Alpha-2b Research Articles

Functional Polymorphism in the ADRB3 Gene, Encoding the Beta-3 Adrenergic Receptor, and Response to Intra-Detrusor Injection of Botulinum Toxin-A in Women with Overactive Bladder.

There are reports suggesting an association between the rs4994 polymorphism in the ADRB3 gene encoding the beta-3 adrenergic receptor and OAB risk in females. The injection of botulinum toxin-A into the bladder wall is recommended as a possible treatment for OAB patients in whom first-line therapies have failed. The aim of our study was to analyze the possible association between the ADRB3:rs4994 polymorphism and the patient-perceived response to a single intra-detrusor injection of botulinum toxin-A in Polish women with overactive bladder. The study group consisted of 115 consecutive female patients with OAB. The response to botulinum toxin-A was evaluated at three months after injection, as absolute or relative reductions in OAB symptoms or in scores from questionnaires ICIQ-OAB (parts A and B) and ICIQ-LUTS-QoL (parts A and B). ADRB3:rs4994 variants were identified by the sequencing of genomic DNA extracted from buccal swabs. There were no statistically significant differences between ADRB3:rs4994 [T];[T] homozygotes and [T];[C]+[C];[C] subjects for absolute or relative reductions in symptoms or in scores from all four questionnaire parts at three months after the injection of botulinum toxin-A. Our results do not support the hypothesis that ADRB3:rs4994 polymorphism is associated with the response to the intra-detrusor injection of botulinum toxin-A in Polish females with overactive bladder.

Off-target pharmacological profiling of synthetic cannabinoid receptor agonists including AMB-FUBINACA, CUMYL-PINACA, PB-22, and XLR-11.

Synthetic cannabinoid receptor agonists (SCRAs) are a diverse class of new psychoactive substances that have been associated with multiple instances and types of toxicity. Some SCRAs appear to carry a greater toxicological burden than others, or compared to the prototypical cannabis-derived agonist Δ9-tetrahydrocannabinol (Δ9-THC), despite a common primary mechanism of action via cannabinoid 1 (CB1) receptors. "Off-target" (i.e., non-CB1 receptor) effects could underpin this differential toxicity, although there are limited data around the activity of SCRAs at such targets. A selection of 7 SCRAs (AMB-FUBINACA, XLR11, PB-22, AKB-48, AB-CHMINICA, CUMYL-PINACA, and 4F-MDMB-BUTINACA), representing several distinct chemotypes and toxicological profiles, underwent a 30 μM single-point screen against 241 G protein-coupled receptor (GPCR) targets in antagonist and agonist mode using a cellular β-arrestin recruitment assay. Strong screening "hits" at specific GPCRs were followed up in detail using concentration-response assays with AMB-FUBINACA, a SCRA with a particularly notable history of toxicological liability. The single-point screen yielded few hits in agonist mode for any compound aside from CB1 and CB2 receptors, but many hits in antagonist mode, including a range of chemokine receptors, the oxytocin receptor, and histamine receptors. Concentration-response experiments showed that AMB-FUBINACA inhibited most off-targets only at the highest 30 μM concentration, with inhibition of only a small subset of targets, including H1 histamine and α2B adrenergic receptors, at lower concentrations (≥1 μM). AMB-FUBINACA also produced concentration-dependent CB1 receptor signaling disruption at concentrations higher than 1 μM, but did not produce overt cytotoxicity beyond CP55,940 or Δ9-THC in CB1 expressing cells. These results suggest that while some "off-targets" could possibly contribute to the SCRA toxidrome, particularly at high concentrations, CB1-mediated cellular dysfunction provides support for hypotheses concerning on-target, rather than off-target, toxicity. Further investigation of non-GPCR off-targets is warranted.

Efficacy of Ropeginterferon Alpha 2b in Inducing Treatment Free Remission in Chronic Myeloid Leukemia - an International, Randomized Phase III Trial (ENDURE, CML-IX) of the German CML-Study Group

