bluuebunny/crossref_metadata_2025_split huggingface dataset:
GPCRomics of cardiac fibroblasts and cardiac fibrosis
Authors: Sriram Krishna , Michkov Alexander V. , Aroonsakool Nakon , Insel Paul A. 
Published: November 2023

G‐protein coupled receptors (GPCRs) are the largest category of cell surface receptors, serving as the most widely used set of targets for pharmacological intervention. Seeking to identify novel targets for cardiac fibrosis, we have used mRNA‐based (GPCRomic) methods to detect and quantify (i.e., profile) non‐chemosensory “endo‐GPCRs” in cardiac fibroblasts (CFs) and, complemented these results by verification of GPCR expression/function using antibody‐based and functional analyses. We used 2D, low passage (passage 1 or 2) cultures of primary isolates of CFs and myofibroblasts (MyoFs) from humans, rats and mice.RESULTSWe found that CFs express ~120 GPCRs, including a large number (~40–50) of orphan GPCRs, and multiple highly expressed adhesion GPCRs. Biological replicates of human CFs correlate closely in terms of identity of GPCRs expressed and the magnitude of their expression. Further, our data show that MyoFs possess a smaller (number and average expression) of GPCRs but a distinct subset of GPCRs have increased expression in MyoFs compared to CFs. Growth of CFs on soft substrates (which minimize the spontaneous transformation of cultured CFs into MyoFs) produces relatively small effects on the profile of GPCRs expressed by CFs. A subset of GPCRs remains highly and robustly expressed in CFs, regardless of substrate stiffness, pro‐fibrotic stimulation, duration of culture, or growth under hypoxic conditions. The GPCR profiles of mouse and rat CFs closely mimic that of human CFs as do the GPCR profiles of rat and human MyoFs. Further, we find that the GPCR expression profile of rat CFs is very different from that of freshly isolated rat cardiomyocytes, implying that there are numerous potential GPCR targets for cardiac fibrosis, which if used therapeutically, are predicted to have minimal on‐target effects on cardiomyocytes. We conclude that a GPCRomic approach provides novel insights regarding the regulation of CFs and MyoFs and identifies numerous potential targets for cardiac fibrosis.Support or Funding InformationSupported by Bristol‐Myers Squibb
Establishment of induced Ovarian Cancer Stem Cells by Defined Factors
Authors: Chen Yu‐An , Lu Chen‐Yu , Cheng Wen‐Fang , Chang Nan‐shan , Ho Hong‐Nerng , Chen Hsin‐Fu 
Published: November 2023

Drug resistance and metastasis are considered to be the major factors leading to tragic prognosis and high rate of mortality in cancer patients. Recent evidences suggest that cancer stem cells (CSCs) possess the essential property of self‐protection from chemotherapy, exhibit the ability to self‐renew and generate cancer cells by asymmetric cell division. Nevertheless, the currently known mechanisms likely explain only the tip of an iceberg about CSC property, acquisition and maintenance. The information from data mining is still limited due to the rarity of CSCs in clinical samples. Ovarian cancer is the most lethal malignancy of the female reproductive system. CSCs in ovarian cancers are considered to acquire the molecular armaments of stem cells, which carry stem cell property and can determine the level of malignant potential of the cancer cells, similar to the findings in other cancer cell types.In this study, we introduced a set of defined factors (OCT3/4, SOX2, KLF4 and c‐Myc) into human serous ovarian cancer cells (OVCAR‐3 cell line) through Sendai virus reprogramming, aiming to induce CSC properties in the cancer cells. We observed that the ovarian cancer cells transduced with the four Yamanaka factors showed significantly enhanced pluripotent stem cell properties and drug resistance. We thus named the cells with CSC properties as induced ovarian CSCs (iOCSCs). These iOCSCs could be maintained and expanded up to at least 40 passages. Consistent with the original ovarian cancer cells, iOCSCs could be differentiated into ovarian lineage cells, displaying markers as CA125 and CK‐7. Intriguingly, iOCSCs also acquired the ability to differentiate into cells belonging to the three embryonic germ layers, similar to human iPS cells or ES cells.This present approach thus makes it feasible to obtain abundant CSCs from ovarian cancer cell line to support our understanding and in‐depth research into CSCs and hopefully can help in developing new therapeutic strategies to treat ovarian cancers.Support or Funding InformationResearch supported by National Science Council of the Republic of China (NSC) and National Taiwan University Hospital (NTUH), Taiwan
Sexual dimorphism in left ventricular and electrocardiographic effects of chronic cyclosporine in rats
Authors: El‐Mas Mahmoud M , El‐Bassossy Hany M. , Banjar Zainy M 
Published: November 2023

