bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1: Oct4 is Re-activated in Smooth Muscle Cells through KLF4/Hif1α and Hydroxymethylation Mechanisms and Plays a Critical Role in Atherogenesis
Authors: Cherepanova Olga A , Gomez Delphine , Shankman Laura S , Williams Jason , Geng Yong-Jian , Connelly Jessica J , Owens Gary K 
Published: March 2022

The pluripotency factor Oct4 is a protein that regulates embryonic stem cell (ESC) totipotency and is the only gene thought to be indispensable for production of induced pluripotent stem (iPS) cells. Although there is evidence that Oct4 may be expressed in non-ESC, this is controversial and as yet there is no direct evidence that Oct4 plays a functional role within somatic cells.
We demonstrate that Oct4 is markedly induced in cells within atherosclerotic lesions from ApoE-/- mice fed a high-fat diet and within human coronary artery atherosclerotic lesions. We show that smooth muscle cell (SMC)-specific conditional knockout (KO) of Oct4 within Western diet-fed ApoE-/- mice resulted in increased atherosclerotic lesion size and other changes consistent with decreased plaque stability. Moreover, using a novel SMC lineage tracing mouse developed in our lab, we also show that lesions from SMC-specific conditional Oct4 KO mice contained far fewer de-differentiated (Acta2-) SMCs likely as the result of impaired SMC de-differentiation and migration from the media into the intima and not to altered proliferation or apoptosis. Further mechanistic studies using in vitro and in vivo Chromatin Immunoprecipitation assays, glycosylation-coupled methylation sensitive qPCR and hMeDIP show that activation of Oct4 within SMCs and within atherosclerotic lsions was associated with hydroxymethylation of the Oct4 promoter and was Klf4- and Hif1α-dependent. In addition, using in situ hybridization/proximity ligation (ISH-PLA) assay developed in our lab we showed hydroxymethylation of the Oct4 promoter within individual SMC-derived cells in atherosclerotic lesions of our SMC-lineage tracing ApoE-/- mice. These results are the first to show that Oct4 plays a direct functional role in regulating phenotypic transitions in somatic cells, and provide molecular mechanisms required for Oct4 re-activation.
Abstract 147: Characterization of the Immune Response to Aortic Aneurysm in a Murine Model
Authors: Byrne John S , Besla Rickvinder , Ensan Sherine , Li Angela , Degousee Norbert , Ouzounian Maral , Lindsay Thomas , Robbins Clint 
Published: March 2022

Our understanding of vascular pathology relies on inducible animal models of disease that remain poorly described, and concern exists regarding indirect effects of the inducing agents. We sought to perform a detailed characterization of the immune response to a commonly used murine aneurysm model.
Aneurysms were generated in 10 adult C57/Bl6 mice using 2 weeks of oral beta-amino propriono-nitrile (BAPN) administration and 4 weeks of angiotensin-2 (AT2) delivered via osmotic pump. FACS analysis was used to characterize progenitor and hematopoietic cell lines in bone marrow, blood and aorta. 3 mice died of aortic rupture between days 8 and 9. The remaining 7 were compared to age-matched controls.
BAPN/AT2 treatment caused aortic dilatation at all aortic regions from the root to the descending thoracic aorta (P<0.01). The suprarenal aorta was aneurysmal in all BAPN/AT2 mice with a maximum diameter of 1.86±0.1mm compared to controls 0.98±0.03mm (P<0.001). In the blood, total cells counts at day 2, 16 and 28 were increased (P<0.001). CD45-2 and B-cell counts were significantly increased in the BAPN/AT2 mice compared to controls (P<0.001). The aorta of BAPN/AT2 mice had significantly increased macrophage counts (P<0.001) with no difference in monocytes, neutrophils, T or B cell counts. Aortic tissue macrophages were predominantly G0 G1 (97±1%) phase. The bone marrow CD45-2 compartment was similar in aneurysmal BAPN/AT2 and control mice but Ly6C lo-inter monocytes were increased in BAPN/AT2 mice. Marrow CD117+/lin- hematopoietic progenitor and stem cells (P=0.4), and differential counts of CD117+/Sca1+ hematopoietic stem cells and CD117+/Sca1- progenitor cells were similar in control and BAPN/AT2 treated mice.
BAPN/AT2 induces aortic disease with aneurysmal degeneration of the suprarenal aorta. We observed a significant decrease in aortic macrophage proliferation suggestive of a mechanism of expansion independent of local macrophage proliferation. We did not observe a significant change in the bone marrow stem or progenitor cell compartment suggesting aortic injury is driving inflammation rather than direct stimulus of the marrow compartment by BAPN/AT2.
Abstract 150: Identification of SYNPO2, SYNM, LMOD1, PDLIM7 and PLN as Novel Markers of Smooth Muscle Cells in Atherosclerosis
Authors: Hedin Ulf , Perisic Matic Ljubica , Razuvaev Anton , Sabater-Lleal Maria , Lengquist Mariette , Miller Clint L , Roy Joy , Kronqvist Malin , Gonzalez Diez Maria , Frebelius Siw , Leeper Nick , Maegdefessel Lars , Quertermous Thomas , Paulsson-Berne Gabrielle , Odeberg Jacob , Hansson Göran K , Hamsten Anders 
Published: March 2022