Background: Only a minority of chronic myeloid leukemia (CML) patients can eventually successfully discontinue tyrosine kinase inhibitor (TKI) therapy after achieving a stable deep molecular remission. Mechanisms underlying a durable treatment free remission (TFR) are not well understood but supposedly involve restoration of anti-CML immunity. It was therefore hypothesized that interferon alpha (IFN) - a potent inducer of Th1-immunity and prior standard therapy in CML - might improve the TFR probability. Randomized trials to improve TFR rates in CML are currently lacking. ENDURE, CML-IX (NCT03117816) is a multicenter, international phase III trial evaluating the role of a novel form of pegylated proline interferon-alpha 2b (ropeg-interferon-alpha, ropeg-IFN) in inducing TFR. Patients and Methods: CML patients in stable deep molecular remission (MR4 or better for at least 12 months prior to screening) and with history of at least 3 years of TKI exposure were eligible to participate in this study. Patients in stable molecular remission could also be included if they had failed a former TKI discontinuation attempt (2nd stopper). Patients were randomized 1:1 to receive either ropeg-IFN 100μg s.c. every 2 weeks for 15 months (ropeg-IFN arm) or no further treatment after TKI stop treatment (no treatment arm). The primary efficacy endpoint of the trial was molecular relapse free survival (MRFS) with molecular relapse being defined as loss of major molecular remission (MMR), which is any increase of the BCR::ABL1 transcript level to >0.1% according to the international scale (IS). Time to relapse is defined as the time from randomization to relapse. Secondary endpoints were MRFS at month 6, 12 and 24 after TKI discontinuation. BCR::ABL1 measurable residual disease (MRD) was monitored from peripheral blood and centrally assessed for all centers in Germany and locally at the French centers. There was a post study follow up to collect clinical and molecular data. Results: Between May 2017 and June 2021, 203 evaluable patients (68 female, 135 male) with a median age of 55 years were randomized at 24 centers in Germany and 3 centers in France to receive ropeg-IFN (n=95) or no treatment (n=108) after TKI stop. For 77 patients in the ropeg-IFN arm (81%) and 86 patients in the no treatment arms (80%) it was their first TKI discontinuation attempt. At the time of data cut off in June 2022, the median observation time for all patients was 36 months. The hazard ratio (HR) of molecular relapse for the ropeg-IFN cohort versus the no treatment cohort was 1.03 (95% CI, 0.68 to 0.1.55; log-rank P=0.89). The Kaplan-Meier probabilities of molecular relapse free survival (MRFS) by 6, 12, and 24 months after TKI discontinuation were 73% (95%-CI: 62-81%), 64% (95-CI: 53-73%) and 56% (95%-CI: 45-66%) for the ropeg-IFN versus 67% (95-CI: 57-75%), 60% (95-CI: 50-69%) and 59% (95%-CI: 49-68%) for no treatment (Figure 1). The MRFS at months 6 and 12 after TKI stop (secondary endpoints) were 70% (95%-CI: 60-79%) and 64% (95%-CI: 53-73%) in 91 patients for ropeg-IFN group versus 65% (95-CI: 56-74%) and 59% (95-CI: 49-68%) in 107 and 104 patients for the no treatment group. Of 90 patients who were candidates for restarting TKI due to MMR loss, 82 patients were actually evaluable for TKI restart. Of those, 77 patients re-achieved at least MMR within 12 months (median time to reestablishing MMR was three months). Of the 5 patients who did not regain MMR, one patient withdrew consent and others were treated for only 1, 3, 4, and 11 months after TKI restart. Ropeg-IFN was well tolerated. Hematological and non-hematological WHO grade 3/4 toxicity occurred in 23 patients in the ropeg-IFN (31 adverse events) and in 20 patients in the no treatment arm (26 adverse events). Three deaths were reported on study which were unrelated to CML (cardiac arrest, fall from stairs, unknown). A detailed safety analysis will be presented at the meeting. Conclusion: Ropeg-IFN maintenance after discontinuing TKI-monotherapy does not increase the proportion of patients, who persistently maintain at least an MMR. The German CML-V (TIGER) trial currently explores the impact of IFN maintenance on TFR when patients receive a combination of TKI plus IFN before TKI stop. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Structure-based drug discovery facilitates future painkiller development.

Structure-based drug discovery facilitates future painkiller development.

Real-life data of adjuvant IFN-α2b and MMC in conjunctival melanocytic lesions

PurposeWe herein compare topical interferon alpha 2b (IFN-α2b) to topical mitomycin C (MMC) in the adjuvant management after excision of primary acquired melanosis with atypia (PAM) and melanoma of the conjunctiva/cornea (CM).MethodsWe included 25 tumors from 25 patients (six with PAM and 19 with CM). After surgical excision, four patients started with adjuvant IFN-α2b (two in combination with radiotherapy), 19 with MMC, and two with radiotherapy alone. Five patients were switched from initial MMC/radiotherapy to IFN-α2b during follow-up. Efficacy was assessed via time to tumor recurrence and initial therapy response.ResultsWith initial IFN-α2b, three patients (3/4, two with additional radiotherapy) showed complete remission (follow-up: 1478–1750 days) and one recurrence (1/4) was noted after 492 days. With initial MMC, no recurrence was recorded in 15 of the 19 patients (follow-up: 99–4732 days). Five patients were switched from MMC or radiotherapy to IFN-α2b: two patients showed complete remission (2/5), while another two (2/5) experienced recurrences and remained without recurrence after repeated courses of IFN-α2b (follow-up: 1798 and 1973 days). Only one patient showed incomplete response. Adverse effects were recorded in five patients, all received MMC.ConclusionTopical IFN-α2b (arguably together with radiotherapy) may be a viable alternative to MMC in PAM and CM. We observed fewer side effects at similar response rates. However, when response to MMC was poor, IFN-α2b may also be of limited utility.

