
Mmp3
and
Cxcl12
expression, and its appearance correlated with higher risk plaque features such as increased positive remodeling and macrophage content. Further, investigation of transcriptional mechanisms by which Smad3 alters SMC cell fate revealed novel roles for Hox and Sox transcription factors whose direct interaction with Smad3 regulate an extensive transcriptional program balancing remodeling and vascular extracellular matrix. These findings have significant implications for atherosclerotic and Mendelian aortic aneurysmal diseases. Together, these data suggest that
Smad3
expression in SMC inhibits the emergence of specific SMC phenotypic transition cells that mediate adverse plaque features, including positive remodeling, monocyte recruitment, and vascular calcification.
Abstract 222: The Molecular Mechanisms Of Coronary Artery Disease Risk At The
Pdgfd
Locus
Authors: Kim Hyun-Jung , Cheng Paul , Kundu Ramendra K , Nguyen Trieu , Kim Juyong B , Sharma Disha , Shi Huitong , Weldy Chad , Quertermous Thomas
Published: November 2022

Background:
Atherosclerotic cardiovascular diseases, including coronary artery disease (CAD), are the worldwide leading cause of death. Recent genome wide association studies (GWAS) have now identified over 250 genetic loci that are associated with CAD, and efforts in this lab and others are investigating the mechanisms of association at these loci. The CAD GWAS locus at 11q22.3 is of potential interest because it encodes the
PDGFD
gene, a member of the platelet-derived growth factor family whose expression is up-regulated in atherosclerosis. But whether disease allelic variation regulates
PDGFD
expression and how this growth factor might influence the risk of CAD has not been studied.

Methods:
The causal variant and gene were investigated with colocalization and
in vitro
transcription studies in cultured human coronary artery smooth muscle cells (SMC). Single-cell transcriptomic profiling (scRNAseq) and histological analyses were conducted in SMC-specific lineage tracing in an
in vivo Pdgfd
knockout mouse model of atherosclerosis.

Results:
The causal variant rs2019090 was linked to
PDGFD
expression in SMC.
In silico
analysis of transcription factor binding site prediction and
in vitro
haplotype-specific chromatin immunoprecipitation combined with qPCR analysis revealed another causal CAD factor forkhead box c1 (FOXC1) preferentially binds to the A-allele of variant rs2019090 and controls the expression level of
PDGFD
. scRNAseq data identified that loss of
Pdgfd
resulted in a marked increase in differentiated SMC but a decrease in fibromyocyte (FMC) and chondromyocyte (CMC) transition populations, as well as a significant decrease in plaque macrophages. Differentially expressed gene analysis in SMC lineage cells was highly enriched for CMC genes such as
Col2a1
and
Ibsp.
Lesion histology analysis identified an increase in medial SMC, but a decrease in fibrous cap SMC, with decreased
Cd68
and
Col2a1
stained area.

Conclusion:
PDGFD
is a causal CAD gene that increases disease risk by promoting SMC transition primarily to the CMC phenotype, and recruitment of monocyte-macrophage lineage cells to disease lesions. These findings support suppression of
PDGFD
as a possible therapy for human atherosclerotic disease.
Abstract 224: Myeloid Cell PKM2 Deletion Enhances Efferocytosis And Reduces Atherosclerosis
Authors: Doddapattar Prakash , Dev Rishabh , Ghatge Madankumar , Patel Rakeshkumar , Jain Manish , Dhanesha Nirav , Lentz Steven R , Chauhan Anil K
Published: November 2022

Rationale:
The glycolytic enzyme pyruvate kinase muscle 2 (PKM2) is upregulated in monocytes/macrophages of patients with atherosclerotic coronary artery disease. However, the role of cell type-specific PKM2 in the setting of atherosclerosis remains to be defined.

Objective:
We determined whether myeloid cell-specific PKM2 regulates efferocytosis and atherosclerosis.