Smooth muscle cell (SMC) activation is a hallmark of vascular remodeling associated with downregulation of SMC specific genes and altered cell function. Our aim was to identify a novel set of genes silenced following SMC activation in vascular disease.
We compared microarrays from carotid plaques (n=177) vs. undiseased arteries and symptomatic vs. asymptomatic patients and found that ‘muscle contraction’, ‘muscle differentiation’ and ‘cytoskeleton based migration’ were the most significantly downregulated pathways (p=6.06e-23; 5.22e-11; 4.62e-11 respectively). Genes clustered in these categories were known SMC markers together with a novel set of genes functionally coupled to actin cytoskeleton: SYNPO2, SYNM, LMOD1, PDLIM7 and PLN. Transcription factors regulating the expression of these genes showed enrichment of SRF, MYOD1 and MYOGENIN, known to control muscle differentiation. Downregulation of these genes was validated in independent microarrays from carotid (n=21) and coronary plaques (n=38). Their expression was positively correlated to typical SMC markers both in human plaques (Pearson r>0.8, p<0.0001) and rat carotid intimal hyperplasia (r>0.9, p<0.0001). In microarrays from rat carotid balloon injury, these genes were downregulated in the early phases of tissue remodeling but gradually reappeared in the mature intima. By immunohistochemistry, the proteins localized to SMCs in normal human vessels, but were mostly absent in plaques and restenosis tissues. In vitro, SYNPO2 and PDLIM7 localized to actin filaments, SYNM to cortical cytoskeleton, PLN to nucleus while LMOD1 was rapidly lost in cultured human and primary rat SMCs. Polymorphisms in some of these genes associated with carotid intima-media thickness in high-risk coronary artery disease subjects (n=3400). By eQTL analyses, rs11746443 influenced PDLIM7 expression in plaques. Interestingly, this polymorphism constitutes the binding site for HEY1 transcription factor implicated in vascular development.
We identified a panel of novel SMC proteins, which are lost in vascular disease and reflect the altered phenotype of SMCs in vascular remodeling. Our results indicate that these could be early and sensitive markers of the SMCs dedifferentiation in atherosclerosis.
Abstract 171: Deficiency of the Co-stimulatory Molecule Cd27 Impairs Treg Development and Exacerbates Atherosclerosis.
Authors: Winkels Holger , Smeets Esther , Meiler Svenja , Beckers Linda , Dandl Angelika , Reim Sigrid , Spitz Charlotte , Bürger Christina , Weber Christian , Lutgens Esther , Gerdes Norbert 
Published: March 2022