Although cardiotoxicity has been recognized as one of the adverse effects of the immunosuppressant drug cyclosporine A (CSA), limited or no information is available regarding the sex specificity of CSA cardiotoxicity. Here we tested the hypothesis that the left ventricular (LV) and electrocardiographic effects of chronic CSA and related inflammatory and histopathological derangements are sex‐related. Adult male and female rats were treated with CSA (15 mg/kg/day, sc) or its vehicle for three weeks. Afterwards, studies were undertaken to assess: (i) LV and electrocardiographic characteristics, and (ii) alterations in LV inflammatory (nuclear factor kappa B, NFκB; angiotensin receptors type 1, AT1) and histopathological profiles. In both male and female rats, CSA significantly reduced LV slope of end‐systolic pressure volume relationship (ESPVR), and isovolumic relaxation constant (Tau), suggesting LV systolic and diastolic dysfunction. Histopathologically, the treatment of either rat sex with CSA resulted in vascular congestion, blood extravasation, and pyknotic or even absent nuclei. We also report sex‐dependent electrocardiographic effects for CSA. Whereas CSA increased QTc and T peak trend intervals in male rats, markers of LV arrhythmogenesis, evidence of prolonged AV conduction was noted in female rats as reflected by the prolonged P duration and PR interval. Other sex‐related effects for CSA included (i) increased blood cholesterol, rate of rise (dp/dtmax) and fall (dp/dtmin) in LV pressure and LV content of NFκB and AT1 expression in male rats, and (ii) increased LV content of adiponectin and elevated systolic and diastolic blood pressures in female rats. Although the two rat sexes are prone to the negative LV influences of CSA, the male rat seems to be more vulnerable to inflammatory and LV adverse effects of CSA.Support or Funding InformationThis work was funded by the Deanship of Scientific Research (DSR), King Abdulaziz University, Jeddah, under grant No. (166 ‐ 1001 ‐D1435). The authors, therefore, acknowledge with thanks DSR technical and financial support
Adipocyte derived factors affect cardioprotective programs initiated by short term high fat feeding
Authors: Haar Lauren , Luther Kristin , Ren Xiaoping , Wang Yang , Jones WK 
Published: November 2023

RationaleHigh fat mediated cardioprotection is a form of protection that utilizes short‐term high fat diet to decrease infarct size post ischemia/reperfusion injury. This protection is dependent on the increase of autophagy related proteins and the activation of NF‐κB mediated gene transcription. This study addresses the hypothesis that signaling from adipose tissue can remotely influence the cardioprotective markers associated with high fat mediated (HFD) cardioprotection.MethodsHormonal activation of cardiac receptors was assessed through treatment with murine recombinant adiponectin and AdipoRon peptide. Isolation of adipocyte derived exosomes was also preformed from 3T3‐L1 (ATCC® CL‐173) cells during pre differentiation, mitotic expansion, and post differentiation states. Viability of cells pretreated with each of the previously described agents was then assessed with Hoescht 33342 and propridium iodide staining before and after the administration of an oxygen glucose assay (OGD) protocol. Autophagy measurements were performed with LC‐3A/B and Beclin‐1 western blot, and the Premo Autohpagy Sensor® transduction for each treatment condition. In vivo studies were preformed in wild type C57/B6J and AdipoQ‐KO murine models, with in vivo LAD occlusion (30m) and reperfusion (24h).ConclusionExosomes isolated from 3 phases during the differentiation of 3T3‐L1 (ATCC® CL‐173) cells into mature adipocytes did not increase the viability of cells after OGD treatment. Post OGD protocol, AdipoRon (25uM) treatment improved viability and cell survival relative to vehicle treated H9c2(2‐1) (ATCC® CRL‐1446) controls. Total serum levels of adiponectin were increased by an average of 15% in 24h HFD wild type animals relative to control fed wild type animals, and AdipoQ‐KO animals lacked cardioprotection at previously reported cardioprotective high fat fed time points. Future studies will focus on the assessment of in vivo hormonal signaling during high fat mediated cardioprotection.
Blocking effect of carvedilol, a beta‐blocker on Na+/Ca2+ exchange current in isolated cardiac ventricular cells of guinea pig
Authors: Watanabe Yasuhide , Kimura Junko 
Published: November 2023

