bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3410: FAK is Required for Neural Crest Cell Remodeling of the Cardiac Outflow Tract
Authors: Vallejo-Illarramendi Ainara , Reichardt Louis F 
Published: July 2021

Background: Cardiac neural crest cells (NCCs) participate in the remodeling of the outflow tract and pharyngeal arch arteries during cardiovascular development. Abnormal growth factor and integrin signaling in cardiac NCCs leads to cardiovascular malformations. Focal adhesion kinase (FAK) is a ubiquitous protein kinase that transduces integrin and growth factor signaling pathways. However, a direct role for FAK in cardiac NCC morphogenesis has not been demonstrated. Here we have used a Cre/loxP method for NCC-specific deletion of FAK in mouse embryos.
Results: We found that targeted deletion of FAK in NCC results in craniofacial and cardiovascular malformations, leading to early postnatal lethality. Mutant mice display cleft palate together with several cardiovascular defects, including persistent truncus arteriosus, transposition of the great arteries, overriding aorta, ventricular septal defect and type B interruption of the aortic arch, all of which resemble common forms of human congenital heart disease. We found that NCC counts in the outflow tract of FAK mutant and control littermates are similar at E11, suggesting that FAK is not required for cardiac NCC migration to the outflow tract. Furthermore, NCCs appear to differentiate normally into smooth muscle cells in the outflow tract of FAK conditional mutant embryos at E11 and E12.5. In contrast, NCCs in the aortic arch arteries of FAK conditional mutants show impaired differentiation into smooth muscle cells in these embryonic stages. Additionally, we observed that rotation of the outflow tract fails to occur by E11 in FAK mutants, resulting in a dextroposed aortic root. Moreover, FAK-deficient NCCs form a less compact mesenchyme in the conotruncal cushions and show disorganized F-actin stress fibers in the aorticopulmonary septum.
Conclusions: Our results indicate that FAK plays an essential role in the development of the cardiac outflow tract and in the remodeling of aortic arch arteries. However, it is dispensable for migration and differentiation of cardiac NCC in the outflow tract. Further studies with this novel model of congenital heart disease may help elucidate the NCC signaling pathways that orchestrate septation of the outflow tract.
Abstract 3433: Cardiac Myocyte-specific Caveolin-3 Overexpression Modulates ERK/EGFR Signaling and Attenuates Cardiac Hypertrophy
Authors: Horikawa Yousuke T , Tsutsumi Yasuo M , Patel Hemal H , Insel Paul A , Roth David M 
Published: July 2021

Introduction: The localization and organization of signaling molecules involved in cardiac hypertrophy and remodeling are poorly understood. Caveolae, invaginations of the sarcolemma and enriched in the protein caveolin (Cav), localize signaling molecules involved in cardiac hypertrophy and remodeling. Cav-3, which is unique to myocyte caveolae, inhibits extracellular signal-regulated kinase (ERK) phosphorylation in vitro and binds the epidermal growth factor (EGF) receptor. We hypothesized that cardiac myocyte-specific overexpression of Cav-3 (Cav-3 OE) would modulate ERK signaling via direct interaction and alteration in EGF receptor signaling and would attenuate cardiac hypertrophy.
Methods: Transgenic mice with cardiac myocyte-specific Cav-3 OE (generated using an α-myosin heavy chain promoter) and littermate controls (C) were subjected to transverse aortic constriction (TAC) for 4 weeks. Echocardiography, histology and molecular analysis were performed. Cardiac myocytes were isolated and stimulated with phenylephrine (Phe) and EGF to assess ERK activity via activation of Gq and EGF receptors, respectively.
Results: Cav-3OE mice had reduced heart weight/tibia length ratio post-TAC compared to C (9.9±0.8 vs. 12.9±0.5 mg/mm; n=8; P<0.05) and decreased myocyte cross sectional area compared to C (203±8 vs. 327±17 μm2/cell; n=7; P<0.001). Cav-3 OE and C mice had similar fractional shortening (FS) pre-TAC but post-TAC, Cav-3 OE had greater FS compared to C (29.2±3.2 vs. 20.8±3.0%; n=8–9; P<0.05). Fibrosis, as assessed by trichrome stain, was decreased in Cav-3 OE compared to C mice. Post-TAC, pERK expression in cardiac homogenates was decreased in Cav3-OE mice vs. C. In vitro, pERK had, similar expression in the two groups in response to Phe, but pERK was expressed at higher levels in Cav3-OE myocytes in response to EGF receptor simulation.
Conclusion: These results show that cardiac-specific Cav-3 OE alters ERK/EGFR signaling in cardiac myocytes, producing a phenotype with blunted cardiac hypertrophy and preserved cardiac function post-TAC. This suggests that cardiac myocyte-specific Cav-3 OE may provide a novel approach to improve cardiac function in pathological cardiac hypertrophy and remodeling.
This research has received full or partial funding support from the American Heart Association, AHA Western States Affiliate (California, Nevada & Utah).
Abstract 3441: Paraoxonase-192 Polymorphism Interaction with Low HDL Cholesterol in Coronary Artery Risk
Authors: Pereira Andreia M , Mendonca Isabel M , Freitas Ana I , Sousa Ana C , Gomes Susana , Faria Paula , Freitas Sonia , Ornelas Ilidio , Brehm Antonio , Cardoso Antonio , dos Reis Roberto Palma 
Published: July 2021

