Adenosine-treated Group Research Articles

Staphylococcus epidermidis Cicaria, a Novel Strain Derived from the Human Microbiome, and Its Efficacy as a Treatment for Hair Loss.

The skin tissue of the scalp is unique from other skin tissues because it coexists with hair, and many differences in microbial composition have been confirmed. In scalp tissues, hair loss occurs due to a combination of internal and external factors, and several studies are being conducted to counteract this. However, not many studies have addressed hair loss from the perspective of the microbiome. In this study, subjects with hair loss and those with normal scalps were set as experimental and control groups, respectively. In the experimental group, hair loss had progressed, and there was a large difference in microbiome composition compared to the group with normal scalps. In particular, differences in Accumulibacter, Staphylococcus, and Corynebacterium were found. From Staphylococcus epidermidis Cicaria, two active components were isolated as a result of repeated column chromatography. Spectroscopic data led to the determination of chemical structures for adenosine and biotin. Fractions were obtained, and ex vivo tests were conducted using hair follicles derived from human scalp tissue. When the microbiome adenosine-treated group was compared to the control group, hair follicle length was increased, and hair root diameter was maintained during the experimental periods. In addition, the Cicaria culture medium and the microbial adenosine- and biotin-treated groups maintained the anagen phase, reducing progression to the catagen phase in the hair growth cycle. In conclusion, it was confirmed that the Cicaria culture medium and the microbial adenosine and biotin derived from the culture were effective in inhibiting hair loss.

Abstract A87: The A2AR antagonist AZD4635 prevents adenosine-mediated immunosuppression in tumor microenvironment and enhances antitumor immunity partly by enhancing CD103+ dendritic cells

Abstract Adenosine signaling via the adenosine 2A receptor (A2AR) is emerging as an important regulatory mechanism of immune responses. Adenosine levels are increased in the tumor microenvironment by the CD39/CD73 axis and adenosine signaling through the adenosine 2A receptor (A2AR) on immune cells elicits a range of immunosuppressive effects that promote tumor growth and limit the efficacy of immune checkpoint inhibitors. We provide evidence that AZD4635, an oral A2AR antagonist, inhibits downstream signaling causing enhanced antigen presentation by CD103+ DCs and increased T-cell activation. In our in vitro assays, NECA (a stable analog of adenosine) caused potent suppression of CD3/CD28 stimulated IFNγ secretion in T cells with 1 μM NECA leading to 80% suppression of IFNγ secretion. Incubation of CD8+ T cells with AZD4635 restored IFNγ secretion back to baseline levels with an observed EC50 of 0.053μM and 0.598 μM at 0.1 and 1 μM NECA, respectively. Treatment of mice bearing CT26 colorectal carcinoma and MC38 colorectal carcinoma with AZD4635 monotherapy led to tumor growth inhibition with (TGI) of 44% and 73%, respectively, and in combination with anti-PD-1 with a TGI of 73% and 91%, respectively. Ex vivo analysis of MC38 OVA tumor-bearing mice showed increase in intratumoral CD8 T cells and CD103+DCs in treated groups compared to control. In particular, combination treatment increased the frequency of OVA antigen specific CD8+ T cells as measured by % of cells expressing the activation marker CD25 (2-fold increase) or the proliferation marker Ki67 (2-fold increase). Aligned with increased tumor antigen-specific T-cell response was a 2-fold increase of intratumoral CD103+ DCs. CD103+DCs play a critical role in tumor antigen trafficking and cross-priming to T cells, which are crucial for successful tumor rejection. In vitro, treatment of CD103+ DCs with NECA suppressed about 50% of pinocytosis (uptake of fluorescent beads) and antigen presentation (OVA peptides), which was almost completely rescued by AZD4635 treatment. We extend these studies into human DCs, and show adenosine promotes a tolerogenic phenotype that can be reversed with AZD4635 restoring antigen presentation and T-cell costimulation by DCs, leading to 50% increase in priming and expansion of antigen-specific T cells compared to adenosine-treated group. Our results support the role of adenosine signaling as an intrinsic negative regulator of CD103+ DC maturation and priming. We show that potent inhibition of A2AR with AZD4635 reduces tumor burden and enhances antitumor immunity. Our preclinical data support the evaluation of AZD4635 as a single agent and in combination with durvalumab (αPD-L1 antibody) in patients with solid malignancies (www.clinicaltrials.gov- NCT02740985, NCT03980821). Citation Format: Dinesh Chandra, Christine M. Barbon, Alexandra Borodovsky, Yanjun Wang, Minwei Ye, Laura Prickett, Kris Sachsenmeier, Wenlin Shao, Carl Barrett, Stephen Fawell, Deanna A. Mele, Alwin Schuller. The A2AR antagonist AZD4635 prevents adenosine-mediated immunosuppression in tumor microenvironment and enhances antitumor immunity partly by enhancing CD103+ dendritic cells [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr A87.