BackgroundCarvedilol has a1, b1, and b2‐adrenergic blocking effects together with the effects such as vasodilating, anti‐ischemic, anti‐oxidant, anti‐apoptotic, anti‐proliferate and anti‐inflammatory activities. Carvedilol inhibited IKr, IKs, ICa‐L and Ito in rabbit cardiac ventricular myocytes. On the other hand, metoprolol, a nonselective b‐blocker also has blocking several channels, including IK1, ICa and Ito in hearts. Murayama et al. (2013) reported that carvedilol and its analogues inhibit delayed after depolarizations (DADs) in Langendorff‐perfused rabbit hearts. It is well known that DADs are caused by the Na+/Ca2+ exchange current (INCX).MethodsWe examined the effects of carvedilol and metoprolol, two b‐blockers, on INCX by using the whole‐cell voltage‐clamp method in isolated guinea pig ventricles and CCL39 fibroblast cells expressing dog cardiac Na+/Ca2+ exchanger (NCX1).ResultsCarvedilol suppressed INCX in a concentration‐dependent manner but metoprolol did not in isolated guinea pig cardiac ventricular myocytes. IC50 values for the Ca2+ influx (outward) and efflux (inward) components of INCX were 69.7 μM and 61.5 μM, respectively. Carvedilol at 100 μM inhibited INCX in CCL39 cells expressing wild type NCX1 similarly to mutant NCX1 without the intracellular regulatory loop. Carvedilol at 30 μM abolished ouabain‐induced DADs in isolated cardiac ventricular myocytes.ConclusionsCarvedilol suppressed bi‐directional INCX in a concentration‐dependent manner with the IC50 value of approximately 65 μM. We conclude that carvedilol inhibits NCX1 at supratherapeutic concentrations.
Intralipid Protects against Hypoxia/Reoxygenation Injury in Human Coronary Artery Endothelial Cells and Cardiomyocytes
Authors: Salzman Michele Marie , Warner Edgewood R. , Riess Matthias L. 
Published: November 2023

INTRODUCTIONHypoxic conditions cause extensive cellular damage, particularly to mitochondria and the plasma membrane. Upon reoxygenation, incomplete reduction of oxygen (O2) can produce reactive oxygen species that can cause additional reperfusion injury to damaged and viable cells. Intralipid (IL) has been demonstrated to be cardio‐protective when delivered as a pre‐ or post‐conditioning agent. We hypothesized that IL delivered during reoxygenation will protect human coronary artery endothelial cells (HCAECs) and human cardiomyocytes (HCMs) from reoxygenation (reperfusion) injury.OBJECTIVESWe compared 1) hypoxic cells vs hypoxic/reoxygenated (H/R) cells vs hypoxic cells reoxygenated with differing concentrations of IL; and 2) cells on the basis of cellular “age” (passage; e.g., P7 vs P15).METHODSCell cultures were placed into complete media/normoxic conditions (FBS & glucose; 95% O2, 5% CO2) or hypoxic media/hypoxic conditions (0% FBS & glucose; 1% O2, 94% N2, 5% CO2) at 37°C for 3 hrs. Samples were then either immediately assayed for cell proliferation, metabolic activity, and cytotoxicity, or they were incubated ± IL (0.1%, 0.5%, 1%, or 2%) in complete media/normoxic conditions for 2 hrs and then assayed for the same parameters.RESULTSYounger cells had a greater proliferative capacity than older cells in both cell types. However, there were equivalent decreases in proliferative capacity in both hypoxic and H/R exposed cells, and IL had no effect on cell proliferation, regardless of concentration. Metabolic activity was noted to be higher in older cells than younger cells in both cell types. When exposed to H/R, HCAECs metabolic activity decreased, while HCMs had slightly increased metabolic activity. In both cell types, significant increases in metabolic activity occurred notably with IL concentrations of 1–2%. LDH release was greater in older than in younger cells of both cell types, and was more extensive in hypoxic only cells compared to H/R cells. The dose‐dependent impact of IL on cytotoxicity was indeterminable.CONCLUSIONSCells differ based on their “age” (passage), and also on what cellular function is affected by H/R. IL increased metabolic activity in both cell types, specifically at concentrations of 1–2%; however, there was no determinable effect on cell proliferation or cytotoxicity. Further studies need be conducted to determine which metabolic pathways are being activated, and whether those pathways are related to cell survival during stressful conditions such as H/R.Support or Funding InformationSupported by the Department of Veterans Affairs (IK2BX001278), NIH (R01 HL095122), and institutional funds.
The Effect of Losartan on Marfan Syndrome Is Angiotensin II Receptor Type 1 (ATR1) Independent
Authors: Sellers Stephanie , Chan Rayleigh , Mielnik Michael , Jermilova Una , Pavlovic Marijana , Hirota Jeremy , Seidman Michael , Hogg James C , Esfrandiarei Mitra , Van Breemen Casey , Bernatchez Pascal 
Published: November 2023