Background: Increased lipid peroxidation is associated with accelerated progression of atherosclerosis. Paraoxonase (PON1) is an antioxidative enzyme in high density lipoproteins (HDL), which protects against lipid per oxidation and Coronary Artery Disease (CAD). PON 1 activity is under genetic control and its molecular basis is a polymorphism in the PON 1 gene that shows two common isoforms: Q (192 Gln) with high ability to protect LDL from lipid peroxidation in vitro, and R (Arg) with lower ability. Aim: To explore the interaction of the R allele paraoxonase gene and low HDL cholesterol concentrations, in CAD risk emergence.
Methods: 818 individuals participated in the study, 298 coronary patients, 55.0±10.3 years, 78.9% male sex, and 520 healthy controls, 47.3±12.6 years, 57.5% male sex, age and sex matched. Low HDL-C was defined as <0.90 mmol/L in men and < 1.11 mmol/L in the women. Comparisons of genotypes between cases and control subjects were performed by a chi-square test. Statistical significance was accepted at p<0.05. Odds ratio as well as their 95% confidence intervals for the RR genotypes and HDL deficient subjects were computed using univariate analysis (2x2 tables). To determine the interaction between the RR paraoxonase genotype and HDL-deficient subjects, we used the epidemiologic tables 4x2 and the synergy measures: the additive model (Rothman’s synergy index) and the multiplicative model (Khoury’s synergy index). We calculated the relative excess risk (RERI) and the attributable proportion (AP) due to interaction (Rothman).
Results: The PON 1 RR192 is associated with coronary heart disease [OR=1.61; p=0.043] in whole population. The HDL-deficient subjects 192 RR homozygotes showed an increased risk of CAD (OR=17.38; p< 0.0001) compared to normal HDL 192 RR (OR=1.39; p=0.348) and HDL-deficient subjects not carrying RR genotype (OR=7.79; p<0.0001). The genotype PON 192 RR increases the risk of CAD in the HDL-deficient populations (SI=2.3, SIM=1.6). The attributable proportion due to this interaction (AP) was 0.53, meaning that 53% of CAD was explained by this interaction.
Conclusion: These data suggest the existence of a synergistic effect of the RR 192 PON 1 genotype (with lower ability) and HDL-deficient subjects in CAD emergence.
Abstract 3625: Targeted Ultra-Small Iron Oxide Particles for In Vivo MR Detection of Atherosclerotic Lesions Using Antibodies against Oxidized Low Density Lipoprotein
Authors: Briley-Saebo Karen , Shaw Peter X , Calcagno Claudia , Choi Seung-Hyuk , Fuster Valentin , Tsimikas Sotirios , Fayad Zahi A 
Published: July 2021