Promotive effects of adenosine on human hair follicle growth and angiogenin expression

Objective To evaluate the promotive effects of adenosine on human hair follicle growth and angiogenin expression. Methods Intact human hair follicles in the anagen phase were isolated from the scalp, and then divided into 6 groups to be treated with adenosine at different concentrations of 0, 0.1, 1, 10, 100 and 1 000 μmol/L respectively. Each adenosine group was further divided into 2 subgroups to be treated either with or without 0.8 mg/L anti-human angiogenin antibody, and hair follicles receiving no treatment served as the control group. The growth length of hair follicles during 6-day culture was measured by using a microscope. Human dermal papilla cells were isolated by a two-step enzyme digestion method, and then classified into the same groups as above. After 48-hour culture, methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate cellular proliferative activity, flow cytometry to analyze cell cycle, reverse transcription (RT)-PCR to measure the mRNA expression of angiogenin in dermal papilla cells, and enzyme-linked immunosorbent assay (ELISA) to measure the protein expression of angiogenin in the culture supernatant of dermal papilla cells. Results Compared with the control group, the 10-1 000 μmol/L adenosine-treated groups showed significantly increased growth of hair follicles in vitro (F = 377.776, P < 0.05), and 100 μmol/L adenosine had the strongest promotive effect (P < 0.05). However, anti-human angiogenin antibody could partly antagonize the promotive effect. Compared with the control group, the proliferative activity of human dermal papilla cells was significantly enhanced in the 0.1-1 000 μmol/L adenosine-treated groups (F = 230.067, P < 0.05), and adenosine at 10 and 100 μmol/L had the strongest enhancing effects (both P < 0.05). Similarly, anti-human angiogenin antibody could partly antagonize the enhancing effects of adenosine on cell proliferation. As flow cytometry showed, compared with the control group, the 10-1 000 μmol/L adenosine-treated groups showed significantly lower proportions of dermal papilla cells in G1 phase (F = 75.5, P < 0.05), but higher proportions of cells in G2 phase (F = 15.7, P < 0.05) and S phase (F = 81.8, P < 0.05). In addition, the mRNA and protein expressions of angiogenin were significantly up-regulated in human dermal papilla cells treated by 10 and 100 μmol/L adenosine compared with the control group (F = 942.630, 148.544, respectively, both P < 0.05). Conclusion Adenosine can promote the growth of human hair follicles and proliferation of dermal papilla cells, likely by up-regulating angiogenin expression. Key words: Adenosine; Angiogenins; Hair follicle; Cell proliferation; Cell cycle; Dermal papilla cells

Cardioprotective effect of high‐dose intragraft adenosine infusion on microvascular function and prevention of no‐reflow during saphenous vein grafts intervention