Atherosclerosis, an inflammatory disease of large arteries, is - through its clinical manifestations stroke and myocardial infarction - the leading cause of morbidity and mortality in the industrialized world. Adaptive immunity and co-stimulatory signals play a pivotal role during all stages of the disease. Recently, regulatory T cells (Treg) were attributed an anti-inflammatory and anti-atherogenic role. The interaction of CD70, a member of the tumor necrosis factor super family with its receptor CD27 modulates Treg development but also affects T cell proliferation, differentiation, and activation at the sites of antigen priming and at effector function. We hypothesized an increased atherosclerotic burden and an exacerbation of disease upon CD27 deficiency.
Cd27+/+Apoe-/- or Cd27-/-Apoe-/- bone marrow was transplanted into Apoe-/- recipient mice. After reconstitution, mice were fed a 0.15% high fat/cholesterol diet for 6 weeks. Plaque area at the aortic sinus was increased 2.3-fold in Cd27-/-Apoe-/- chimeras. Plaques contained high amounts of macrophages, which was accompanied by a reduced frequency of aortic (54.1%) and splenic (17.7%) Tregs. Similar results were obtained in 18 wk old Cd27-/-Apoe-/- on a normal chow diet, that also displayed an increase in atherosclerotic plaque area and a 2.5-fold higher plaque macrophage content. In addition, Cd27-/-Apoe-/-mice showed a significant decrease in the abundance of splenic (26%) and aortic (27%) Tregs. Remarkably, 28 week-old Cd27-/-Apoe-/- mice did not exhibit this phenotype, suggesting that CD27 only plays a role in earlier stages of atherosclerosis.
Although Cd27-/-Apoe-/- Tregs did not show changes in suppressive or migratory capacity compared to Cd27+/+Apoe-/-, Cd27-deficiency did result in a marked increase in thymic Treg apoptosis.
Taken together, our data reveal that deficiency for CD27 impairs thymic Treg development, which thereby exacerbates early and intermediate atherosclerosis by increasing plaque inflammation.
Abstract 220: Deficiency in Triggering Receptor Expressed in Myeloid Cells-1 Substantially Attenuates Atherosclerotic Lesion Development
Authors: Zysset Daniel , Weber Benjamin , Rihs Silvia , Freigang Stefan , Banz Yara , Saurer Leslie , Mueller Christoph 
Published: March 2022

TREM-1 is a potent amplifier of pro-inflammatory responses in myeloid cells. While the significance of TREM-1 in acute microbial-induced septic shock has long been acknowledged, our previous studies in experimental models of colitis have also suggested an important role for TREM-1 in driving non-infectious and chronic inflammatory diseases.
Here, we aim to define the role of TREM-1 in atherosclerosis, which is recognized as a chronic inflammatory disorder of the arteries with a major involvement of monocytes and macrophages. TREM-1-deficient (Trem1-/-) mice were backcrossed with apolipoprotein E-deficient (Apoe-/-) mice and were fed a high cholesterol/fat diet for 16 weeks. Compared with Trem1+/+ Apoe-/- mice, Trem1-/- Apoe-/- mice were substantially protected from diet-induced atherosclerosis. Interestingly, while the number of individual atherosclerotic plaques did not differ between the two groups of mice, the average lesion size and total surface area of atherosclerotic lesions were significantly reduced in Trem1-/- Apoe-/- mice. Moreover, analysis of early atherosclerotic lesions revealed no difference between the two groups. Hence, TREM-1 appears to be involved in the progression, rather than the initiation of atherosclerotic lesions. In line with the reduced lesion size, FACS analysis of aortic cell preparations revealed substantially lower frequencies of infiltrating CD11b+ myeloid cells in Trem1-/- Apoe-/- mice. To gain insight into the underlying mechanisms of TREM-1-mediated effects in atherogenesis, we are currently analyzing mRNA expression profiles of aortic segments derived from Trem1+/+ Apoe-/- and Trem1-/- Apoe-/- mice. These studies will be complemented by immunohistochemistry stainings and by in vitro studies that are aimed to directly determine the impact of TREM-1 on foam cell generation and cholesterol efflux.
Abstract 251: Selective Deletion of Smad4 in Smooth Muscle Cells Causes Thoracic Aortic Aneurysm in Mice
Authors: Carnevale Daniela , Da Ros Francesco , Carnevale Raimondo , Cifelli Giuseppe , Bizzotto Dario , Braghetta Paola , Bressan Giorgio , Lembo Giuseppe 
Published: March 2022