Deployment of metabolic heat rate based soft sensor for estimation and control of specific growth rate in glycoengineered Pichia pastoris for human interferon alpha 2b production

Lack of appropriate process models, reliable online sensors, and process variability in bioprocess systems are poising challenges in real-time monitoring and control of critical process parameters (CPPs). This present investigation deals with the development of a non-invasive soft sensor by utilizing metabolic heat rate as input signal for online estimation of specific growth rate (μest) during the induction phase of glycoengineered Pichia pastoris for human interferon-alpha 2b (huIFNα2b) production. Feedforward strategy employing a predetermined exponential feeding of methanol during the induction phase was dealt at defined setpoint values (μSP). Standard PID controller with predetermined gain values regulated methanol feeding in accordance with the deviation from the μSP value. An adaptive PID (gain scheduling) significantly minimized the deviation of μ from its μSP value, reduced the amplitude of oscillation and achieved long-term controller stability. Robust control of methanol feeding by adaptive PID resulted in a 1.5 and 2.2-fold increase in productivity of huIFNα2b compared to standard PID and feedforward controls respectively. Moreover, adaptive PID control facilitated narrow range control of μ for longer durations (> 20 h) with a low average tracking error (< 6%) enumerating its scope of application in therapeutic protein production in near future.

The use of tamsulosin in the treatment of 10–15 mm lower ureteral stones in adults: a double-blinded randomized controlled trial

BackgroundThe discovery of alpha-adrenergic receptors in the ureteral smooth muscle cells led to a thorough investigation of the therapeutic potential of alpha-blockers for ureteral calculi. Tamsulosin is a selective blocker of alpha-1A and alpha-1B adrenoceptors. It significantly improved the expulsion of distal ureteral stones measuring 3–10 mm in most randomized trials. To the best of our knowledge, tamsulosin was not tested before in the management of ureteral stones 10–15 mm. Hence, the present study aimed at estimation of the efficacy of tamsulosin in the expulsion of lower ureteral stones 10–15 mm in length compared to placebo in adult patients (primary goal) and the need for ureteral dilatation at scheduled ureteroscopy.MethodsBetween November 2017 and November 2019, 80 patients with distal ureteral stones 10–15 mm were divided into two equal groups. One group received tamsulosin 0.4 mg/day and the other received a placebo. Patients were followed-up for 8 weeks.ResultsSix patients of the tamsulosin group (15%) passed their stones spontaneously compared to none of the placebo group (p = 0.026). Two of the 6 patients who passed the ureteral stones developed urinary retention and required endoscopic treatment of urethral stones. So, the adjusted spontaneous ureteral stone passage ratios for the tamsulosin and the control groups were 10 and 0%, respectively (p = 0.12). Overall, 37.5% reported adverse effects in the tamsulosin group, and 30% in the placebo group (p = 0.7). The most common adverse effect reported in both groups was dizziness, which occurred more frequently with tamsulosin (25%) than placebo (22.5%) (p = 0.9). We noticed increased ejaculatory dysfunction among men in the tamsulosin group versus placebo group [17.9% vs. 3.5% (p = 0.1)]. Among patients who needed ureteroscopy, ureteral dilatation was always needed in the control group versus 85.3% of the tamsulosin group (p = 0.015). Although 23.5% of the treatment group didn’t need stents after ureteroscopy compared to only 12.5% of the control group, this difference was not statistically significant (p = 0.2).ConclusionsAlthough tamsulosin significantly increased spontaneous passage of 10–15 mm lower ureteral stones in adults, it did not decrease the need for operative intervention. Preoperative tamsulosin significantly facilitated ureteral dilations during ureteroscopic management.