Despite the use of embolic protection devices, no-reflow can still occur during saphenous vein grafts (SVGs) intervention. High-dose intracoronary adenosine infusion preconditions the myocardium, improves coronary flow, and prevents no-reflow. The role of high-dose intragraft adenosine infusion on protection of microvascular function and prevention of no-reflow has not been investigated We investigated the cardioprotective effect of high-dose intragraft adenosine infusion, compared with placebo, on microvascular function and prevention of no-reflow during SVGs intervention. We randomized 22 patients with SVGs stenoses to receive either a 10-min intragraft adenosine infusion (200 μg/min; total dose = 2,000 μg) or normal saline prior to stenting. Average peak velocity (APV), coronary flow velocity reserve (CVR), thrombolysis in myocardial infarction (TIMI) frame count (TFC), TIMI myocardial perfusion grade (TMPG), and the rate of no-reflow were compared between the two groups before adenosine or saline infusions and after stenting After stenting, hyperemic APV, CVR, and TMPG were significantly higher in the adenosine-treated group than in the control group (60 ± 18 vs. 35 ± 10 cm/sec; 2.6 ± 0.54 vs. 1.8 ± 0.47; and 2.8 ± 0.90 vs. 2.1 ± 0.80, respectively; P < 0.05. TFC was significantly lower in the adenosine-treated group than in the control group (14 ± 3.0 vs. 26 ± 13; P < 0.05). In the control group, four patients (36%) developed no-reflow compared to none in the adenosine-treated patient; P < 0.05 CONCLUSIONS: This study provides the first evidence that high-dose intragraft adenosine infusion compared with placebo protects microvascular function and prevents no-reflow during SVGs intervention.

Intracoronary infusion of adenosine reduces infarct size and no-reflow in ST-segment elevation myocardial infarction

Purpose: Adenosine exerts a variety of actions that may attenuate many of the proposed mechanisms of reperfusion-mediated injury, including inhibition of neutrophil-mediated microvascular obstruction. Yet, attempts to achieve cardioprotection with administration of adenosine at reperfusion have yielded conflicting results in both pre-clinical and clinical settings. We sought to determine the effects of adenosine on infarct size (IS) and no-reflow in an appropriate translational porcine model, taking into account the predicted terms for efficacious intervention (i.e., prolonged intracoronary infusion starting just prior to reperfusion). Methods: Thirty-six open-chest swine of either sex were subjected to a 45-min left anterior descending coronary artery occlusion followed by 2 h of reperfusion. Animals received either intracoronary adenosine (50 μg/kg/min; n=15) infused for 2 h starting 5 min prior to reperfusion or an equal volume of saline (controls; n=21). The area-at-risk was delineated with Evans Blue, IS was determined histochemically with triphenyltetrazolium staining and no-reflow measured with thioflavin S staining. Results: Adenosine infusion significantly increased coronary blood flow throughout the 2-h reperfusion period compared with controls (p<0.001), with a maximum fourfold increase relative to baseline at 30 min of reperfusion (397±62% vs. 217±18% in controls; p<0.001). In contrast, adenosine did not affect systemic hemodynamic variables. Area-at-risk was similar between groups (p=NS). Administration of adenosine resulted in a significant reduction in IS (% area-at-risk; 46±4%) compared with controls (59±3%; p=0.02). Likewise, no-reflow (% IS) was significantly reduced in the adenosine-treated group (26±6%) compared to controls (49±6%; p=0.03). Conclusion: High and prolonged intracoronary adenosine infusion during early reperfusion limits infarct size and no-reflow in a porcine preparation of ST-segment elevation myocardial infarction. Further studies are warranted to confirm this effect, especially, when considering that there is no successful clinical pharmacological treatment for prevention of reperfusion-mediated injury at present.

EFFECTS OF FERTILIZATION PROMOTING PEPTIDE, ADENOSINE, AND PENTOXIFYLLINE ON THAWED HUMAN SPERM

Fertilization promoting peptide (FPP) and adenosine were demonstrated to be potential modulators of sperm capacitation in mammals. Both FPP and adenosine, by modulating the adenylate cyclase (AC)/cAMP signaling pathway, elicit similar biphasic responses in mammalian sperm (i.e., stimulating capacitation and inhibiting spontaneous acrosome loss). Pentoxifylline, an artificial sperm stimulant, is clinically used to enhance motility of sperm from infertile men. By inhibiting phosphodiesterase, pentoxifylline increases the intracellular cAMP level of sperm, and thus contributes to capacitation, hyperactivation, and acrosome reaction in animal studies. The effects of FPP, adenosine, and pentoxifylline on thawed human sperm are stressed. Chlortetracycline (CTC) fluorescence assessment revealed that none of the 3 reagents improved fertilization ability of post-thawed sperm. Motility studies with computer-aided sperm analyzer (CASA) showed significantly smaller STR (straight-line velocity) and LIN (linearity) in the FPP-treated group at 4 h of incubation p\leqq. 005, significantly larger VCL in the adenosine-treated group p\leqq. 05, and significantly larger VCL (curvilinear velocity) and smaller LIN in the pentoxifylline-treated group p\leqq. 05. Significant decreases in percentage motility were also noted in both FPP and adenosine-treated groups p\leqq. 05. It would appear that FPP potentiates fertilizing ability and prevents spontaneous acrosome loss, via regulating membrane-bound Na + -K + ATPase, and/or Ca 2+ ATPase, by keeping the sperm intracellular Ca 2+ concentration within the physiological range optimal for fertilization.

