Increasing interest in psychedelic and related drugs as potential therapies for a wide spectrum of difficult to treat conditions that extend beyond neuropsychiatric disorders provided the impetus for this Themed Issue. This collection of reviews and original articles includes the mechanistic basis of how these drugs act, the current status of preclinical research and progress in clinical trials, and insight into the regulatory processes that determine clinical approval. In this Editorial we introduce these aspects and provide an overview of current controversies and challenges in the field, as well as highlighting the exciting potential that these drugs offer.

Incretin-based therapies are gaining momentum as a key strategy for reducing cardiovascular risk in individuals with obesity and/or type 2 diabetes (T2D). As therapeutic approaches extend beyond classical GLP-1 agonism, it remains unclear whether the cardiovascular benefits primarily reflect direct reductions in atherogenic lipoproteins—namely LDL, VLDL, and triglycerides—or broader effects mediated by weight loss, improved insulin sensitivity, and reduced ectopic adiposity. This review examines these relationships in depth. We summarize molecular, cellular, and physiological evidence describing how GLP-1, GIP, glucagon, and amylin signaling regulate intestinal lipid absorption, hepatic apoB-lipoprotein assembly, and peripheral lipoprotein catabolism. These mechanistic insights are integrated with available clinical data on single, dual, and triple incretin-based agonists and related compounds. Overall, incretin-based therapies predominantly reduce triglycerides and VLDL cholesterol, with these changes closely linked to reductions in hepatic ectopic adiposity and paralleling improvements in glycemic control, insulin sensitivity, and body weight. Agents that additionally activate glucagon or amylin pathways consistently produce greater lipid-lowering effects, supporting the concept that lipid benefits scale with global metabolic reprogramming rather than isolated receptor activation. Despite consistent lipid improvements, the short duration of most clinical trials limits assessment of long-term cardiovascular risk reduction. Moreover, the absence of apolipoprotein B data precludes definitive conclusions regarding changes in atherogenic lipoprotein burden. Uncertainty also remains regarding the preservation of lean mass during substantial weight loss. This review provides an up-to-date synthesis linking incretin pharmacology to lipid metabolism and identifies priorities for future cardiometabolic research.

Objective: Valproate medications are a leading cause of drug-associated acute pancreatitis (DAAP), yet the underlying mechanisms remain unclear. This study aimed to determine how sodium valproate (Na-VPA) induces pancreatic injury and contributes to acute pancreatitis (AP). Methods: Freshly isolated murine pancreatic acinar cells (PACs) were treated with Na-VPA or major VPA metabolites, and cytotoxicity was assessed by spectrofluorometry and confocal imaging. In vivo, C57BL/6 mice were administered Na-VPA with or without caerulein-induced pancreatitis (CER-AP), and pancreatic injury was evaluated by biochemical and histological analyses. Pancreatic mRNA-sequencing and pseudo-targeted metabolomics were conducted to identify dysregulated pathways. Intracellular methionine-cycle metabolites were quantified by LC-MS/MS, while expression of one-carbon metabolism enzymes was determined by RT-qPCR and western blotting. AutoDock Vina was employed to predict binding affinities of VPA to key metabolic enzymes. The therapeutic effect of S-adenosylmethionine (SAM) supplementation on Na-VPA-exacerbated CER-AP was also tested. Results: Na-VPA and its metabolites induced concentration- and time-dependent PAC death, independent of toxic calcium signaling. In vivo, Na-VPA aggravated CER-AP, increasing pancreatic histology scores and biochemical parameters. Transcriptomic and metabolomic analyses showed dysregulated one-carbon metabolism, validated by altered mRNA and protein expression analysis of key rate-limiting enzymes. Molecular docking indicated direct interactions between VPA and several metabolic enzymes. Na-VPA also activated the pancreatic endoplasmic reticulum (ER) stress pathway. In PACs, Na-VPA reduced methionine and SAM levels, while supplementation with methionine or SAM markedly attenuated cell injury; conversely, ethionine exacerbated it. Moreover, SAM supplementation in vivo significantly ameliorated pancreatic damage and biochemical alterations in Na-VPA–exacerbated CER-AP. Conclusion: Na-VPA disrupts one-carbon metabolism, triggering ER stress and acinar cell injury and exacerbating experimental pancreatitis. These findings provide new mechanistic insight into valproate-associated AP and identify metabolic targets for potential prevention or therapy.