Background: Oxidized low-density lipoprotein (OxLDL) plays a key role in the progression and destabilization of atherosclerotic plaque. Preliminary studies indicate that gadolinium (Gd) micelles labeled with oxidation-specific antibodies allow for specific targeting of vulnerable plaque. Due to issues associated with Gd toxicity (caused by retention and bio-transformation of the micelles), clinical translation of this platform may be limited. Iron oxides are recognized as safe and effective contrast agents for MRI. As a result, it was hypothesized that lipid coated ultra-small iron oxide particles (LUSPIOs) containing murine antibodies that bind malondialde-hyde lysine (MDA2) or oxidized phospholipids (EO6) associated with oxLDL may allow for the development of a safe platform for in vivo detection of vulnerable plaque.

Materials and Results: Untargeted, MDA2, and EO6 labeled LUSPIOs were prepared and characterized with respect to efficacy, pharmacokinetics, and biodistribution in Apolipoprotein deficient mice (ApoE−/−). MRI was performed at 9.4T prior to and 24 hrs after LUSPIO administration (4 mg Fe/kg) using both gradient echo and positive contrast GRASP sequences. MDA2 and EO6 increased the particle size, blood half-life, and MR efficacy of the LUSPIOs, relative to untargeted particles. Only limited signal attenuation was observed following administration of untargeted LUSPIO. MRI and microscopy revealed that MDA2 and EO6 LUSPIOs accumulate within oxLDL rich foam cells (Fig.1

).

Conclusions: This study suggests that biocompatible LUSPIOs may provide a clinically translatable platform for the MR detection of oxLDL rich foam cells in humans.
This research has received full or partial funding support from the American Heart Association, AHA National Center.
Abstract 3682: Heterozygous Knockout of the Insulin-like Growth Factor 1 Receptor in Mice Leads to Enhanced Insulin Sensitivity and Increased Endothelial Nitric Oxide Production
Authors: Abbas Afroze , Imrie Helen , Rajwani Adil , Viswambharan Hema , Cubbon Richard M , Kahn Matthew B , Wheatcroft Stephen B , Grant Peter J , Kearney Mark T 
Published: July 2021

Introduction Insulin like growth factor-1 (IGF-1) enhances glucose uptake and nitric oxide (NO) production, acting via similar signalling pathways to insulin. Several cross sectional and longitudinal studies have demonstrated an inverse relationship between IGF-1 bioactivity and cardiovascular disease. We assessed the hypothesis that a putative mechanism for these findings may be the modulation of endothelial NO bioavailability by IGF-1.
Methods and Results We investigated the effect of reduced IGF-1 bioactivity on metabolic and endothelial function in male mice, heterozygous for knockout of the IGF-1 receptor (IGF1RKO) and their wildtype littermates (WT). Glucose tolerance tests revealed impaired glucose handling in IGF1RKO mice compared to WT (mean AUC for IGF1RKO mice=1086 ± 26.82(mmol/L)minutes, mean AUC for WT mice=950.5 ± 22.5(mmol/L)minutes, n=5 in each group, p=0.005). However, using an insulin tolerance test, IGF1RKO mice were more insulin sensitive than controls (mean AUC for IGF1RKO=552.2 ± 25.18(mmol/L)minutes, n=10; mean AUC for WT=651.4 ± 22.26(mmol/ L)minutes, n=11, p=0.008). Ex-vivo assessment of vascular function was performed using thoracic aortic rings in an organ bath. Aortic rings from IGF1RKO mice were hypocontractile to phenylepherine compared to those from WT (mean Emax for IGF1RKO mice=0.60 ± 0.05g n=11 versus mean Emax for WT mice=0.79 ± 0.06g, n=10, p=0.03). Addition of the nitric oxide synthase inhibitor, NG-monomethyl-L-arginine (L-NMMA), led to a 65.40 ± 13.40% (n=9) increase in mean Emax in IGF1RKO mice compared to a 15.78 ± 12.00% (n=9) increase in WT mice (p=0.01). In addition IGF1RKO aortic rings were resistant to IGF-1 exposure compared to WT (percentage change in mean Emax for IGF1RKO mice=32.80 ± 11.01%, n=9 versus 66.28 ± 10.60% in WT mice, n=8, p=0.05), but maintained sensitivity to insulin.
Conclusion IGF1RKO mice demonstrate impaired glucose handling but enhanced sensitivity to insulin compared to WT. Aortic rings from IGF1RKO mice are hypocontractile to phenylepherine and have increased basal NO production. These data raise the possibility that reduced IGF-1 action in the endothelium may have a favourable effect on NO bioavailability, possibly by a novel interaction with insulin signalling.
Abstract 3905: MSCs
Wnt11
Derived Paracrine Factors Are Critical to the Repair of Infarcted Myocardium
Authors: Zuo Shi , Dai Ying , Ning Xu , Wang Yigang , Ashraf Muhammad , Xu Meifeng 
Published: July 2021