Effects of fertilization promoting peptide, adenosine, and pentoxifylline on thawed human sperm.

Fertilization promoting peptide (FPP) and adenosine were demonstrated to be potential modulators of sperm capacitation in mammals. Both FPP and adenosine, by modulating the adenylate cyclase (AC)/cAMP signaling pathway, elicit similar biphasic responses in mammalian sperm (i.e., stimulating capacitation and inhibiting spontaneous acrosome loss). Pentoxifylline, an artificial sperm stimulant, is clinically used to enhance motility of sperm from infertile men. By inhibiting phosphodiesterase, pentoxifylline increases the intracellular cAMP level of sperm, and thus contributes to capacitation, hyperactivation, and acrosome reaction in animal studies. The effects of FPP, adenosine, and pentoxifylline on thawed human sperm are stressed. Chlortetracycline (CTC) fluorescence assessment revealed that none of the 3 reagents improved fertilization ability of post-thawed sperm. Motility studies with computer-aided sperm analyzer (CASA) showed significantly smaller STR (straight-line velocity) and LIN (linearity) in the FPP-treated group at 4 h of incubation p<or=005, significantly larger VCL in the adenosine-treated group p<or=.05, and significantly larger VCL (curvilinear velocity) and smaller LIN in the pentoxifylline-treated group p<or=.05. Significant decreases in percentage motility were also noted in both FPP and adenosine-treated groups p<or=05. It would appear that FPP potentiates fertilizing ability and prevents spontaneous acrosome loss, via regulating membrane-bound Na(+)-K(+) ATPase, and/or Ca(2+) ATPase, by keeping the sperm intracellular Ca(2+) concentration within the physiological range optimal for fertilization. adenosinefertilizationFPPpentoxifyllinespermthawing

Protective effect of adenosine against neuronal injury induced by middle cerebral artery occlusion in rats as evidenced by diffusion-weighted imaging

In the present study, adenosine, an inhibitory neuromodulator, was studied in male Wistar rats subjected to 2 h of transient middle cerebral artery (MCA) occlusion. Adenosine (500 mg/kg ip) was administered twice—once at the time of MCA occlusion and again at the time of reperfusion—and evaluated for its protective effect by using diffusion-weighted imaging (DWI) (30 min after reperfusion). After the DWI experiments, one group of animals was euthanized 2 h after reperfusion for the estimation of oxidative stress markers, while in another group, neurological deficit was assessed 24 h after MCA occlusion. In the adenosine-treated group, percent hemispheric lesion area (%HLA) in DWI was significantly attenuated (11.7±5.2) as compared to vehicle-treated group (21.4±4.7). The level of malondialdehyde (MDA) (301.8±22 nmol/g wet tissue) in the adenosine-treated group was significantly decreased as compared to that in the vehicle-treated MCA-occluded rats (420±20 nmol/g wet tissue). An insignificant change was observed in the levels of glutathione in both the vehicle-treated MCA-occluded and the adenosine-treated groups. The neurological deficit was significantly improved in the adenosine-treated group (1.8±0.06) as compared to the vehicle-treated (2.9±0.38) group. This is the first study to demonstrate the effectiveness of adenosine using DWI in the MCA-occluded rats.

