bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P044: Activation of Interferon Regulatory Factor-3 Prolongs Stress-Induced Cardiomyocyte Apoptosis and Decreases Survival Post Myocardial Infarction
Authors: de Couto Geoffrey , Dawood Fayez , Sun Mei , Chen Manyin , Heximer Scott , Stanford William , Medin Jeffrey , Liu Peter 
Published: March 2022

Background. The innate immune response is critical in mediating host protection from foreign antigen or stress. It is evident that multiple innate immune pathways also tightly regulate the remodeling process post-myocardial infarction (MI), however it is not clear how this process is mediated. In order to further elucidate the underlying mechanism of this process, we screened both mouse and human post-MI microarray databases and identified a common, significantly upregulated transcription factor of interest, interferon regulatory factor-3 (IRF3). To assess its function in vivo, we tested the hypothesis that activation of IRF3 post-MI coordinates maladaptive repair by promoting cardiomyocyte apoptosis.
Methods/Results. We randomly allocated IRF-3 knockout (IRF-3-/-) mice and their wild-type (WT) counterparts to either left anterior descending (LAD) coronary artery ligation or sham operation. Four weeks post-MI, deficiency in IRF3 led to improved survival [88% (44/50) vs. 29% (14/49) in WT, p<0.0001], preserved cardiac function via echocardiography [Fractional Shortening: 37% vs. 22.9% in WT, LVEDD: 5.15mm vs. 5.85mm in WT, LVESD: 4.15mm vs. 5.14mm in WT; p<0.057#x005D;, and attenuated infarct size [27% vs. 44% in WT, p<0.05]. Assessing the stress response 1-week post-MI, we found that NFkB signaling (IKKa, Ikba, p65) as well as pan-ubiquitination of proteins, were both significantly downregulated in the border infarct zone of IRF3-/- mice. These findings were complemented with increased protein and mRNA expression of de-ubiquitinating enzyme A20. Interestingly, in IRF3-/- mice these results were linked with decreased apoptosis (Caspases, TUNEL staining), however with a more specific attenuation of the mitochondrial-specific apoptotic pathway (Bid, Cytochrome C, Caspase9).
Conclusions. These data establish a novel link between the innate immune transcription factor IRF3 and maladaptive regulation of cardiomyocyte apoptosis through mitochondrial apoptotic pathway activation. As an acute responder coordinating maladaptive remodeling, IRF3 may prove to be an efficacious therapeutic target to improve outcome following MI.
Abstract P131: An Akt-Phosphomimetic Sequence of the Cavb2 C-Terminal Region Protects L-Type Calcium Channels from Protein Degradation
Authors: Rusconi Francesca , Cazade Magali , Caprari Silvia , Polticelli Fabio , Chemin Jean , Condorelli Gianluigi , Catalucci Daniele 
Published: March 2022

Alteration in the density or function of L-Type Calcium Channels (LTCCs) has been related to cardiovascular diseases such as heart failure and diabetic cardiomyopathy. It could therefore be envisioned that increasing LTCC density may improve cardiac function in heart failure. Recently, we determined that the Ser-Thr kinase Akt plays a key role in regulating cardiac inotropism through the modulation of LTCC density and function. Specifically, we found that the LTCC pore-forming channel subunit Cava1.2 contains highly evolutionary conserved PEST sequences (signals for rapid proteolytic degradation) that are responsible for direct Cava1.2 protein degradation. Phosphorylation of the C-terminal coiled coil of the Cavb2 chaperone subunit enhances LTCC protein stability by preventing PEST-mediated Cava1.2 degradation.
The aim of this study was to further dissect this Akt-dependent fine-tuning mechanism regulating LTCC density, searching for potential Akt-phosphomimetic (APM) molecules that could enhance LTCC density.
Using yeast two-hybrid screening, we found that APM Cavb2 sequences interact with the globular domain of Cavb2 itself in a solvent-exposed region that we named Tail Interacting Domain (TID). Biochemical and functional assays as well as site-specific mutagenesis in TID identified the minimal aminoacid sequence responsible for the TID-tail interaction.
In addition, through an approach comprising western blot analyses, fluorescent-based calcium assays, calcium current (ICaL) measurements, molecular modeling and peptide arrays, we identified the minimal APMs that efficiently protects Cava1.2 from protein degradation.
Based on our in vitro results, we suggest that the identified APM sequence/peptide could be used as a “therapeutic approach”; for increasing or reestablishing impaired cardiac contractility in mouse models of cardiomyopathy in which the LTCC density is altered, thus enhancing inotropism.
Abstract P146: Fas-ligand Induces Cardiomyopathy via the Activation of the Erk1/2 Pathway
Authors: Huby Anne-Cecile , Purevjav Enkhsaikhan , Towbin Jeffrey A 
Published: March 2022