TGFβ signaling have been implicated in the onset of thoracic aortic aneurysms (TAA). Recently, it has been described that mutations in SMAD4, an intracellular transducer required in the TGFβ canonical pathway, present aortic disease in humans. Although transgenic models recapitulating mutations of TGFβ pathway have been studied, Smad4 role in the onset of TAA has never been explored.
Smad4 homozygotes mice are embryonically lethal, thus, we postnatally targeted the gene, using a tamoxifen (TAM)-inducible Cre recombinase under the smooth muscle myosin heavy chain promoter. Activation by TAM suppressed Smad4 in aorta SMCs of Smadf/f;Myh11-CreERT2 mice. We found that, most of these mice died 4/6 months later from hemothorax due to rupture of thoracic aorta, while untreated mice survived. Echocardiography showed a progressive dilation of aorta and TAA formation. Histology revealed progressive fragmentation of elastic lamellae with increased inflammatory infiltrates at sites of disarrangement, rich in macrophages. At the molecular level, we found that lack of Smad4 was followed by increased levels of PSmad2 and PSmad1/5/8 early after TAM, suggesting an enhancement of TGFβ receptors activity, a condition that could increase non-canonical TGFβ signaling, the canonical one being depressed. We actually found a significant increase in one of the non-canonical TGFβ signaling, i.e. the p65 subunit of NFkB, a protein complex controlling DNA transcription and involved in regulating responses to different stress stimuli, including cytokines. Given the particular relevance that the latter molecules may have in TAA, we tested expression of some of them, finding a selective IL-1β upregulation, produced and stored in cells as an inactive precursor and released in its active form by caspase-1 cleavage. In turn, caspase-1 requires NLRP3 inflammasome, a multiprotein platform that assembles in response to infection and tissue damage and is responsible for activation of inflammatory caspases. Therefore, we also tested NLRP3 mRNA levels and found them to be significantly upregulated.
We demonstrated for the first time that selective Smad4 deletion in the SMC of adult mice is sufficient to induce ultrastructural damage and immune reaction culminating in the onset of TAA.
Abstract 264: Circulating Tnni3k Levels Are Useful Diagnosis Biomarker for Acute Ischemic Myocardial Diseases
Authors: Lai Zhong-fang 
Published: March 2022