The impact of adrenoceptor-1 gene polymorphism on structural and functional features in patients with hypertrophic cardiomyopathy

Abstract Background The adrenergic system plays a central role during the clinical course of hypertrophic cardiomyopathy (HCM) and hence beta-blocker therapy is the first-line treatment option according to current guidelines. Previous studies demonstrated variable cellular response to an adrenergic stimulus due to adrenoceptor-1 (ADRB-1) gene polymorphism in several patient groups including hypertension and dilated cardiomyopathy. Moreover, the impact of beta-blockers is also variable in these patients. Purpose We aimed to investigate the impact of ADRB-1 gene polymorphism on structural and functional features among patients with HCM. Methods In this study, we investigated the roles of Arginin389Glycine and Glycine49Serine polymorphism. Patients with a clear diagnosis of HCM according to current guidelines were included. Structural features including maximal wall thickness, interstitial fibrosis and left ventricular/atrial volumes were obtained using cardiac MRI and transthoracic echocardiography. Functional features including heart rate, blood pressure, left ventricular outflow obstruction were also recorded. The risk of sudden cardiac death was calculated using the HCM risk score. The impact of beta-blocker therapy on heart rate and blood pressure were also compared. Results After the exclusion of patients, 147 patients were included in the study. 77% of the study population were male and the mean age was 49.5 years. The hypertrophic segment was septum in 83% of the study population. The mean maximum wall thickness was 20 mm (19–23). With respect to Ser49Gly polymorphism, Serine homozygotes demonstrated higher late-gadolinium enhancement and indicator of interstitial fibrosis (p: 0.007). In concordance with higher LGE, the presence of fragmented QRS on surface ECG (p: 0.026) and a higher prevalence of non-sustained ventricular tachycardia (p: 0.016) in these patients were observed. The risk of sudden cardiac death was also higher in Ser49 homozygotes (p: 0.041). However, with respect to Arg389Gly gene polymorphism, there was no significant difference between Arg389 homozygotes, Arg389Gly heterozygotes and Gly389 homozygotes in terms of structural and functional features. The vast majority of patients reported improved functional capacity and decreased shortness of breath during the follow-up regardless of ADRB-1 gene polymorphism which is objectively validated by decreased pro-BNP levels in all patients. Conclusion Our results indicated that ADRB-1 gene polymorphism, particularly Ser49Gly gene polymorphism may have a significant role during the clinical course of HCM. Since Ser49 homozygote patients demonstrated higher interstitial fibrosis and a higher risk of sudden cardiac death, further studies are required to determine the significance of Ser49Gly gene polymorphism in HCM patients. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Bilimsel Arastırma Projesi (BAP) - Istanbul University - Cerrahpasa, Cerrahpasa Medical School

КЛИНИКО-ГЕНЕТИЧЕСКИЕ АСПЕКТЫ РЕГРЕССА ГИПЕРТРОФИИ МИОКАРДА ЛЕВОГО ЖЕЛУДОЧКА. РЕЗУЛЬТАТЫ ПРОСПЕКТИВНОГО ИССЛЕДОВАНИЯ В ГОРНОЙ ШОРИИ

Abstract. Introduction. Left ventricular hypertrophy is an independent predictor of adverse outcomes in patients with arterial hypertension. Aim. To evaluate clinical and genetic factors, as well as the effectiveness of antihypertensive drugs classes in terms of regression of left ventricular hypertrophy in the indigenous population of Mountain Shoria within 5-year prospective observations. Material and methods. The study was based on the results of a 5-year observation of the indigenous population of Mountain Shoria. The study included 901 people aged 18 and over. At the one-stage stage (from 2013 to 2017), an anamnesis and complaints were collected, a clinical examination by a cardiologist, anthropometry, a biochemical blood test was performed, and genetic testing of polymorphisms of arterial hypertension candidate genes (ACE (I/D, rs 4340), AGT (c.803T&gt;C, rs699), AGTR1 (А1166С, rs5186), ADRB1 (с.145A&gt;G, Ser49Gly, rs1801252), ADRA2B (I/D, rs28365031), MTHFR (С677Т, rs1801133), eNOS (VNTR 4b/4a)), performed electrocardiography, echocardiography. The prospective stage included 263 patients with hypertension without previous antihypertensive therapy. Blood pressure figures were monitored and corrected annually, and left ventricular hypertrophy dynamics was assessed 5 years after inclusion in the study. Statistical processing of the results of the study was carried out using the program "Statistica 10.0". Results and discussion. In the Shors cohort, left ventricular hypertrophy regression was associated with a lower degree of arterial hypertension (odds ratio 1.39) and with the achievement of target blood pressure levels (odds ratio 1.71), as well as with the addition of a diuretic, indapamide retard, to renin-angiotensin-aldosterone system blockers (odds ratio 2.65). In the present study, an association was found between carriage of the D/D genotype of the ACE gene (odds ratio 0.43) and 4a/4a of the eNOS gene (odds ratio 0.35) with negative dynamics in relation to myocardial mass of the left ventricle. With respect to genotype I/I of the ADRA2B gene (odds ratio 2.00) and 4b/4b of the eNOS gene (odds ratio 2.93), an associative relationship with regression of left ventricular hypertrophy was demonstrated. Conclusion. The results obtained once again prove the promise of using genetic approaches for the early diagnosis of left ventricular hypertrophy.

