bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 579: De-differentiation of Somatic Fibroblasts by Mouse Embryonic Stem Cell-free Extracts: Multi-lineage Re-differentiation and Therapeutic Efficacy of Re-programmed Cells in Mouse Models of Hind limb and Acute Myocardial Ischemia
Authors: Rajasingh Johnson , Lambers Erin M , Bord Evelyn , Hamada Hiromichi , Goukassian Ilona , Qin Gangjian , Losordo Douglas W , Kishore Raj 
Published: July 2021

Background: De-differentiation of adult somatic cells into multipotent progenitor cells might provide an attractive, oocyte-independent alternate source for therapeutic cloning to generate pluripotent, autologous stem cells for regenerative medicine. We tested the hypothesis that exposure of reversibly permeabilized NIH3T3 fibroblasts to mouse embryonic stem cell (mES) extracts may provide regulatory molecules required for epigenetic changes leading to re-programming of terminally differentiated somatic cells and that such reprogrammed cells may display multi-lineage differentiation.
Methods and Results: Streptolysin O-permeabilized NIH3T3 cells were exposed to mESC extracts in ATP-regenerating system. Plasma membrane was resealed by culturing in medium 2mM CaCl2. We report that upon exposure to mES extracts 3T3 cells undergo dedifferentiation as evident from morphological changes, induction of mES specific and the loss of lamin A/C, a specific marker of the soma. At epigenetic level, mES extract treatment induced de-methylation of Oct4 promoter and histone3 hyperacetylation at lysine 9 residue, as evident from restriction enzyme mapping, bisulphite genomic sequencing and chromatin immuno-precipitation experiments. Under defined culture conditions in vitro, reprogrammed 3T3 cells can be differentiated into neuronal, adipocyte, cardiomyocyte (CMC) and endothelial cells (EC). Moreover, transplantation of labeled reprogrammed 3T3 cells into mouse hind limb ischemia and acute myocardial infarction models resulted in the gain of physiological functions and showed evidence of acquiring the expression of muscle, EC and CMC specific proteins, in vivo. Moreover, when injected in SCID mice, reprogrammed cells formed teratomas.
Conclusion: Taken together our biochemical, molecular and functional data provide a novel and oocyte- independent approach for the generation of functional autologous stem like cells from terminally differentiated somatic cells which may potentially be of therapeutic application in regenerative medicine.
Abstract 640: Common Genetic Variation in the Promotor of the Ankyrin-2 Gene Modifies QT Interval
Authors: Sedlacek Kamil , Stark Klaus , Weber Stefan , Berger Iris , Pfeufer Arne , Wichmann H.-Erich , Hengstenberg Christian , Jeron Andreas 
Published: July 2021

Cardiac QT interval represents both ventricular depolarization and repolarization events. Previous studies suggested significant genetic contribution to the normal QT interval length variation. In a candidate gene approach, we sought to identify novel genetic determinants of the normal QT interval length within the Ankyrin-2 (Ank-2) gene.
Population and Methods: The study population consisted of 1492 participants of the WHO MONICA population survey. A standard measurement of the QT interval from the 12-lead ECG was available in 1212 individuals. The QT interval was corrected for covariates using different general and population specific formulas (QTc). Altogether, 22 SNPs within the Ank-2 gene were genotyped using the TaqMan technology. Statistical analyses included ANOVA and linear regression based trend test for comparisons of the QTc means.
Results: In the promotor region of the alternative splicing Ank-2 variant, several SNPs and haplotypes were highly significantly associated with the QTc interval length. Specifically, a SNP rs1979086 (minor allele frequency f=0.44) was associated with the QTc length in the whole population (P=0.004 for ANOVA and P<0.001 for the trend test). The QTc difference between the genotype groups was 2.87 ms (95%CI 1.18 – 4.56) and more pronounced in men where the QTc difference between the genotype groups was 4.65 ms (95% CI 2.18 – 7.12; P=0.0006 for ANOVA and P=0.0004 for the trend test). The results remained robust when QTc extreme quartiles and quintiles, respectively, were tested in a case-control pattern. Highly significant association results were obtained for all genetic models used (OR 1.58; P=0.00009; trend test for extreme QTc quartiles). A frequent haplotype (f=0.28) spanning approximately 20 kb in this region was highly significantly associated with the QTc length in the whole study population (QTc difference vs. most prevalent haplotype 3.66 ms, 95%CI 3.14 – 4.17, P<0.0001).
Conclusion: A small genomic region in the promotor of the Ank-2 gene contains SNPs and haplotypes that significantly influence the QTc interval in the normal population of German descent. A gender-specific effect with greater influence in male participants requires further evaluation.
Abstract 680: KC706, an Oral p38 MAP Kinase Inhibitor, Increases HDL-C
Authors: Miller Stephen G , Crowley Constance , Lundstrom Jan , Larson Christopher J , Prior Kent E , King Bernard D 
Published: July 2021