Fas receptor and its ligand, Fas-L, are the most well characterized co-stimulatory molecules playing an essential role in the induction of programmed cell death or apoptosis. Increase of circulating Fas-L in serum and in cardiomyocytes are described in different models of cardiac diseases such as myocarditis, pressure overload, AND myocardial infarction/ischemia. In addition, associated activation of inflammatory pathwayshas been observed. However, no studies have yet revealed mechanisms underlying the pathogenecity of FasL in the heart.
Transgenic mice (C57/Bl6) with cardiac-specific over-expression of Fas-L were investigated. We characterized cardiac function by echocardiography. Cellular and molecular studies were performed on heart tissue using genomic, histologic, immunological, and protein analysis techniques.
Fas-L transgenic mice had a significant rate of mortality (p<0.0001) compared to WT due to heart failure. Histological studies of Fas-L hearts revealed predominant perivascular fibrosis associated with inflammation. Expression studies of secondary messengers of the activated Fas/FasL system demonstrated up-regulation of an active CASP-3 confirming induction of apoptosis in myocardium. Down-regulation of FLIP, NIK and NF-κB was evident. Interestingly, RIP was up-regulated leading to the activation of ERK1/2 (4.27±0.158 vs 10.086±1.591 in WT and FasL mice hearts, respectively, p=0.022). Analysis of inflammatory mediators revealed increase in secretion of interleukines (IL-17, IL-6, IL-1β and TNF-α) and diffuse infiltration of CD3 positive cells. Studies of fibrotic pathways showed an increased expression of periostin and osteopontin, but not activation of TGFβ1, showing that periostin and osteopontin can be activated independently of TGFβ1 via the ERK1/2 pathway,
Our findings suggest that Fas-L induces cardiac failure, via an activation of the ERK1/2 pathway, which is an important player involved in the hypertrophic response in the heart. Targeting the ERK1/2 pathway in cardiac diseases associated with Fas-L expression may therefore be able to prevent progression to cardiomyopathy.
Abstract P298: Probucol Inhibits Oxidized Low-Density Lipoprotein--Induced Immune Maturation of Dendritic Cells via Heme Oxygenase 1
Authors: Sun Aijun , Jin Xueting , Wang Keqiang , Zou Yunzeng , Ge Junbo 
Published: March 2022

The anti-atherosclerotic mechanism of Probucol remains to be elucidated. Studies confirmed the key role of Dendritic Cells (DCs) and cytoprotective role of heme oxygenase 1(HO1) in the development of atherosclerosis. Here, we tested the hypothesis that probucol exerts is anti-atherosclerotic effect by inhibiting maturation of DCs via HO1.
[Method]Immature DCs were derived from purified human monocytes; HO1 siRNA, probucol ,ox-LDL or LDL was added to different groups of medium. The maturation markers of DCs were analyzed by Flow cytometric analysis, including CD1a, CD40, CD86, HLA-DR, endocytosis function and the cytokines secretions of culture supernatants (IL-4,TNF-α) by ELISA. The expression of HO1, STAT1 (signal transducers and activators of transcription 1) and STAT1(Tyr 701) phosphorylation were analyzed by Westernblot and the expression of CIITA(MHC II transactivator) by Real Time PCR.
[Results] Ox-LDL promoted immune maturation of DCs : MIF of CD86,CD 40, CD1a and HLA-DR was significantly increased after ox-LDL stimulation (7697±672 vs 2120±662, 7844±754 vs 843±109 ,323±31 vs 272±14,2636±173 vs 1443±45,respectively); decreased endocytosis function (26256±354 vs 34511±3078); increased TNF-α(43±5 vs 30±0.3 pg/ml), suppressed IL-4 (852±9 vs 793±11 pg/ml)2. Probucol inhibited immune maturation of DCs ,which was most obviously in ox-LDL treated group : MIF of CD 86 CD 40, CD1a and HLA-DR was significantly decreased after probucol treatment (7697±672 vs 3202±660,7844±754 vs 843±109,323±31 vs 56±31, 2636±173 vs 304±60), endocytosis (26256±1689 vs 109568±4195), TNF-α(43±5 vs 14±0.6 pg/ml), IL-4(793±11 vs 1037±17 pg/ml) (P<0.05). 3. After silencing of the expression of HO-1 by siRNA, immune maturation markers of DCs were increased than respective non-silencing group ,such as CD86, CD 40, HLA-DR (549±19 vs 353±23, 2645±307vs 524±119,1179±66 vs 766±23,respectivly), endocytosis function were suppressed (7408±229 vs 10860±706), TNF-αwas increased (22±0.2 vs 14±0.6 pg/ml), IL-4 was decreased (926±2 vs 1037±17 pg/ml) (P<0.05), STAT1(701) phosphorylation and CIITA expression level were increased.
[Conclusion]Probucol significantly inhibited immune maturation of DCs and is associated with HO1, STAT1/CIITA signaling pathway.
Abstract P304: Salvianolic Acid B Suppresses Human Monocyte-Derived Dendritic Cell Maturation via PPARγ Activation
Authors: Sun Aijun , Liu Hongying , Wang Shijun , Shi Dazhuo , Xu Lei , Cheng Yong , Wang Keqiang , Chen Keji , Zou Yunzeng , Ge Gunbo 
Published: March 2022