Our recent data showed that TNNI3K, a MAP kinase specifically expressed in cardiac muscles and interacted with cardiac troponin I, TNNI3K high-expression enhanced cardiac performance, attenuated ischemia-induced ventricular remodeling in mice with myocardial infarction, and protected the myocardium from ischemic injury.
In the present study, availability of circulating TNNI3K level was investigated using anti-TNNI3K polyclonal antibodies with ELISA, in patients diagnosed as AMI, chronic heart failure and acute renal failure; and in healthy volunteers groups. Data shown that circulating TNNI3K levels were significantly higher in AMI when compared with other two groups (p<0.001), indicating that measurement of circulating TNNI3K is a novel and useful diagnosing tool for AMI. Then, we made an anti-TNNI3K monoclonal antibody to investigate whether circulating TNNI3K levels play the same role for diagnosis of ischemic myocardial diseases, the availability of serum TNNI3K level was investigated with anti-TNNI3K monoclonal-antibodies in AMI, chronic heart failure and acute renal failure patients, and in healthy volunteers respectively. Data shown that circulating TNNI3K levels were significantly higher in AMI (p<0.001) and sensitivity and specificity are higher, ROC curve is more best than that of polyclonal antibodies, indicating that measurement of circulating TNNI3K level using the monoclonal-antibodies may be a novel and useful tool to diagnose AMI. Furthermore, both ELISA and gold colloid methods are used to observe their sensitivity and specificity. Results showed that the sensitivity and specificity of anti-TNNI3K monoclonal antibody were higher than other biomarkers anti-TNNI3K monoclonal antibody and the measurment time in the gold colloid method is significantly shortened when compared with using ELISA method. In conclusion, using TNNI3K as a molecular target is a new potential approach for developing therapeutic or diagnostic agents for ischemic cardiac diseases by means of molecular biological techniques.
Abstract 271: Id3 Promotes Obesity-Induced Inflammatory Macrophage Accumulation Through Proliferation of MCP-1-Producing Adipocyte Progenitor Cells
Authors: Kaplan Jennifer L , Marshall Melissa A , McSkimming Chantel , Harmon Daniel B , Garmey James , Oldham Stephanie , Hallowell Peter , McNamara Coleen A 
Published: March 2022

High fat diet (HFD)-induced increases in MCP-1 are a hallmark feature of adipose tissue inflammation during obesity. Macrophages are the main producers of MCP-1 in established obesity; however, early increases in obesity-induced production of MCP-1 precede migration of M1 macrophages to adipose tissue. Global deletion of transcription factor Inhibitor of Differentiation 3 (Id3) attenuates HFD-induced obesity and adipose tissue inflammation. Specifically, Id3-/- mice have less visceral adipose tissue (VAT), fewer M1 adipose tissue macrophages, and attenuated HFD-induced MCP-1 production. The source of initial obesity-induced MCP-1 in vivo, as well as the role that Id3 plays in regulation of MCP-1, is currently unknown.
Flow cytometry demonstrated that committed CD45-CD31-Ter119-CD29+CD34+Sca-1+CD24- adipocyte progenitor cells (AdPCs) express high levels of MCP-1. HFD increased the number of AdPCs in an Id3-dependent manner. Loss of Id3 increased levels of p21Cip1 and attenuated HFD-induced proliferation of AdPCs, resulting in lower levels of MCP-1 and reduced M1 macrophage numbers in VAT, compared to Id3+/+ littermate controls. Adoptive transfer of 50,000 Id3+/+ AdPCs into Id3-/- recipient mice restored adipose tissue inflammation via increased MCP-1 levels and M1 macrophage accumulation in VAT. Notably, the M1:M2 macrophage ratio significantly increased due to injection of AdPCs. Additionally, flow cytometry identified MCP-1-producing CD45-CD31-CD34+CD44+CD90+ AdPCs in human omental and subcutaneous adipose tissue. The percentage of AdPCs expressing MCP-1 was higher in omental adipose compared to subcutaneous adipose. Furthermore, high surface expression of CD44 marked abundant MCP-1 producers, only in omental adipose tissue.
The present study provides the first in vivo evidence that committed AdPCs are the source of early obesity-induced MCP-1. Id3 is a critical regulator of p21Cip1 expression and proliferation in AdPCs, and in this manner, promotes overall MCP-1 expression and M1 macrophage accumulation in VAT. Inhibition of Id3 and AdPC expansion, as well as CD44 expression in human AdPCs, may serve as unique therapeutic targets for the regulation of adipose tissue inflammation.
Abstract 339: Hyperhomocysteinemia Reduces Large HDL Particle and EL Expression via Hypomethylation Related Mechanism in Mice
Authors: Xi Hang , Choi Seungbum , Dipaul James M , Zhang Yuling , Yang Xiaofeng , Wang Hong 
Published: March 2022

Hyper-homocysteinemia (HHcy) has been identified as an independent risk factors of cardiovascular diseases (CVD). We previous reported HHcy is associated with reduced HDL-cholesterol and decreased large HDL particle in mouse and human CVD. Here, we investigated underlying mechanism of homocysteine (Hcy)-related HDL biosynthesis and degradation.