Impact of pharmacotherapy with tiotropium/olodaterol on the pulmonary function in COPD patients depending on the Arg16Gly polymorphism of ADRB2 gene

Annotation. Chronic obstructive pulmonary disease (COPD) is a prevalent chronic disease. Endogenic or genetic factors play a significant role in the COPD development. The aim of our study was to investigate a response to the tiotropium/olodaterol treatment in patients with different ADRB2 gene polymorphism. 100 patients with the diagnosis of COPD were included into the study. Pulmonary function testing was performed during all study visits. All patients received treatment according to GOLD 2019, which included a combination of long-acting beta-2 agonists (LABA) and long-acting muscarinic antagonists (LAMA). According to the results of genetic testing all patients were divided into 3 groups: Arg16Arg – 23 patients (23%), Arg16Gly – 39 patients (39%), Gly16Gly – 38 patients (38%). Statistical processing of research materials was carried out using the SPSS statistical program package for Windows version 26.0. Descriptive statistics were determined for each interval variable and presented as mean ± standard deviation (SD). Kolmogorov-Smirnov test, Mann-Whitney test and T-test, Wilkson test, t-test, Kruskal-Wallis test were used as part of the one-way ANOVA analysis, as well as with the analysis of covariance (ANCOVA) followed by pairwise comparison of groups using Bonferroni corrections. Groups did not differ by sex, age, part of smokers, COPD duration. There was significant difference in RLM at the time of inclusion (p=0.01). There was a significant difference in the Pain value at the time of study completion (р=0.024). When comparing values of SGRQ questionnaire, there was a significant difference in the scales of Impact (р=0.006) and Total score (р=0.021). Thus, group Gly16Gly had better dynamics of SF-36 and SGRQ values during the study. Despite the absence of significant difference we can state, that Gly16Gly genotype has better response to treatment when compared to Arg16Arg, which is probably conditioned by the development of sensitization of ADRB2 receptor in Arg16Arg group.

Vasovagal Syncope Is Associated with Variants in Genes Involved in Neurohumoral Signaling Pathways.

Vasovagal syncope (VVS) is the most common cause of sudden loss of consciousness. VVS results from cerebral hypoperfusion, due to abnormal autonomic control of blood circulation, leading to arterial hypotension. It is a complex disease, and its development is largely associated with genetic susceptibility. Since abnormal neurohumoral regulation plays an important role in VVS development, we analyzed the association of VVS with polymorphic variants of ADRA1A, ADRB1, HTR1A, ADORA2A, COMT, and NOS3 genes, the products of which are involved in neurohumoral signaling, in patients with a confirmed VVS diagnosis (157 subjects) and individuals without a history of syncope (161 subjects). We were able to identify the associations between VVS and alleles/genotypes ADRA1A rs1048101, ADRB1 rs1801253, ADORA2A rs5751876, and COMT rs4680, as well as NOS3 rs2070744 in biallelic combination with COMT rs4680. Thus, we are the first to observe, within a single study, the role of the genes that encode α- and β-adrenergic receptors, catechol-O-methyltransferase, adenosine receptors and nitric oxide synthase in VVS development. These findings demonstrate that the genes involved in neurohumoral signaling pathways contribute to the formation of a genetic susceptibility to VVS.

Clinical course of COPD in patients with Arg16Gly (rs1042713) polymorphism of ADRB2 gene.

The ADBR2 gene has been studied for its possible relationship with the development and clinical course of chronic obstructive pulmonary disease (COPD), including response to beta-2 agonists, with existing data being contentious on the subject. So, the purpose of this study was to look into the potential impact of the arginine-16-glycine (Arg16Gly) polymorphism on the clinical course and drug utilization in COPD patients. Data show that patients with Arg16Arg have a lower number of hospital admissions for exacerbations (p=0.048), but only in the total number of exacerbations, including those treated out-patients (p=0.086). Each glycine (Gly) copy was associated with a higher number of exacerbations (OR: 0.25; 95% CI: 0.00-055; p=0.048). The number of exacerbations after LABA/LAMA treatment was similar across groups, indicating that all ADRB2 variants responded well to the treatment. Furthermore, there were no statistically significant differences in mMRC and CAT values across all study visits. Interestingly, groups differed in their use of antibiotics (AB) at all visits, with Arg16Arg being associated with the least amount of AB use. There was also a link discovered between clycine copies and increased use of glucocorticoids. As a result, Arg16Gly is involved in the clinical course of COPD as well as the utilization of drug groups. Based on the findings, we can speculate that the cross-talk between the ADRB2 gene and the corticosteroid receptor is altered in patients with the Gly16Gly genotype.