Methods and Results:
We generated novel myeloid cell-specific PKM2
-/-
mice on Ldlr-deficient background (PKM2
mye-KO
Ldlr
-/-
). Controls were littermate PKM2
WT
Ldlr
-/-
mice. To rule out sex-based differences, male and female mice were placed on a high-fat "Western" diet for 14 weeks, starting at eight weeks. PKM2 was upregulated in macrophages of Ldlr
-/-
mice fed the Western diet compared with a control chow diet. Myeloid cell-specific deletion of PKM2 led to a significant reduction in lesions in the whole aorta and aortic sinus despite high cholesterol and triglyceride levels. Furthermore, we found decreased macrophage content in the lesions of myeloid cell-specific PKM2
-/-
mice associated with decreased MCP-1 levels in plasma, reduced transmigration of macrophages in response to MCP-1, and impaired glycolytic rate. Macrophages isolated from myeloid-specific PKM2
-/-
mice fed the Western diet exhibited reduced expression of pro-inflammatory genes, including MCP-1, IL-1β, and IL-12. Myeloid cell-specific PKM2
-/-
mice exhibited reduced apoptosis concomitant with enhanced macrophage efferocytosis and upregulation of LRP1 in macrophages
in vitro
and atherosclerotic lesions
in vivo
. Silencing LRP1 in PKM2-deficient macrophages restored inflammatory gene expression and reduced efferocytosis. As a therapeutic intervention, inhibiting PKM2 nuclear translocation using a small molecule reduced glycolytic rate, enhanced efferocytosis, and reduced atherosclerosis in Ldlr
-/-
mice.

Conclusion:
Genetic deletion or limiting PKM2 nuclear translocation in myeloid cells reduces atherosclerosis by suppressing inflammation and enhancing efferocytosis.
Abstract 226: Novel Athero-protective Role Of Chk1-induced Senp2 S344 Phosphorylation Under Laminar Flow
Authors: Nguyen Minh T.H. , Li Shengyu , Masaki Imanishi , Velatoorua Loka reddy , Banerjeea Priyanka , Abe Rei , Ko Kyung Ae , Kotla Sivareddy , Gi Young Jin , Cooke John J. , Fujiwarab Keigi , Borchrdt Mae K , Wang Guangyu , Abe Jun-ichi , Le Nhat-Tu
Published: November 2022

Atherosclerosis (AthS) tends to form at arterial regions exposed to disturbed (DF) but not laminar (LF) flow. Previously, we reported the significant role of SUMOylation in DF-induced endothelial (EC) activation and subsequent AthS formation via upregulating p53 and ERK5 SUMOylation. LF can inhibit the basal levels of p53 and ERK5 SUMOylation, but how LF inhibits SUMOylation remains unknown. A chemical genetics approach with high-resolution mass spectrometry revealed that the cell-cycle checkpoint kinases CHK1 can phosphorylate SENP2 S344 site, but the functional role remains largely unknown. We aimed to study the functional role of CHK1 in SENP2 function and subsequent AthS. First, we generated phospho-specific SENP2 S344 antibody, and found that LF, but not DF, increases SENP2 S344 phosphorylation. We found the increase of CHK1 S280 phosphorylation (related to nuclear translocation) after LF, suggesting LF-induced nuclear translocation of CHK1. The CHK1 specific inhibitor of GDC0575 and the depletion of CHK1 inhibited LF-induced SENP2 S344 phsophorylation and increased both p53 and ERK5 SUMOylation in ECs. These data suggested the crucial role of CHK1 in LF-induced SENP2 S344 phosphorylation and upregulating de-SUMOylation activity. Next, we generated CRISPR/Cas9-induced
Senp2
S344A knock-in (KI) and found that the mutation of SENP2 S344A accelerated p53 and ERK5 SUMOylation. LF-induced reduction of p21, cleaved caspase 3, and ICAM-1 were all reversed by SENP2 S344A mutation. Lastly, we found that the significant acceleration of AthS formation in both ascending aorta/arch (DF area) and descending aorta (LF area) in KI mice compared to wild type (WT) mice after receiving AAV-PCSK9 and high fat diet. We also performed bone marrow transplantation (BMT) after 13 Gy whole body radiation, and larger AthS but only in the aortic arch was observed in BMT mice from WT donor to KI recipient, but not in BMT mice from WT donor to WT recipient, supporting the role of endothelial SENP2 S344 phosphorylation on AthS. The radiation reduced CHK1 expression in ECs, which may explain the different regulatory pattern under BMT or non-BMT. Taken together, these results suggest the critical role of CHK1-mediated SENP2 S344 phosphorylation on LF-induced anti-AthS effects.
Abstract 232: Qualitative Single-cell Assessment By Imaging Mass Cytometry (imc) Analysis Reveals Senescence-associated Stemness (sas) Induced By Erk5 S496 Phosphorylation In Atherosclerotic Plaque
Authors: Kotla Sivareddy , Imanishi Masaki , Li Shengyu , Ko Kyung Ae , Samanthapudi Venkatasubrahman K , Savage Hannah , Schadler Keri , Shen Ying H , Deswal Anita M , Lin Steven , Reyes-Gibby Cielito , Yeung Sai-Ching , Pownall Henry J , Fujiwara Keigi , Burks Jared K , Le Nhat-Tu , Wang Guangyu , Abe Jun-ichi
Published: November 2022