Effects of adenosine pretreatment on detection of free radicals in ischemic and reperfused canine gracilis muscle flaps by use of spin-trapping electron paramagnetic resonance spectroscopy

To determine whether adenosine pretreatment attenuates free radical production and muscle damage in ischemic and reperfused canine skeletal muscle. 9 healthy mixed-breed dogs. Dogs were anesthetized, and both gracilis muscles were isolated, leaving only the major vascular pedicle intact. Saline (0.9% NaCl) solution was injected into the artery supplying the control flap, whereas adenosine (10 mg) was injected into the contralateral artery. Ischemia was induced in both flaps for 4 hours. alpha-Phenyl-N-tert-butylnitrone was administered IV to each dog 1 hour prior to reperfusion. Following 15 minutes of reperfusion, effluent blood samples from each muscle flap were obtained and processed for spin-trapping electron paramagnetic resonance (EPR) spectroscopy. Muscle biopsy specimens were obtained for histologic evaluation, and dogs were euthanatized. EPR spectra of strong intensity were obtained from analysis of 5 of 9 paired samples. Signals identified were characteristic of oxygen- and carbon-centered free radical adducts. Signal intensity of spectra from adenosine-treated flaps was significantly less than that of control flaps; mean signal attenuation was 36% in the adenosine-treated group. Histologic evaluation of muscle flaps did not reveal significant differences between groups. Treatment of canine muscle flaps with adenosine prior to a period of ischemia reduced but did not completely attenuate free radical production after reperfusion. However, adenosine pretreatment did not affect histologic abnormalities.

The gastric cytoprotective action of adenosine and prostaglandin E2 in rabbits.

The direct protective action of adenosine and prostaglandin E2 (PGE2) was examined in an isolated gastric gland preparation in rabbits. Ethanol (8%, v/v) incubation markedly increased the release of lactate dehydrogenase (LDH) and number of non-viable glands in the preparation. Both effects were prevented by PGE2 preincubation in a concentration (10(-6), 1.4 x 10(-5) or 2.8 x 10(-5) M)-dependent manner. The protective action was smaller in adenosine-treated groups, and yet the highest concentration (10(-4) M) of the compound also significantly inhibited the cytotoxic effects of ethanol. These findings indicate that both adenosine and PGE2 possess cytoprotective action on gastric glands in rabbits, but the former compound exerts its action beyond physiological concentrations. It is concluded that endogenous PGE2, but not adenosine may act as an ulcer modulator in the stomach.

Effects of low‐dose adenosine on myocardial performance after coronary artery bypass surgery

The effect of a non-hypotensive dose of adenosine infusion on myocardial performance after coronary artery bypass surgery was examined. Upon arrival at the intensive care unit, 16 patients (14 males, 2 females; mean age 64.5, range 46-71) were randomized to a blinded infusion of either low-dose adenosine (n = 8) or placebo (n = 8). The infusion continued at a rate corresponding to 30 micrograms.kg-1.min-1 of adenosine into the right ventricle over 4 h. Data were collected from the arterial line, thermodilution pulmonary artery catheter, transoesophageal echocardiogram (TEE), and 12-lead ECG on six occasions: before infusion, hourly during the infusion, and 1 h after terminating the infusion. Mean arterial blood pressure did not differ between the adenosine and placebo groups at any measurement point. Heart rate increased by approximately 15% during the first hour of adenosine infusion. Cardiac index increased by approximately 50% during infusion of adenosine and cardiac index remained higher while systemic vascular resistance remained lower in the adenosine-treated group during infusion. The E/A ratio (ratio between peak left ventricular inflow blood velocities during early filling and atrial contraction) was significantly higher in the adenosine-treated group after treatment for 1 h while the area injection fraction did not differ between groups at any time. The number of patients with ischaemic events as judged from ECG and from left ventricular regional wall motion abnormalities (RWMA) as visualized by TEE did not differ between groups (ECG: one patient in the adenosine group and one patient in the placebo group-RWMA: four patients in the adenosine group versus three in the placebo group).(ABSTRACT TRUNCATED AT 250 WORDS)

Reduction of myocardial reperfusion injury by intravenous adenosine administered during the early reperfusion period.