Genetically engineered MSCs have been shown to serve as an effective vehicle for the delivery of protective proteins. We hypothesized that Wnt11 enhanced MSCs induced cardioprotection of ischemic myocardium via paracrine effect. We have successfully transfected Wnt11 into MSCs.
Method: Conditioned medium (con-M) was obtained from various MSCs which were exposed to normoxia or hypoxia for 24 hours and was concentrated 100 times by ultrafiltration. Con-M (50μl) was injected into the border zone of infarcted myocardium immediately after rat underwent left anterior descending coronary artery (LAD) occlusion. Cultured cardiomyocytes (CM) were also treated with Con-M before cells were exposed to hypoxia.

Result: A marked improvement in cardiac function was observed in the con-M from MSCs
Wnt11
compared to the con-M from MSCs
GFP
(Table 1

). Microarray analysis revealed that Wnt11 was 204 fold higher in MSCs
Wnt11
than that in MSCs
GFP
. Moreover, many growth factors and cytokines were upregulated when MSCs
Wnt11
were exposed to hypoxia for 24 hours (e.g. BMP4, 1.64; FGF1, 1.61 and TGFμ2, 2.21 fold higher than that in MSCs
GFP
, respectively; all p<0.05). Con-M significantly improved the survival of CM as expressed by an increased intake of MTT when CM were exposed to hypoxia. Neutralizing antibodies, including anti-Wnt11 (6 and 12 μg/ml) (Fig. 1

), anti-BMP4 (3μg/ml), anti-FGF-1 (6μg/ml) and anti-TGFβ-2 (12μg/ml), abrogated the protection of CM induced by con-M, especially anti-FGF-1 (Fig. 2

). It is concluded that MSCs
Wnt11
protect cardiomyocytes by releasing cytoprotective factors in their environment.

Table 1.

Effects of condition medium on cardiac function

Figure 1.

Anti-Wnt11 antibody increased CM injury and abrogated the protective effect of conditioned medium against hypoxic damage. N = 4 in each group

Figure 2.

Neutralizing antibodies abolished the CM protection on cultured CM. #, p<0.05
vs
normal control; *, p<0.05
vs
hypoxia; †, p<0.05
vs
Con-M-MSC
GFP
, ‡, p<0.05
vs
Con-M-MSC
Wnt11
respectively.
Abstract 5272: Perinatal Loss of Nkx2–5 Results in Rapid Conduction and Contraction Defects
Authors: Briggs Laura E , Takeda Morihiko , Cuadra Adolfo E , Wakimoto Hiroko , Lu Jonathan T , Yasui Kenji , Ikeda Yasuhiro , Chien Kenneth R , Kasahara Hideko 
Published: July 2021