Background and purpose: Salvianolic acid B (Sal B), a water-soluble antioxidant derived from a medicinal herb, is known to be effective in the prevention of atherosclerosis. Here, we tested the hypothesis that the anti-atherosclerotic effect of Sal B might be mediated by suppressing human monocyte-derived dendritic cells (DCs) maturation.
Experimental approach: DCs was derived by incubating purified human monocytes with GM-CSF and IL-4. DCs was pre-incubated with or without Sal B and stimulated by oxidized low density lipoprotein (ox-LDL) in the presence or absence of PPAR-γ siRNA. Expression of DCs membrane molecules were analyzed by FACS, cytokines were measured by ELISA, and TLR4 associated signaling pathway was determined by Western blotting. Key results: Ox-LDL promoted DCs maturation, stimulated CD40, CD86, CD1a, HLA-DR expression and IL-12, IL-10, TNF-α production and upregulated TLR4 signaling. These effects could be significantly inhibited by Sal B. Sal B also triggered PPAR-γ activation and promoted PPAR-γ nuclear translocation, attenuated ox-LDL-induced upregulation of TLR4 and myeloid differentiation primary-response protein 88 (MyD88), and inhibited downstream p38-MAPK signaling cascade. Knocking down PPAR-γ through siRNA transfection significantly blocked above mentioned effects of Sal B. Conclusions and implications: Our data suggested that Sal B effectively suppressed ox-LDL-induced DCs maturation through PPAR-γ activation.
Abstract 111: Reduced MK5 Expression Alters Migration, Proliferation, and Collagen Biosynthesis in Murine Ventricular Fibroblasts
Authors: Sahadevan Pramod , Nawaito Sherin A , Calderone Angelo , Gaestel Matthias , Allen Bruce G 
Published: March 2022

MAP kinase-activated protein kinase-5 (MK5), a protein serine/threonine kinase expressed in the heart, has been identified as a substrate for p38α/β and ERK3/4 MAPKs. However, the interacting partners and physiological function of MK5 are just beginning to be understood. This study examined the role of MK5 in murine cardiac ventricular fibroblasts.
Confocal immunocytofluorescence microscopy (CIFM) revealed that MK5 immunoreactivity localized primarily to the nucleus whereas ERK3 immunoreactivity was in the cytoplasm. Following serum stimulation, phospho-MK5 immunoreactivity was observed in the cytoplasm and appeared to be associated with the cytoskeleton and pseudopodia. ERK3 immunoreactivity redistributed to membrane ruffles and/or lamellipodia. ERK3 has been reported to be unstable in the absence of MK5: in cardiac fibroblasts, ERK3 immunoreactivity was unaffected by acute knockdown of MK5 with siRNA (MK5-kd), suggesting an as-yet unidentified binding partner may stabilize ERK3 in these cells.