Introduction: KC706 is a novel p38 MAP kinase inhibitor with anti-inflammatory properties. In rats and monkeys, KC706 administration was associated with a 50 – 60% ↑ in HDL-C. Because HDL-C is an important target for the treatment of CV disease a Phase 2a study was designed to determine whether KC706 administration would elevate HDL-C levels in patients with dyslipidemia.
Methods: A double-blind, placebo-controlled, 6-week study was conducted in patients with ↓ HDL-C and ↑ triglyceride (TG) levels. Patients (N = 115) were randomized to placebo or to KC706 (150 or 300 mg po qd). Standard chemistry and hematology parameters and lipid, metabolic, inflammatory, and thrombotic markers were monitored. The primary end point was an increase in HDL-C in treatment v. placebo groups.
Results: Patients were primarily male (59%) and white (97%). At baseline, 35% were receiving statins, 44% were hypertensive, 15% had CV disease, 70% fulfilled ATPIII criteria for metabolic syndrome, and 43% had CRP > 3 mg/L. The primary endpoint of the study was met at both dose levels. Compared to baseline, HDL-C ↓ 2.9% in the placebo group, ↑ 18.6% in the 150 mg group (p < 0.0001 v. placebo) and ↑ 11.6% in the 300 mg group (p = 0.0003 v. placebo) after 6 wks of treatment. Effects were noted in men and women, whether patients were receiving statins or not, and whether CRP was elevated or not at baseline. ApoAI ↑ significantly in the 150 mg group (p = 0.0056 v. placebo). TG, LDL-C, and VLDL-C were unchanged. CRP levels trended downward in patients with ↑ baseline CRP levels. HDL/Total cholesterol (+22%), ApoA1/ApoB100 (+14%), and HDL/LDL (+31%) increased in the KC706 150 mg group. Mean HDL particle size (measured in a subset of patients) increased from 8.4 to 8.7 nm in the KC706 150 mg group (p = 0.04), from 8.5 to 8.7 nm in the KC706 300 mg group (p = 0.01) but not in the placebo group (8.3 v. 8.4 nm, p = 0.66). No increases in either systolic or diastolic blood pressures were observed in any group. One patient in the 150 mg group and 4 in the 300 mg group had a reversible elevation in either AST and/or ALT of > 3X ULN. Adverse events were similar in the 3 treatment groups.
Conclusions: KC706, an oral p38 MAP kinase inhibitor with anti-inflammatory properties, increased HDL and ApoA1 in patients with dyslipidemia and was well tolerated.
Abstract 925: Rit GTPase Protects Cells from Oxidative Damage
Authors: Rudolph Jennifer L , Shi Geng-Xian , Harrison Susan M , Harrison Douglas A , Andres Douglas A 
Published: July 2021