Homeobox transcription factor Nkx2–5, highly expressed in heart, is known to be a critical factor during early embryonic cardiac development. Having previously demonstrated the persistent expression of Nkx2–5 in cardiac myocytes from the precardiac mesoderm through to the adult heart, we hypothesized that Nkx2–5 actively regulates a critical set of genes in postnatal cardiomyocytes to maintain proper cardiac function. We generated tamoxifen-inducible Nkx2–5 knockout mice that carry homozygous floxed-Nkx2–5 alleles and a heterozygous Cre-ERTM transgene under the control of CMV enhancer and chicken-beta-globin promoter. A single injection of tamoxifen into pregnant mice on gestation day 19 almost completely deleted Nkx2–5 genes, as demonstrated by Southern and Northern blotting. These mice demonstrated rapid conduction and contraction defects within 4 days of tamoxifen-injection. Conduction defect was accompanied by reduction of the cardiac voltage-gated Na+ channel pore-forming a-subunit (Nav1.5-alpha) in ventricles, the largest ion channel in the heart responsive for rapid depolarization of the action potential. In addition, Ca2+ release from the sarcoplasmic reticulum was diminished accompanied by substantial reduction in ryanodine receptor 2. Because excitation and contraction is tightly coupled in hearts, reduced handling of Na+ and Ca2+ demonstrated in Nkx2–5 knockout hearts may mutually lead to rapid conduction and contraction defects. Survival studies demonstrated that approximately 50% of Nkx2–5 knockout mice died before 3 weeks of age, and all the knockout mice that survived demonstrated conduction defects and enlarged hearts at 4 weeks of age. These defects appeared sooner and progressed more rapidly than those seen in mice with embryonic ventricular-restricted deletion of Nkx2–5, likely due to nearly complete deletion of Nkx2–5 in the entire hearts, without compensatory mechanisms during embryonic stages after loss of Nkx2–5. Our results demonstrate that Nkx2–5 is not limited to embryonic heart development; it is critical for both conduction and contraction in the perinatal heart by regulating expression of several important gene products in conduction and contraction.
This research has received full or partial funding support from the American Heart Association, AHA National Center.
Abstract 5279: MCPIP1 is an Inducible Corepressor of NF-KappaB
Authors: Liang Jian , Kolattukudy Pappachan E , Fu Mingui 
Published: July 2021

The transcription factor NF-κB is a central regulator of host immune and inflammatory response. However, excessive and prolonged activation of NF-κB can cause massive damage to host tissues and can result in human inflammatory diseases, such as atherosclerosis and arthritis. Thus, NF-κB activation must be terminated at the appropriate time points. MCPIP1 is a recent identified CCCH-type zinc finger containing protein, which is significantly induced by MCP-1 treatment of human monocytes and thus designated as MCP-induced protein1 (MCPIP1). Here we report that MCPIP1 function as an inducible corepressor of NF-κB to feedback control NF-κB activation. In the transfection experiments, MCPIP1 dose-dependently inhibits inflammatory stimuli-induced NF-κB reporter activity. Overexpression of MCPIP1 does not affect TNFα-initiated IKK activation, NF-κB translocation and DNA binding. Further study reveals that MCPIP1 can directly bind to p65 of NF-κB and recruit NCoR and HDAC1 to form repressing complex, which blocks NF-κB target gene expression. Interestingly, the same stimuli that induce NF-κB activation also can induce MCPIP1 expression and translocation into nucleus. Taken together, our results suggest that MCPIP1 is a novel inducible corepressor of NF-κB and may be a critical player in host immune and inflammatory response.
This research has received full or partial funding support from the American Heart Association, AHA Greater Southeast Affiliate (Alabama, Florida, Georgia, Louisiana, Mississippi, Puerto Rico & Tennessee).
Abstract 5362: Inhibition Of The Epidermal Growth Factor Receptor Transactivation By The Angiotensin II Type I Receptor Prevents Compensatory Cardiac Hypertrophy And Worsens Cardiac Function Following Myocardial Infarction
Authors: Galeotti Jonathan , Hsu Chiao Po , Zhai Peiyong , Sadoshima Junichi 
Published: July 2021