MK5 immunoprecipitates from fibroblast lysates contained ERK3 immunoreactivity and proximity ligation assays indicated the presence of ERK3-MK5 complexes in the cytoplasm: these complexes were less abundant in MK5-kd fibroblasts. Cell migration, in response to serum and/or Ang-II was reduced significantly in MK5-kd fibroblasts. In addition, cell proliferation was decreased in fibroblasts isolated from MK5
-/-
mice compared to fibroblasts from wild-type litter mate mice (MK5
+/+
). Surprisingly, both the abundance of type 1 collagen (COL1A1) mRNA and the secretion of soluble COL1A1 were increased in MK5-kd fibroblasts whereas the ability of Ang-II to increase collagen secretion was unaffected. CIFM revealed staining for COL1A1 was diffuse in MK5
+/+
fibroblasts but condensed in the perinuclear region of MK5
-/-
cells. A similar pattern of subcellular distribution of COL1A1 immunoreactivity was observed in MK5-kd fibroblasts and was observed in passages 0 through 3. Taken together, these data suggest 1) one or more proteins in addition to MK5 serve to stabilize ERK3 in fibroblasts and 2) MK5 may be involved in cardiac fibroblast proliferation, migration, collagen biosynthesis and, possibly, myocardial remodelling.
Abstract 167: Inhibition of Endoplasmic Reticulum Stress Improves Vascular Function in Type II Diabetic Mice
Authors: Lee Dustin M , Battson Micah L , Jarrell Dillon K , Hou Shuofei , Ecton Kayl , Gentile Christopher L 
Published: March 2022

Cardiovascular disease is the leading cause of death in the United States with type 2 diabetes (T2D) representing a major risk factor in its development. Vascular dysfunction, characterized by arterial stiffness and endothelial dysfunction occur prior to overt cardiovascular disease and predict future cardiovascular events and mortality in diabetic individuals.
Dysfunction of the endoplasmic reticulum (ER), or ER stress, is associated with the development and progression of chronic metabolic diseases such as obesity and T2D. However, the role of ER stress in the development of vascular dysfunction observed in T2D is unclear. We hypothesized that inhibiting ER stress would improve both measures of vascular dysfunction observed in diabetic mice.
Male C57BL/6J Leprdb (DB) male mice lacking the leptin receptor were used as a model of T2D. Starting at 4 months of age DB mice were given intraperitoneal injections of ER stress inhibitor, tauroursodeoxycholic acid (TUDCA) at 250mg/kg/day for 4 weeks. C57BL/6J mice were used as controls (n=8 for all groups). Pulse wave velocity (PWV) was measured at baseline (prior to treatment) and after TUDCA treatment. Secondary order mesenteric resistance arteries (MRA) were used to determine endothelial dependent dilation (EDD). DB mice not treated with TUDCA were used for acute studies of EDD by incubating the MRA for 1hr with 0.5mM TUDCA.
At baseline, DB mice displayed increased arterial stiffness compared to C57BL/6J controls as measured by PWV (457±25 vs 348±26 cm/s, p<0.05). TUDCA treatment significantly decreased PWV (402±12 cm/s, p<0.05). Both acute and chronic TUDCA treatment improved DB EDD compared to untreated controls (58.4±13.8 and 49.1±6.1%, vs 19.8±9.6%, p<0.05). Expression of ER stress-associated genes CHOP and ATF4 were significantly elevated in DB perivascular adipose tissue and treatment with TUDCA significantly decreased ATF4 compared to DB. In the aorta, TUDCA significantly decreased GRP78 and XBP1 compared to DB.
These data support the hypothesis that ER stress contributes to the vascular dysfunction observed in T2D and suggest that ER stress may be a potential target in the treatment of T2D related vascular stiffening and dysfunction.
Abstract 324: Identification of Regulators of Cardiac DNA Methylation Using a Systems Genetics Approach in Mice
Authors: Rau Christoph D , Romay Milagros , Wong Eleanor , Tan Wilson , Lusis Aldons J , Foo Roger , Wang Yibin 
Published: March 2022

Systems biology approaches to studying heart failure in humans are hampered by numerous environmental factors and inciting incidents which combine to complicate the application of standard population-based tools and algorithms to the syndrome. We recently reported a study of heart failure in a large cohort of inbred mouse strains (the Hybrid Mouse Diversity Panel; HMDP) treated with the beta-adrenergic agonist isoproterenol. We demonstrated that it was possible to combine systems-level tools with the benefits of an animal model to capture and identify numerous genetic variations which drive heart failure.
Recent research on the epigenome has identified changes to DNA methylation which affect the progression of cardiovascular diseases and suggests that DNA methylation may serve as an important additional marker or even driver of genes involved in heart failure-associated phenotypes. We have supplemented our prior study by completing reduced representational bisulfite sequencing on 88 strains from our panel both with and without catecholamine challenge.
We have identified 75,000 CpGs which vary among the strains of the HMDP across the genome. Each CpG was mapped to the genome using GWAS to identify methylation quantitative trait loci (mQTLs). These mQTLs were then grouped together to identify mQTL hotspots, regions on the genome which regulate a sizable fractions of all observed CpGs. We then overlapped these hotspots with hotspots which regulate the expression of gene transcripts.