The central role of ERK5 S496 phosphorylation in promoting senescence has been reported. Recently the conventional model of senescence as an event of cell cycle arrest has been challenged, given that senescence induced by various stresses reprograms cancer cells to acquire senescence-associated stemness (SAS), which allows them to escape senescence-induced cell cycle arrest with enhanced growth potential. To determine the role of ERK5 S496 phosphorylation in SAS, we utilized imaging mass cytometry (IMC) by using antibodies against various cell surface markers, senescence, and proliferation. We prepared tissue sections from the plaques in wild-type (WT) and ERK5 S496A knock-in (KI) mice, and the ablated plaque materials were evaluated by the CyTOF mass cytometer. We used the VISIOPHARM program, and single-cell features were computationally segmented using a watershed algorithm. Nine clusters were detected in WT (total 7903 cells) and ERK5 S496A KI (total 6166 cells) plaques. Single-cell analysis of p53 and Ki67 revealed three groups of the cells with different pattern of p53 and Ki67 expression (Fig.A). We calculated log scaled ratio of Ki67 and p53 of selected cells by log
10
(expression of Ki67/ expression of p53), and further calculated one-dimension density of this log scaled ratio to cluster cells into three groups by using cutoff of ration with log
10
(-1.01) and log
10
(-0.024) (Fig.B). Group 2 myeloid cells showed that p53 and Ki67 were co-expressed lineally, suggesting that this group of cells escaped from the growth-suppression effects of p53, which is also known as a senescence marker. Of note, the % of ERK5 S496A KI cells in group 2 were significantly lower than those in WT (Fig.C). Also, in vitro oxidized LDL remarkably increase β-gal
+
Ki67
+
cells, which was significantly inhibited in ERK5 S496A KI macrophage compared to those in WT. Collectively, these data implicates ERK5 S496 phosphorylation in a unique role-regulation of SAS during atherogenesis.
Abstract 238: Serum Opacity Factor Normalizes Dysfunctional High Density Lipoproteins In Srbi -/- Female Mice And Rescues Fertility
Authors: Yelamanchili Dedipya , Gillard Baiba K , Gotto Antonio M , Pownall Henry J , Rosales Corina
Published: November 2022

Although plasma HDL-C levels negatively correlate with atherosclerotic cardiovascular disease (ACVD), attempts to reduce ACVD risk by raising plasma HDL have disappointed. Thus, hypotheses about salutary HDL effects have shifted from higher-is-better to function-is-more-important. The SRBI-/- mouse is an extreme model of HDL dsyfunctionality; compared to WT mice, SRB1-/- mice have higher plasma HDL levels and an HDL surface that is free cholesterol (FC)-rich (60 vs. 15 mol%). This would be expected to increase HDL-FC bioavailability to cytotoxic levels. HDL dysfunctionality among SRB1-/- mice is associated with multiple metabolic abnormalities—impaired cell membrane structure and function and atherosusceptibility, despite having high plasma HDL-C levels, and infertility among female SRB1-/- mice. Dietary probucol, an HDL-lowering drug, partially reverses infertility in SR-B1-/- mice. The mechanisms underlying ovaritoxicity is not known and interventions that fully reverse this state has not been identified. We hypothesized that a bacterial protein, serum opacity factor (SOF), which acts on HDL and lowers cholesterol in WT mice by 43% in 3 hours, would normalize HDL functionality and rescue infertility in SRBI-/- female mice. SOF DNA was cloned into a TBG-AAV8 plasmid and virus produced by UPENN Vector Lab. The AAV-SOF was delivered by IP injection leading to the endogenous expression of SOF. Probucol-fed and AAV-GFP mice were used as controls. SOF expression drastically decreased plasma cholesterol and normalized the size of HDL in SRBI-/- females comparable to that of WT mice. Fertility was restored in female mice injected with SOF while GFP controls remained infertile. Compared to Probucol-fed mice, AAV-SOF females had a higher percentage of fertile females and shorter time to first litter drop. We concluded that SOF performed equally or better than probucol in rescuing fertility and normalizing HDL.
Abstract 246: High Density Lipoprotein Cholesterol Efflux Capacity And Delirium After Surgery
Authors: Smith Loren E , Yancey Patricia G , Linton MacRae F
Published: November 2022