We examined protein/gene expression, activities of four HDL-degradation related lipases including endothelial lipase (EL), lipoprotein lipase (LPL), hepatic lipase (HL), and pancreatic lipase (PL). Severe HHcy (98.4±22 μM) was established in cystathionine betasynthase-gene mutant mice (
Cbs
+/-
), moderate HHcy (23.5±5 μM) was also developed in
Cbs
+/+
mice, both fed a high methionine (HM) diet for 8-week. Moderate and severe HHcy increased protein levels of EL in aorta (204% and 319%), lung (201% and 264%), and decreased LPL in aorta (98% and 34%), lung (99% and 61%), spleen (98% and 51%). HL and PL protein levels were not changed by HHcy. Activities of overall lipases were increased in moderate and severe HHcy in aorta (19.8±5.2 and 27.5±7.1, arbitrary unit), plasma (64.3±5.5 and 126.5±26.2), adipose (147.0±8.9 and 210.9±38.9) and liver (160.5±41.3 and 276.2±51.3).

Next, we examined tissue levels of Hcy, and its metabolites (SAM, SAH) in heart, lung, brain, spleen, kidney and liver from wild type mice. Tissue expression profile of Hcy degradation and synthesis-related enzymes were established by database mining analysis. We found that mRNA levels of EL, HL, and PL were positively correlated with Hcy and negatively correlated with SAM/SAH ratio, which indicated of methylation status. In contrast, LPL mRNA levels were negatively correlated with Hcy and positively correlated with SAM/SAH ratio. These data suggest that EL, HL, PL gene expression may be induced by Hcy-related hypomethylation.
We identified a large CpG island with 113 CG dinucleotide pairs in human EL promoter region, where four HDL-related single nucleotide polymorphisms (SNPs) was overlapping with CpG containing transcription factor consensus element.
Our study suggests that HHcy inhibits HDL biosynthesis, promotes HDL degradation via EL induction and DNA hypomethylation related mechanisms.
Abstract 352: Adventitial Stem Cell Progenitors Express Neural Stem Cell Markers
Authors: Fitzpatrick Emma , Kennedy Eimear , Hakimjavadi Roya , Cahill Paul A 
Published: March 2022

Many forms of cardiovascular disease (CVD) are charaterised by intimal medial thickening (IMT) due to the formation of a neointimal layer as a result of the accumulation of vascular smooth muscle cells (vSMC) within the vessel wall. There is strong evidence, albeit controversial, that indicates a putative role for both adventitial (Sca1
+
, CD34
+
and Lin
-
) and medial multipotent vascular progenitor stem cell-derived [Sox10, Sox17 and S100β] vSMC in contributing to IMT.

The objective of this study was to characterise adventitial progenitor stem cells (APCs) from aortic explants and compare them to medial MVSCs. APC and MVSC phenotype was determined by immunocytochemistry using antisera specific for adventitial stem cell marker Sca1 and stem cell markers Sox10, Sox 17 and S100β respectively. Differentiated vSMC markers, α-actin, calponin 1 and smooth muscle myosin hevy chain II (Myh11) were used to detect SMCs. Discrete APC populations were positive for neural stem cell markers [Sox 10, Sox17 and S100β] as well as marker α-actin. Notably a significant number of intial cells migrating from the adventitial explant were Sox 10, Sox 17 and S100β positive respectively. Similarly, MVSC from medial explants were positive for neural stem cell markers [Sox 10, Sox17 and S100β] and Sca1 (when grown in maintenance media). There was a distinct absence of smooth muscle myosin heavy chain II filaments (differentiated vSMC markers) in both the APC and MVSC cell populations.
We conclude that APCs and MVSCs may be derived from a similar source.