Expression of the β1-adrenergic receptor (ADRB1) gene in the myocardium and β-adrenergic reactivity of the body in patients with a history of myocardium infraction

β1-adrenergic receptors (β1-AR) directly affect on intracardiac hemodynamic and the ability of the heart to tolerate physical activity by regulating its inotropic and chronotropic functions. Severe hypersympathicotonia, specific to coronary artery disease (CAD) and chronic heart failure (HF), leads to impaired functioning of β1-AR. The aim of this research was to assess the expression level of the β1-AR ADRB1 gene in the myocardium, to evaluate the β-adrenergic reactivity of the membrane (β-ARM) of erythrocytes, and to analyze the association of these parameters with myocardial contractile dysfunction in patients with a myocardial infarction (MI) in the past and without it. The study included 126 patients with chronic CAD. Among the patients, 55.6 % had a history of MI at least 6 months ago. The expression of the ADRB1 gene was assessed using real-time polymerase chain reaction. With this purpose, we isolated RNA from the right atrial appendage, which was excised when a heart–lung machine was connected during a planned coronary bypass surgery. β-ARM was evaluated in 57 patients. This method is based on the fact of inhibition of hemolysis of erythrocytes, placed in a hyposmotic medium, in the presence of a β-blocker. Within the whole sample of patients, the expression of the ADRB1 gene is comparable in different functional classes of HF. There was no linear correlation between the expression of the ADRB1 gene and left ventricle ejection fraction (LVEF). In patients with a history of MI, the expression of the ADRB1 gene was elevated when compared to a group of patients without MI (p = 0.017). Patients with a history of MI had higher values of β-ARM than those without MI (p = 0.017). The reverse correlation between β-ARM and LVEF (r = -0,570, p = 0,002) was revealed in the group of patients without MI but not in the group of patients with a history of MI (r = -0,137, p = 0,479). In the sample of patients with chronic CAD, in the myocardium of subjects with a history of MI, the relative expression of ADRB1 gene was higher compared to the group of patients without MI. In patients with different functional classes (FC) of HF and with different ejection fraction, both with MI and without it, ADRB1 gene expression was comparable. In the group of patients with a history of MI, an increase in β-ARM was observed, i.e. decrease in the number or sensitivity of β-AR. Among patients without MI, an inverse correlation was found between β-ARM and LVEF.

In vivo hypotensive effect of aminosilanol-based nanocomposites bearing antisense oligonucleotides

The search for new effective drugs for the treatment of arterial hypertension continues to be an urgent task. We studied the effect of aminosilanol nanocomposites (Si∼ODN) that contained antisense oligonucleotides on the blood pressure in the hypertensive ISIAH rat strain, an experimental model of stress-sensitive arterial hypertension. The intraperitoneal, intravenous, and inhalation administration of Si∼ODN targeted to mRNAs of the AT1A, ADRB1, and ACE1 genes decreased systolic blood pressure by 12–40 mm Hg. The use of scrambled oligonucleotides led to no antisense effect. The proposed in vivo delivery system was shown to be suitable for oligonucleotides with both natural and modified internucleotide bonds. The Si∼ODN nanocomposites demonstrated the absence of toxicity under the studied conditions. The intravenous administration of a fluorescently labeled nanocomposite led to its accumulation in the animal's liver and kidneys, and when inhaled, a certain amount was detected in the brain. The content of the label reaches a maximum one day after the treatment and is almost eliminated from the body in two weeks. The best way to correct hypertension in ISIAH rats using the studied nanocomposites may be periodic inhalation of the nanocomposite that carries an oligonucleotide targeted to ACE1-mRNA.

FTO promoted adipocyte differentiation by regulating ADRB1 gene through m6A modification in Hycole rabbits

N6-methyladenosine (m6A), the most abundant internal mRNA modification in eukaryotes, plays a vital role in regulating adipogenesis. However, its underlying mechanism remains largely unknown. Our previous study found that ADRB1 gene has m6A modification in both muscle and fat tissue. In this study, we interfered with FTO and ADRB1 genes After we cultured rabbit preadipocytes respectively. Oil red O staining and triglyceride assay were used to detect adipocyte differentiation. RT-qPCR was used to detect gene expression level and MeRIP-qPCR was used to detect the m6A modification level of gene. The results showed that FTO promoted the differentiation of adipocytes. At the same time, FTO up regulated the expression of ADRB1 gene and down regulated the m6A modification level of ADRB1 gene. Finally, we found that ADRB1 inhibited adipocyte differentiation. Together, we showed that FTO promoted adipocyte differentiation by regulating ADRB1 gene through m6A modification.