Background:
Postoperative delirium is associated with increased perioperative morbidity and mortality. Poor cerebrovascular health may increase delirium risk. We hypothesized that higher HDL concentration is associated with less postoperative delirium.

Methods:
After IRB approval, we prospectively recruited 65 adults undergoing major, elective cardiac and vascular surgery. Research staff performed preoperative dementia screening using the Mini Mental Status Exam (MMSE) and the Trail Making Test Part B. Postoperative Richmond Agitation Scale Scores and Confusion Assessment Method tool scores were collected from the medical record on postoperative days 1 and 2. Because HDL function often correlates with disease better than cholesterol level, HDL cholesterol efflux capacity (CEC) was measured in J774 macrophages loaded with
3
H-cholesterol in acetylated-LDL and then incubated with ApoB-depleted patient serum for 24 hours. Multivariate logistic regression was used to assess the association between HDL-C and HDL CEC and postoperative delirium, adjusted for age, preoperative MMSE score, preoperative Trail making score, and cardiopulmonary bypass exposure.

Results:
HDL-C was correlated with HDL CEC (Spearman’s R=0.37, p=0.002). One patient in the first (lowest) tertile, one patient in the second, and six patients in the third HDL-C tertile developed delirium. In contrast, four patients in the first tertile, three patients in the second, and one patient in the third tertile of CEC developed delirium. A higher preoperative HDL-C was independently associated with a higher odds of postoperative delirium (odds ratio 1.14 (95% CI 1.03, 1.26), p=0.01). In contrast, a higher preoperative HDL CEC was associated with a lower odds of delirium (odds ratio 0.82 (95% CI 0.68-0.99), p=0.04).

Conclusions:
Higher HDL CEC, but not HDL-C, was associated with less postoperative delirium. In this patient population, HDL function may play an important role in the development of postoperative delirium.
Abstract 272: Identifying Vascular Determinants Of Optimal Cerebral Perfusion Pressure In Traumatic Brain Injury Using A Continuous Measurement Algorithm.
Authors: Cronin Michael F , Teichmann Daniel , Stafford Rebecca , Heldt Thomas , Holsapple James
Published: November 2022

Introduction:
Individualized therapy targeting certain cerebral perfusion pressure (CPP) values in patients with traumatic brain injury may improve outcomes. One method to obtain “optimal” CPP (CPPopt) uses a measurement algorithm with continuous arterial blood pressure (ABP) and intracranial pressure (ICP) to estimate CPPopt where the pressure reactivity index is lowest. Yet, there is a knowledge gap regarding the cerebrovascular conditions that determine the algorithm’s success and validity. Here, we use a validated brain vasculature model to assess the algorithm’s performance at varying simulated vascular conditions.

Methods:
We used a convenience sample of 12 patient records with 4 hours of high-resolution ABP from the MIMIC III database. Each ABP set generated a synthetic ICP from our model. The algorithm then performed 38,815 CPPopt estimates across 12 patient records based on given input parameters. We examined “G”, an autoregulation parameter representing the sensitivity of ICP response to ABP change, and “Tau”, a time parameter representing the delay in ICP response to ABP change. We varied G and Tau independently and measured changes in frequency of algorithm success in finding CPPopt (CPPopt coverage), pressure reactivity index (PRx), and CPPopt itself.

Results:
1.As G increases, CPPopt coverage, and mean minimum PRx decrease.2.As Tau increases, CPPopt coverage, and mean minimum PRx increase.3.CPPopt coverage had a median success rate of 27.111% and IQR of 29.118% across G and median success rate of 45.166% and IQR of 35.3362% across Tau for 38,815 algorithm estimates across the 12 patient records.