Three genomic regions were identified as multi-omic master regulators which affect both DNA methylation and gene expression for over 5% of the expressed transcripts and varying CpGs. Closer examination of these loci identified the serine peptidase inhibitor
Serpina3n
as a likely candidate gene for the regulation of DNA methylation, gene expression and cardiac phenotypes. Subsequent
in vitro
and
in vivo
work demonstrate that
Serpina3n
knockdown results in reduced cellular hypertrophy and changes to hypertrophy-related gene expression. Further analysis of the other master regulatory regions will likely reveal additional genes which will improve our understanding of how DNA methylation acts to regulate pathways underlying heart failure.
Abstract 436: Targeting Fatty Acid Oxidation by Acetyl-CoA Carboxylase 2 Deletion in Pathological Hypertrophy
Authors: Ritterhoff Julia , Villet Outi , Kolwicz Stephen C , Shao Dan , Tian Rong 
Published: March 2022

We have previously shown that preserving fatty acid oxidation (FAO) by cardiac-specific deletion of Acetyl-CoA Carboxylase 2 (ACC2) prevents the shift of substrate preference towards glucose, reduces cardiomyocyte hypertrophy and preserves cardiac function during chronic pressure overload.
To determine whether maintaining FAO specifically prevented cardiomyocyte hypertrophy, we treated adult rat cardiomyocytes (CMs) with and without adenoviral-mediated ACC2 knock-down (KD) with phenylephrine (PE, 10 μM). ACC2 KD effectively prevented CM hypertrophy after PE stimulation compared to control CMs (+9±6% vs. 42±6%) in medium supplemented with fatty acids (FA) (5.5 mM glucose, 0.4 mM mixed long-chain FAs and 0.1 mU/ml insulin). Whereas PE stimulation in control CMs increased glucose uptake (+28±8%) this was normalized after ACC2 KD. Inhibiting FAO by etomoxir or increasing glucose utilization by dichloroacetate abolished the beneficial effects of ACC2 KD after PE stimulation. When cultured in glucose-free medium supplemented with FA, ACC2 KD was incapable of preventing cardiomyocyte hypertrophy. However, replacing glucose with pyruvate or propionate restored the anti-hypertrophic effect of ACC2 KD.

To determine the therapeutic effects of increasing FAO
in vivo
, male mice were subjected to transverse aortic constriction (TAC) and sham surgery. Three weeks after surgery, TAC mice had a significant increase in left ventricular (LV) mass as determined by echocardiography compared to sham operated mice (130 vs. 92 mg). At this time point, cardiac-specific ACC2 deletion (iKO) was induced by tamoxifen (tam) administration. ACC2 protein was effectively deleted in iKO sham and TAC hearts compared to controls (CON) 2 weeks post tam injection. FAO was 2-fold higher in iKO TAC vs. CON TAC hearts as assessed by isolated heart perfusion and 13C NMR spectroscopy. CON and iKO mice will be followed until 12 weeks after TAC to determine cardiac function and assess hypertrophy.

Together, these data indicate that increasing FAO via inactivation of ACC2 exerts anti-hypertrophic effect in adult cardiomyocytes. Deleting ACC2 in the hypertrophic heart
in vivo
can increase FAO and represents a valid target to treat pathological cardiac hypertrophy.
Studies of a Humoral Sodium Transport Inhibitory Activity in Normal Dogs and Dogs with Ligation of the Inferior Vena Cava
Authors: BUCKALEW VARDAMAN M. , LANCASTER CARLTON D. 
Published: March 2022

Plasma ultrafiltrates of dogs acutely expanded with saline inhibit toad bladder sodium transport. The present studies were performed to delineate further the chemical and physiological characteristics of this inhibitory activity.
Column chromatography with Sephadex G-10 showed the activity to be present in the fraction ahead of the salt peak. Sodium balance studies in normal dogs were performed to determine the relationship between factor activity and the daily fluctuations of renal sodium excretion. A significant correlation was observed between the presence of humoral inhibitory activity and spontaneous antinatriuresis. In the absence of antinatriuresis, inhibitory activity was absent, regardless of the level of sodium intake. In normal dogs given saline loads acutely, inhibitory activity was present 60 and 120 minutes after the beginning of saline loading. In dogs with constriction of the inferior vena cava, no activity was observed during saline loading for up to 120 minutes.
The data are compatible with the interpretation that the humoral sodium transport inhibitory activity is a natriuretic hormone released in response to expansion of the extracellular fluid volume.