The angiotensin II type I receptor (AT1R) has been shown to activate the epidermal growth factor receptor (EGFR). However, the downstream effects of this activation have not yet been elucidated in the heart. To examine the function of the AT1R-EGFR pathway, we have created an AT1R mutant in which amino acid 319 is mutated from tyrosine to phenylalanine (Y319F). The Y319F mutant lacks the ability to transactivate the EGFR. We generated transgenic lines overexpressing either wild type AT1R (Tg-WT) or Y319F (Tg-Y319F) only in the heart with similar levels of overexpression, and evaluated the contribution of the AT1R-EGFR pathway to cardiac responses against stress. Under baseline conditions at 3 months of age, cardiac function of Tg-WT and Tg-Y319F was not significantly different from non-transgenic mice (NTg) except that Tg-WT showed mild left ventricular hypertrophy. To examine the role of the AT1R-EGFR pathway under stress, we induced myocardial infarction (MI) by permanently ligating the left anterior descending coronary artery. Four weeks after MI, increases in heart weight/tibia length (Tg-WT, Tg-Y319F, NTg: 12.57, 10.11, 9.96, p<0.05 vs Tg-WT) and left ventricular myocyte cross sectional area (35, 5, 29% vs sham, p<0.05) were significantly attenuated in Tg-Y319F compared to Tg-WT. Fibrosis was also milder in Tg-Y319F than in Tg-WT (10.7% vs 22.9%). These results suggest that the AT1R-EGFR pathway plays an important role in mediating LV hypertrophy and fibrosis after MI. Interestingly, however, echocardiographic measurement showed that Tg-Y319F have impaired LV ejection fraction (Tg-WT, Tg-Y319F, NTg: 51, 19, 44%, p<0.05) and %fractional shortening (21.4, 6.7, 17.7%, p<0.05) compared with Tg-WT or NTg. Tg-Y319F showed an increase in mortality in days 0–28 following MI, when compared to Tg-WT and NTg (35, 67, 30%, p<0.5). Additionally, lung weight/tibia length was increased in Tg-Y319F (11.3, 24.7, 16.1, p<0.05) These results suggest that the lack of EGFR activation causes cardiac dysfunction after MI. In summary, transactivation of the EGFR following MI may play a compensatory role, thereby protecting the heart from further deterioration of cardiac function.
This research has received full or partial funding support from the American Heart Association, AHA Founders Affiliate (Connecticut, Maine, Massachusetts, New Hampshire, New Jersey, New York, Rhode Island, Vermont).
Abstract 5542: Role of Scavenger Receptor BI in Sepsis
Authors: Guo Ling , Smart Eric , Li Xiangan 
Published: July 2021

Scavenger receptor BI (SR-BI) is a well-established HDL receptor that regulates plasma HDL cholesterol levels by selectively uptake cholesterol ester from HDL. We recently reported a novel function of SR-BI namely protection against endotoxin-induced animal death. Using a lipopolysaccharides (LPS) induced endotoxic animal model, we found that LPS challenge induced 90% fatality of SR-BI null mice while all the wild type littermates survived, indicating that SR-BI plays a pivotal role in protection against LPS toxicity in vivo. In the current studies, we determined whether SR-BI plays a protective role in sepsis using an established septic animal model (cecal ligation and puncture, CLP). We demonstrate that CLP treatment induced 100% fatality of SR-BI null mice (n=20) whereas only 21% fatality of wild type littermates (n=27) and 36% fatality of heterozygous littermates (n=30). Moreover, SR-BI null mice exhibited exaggerated and prolonged inflammatory response as shown by elevated proinflammatory cytokine levels in serum upon CLP treatment compared to wild type littermates. Further studies revealed that the plasma LPS levels were significantly higher in SR-BI null mice compared to wild type control mice, indicating that SR-BI is required for LPS clearance in sepsis. Our findings clearly demonstrate that SR-BI is a critical protective modulator in sepsis in mice, which may reveal a novel target for the intervention of sepsis.
(Supported by grant from American Heart Association 0530241N)
This research has received full or partial funding support from the American Heart Association, AHA National Center.