Conclusion:
The CPPopt algorithm may have a higher likelihood of finding CPPopt in pathological conditions where G is expected to be low and Tau to be large. The low and variable algorithm success rate poses a challenge for clinical use. More work is needed to understand the algorithm’s success and validity to determine its clinical viability.
Abstract 290: Ccr2 Expression Is Increased In Patients With Symptomatic Carotid Arterial Occlusive Disease
Authors: Engel Connor , Zaghloul Mohamed , Meade Rodrigo , Woodard Pamela K , Gropler Robert J , Liu Yongjian , Zayed Mohamed A
Published: November 2022

Objectives:
Chemokines and their receptors play critical roles in the progression of atherosclerosis. The chemokine receptor, C-C chemokine receptor type 2 (CCR2), mediates an essential role in the differentiation of pro-inflammatory monocytes into macrophages. Recent studies suggest that proinflamatory macrophages in plaques predict plaque progression. While there is no clinical consensus on which patient with asymptomatic carotid artery stenosis would benefit from carotid endarterectomy (CEA), we hypothesized that CCR2 plaque content may predict plaque progression.

Methods:
Carotid plaque of ten patients undergoing elective CEA were harvested from the operating room, paraffin-embedded, sectioned into 5μm segments, immuno-stained for CCR2 and CD68, and imaged. Three 200х500μm regions-of-interest (ROIs) were randomly selected within 200μm of the lumen (superficial intima), outside 200μm of the lumen (deep intima), and the arterial media. Two independent observers evaluated the number of CCR2 and/or CD68 positive cells. Counts in each ROI were averaged and analyzed with Student’s t-test.

Results:
Patients with symptomatic carotid artery stenosis (6/10) demonstrated higher CCR2 and CCR2-CD68 double positive cells within the superficial intima of carotid artery plaques (A, p<0.05, n=10). Similarly, patients with symptomatic disease also showed increased CCR2 in the plaque media (B, p<0.05, n=10). Interestingly, we also observed a higher number of CCR2 positive cells in the media layer of smokers (6/10) compared to non-smokers (C, p<0.05, n=10).

Conclusions:
Our study demonstrates the close association between CCR2 cellular content and the incidence of symptomatic carotid artery disease. Additionally, CCR2 plaque content appears to correlate with risk-factors such as smoking. These studies have important implications regarding the impact of immune modulation on carotid plaque vulnerability in patients with asymptomatic carotid artery stenosis.
Abstract 291: Phosphodiesterase 10A Regulates Medial Artery Calcification
Authors: Cai Yujun , Xie Yangzhouyun , Berezowitz Alexa , Flores Alyssa M , Wang Xue-Lin , Guzman Raul J
Published: November 2022

Vascular calcification results from deposition of calcium hydroxyapatite crystals in the vessel wall. It is highly prevalent in patients with chronic kidney disease (CKD), diabetes, and peripheral artery disease (PAD). In lower extremity arteries, elevated calcification levels are associated with an increased risk of ischemic events including amputation. The cyclic nucleotides cAMP and cGMP, controlled by distinct cyclic nucleotide phosphodiesterase (PDE) isozymes, play important regulatory roles in a variety of human diseases. Using a qPCR PDE array, we found that PDE10A was the most highly induced isoform in a rodent model of arterial calcification. PDE10A was also markedly increased in calcifying vascular smooth muscle cells (VSMCs)
in vitro
, calcified arteries from rodents with CKD, and calcified human tibial arteries. Knockdown or inhibition of PDE10A markedly attenuated high phosphate-induced VSMC osteogenic transformation and calcification in both rat and human VSMCs. Importantly, deficiency of PDE10A significantly decreased arterial calcification in both
ex vivo
aortic ring and
in vivo
vitamin D
3
medial calcification models. Several labs including our own have previously demonstrated that matrix metallopeptidases (MMPs) are involved in vascular calcification. Using a loss-of-function strategy and bioinformatics analysis, we found that MMP-3 expression was regulated by PDE10A in calcifying VSMCs. Our further mechanistic studies showed that PDE10A could regulate vascular calcification by controlling p38 MAPK signaling and MMP-3 activity through cGMP/PKG signaling. These findings suggest that PDE10A plays a critical role in the development of medial artery calcification, and that targeting it may provide a novel therapeutic strategy to reduce medial calcification in patients with PAD and CKD with the ultimate goal of preventing major limb amputation.