Signaling cascades that contribute to the regulation of cellular survival influence many cardiovascular diseases. Elucidating the mechanisms of these cascades is thus important for understanding basic biology and for therapeutic intervention. We have identified an evolution-arily conserved group of Ras-related GTPases, including two mammalian genes (Rit and Rin) and a single Drosophila ortholog (Ric), that we hypothesize plays a critical role in trophic factor-mediated anti-apoptotic signaling. Expression of activated Rit in PC6 cells induces potent activation of the MAP kinases ERK1/2 and p38, and promotes neurite outgrowth and cell survival. Furthermore, RNAi-mediated Rit silencing sensitized PC6 cells to a wide variety of stresses, in part by inhibiting NGF-mediated activation of ERK and p38 MAP kinases and CREB. Here we present studies from two genetic models designed to further assess our hypothesis. First, we generated a transgenic mouse over-expressing constitutively activated Rit (RitL79). To begin to assess the contribution of Rit signaling to cellular survival, we examined the ability of cortical neuronal cultures isolated from Rit-TG versus wild-type littermates to survive reactive-oxygen-species (ROS)-mediated cell death. Cell viability for wild-type cortical neurons following a 4 h exposure to H2O2 (60 μM) was reduced approximately 90%, while cortical neurons isolated from Rit-TG mice were largely unaffected by this treatment. Thus, RitL79-mediated signaling protects cortical neurons against ROS-mediated apoptosis. As further confirmation of the important role this protein family plays in survival from stress, we generated a Drosophila strain null for D-Ric. Mutants are homozygous viable and show no obvious developmental abnormalities. However, D-Ric mutants are susceptible to environmental stresses, including heat stress and dry starvation. Taken together, these studies suggest a conserved role for Rit/Ric in promoting cellular survival and suggest that Rit signaling is a potential target for pharmaceutical intervention. We look forward to assessing Rit’s role in other systems including cardiac and vascular cell types.
This work was supported by a Predoctoral Fellowship from the AHA.
Abstract 939: Galpha-i Signaling in Neural Crest Cells is Required for Normal Migration of Cardiac Neural Crest Cells, Cardiac Development, and Survival
Authors: Ruppel Kathleen M , Kataoka Hiroshi , Iwaki Michelle , Cornelissen Ivo , Coughlin Shaun R 
Published: July 2021

G protein coupled receptors (GPCRs) have long been known to play crucial roles in transducing environmental signals to the adult cardiovascular system. In recent years, the roles of G protein-mediated signaling pathways in orchestrating the interactions of different tissues during cardiovascular development have become increasingly evident. To analyze the role of G protein signaling pathways in vivo we have generated mice where the function of the heterotrimeric G alpha subunit Gai can be ablated in a cell type specific manner utilizing the Cre-loxP system. We have mated these mice to two different neural crest-specific Cre lines in order to probe the effects of loss of Gai mediated signaling on the ability of neural crest cells (NCC) to contribute to the developing outflow tract and aortic arch arteries.
METHODS: We have generated mice that express the Gai-inhibiting pertussis toxin S1 subunit (PTX) from the ROSA26 locus in a Cre recombination dependent manner (ROSA-PTX mice). These were mated to mice expressing either the Wnt1 Cre or P0 Cre transgene. Wnt1Cre is active in both premigratory and migratory NCC, whereas P0Cre is active only in migratory NCC and their derivatives.
RESULTS: P0Cre-ROSA-PTX mice were normal at birth and demonstrated no structural heart defects. In contrast, Wnt1Cre-ROSA-PTX mice were present in normal numbers at late gestation but died perinatally due in part to cardiac outflow tract defects. Excision reporter and in situ hybridization studies suggest this is secondary to a delay/blockage of cardiac NCC migration into the developing outflow tract. NCC migration into the pharyngeal arches was unaffected in these mice and no craniofacial, thymic, or aortic arch abnormalities were observed.
CONCLUSIONS: These results indicate that Gai-mediated signaling is required in premigratory or early migratory cardiac NCC for normal development of the outflow tract. In contrast, endothelin A receptor knockout mice (currently the only GPCR knock out with a neural crest phenotype) are thought to exhibit defects of postmigratory NCC function. RNA profiling of NCC for GPCRs involved in this Gai-dependent pathway has revealed several potential candidate receptors, including orphan receptors. Further analysis of these receptors is underway.
Abstract 1105: Gadofluorine M-Cy 3, a Novel Paramegnetic Contrast Agent for the in vivo and Histolgical Identification of Transplanted Cells.
Authors: Adler Eric , Bystrup Anne , Saebo Karen , Mani Venketesh , Giovannone Steve , Young Wilson , Frank Joseph , Weinmann Joachim , Keller Gordon , Fayad Zahi 
Published: July 2021

PURPOSE: Gadofluorine M with a fluorescent dye (GdFMCy3) is a lipophilic paramegnetic contrast agent that is readily absorbed by cultured cells. We hypothesized that this agent would be superior to iron oxide based techniques for cell tracking post cell transplantation.
METHODS: Embryonic Stem Cell derived cardiac progenitor cells (ES-CPCs) were generated using previously established methods and incubated for 12 hours with 5 mM GdFMCy3, or transfected with iron oxide using published protocols. Cell survival was >95% for cells incubated with both GdFMCy3 and iron. 500,000 cells labelled with GdFMCy3, iron oxide or control (no contrast agent) were directly injected into the myocardium of mice (n=5/group). Mice were scanned over a two week interval post injection at 9.4T using gated T1-weighted sequences (GdFMCy3), T2* weighted GRE sequences (iron oxide) or the positive contrast sequence GRASP (iron oxide). Mice were sacrificed and the hearts sectioned for microscopy. Perl staining and fluorescence microscopy were used to identify iron oxide and GdFMCy3 within the myocardium, respectively.