Adenosine influences the function of several cell types thought to be involved in the pathogenesis of myocardial reperfusion injury. We have previously demonstrated that intracoronary administration of adenosine enhances myocardial salvage 24 hours after reperfusion. To determine if these beneficial effects could be obtained during a prolonged period of reperfusion using an intravenous route of administration, 22 closed-chest dogs were subjected to 90 minutes of proximal left anterior descending coronary artery occlusion and 72 hours of reperfusion. Animals randomly received either intravenous adenosine (0.15 mg/kg/min) or an equal volume of Ringer's lactate during the first 150 minutes of reperfusion. The area at risk was defined in vivo with Monastral blue, and infarct size was measured histologically with Mallory's trichrome stain. Serial global and regional ventricular function were determined with contrast ventriculography and analyzed using a computerized radial shortening method. Biopsies were obtained from the central ischemic zone to assess endothelial ultrastructure and capillary obstruction. No significant effects in heart rate or blood pressure were noted during adenosine infusion. Transmural collateral blood flow during ischemia was similar in the groups. Infarct size expressed as a percentage of the anatomical area at risk was significantly less in the adenosine-treated group (35.3 +/- 4.3% in controls versus 17.1 +/- 4.3% in treated animals, p less than 0.01). A progressive decrease in transmural blood flow was noted in control animals during reperfusion, resulting in a significant reduction at 3 hours compared with the preocclusion value (0.69 +/- 0.11 ml/min/g [at baseline versus 0.45 +/- 0.10 ml/min/g at 3 hours, p less than 0.05]). In contrast, flow in adenosine animals at 3 hours was similar to baseline values (0.91 +/- 0.15 ml/min/g at baseline versus 0.98 +/- 0.14 ml/min/g at 3 hours, p = NS) and was significantly higher (p less than 0.05) than the control group. Radial shortening in the ischemic zone was significantly improved at 3 (-2.6 +/- 2.8% in controls versus 11.6 +/- 3.3% in treated animals, p less than 0.01) and 72 hours (5.5 +/- 2.0% in controls versus 17.3 +/- 3.5% in treated animals, p less than 0.01) after reperfusion in treated animals. Electron microscopy showed reduced neutrophil and erythrocyte plugging of capillaries with relative preservation of endothelial cell structure in the adenosine group.(ABSTRACT TRUNCATED AT 400 WORDS)

Functional and metabolic evidence of enhanced myocardial tolerance to ischemia and reperfusion with adenosine

An isolated, isovolumetrically contracting rat heart preparation, perfused at constant flow, was used to test the hypothesis that adenosine treatment (100 microM) throughout the experiment could enhance the repletion of adenosine triphosphate and the recovery of ventricular function following 10 minutes of global, normothermic (37 degrees C) ischemia. Left ventricular developed pressure was measured with an intraventricular balloon, and myocardial adenine nucleotides were measured from freeze-clamped tissues in a parallel series of experiments. The adenosine triphosphate level in the adenosine-treated hearts was not different from that of the untreated control hearts at the end of 30 minutes of equilibration but was significantly (p less than 0.05) higher at the end of 10 minutes of ischemia and at 15, 30, and 60 minutes of reperfusion. Left ventricular developed pressure in the adenosine-treated group at the end of 30 minutes of equilibration (92 +/- 3 mm Hg) was not significantly different from that of the control hearts (101 +/- 10 mm Hg). During the reperfusion period the control group returned to 75% +/- 7%, 73% +/- 6%, and 73% +/- 6% of the preischemic control function at 15, 30, and 60 minutes of reperfusion, respectively. The adenosine-treated group had significantly greater return of function to 86% +/- 3%, 96% +/- 3%, and 95% +/- 3% of the preischemic control at 15, 30, and 60 minutes of reperfusion, respectively. In a protocol to assess the effect of adenosine during ischemia, we found that adenosine (100 microM) increased the time to onset of ischemic contracture by 50% from 12 +/- 3 to 18 +/- 3 minutes and decreased the rate of net adenosine triphosphate degradation. Our data suggest that under these experimental conditions, adenosine enhances myocardial preservation by reducing the net degradation of adenosine triphosphate during ischemia and facilitating the repletion of adenosine triphosphate during reperfusion.