BackgroundMarfan Syndrome (MFS) leads to life threatening cardiovascular and pulmonary complications including aortic root dilation and emphysema‐like lung destruction. The new drug of choice to treat MFS is Losartan as it has been shown to reduce aortic and lung pathology in mouse models of MFS and is the focus of on‐going clinical trials. Losartan selectively blocks angiotensin receptor type I (ATR1), which is a key player in MFS pathogenesis. However, despite its promising therapeutic activity, Losartan has never been evaluated in the absences of its proposed target, and therefore genetic loss of ATR1 should protect against MFS and reveal the full potential of LosartanHypothesisA model of MFS lacking ATR1 will prevent the cardiovascular and pulmonary phenotype of MFS and thereby confirm the mechanism of action of Losartan and reveal the full therapeutic potential of complete ATR1 blockade in MFS.MethodsA MFS model lacking ATR1 was generated using a commonly used MFS model (Fibrillin C1039G) and ATR1 knockout mice (ATR1−/−). The resulting MFS ATR1−/− mice, both treated and untreated with Losartan, were used to evaluate the impact of ATR1 knockout on MFS. Aortic and lung pathology was evaluated through echocardiogram and histology while MFS skeletal manifestation were analyzed using radiography.ResultsEchocardiograms and histology show that both MFS and MFS ATR1−/− mice have similar aortic root dilation, aortic wall thickening and elastic fibre fragmentation in stark contrast to our hypothesis. Additionally, both MFS and MFS ATR1−/− groups demonstrate emphysema‐like airspace widening and significant changes in skeletal structure. Finally, when treated with Losartan MFS ATR1−/− mice showed similar reduction in aortic root dilation and airspace widening as MFS mice.ConclusionData from our novel model shows that loss of the predominant ATR1 isoform fails to prevent MFS complications. Furthermore, Losartan treatment in the absence of ATR1 is still an effective therapeutic for MFS aortic and lung pathology. This suggests that the therapeutic utility of Losartan in MFS is off‐target and completely independent of ATR1.Support or Funding InformationThis work was supposed by funds from the Canadian Institutes of Health Research (CIHR) and fellowships to SS from CIHR and the University of British Columbia.
Bone Marrow‐Derived Stromal Cells (BMSCs) Improve Microvascular Perfusion in Skin Flaps Subjected to Ischemia Reperfusion Injury (IRI)
Authors: Tang Ya Hui 
Published: November 2023

Skin flaps are frequently used for the functional and cosmetic repair of wounds and defects left behind after trauma, burns, and tumors. Flap survival mainly depends on the blood supply provided through re‐established vessels of the dermal and subdermal plexus. We have previously demonstrated the efficacy of BMSCs in the enhancement of wound healing and in improving flap survival.ObjectiveTo determine if the intravascular delivery of BMSCs improve blood perfusion in the skin flaps following IRI.MethodsA cutaneous flap model perfused via the inferior epigastric artery was created in wild type (C57Bl/6), which underwent 3.5 hours of ischemia, followed by reperfusion and BMSCs transplantation. Blood perfusion in the flap was monitored using a noncontact, real‐time and noninvasive Laser Speckle Contrast Analysis (LSCA) system. Flap survival and BMSCs recruitment were also assessed.ResultsThe numbers of trans‐migrated BMSCs appearing in the flap interstitial spaces were increased, compared to control (sham) mice, within 4h after IRI. Blood flow, both proximal and distal to the vascular pedicle was marked increased (p<0.01) in postischemic flaps treated with BMSCs treatment, when compared to IRI controls at 3 days of reperfusion.ConclusionThe results of this study indicate that the enhanced wound healing and improved flap survival observed following BMSC treatment may be related to an improvement in tissue blood flow.Support or Funding InformationSupported by a Malcolm Feist Postdoctoral Fellowship
Attenuation of EPA and DHA‐Mediated Platelet Inhibition Effects Following Heterogeneous Agonist Treatment
Authors: Bormann Jordan L. , Meyer Evan S. , Voigt Ellen M. , Whiteheart Sidney W. , Larson Mark K. 
Published: November 2023