Impact of pharmacotherapy with tiotropium/olodaterol on the pulmonary function in COPD patients depending on the Arg16Gly polymorphism of ADRB2 gene

BACKGROUND. Chronic obstructive pulmonary disease (COPD) is a widespread chronic disease, that can be prevented and treated. Internal or genetic factors play a significant role in the COPD development.&#x0D; OBJECTIVE. To investigate a response to the tiotropium/olodaterol treatment in patients with different ADRB2 gene polymorphism.&#x0D; MATERIALS AND METHODS. 100 patients with the diagnosis of COPD were included into the study. Pulmonary function testing was performed during all study visits. All patients received treatment according to GOLD 2019, which included a combination of long-acting b2-agonists and long-acting muscarinic antagonists. According to the results of genetic testing all patients were divided into three groups: Arg16Arg – 23 patients (23 %), Arg16Gly – 39 (39 %), Gly16Gly – 38 (38 %).&#x0D; RESULTS. Groups did not differ by sex, age, part of smokers, COPD duration. There was a significant difference in force vital capacity (FVC), which was 66.92±4.33 % in Arg16Arg group, 79.75±3.51 % in Arg16Gly group and 76.59±2.73 % in Gly16Gly group (p=0.05). There was a weak negative correlation between Arg16Arg genotype and FVC (r=-0.204; p=0.043). In regression analysis Arg16Arg genotype was associated with 10.52 % lower FVC on average (odds ratio -10.523; 95 % confidence interval from -20.502 to -0.544; p=0.039). A statistically significant improvement of forced expiratory volume in the first second (FEV1) and FVC was observed in all groups during the study (p&lt;0.001).&#x0D; CONCLUSIONS. Arg16Arg genotype was associated with the lowest FVC values during all study visits. Combination of tiotropium/olodaterol was effective in improving pulmonary function in patients with different ADRB2 genotypes, which was demonstrated by a significant improvement of FEV1 and FVC in study groups during the study.

Yohimbine, an α2-Adrenoceptor Antagonist, Suppresses PDGF-BB-Stimulated Vascular Smooth Muscle Cell Proliferation by Downregulating the PLCγ1 Signaling Pathway.

Yohimbine (YOH) has antiproliferative effects against breast cancer and pancreatic cancer; however, its effects on vascular proliferative diseases such as atherosclerosis remain unknown. Accordingly, we investigated the inhibitory mechanisms of YOH in vascular smooth muscle cells (VSMCs) stimulated by platelet-derived growth factor (PDGF)-BB, a major mitogenic factor in vascular diseases. YOH (5–20 μM) suppressed PDGF-BB-stimulated a mouse VSMC line (MOVAS-1 cell) proliferation without inducing cytotoxicity. YOH also exhibited antimigratory effects and downregulated matrix metalloproteinase-2 and -9 expression in PDGF-BB-stimulated MOVAS-1 cells. It also promoted cell cycle arrest in the initial gap/first gap phase by upregulating p27Kip1 and p53 expression and reducing cyclin-dependent kinase 2 and proliferating cell nuclear antigen expression. We noted phospholipase C-γ1 (PLCγ1) but not ERK1/2, AKT, or p38 kinase phosphorylation attenuation in YOH-modulated PDGF-BB-propagated signaling pathways in the MOVAS-1 cells. Furthermore, YOH still inhibited PDGF-BB-induced cell proliferation and PLCγ1 phosphorylation in MOVAS-1 cells with α2B-adrenergic receptor knockdown. YOH (5 and 10 mg/kg) substantially suppressed neointimal hyperplasia in mice subjected to CCA ligation for 21 days. Overall, our results reveal that YOH attenuates PDGF-BB-stimulated VSMC proliferation and migration by downregulating a α2B-adrenergic receptor–independent PLCγ1 pathway and reduces neointimal formation in vivo. Therefore, YOH has potential for repurposing for treating atherosclerosis and other vascular proliferative diseases.

Molecular Variants in Genes related to the Response to Ocular Hypotensive Drugs in an Afro-Colombian Population