RESULTS: GdFMCy3 labelled cells were successfully identified in vivo at 9.4T (figure

panel B). Good correlation between MRI and histology was observed for both cell labels (figure

). Contrast to noise ratios were significantly higher in the GdFMCy3 group relative to the iron oxide group (figure

).

CONCLUSIONS: GdFM-Cy3 is readily taken up by stem cells and easily identified by both MRI and fluorescence microscopy. Given its superior contrast to noise ratio GdFM-Cy3 may be an excellent alternative to iron oxide for in vivo detection and tracking of transplanted cells.
Abstract 1111: Embryoinc Stem Cell Derived Cardiovascular Progenitor Cells Improve Function More Than Hemangioblasts in a Mouse Model of Myocardial Infarction
Authors: Adler Eric , Chen Vincient , Bystrup Anne , Young Wilson , Giovannone Steve , Kattman Steve , Fayad Zahi , Keller Gordon 
Published: July 2021

BACKGROUND: Intramyocardial transplantation of stem cells improves left ventricular ejection fraction (EF) in animal studies and preliminary clinical trials. The mechanism may involve either replacement of myocytes or improved vascular supply to existing myocytes. We recently identified an Embyronic Stem cell derived cardiovascular progenitor cell (ES-CPC) that is the common precursor of cardiomyocyte and vascular cell lineages. To determine whether myocyte transplantation improves myocardial function more than angiogenesis alone does, we compared the effect of ES-CPCs to hemangioblasts (vascular/hematopoetic progenitor cells) on EF in a mouse model of myocardial infarction.
METHODS: ES-CPC and hemangioblasts were isolated from a doxycycline-responsive, Notch-inducible ES cell line containing Notch 4 cDNA under the control of a tetracycline-inducible promoter. Notch induction of mesoderm-derived ES cells resulted in a CPC phenotype, whereas non-induced cells developed into hemangioblasts. Mice underwent transplantation of 500,000 ES-CPC (n=20), hemangioblasts (n=16), or an equal volume of serum-free media (n=12) 30 minutes after surgically-induced myocardial infarction. All cell lines constitutively expressed green fluorescent protein (GFP). EF was assessed two weeks post-transplantation using 9.4 Tesla MRI. Mice were then euthanized and frozen heart sections were examined using fluorescent microscopy.
RESULTS: The mean EF was 59Â ± 15, 46Â ± 17, and 39Â ± 13% in the ES-CPC, hemangioblast, and control groups, respectively (p<0.05 for the differences among all 3 groups; ANOVA). GFP + cells were detected in frozen sections of both the ES-CPC and hemangioblast groups. GFP + cells in ES-CPC treated hearts expressed markers associated with both cardiomyocyte and vascular phenotypes, whereas the GFP + cells in the hemangioblast group expressed markers associated with vascular phenotypes.
CONCLUSIONS: Both hemangioblast and ES-CPC transplantation improves EF in a mouse model of myocardial infarction, but ES-CPC transplantation was more effective. This suggests that enhancement of myocardial function by transplantation of both cardiomyocyte and vascular phenotypes exceeds that with vascular phenotypes alone.
Abstract 1117: Genetic Modification with Sonic Hedgehog Mitigates Aging-induced EPC Dysfunction and Augments Therapeutic Angiogenesis of Endogenous EPCs following Acute Myocardial Infarction
Authors: Ii Masaaki , Horii Miki , Akimaru Hiroshi , Kusano Kengo F , Thorne Tina , Asahara Takayuki , Losordo Douglas 
Published: July 2021