Agonist‐induced platelet activation following vascular damage is a critical event for the propagation of thrombi formed as a part of either hemostasis or thrombosis. Several studies have indicated this activation can be impaired after platelet membrane enrichment with the polyunsaturated omega‐3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). This attenuated platelet function represents a potential mechanism behind the anti‐thrombotic benefits of EPA and DHA observed in many human trials. However, nearly all platelet studies with EPA and DHA examined the alterations to platelet activation in response to a single agonist, rather than with multiple agonists that mimic the more complex heterogeneous environment of a developing arterial clot. We therefore asked if high levels of EPA and DHA could still inhibit platelet activation if platelets were stimulated with a combination of agonists designed to simulate the activating conditions found in the thrombus inner core (using thrombin and collagen) or outer shell (using ADP and thromboxane A2). To test this question, we treated platelets with EPA and DHA concentrations in vitro that raise the membrane content of both fatty acids to levels analogous to subjects taking EPA and DHA supplements. After this modification, the platelets were stimulated with the agonist combinations and measured for an activation response. Surprisingly, inherent measures of platelet activation using these agonist combinations, including adhesion, aggregation, granule release, and integrin activation were either only mildly affected or non‐significantly altered after EPA and DHA membrane enrichment. We also tested the ability of the core agonist combination to stimulate platelet‐derived thrombin generation, a catalytic process thought to be necessary for rapid thrombus core development. Contrary to the other measures of platelet function, thrombin generation was still consistently impaired by elevated EPA and DHA levels. We conclude that elevated levels of EPA and DHA in the developing clot environment do not readily inhibit most events of platelet activation. Instead, the impairment of amplified thrombin generation in the platelet‐rich arterial clot may provide a more likely candidate for how EPA and DHA prevent detrimental cardiovascular events.Support or Funding InformationResearch reported in this abstract was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103443, a South Dakota/National Science Foundation EPSCoR RII Track 1 grant, the Sanford Program for Undergraduate Research, and an American Physiological Society Undergraduate Summer Research Fellowship.
Effects of SIRT3 deficiency on vasomotor function and atherosclerotic plaque composition in mice
Authors: Roos Carolyn M , Hagler Michael A , Zhang Bin , Miller Jordan D 
Published: November 2023

Excess mitochondrial reactive oxygen species can drive endothelial dysfunction and vascular calcification in atherosclerosis. Recently, SIRT3 has been linked to the activity of the ROS‐degrading enzyme MnSOD through regulation of protein acetylation levels. It remains unclear, however, whether SIRT3 protects against endothelial dysfunction and osteogenic signaling in hypercholesterolemia. We hypothesized, that reduction of SIRT3 will impair endothelial function, decrease mitochondrial antioxidant capacity, and increase osteogenic signaling in hypercholesterolemic mice. We used Ldlr−/−/ApoB100/100 (LA) mice that were either wild‐type (LA‐SIRT3+/+) or null for SIRT3 (LA‐SIRT3−/−). After 12 months of western diet feeding, we assessed endothelial function in aorta using isolated organ chamber baths and measured changes in gene expression using qRT‐PCR. As expected, SIRT3 gene expression was significantly reduced in LA‐SIRT3+/− mice compared to LA‐SIRT3+/+ mice, and undetectable in LA‐SIRT3−/−. Surprisingly, endothelial‐dependent relaxation to acetylcholine (MRAch) was unchanged between LA‐SIRT3+/+ and LA‐SIRT3−/− (37.0±4.4%, 38.2±4.4%). Endothelial‐independent relaxation (sodium nitroprusside) was also unchanged. MnSOD expression was reduced in LA‐SIRT3−/− compared to LA‐SIRT3+/+ (0.80 ± 0.02 vs 1.0±0.08, p < 0.05). In contrast, the late‐stage osteoblast marker, osterix, tended to increase in LA‐SIRT3−/− (1.3 fold) but failed to reach statistical significance. Collectively, these results suggest that losses of SIRT3 do not impact endothelial function in advanced atherosclerosis, but may contribute to augmentation of osteogenic signaling and accelerated progression of vascular calcification.Support or Funding InformationNHLBI