Aims: This study aimed to conduct an exploratory analysis of the pharmacogenomic variants involved in ocular hypotensive drugs to understand the individual differential response in an Afro-descendant population. Background: Glaucoma is the leading cause of irreversible blindness worldwide. The pharmacologic treatment available consists of lowering intraocular pressure by administering topical drugs. In Asian and Caucasian people, pharmacogenomic variants associated with the efficacy of these treatments have been identified. However, in Afro-descendant populations, there is a profound gap in this knowledge. Objective: This study identified the pharmacogenomic variants related to ocular hypotensive efficacy treatment in Afro-descendant individuals from the Archipelago of San Andres and Providence, Colombia. Methods: An analysis of whole-exome sequencings (WES), functional annotation, and clinical significance was performed for pharmacogenomic variants reported in PharmGKB databases; in turn, an in silico available prediction analysis was carried out for the novel variants. Results: We identified six out of 18 non-synonymous variants with a clinical annotation in PharmGKB. Five were classified as level three evidence for the hypotensive drugs; rs1801252 and rs1801253 in the ADRB1 gene and rs1042714 in the ADRB2 gene. These pharmacogenomic variants have been involved in a lack of efficacy of topical beta-blockers and higher systolic and diastolic pressure under treatment with ophthalmic timolol drug. The rs1045642 in the ABCB1 gene was associated with greater efficacy of treatments with latanoprost drug. Also, we found the haplotypes *17 for CYP2D6 and *10 for CYP2C19; both related to reducing the enzyme activity to timolol drug metabolization. In addition, we observed 50 novel potentially actionable variants; 36 synonymous, two insertion variants that caused frameshift mutations, and 12 non-synonymous, where five were predicted to be pathogenic based on several pathogenicity predictions. Conclusion: Our results suggested that the pharmacogenomic variants were found to decrease the ocular hypotensive efficacy treatment in a Colombian Afro-descendant population and revealed a significant proportion of novel variants with a potential to influence drug response.

The role of association of ADRB2 gene polymorphism with therapeutic response to β2-agonists in children with recurrent bronchial obstruction

Background. The study of the effect of drugs at the molecular genetic level in the recurrent course of bronchial obstruction (RBО) in children improved the understanding of the pathogenesis of the disease and made it possible to better characterize the reliability of use in therapy. The aim of the study was to assess the clinical significance of the polymorphism of the Arg16Gly loci of the ADRB2 gene with a therapeutic response to β2-agonists in children with RBО of Uzbek ethnicity. Materials and methods. Tested 88 patients aged 6 to 15 years with recurrent bronchial obstruction and bronchial asthma with the polymorphism of the ADRB2 gene loci and determined the effectiveness of salbutamol in these groups of patients. Results. Results of the study of the significance of the polymorphism of the Arg16Gly locus of the ADRB2 gene with a therapeutic response to β2-agonists in children with RBO showed high efficacy of salbutamol in 72.9 % of children, among whom representatives of the A/G and A/A genotypes were found in the largest number. At the same time, the low efficacy of salbutamol was found in the greatest amount in carriers of the mutational genotype G/G of the Arg16Gly locus of the ADRB2 gene. Conclusions. The predictor role of the polymorphic locus rs1042713 (Arg16Gly) was determined in carriers of the unfavorable 46G allele of the G/G genotype of the ADRB2 gene, which was a genetic marker of children’s predisposition to RBO and BA in children. Children with the G/G genotype of the Arg16Gly locus of the ADRB2 gene are at risk for severe RBO and BA in children. Children with RBO carriers (Arg16Gly) of the homozygous G/G genotype have a high incidence of RBO recurrence against the background of ARVI and are at risk of developing BA.

Regulation of heart rate following genetic deletion of the ß1 adrenergic receptor in larval zebrafish.

AimAlthough zebrafish are gaining popularity as biomedical models of cardiovascular disease, our understanding of their cardiac control mechanisms is fragmentary. Our goal was to clarify the controversial role of the ß1‐adrenergic receptor (AR) in the regulation of heart rate in zebrafish.MethodsCRISPR‐Cas9 was used to delete the adrb1 gene in zebrafish allowing us to generate a stable adrb1 −/− line. Larval heart rates were measured during pharmacological protocols and with exposure to hypercapnia. Expression of the five zebrafish adrb genes were measured in larval zebrafish hearts using qPCR.ResultsCompared with genetically matched wild‐types (adrb1 +/+ ), adrb1 −/− larvae exhibited ~20 beats min−1 lower heart rate, measured from 2 to 21 days post‐fertilization (dpf). Nevertheless, adrb1 −/− larvae exhibited preserved positive chronotropic responses to pharmacological treatment with AR agonists (adrenaline, noradrenaline, isoproterenol), which were blocked by propranolol (general ß‐AR antagonist). Regardless of genotype, larvae exhibited similar increases in heart rate in response to hypercapnia (1% CO2) at 5 dpf, but tachycardia was blunted in adrb1 −/− larvae at 6 dpf. adrb1 gene expression was abolished in the hearts of adrb1 −/− larvae, confirming successful knockout. While gene expression of adrb2a and adrb3a was unchanged, adrb2b and adrb3b mRNA levels increased in adrb1 −/− larval hearts.ConclusionDespite adrb1 contributing to the setting of resting heart rate in larvae, it is not strictly essential for zebrafish, as we generated a viable and breeding adrb1 −/− line. The chronotropic effects of adrenergic stimulation persist in adrb1 −/− zebrafish, likely due to the upregulation of other ß‐AR subtypes.