Background: Clinical trials of autologous stem/progenitor cell transplantation in ischemic heart diseases are now on-going. Simultaneously, mounting evidence suggests that age related decreases in EPC number or function may limit this therapeutic approach. We tested the hypothesis that gene modification of aged EPCs, using the embryonic signaling molecule, sonic hedgehog (Shh), may ameliorate aging induced dysfunction of EPCs and thereby enhance the reparative effect of autologous EPCs.
Methods and Results: EPCs from aged mice (24 m.o.), previously shown to exhibit impaired potency, were used and compared to EPCs from young (3 m.o.) mice. Shh gene transfer significantly improved proliferation and anti-apoptosis activities vs. empty vector transfected (EV) aged EPCs, up to a similar level of EV young EPCs. Next, we evaluated the therapeutic effect of Shh-gene-modified aged EPCs (Shh-EPC) vs. EV aged EPCs (EV-EPC) vs. PBS (Ctrl) injection in a mouse AMI model. Shh-EPCs or EV-EPCs (1000 cells) or PBS was injected into the ischemic myocardium immediately following MI. Scar length in the infarcted LV 14 days after surgery was significantly reduced in the Shh-EPC group compared to the EV-EPC group and the PBS group associated with increased capillary density in the peri-infarct area. In addition, Ki67 positive cardiomyocytes were observed with increased frequency in the Shh-EPC group. In Tie2-LacZ BMT mice, beta-gal positive-endogenous BM-derived EPC recruitment was significantly greater in ischemic myocardium in the Shh-EPC group than that in the EV-EPC group. Differentiation of recruited EPCs into endothelial cells or cardiomyocytes was also observed more frequently in the Shh-EPC group compared to the EV-EPC group. These data suggest that cardiac repair was induced by a small number of transplanted Shh gene modified aged EPCs with the increased paracrine effects following MI.
Conclusion: Shh gene transfer to aged EPCs ameliorates aging-induced EPC dysfunction. Intramyocardial transplantation of Shh-gene-modified aged EPCs augments therapeutic efficacy for myocardial repair following MI by promoting endogenous EPC recruitment. Shh gene modified EPCs may represent a second-generation EPC-based therapeutic strategy in ischemic heart disease.
Abstract 1126: Adiponectin Prevents Neointimal Hyperplasia After Vascular Injury In Adiponectin Transgenic Mice
Authors: omori yasutoshi 
Published: July 2021

Background: Adiponectin has been reported to accumulate in the human injured artery and regulate the development of atherosclerosis, decreasing the expression of adhesion molecules in vascular endothelial cells, and inhibiting the proliferation of vascular smooth muscle cells. However, the role of adiponection after vascular injury is not fully elucidated. Therefore, we investigate whether adiponectin prevents neointimal hyperplasia after vascular injury in adiponectin transgenic mice (TG).
Methods: C57/BL6 mice (WT) and TG of 6 –7 week age were used. We inserted a large wire (0.38 mm in diameter) into femoral artery from distal side to proximal side to make the vascular endothelium damaged model. Mice were sacrificed at 1, 2 and 4 weeks, and non injured mice were used as control. Tissue concentration of fat and liver and serum concentration of adiponectin was measured using ELISA. Intima/ media ratio was measured in morphometrical analysis. Immunohistochemical staining of anti-adiponectin, anti-α-smooth muscle embryo (α-SMemb), anti-α-smooth muscle cell actin, von Willebrand Factor (vWF) and intercellular adhesion molecule-1 (ICAM-1) was performed using paraffin embedded sections.
Result: In immunohistochemical analysis at 1 week, α-SMemb positive cells, which were also positive for ICAM-1 were observed in WT, but were not seen in TG. Adiponectin positive cells, which were also positive for vWF were observed in the endothelial cells in TG, but were not detected in WT at 1 week, but those cells were observed in th endothelial cells in TG and WT at 2 and 4 weeks. In α-SMA staining at 4 weeks most neointimal area was stained with this anti-body in TG. On the other hand, in neointima of WT, only some area was stained with this anti-body. In morphometrical analysis, intima/media ratio was significantly smaller in TG than in WT at 1, 2 and 4 weeks. Adiponectin concentration of fat, liver and serum was significantly higher in TG than in WT in non-injured model and at 4 weeks. At 1 week liver and fat adiponectin level is significantly higher in TG than in WT, and serum adiponectin level in TG tended to be high as compared to that of WT.
Conclusions: Adiponectin prevents neointimal hyperplasia after vascular injury, probably through the inhibition of inflammatory reaction.