Similar to NINJ1, SIGLEC12 might also be involved in acinar cell plasma membrane rupture in pancreatitisCaglar BERKEL 1*1 Tokat Gaziosmanpasa University, Department of Molecular Biology and Genetics, Tokat, Türkiye*correspondence to: caglar.berkel@gop.edu.tr; 0000-0003-4787-5157Dear Editor,A very recent study published in British Journal of Pharmacology by Xin et al. [2025] showed that knocking out NINJ1 reduces acinar cell necrosis and the release of ox-mtDNA (oxidised mitochondrial DNA) from these cells in cell models of acute and mild acute pancreatitis, and protects against acute pancreatitis in mouse models. Authors used sodium taurocholate (STC) and ceruletide (caerulein, CER) to induce acute pancreatitis (AP) and mild acute pancreatitis (MAP), respectively [Xin et al., 2025]. They showed that both STC and CER significantly increase the extracellular ox-mtDNA levels (ox-mtDNA released from acinar cells) and necrosis of acinar cells. In contrast, knocking out NINJ1 in these cells was shown to markedly decrease both necrosis and the release of ox-mtDNA, following the treatment of cells with STC or CER, pointing to the involvement of NINJ1 in both acinar cell necrosis and the release of ox-mtDNA from acinar cells, in cell models of both acute pancreatitis and mild acute pancreatitis [Xin et al., 2025]. However, knocking out NINJ1 did not completely reverse these events (i.e. STC- or CER-induced acinar cell necrosis and the release of ox-mtDNA to the extracellular space), suggesting that another player like NINJ1 might exist [Xin et al., 2025]. Authors also reported that inhibition of ferroptosis, pyroptosis, apoptosis and necroptosis of pancreatic acinar cells alone could not completely block STC-induced acinar cell death (necrosis), showing that STC-induced acinar cell death is not attributable to a single regulated cell death mechanism [Xin et al., 2025].NINJ1 (Ninjurin 1), a transmembrane protein, mediates plasma membrane permeabilization and rupture during the final lytic phase of multiple regulated cell death mechanisms such as pyroptosis, ferroptosis, parthanatos, H2O2-induced necrosis, secondary necrosis and cuproptosis [Kayagaki et al., 2021; Dondelinger et al., 2023]. However, NINJ1 does not contribute to plasma membrane rupture during necroptosis, and therefore, it dispensable for PMR during necroptosis [Kayagaki et al., 2021; Dondelinger et al., 2023]. Another very recent study published this year reported that another protein, SIGLEC12, mediates plasma membrane rupture during necroptotic cell death [Noh et al., 2025]. Authors identified SIGLEC12 as a main mediator of necroptosis downstream of MLKL at the plasma membrane rupture step, and showed that cells without SIGLEC12 are defective in necroptosis-induced plasma membrane rupture [Noh et al., 2025]. Besides, they found a high amino acid similarity between SIGLEC12 and NINJ1, pointing to possible functional homology [Noh et al., 2025].Since Xin et al. observed that the deletion of NINJ1 does not completely reverse STC- or CER-induced acinar cell death and the release of ox-mtDNA to the extracellular space, and that the inhibition of different regulated cell death mechanisms alone could not completely block STC-induced acinar cell death [2025], we hypothesized that in addition to NINJ1, SIGLEC12, another recently identified membrane-rupturing protein, might be involved in acinar cell necrosis and the release of ox-mtDNA following the treatment of cells with STC and CER (i.e. in cell models of acute pancreatitis and mild acute pancreatitis). The inhibition of necroptosis by necrostatin-1 decreases cell death in STC-induced pancreatic acinar cells, but only to certain extent [Xin et al., 2025; Ouyang et al., 2021], showing that necroptosis in addition to other regulated cell death programs are involved in STC-induced acinar cell death. Since SIGLEC12 mediates plasma membrane rupture and lytic cell death during necroptosis [Noh et al., 2025], similar to NINJ1 in most other cell death programs [Kayagaki et al., 2021; Dondelinger et al., 2023], SIGLEC12 might also contribute to plasma membrane permeabilization and rupture in acinar cells following the induction with STC, promoting acinar cell death and the release of ox-mtDNA from acinar cells to the extracellular space. This explains why the deletion of NINJ1 fails to completely reverse STC- or CER-induced acinar cell death and the release of ox-mtDNA [Xin et al., 2025]. In other words, both of these membrane rupturing proteins might contribute to acinar cell plasma membrane rupture in response to STC, suggesting that both NINJ1 and SIGLEC12 might be involved in pancreatitis (both acute and mild acute forms).To test these hypotheses, further research is needed where SIGLEC12 knockout pancreatic acinar cells are treated with STC- or CER, and subsequently the levels of cell death (necrosis) and the release of ox-mtDNA are quantified in these cells. Besides, NINJ1 and SIGLEC12 double knockout (DKO) acinar cells should be used in comparison with NINJ1 single knockout and SIGLEC12 single knockout acinar cells in order to determine the relative importance of these two proteins in acute and mild acute pancreatitis. Since STC (sodium taurocholate) induces multiple forms of regulated cell death (including ferroptosis, pyroptosis, apoptosis and necroptosis) in acinar cells in cell models of acute pancreatitis, NINJ1 might particularly mediate plasma membrane rupture if ferroptotic, pyroptotic and apoptotic events are more dominant; however, SIGLEC12 might mediate plasma membrane rupture if necroptotic events are more common.

Background: A drug designed for a specific target often interacts with multiple targets, either unintentionally or as part of its intended mechanism of action. This phenomenon has been called pharmacological pleiotropy or polypharmacology. There are key endogenous ligands such as ATP, GABA and glutamate, that act on various proteins in humans. Furthermore, several drugs act on multiple proteins without apparent structural similarity. G protein-coupled receptors (GPCR) and protein kinases are among the most important families of drug targets. The aim of this review analysis is to identify drugs with dual actions on GPCRs and kinases and clarify what is known about these actions. Approach: Data searches for ligands with affinity for both kinases and GPCRs were conducted in the Drugbank and Pharos databases. Physiochemical properties of selected compounds were identified using the rdMolDescriptors module from RDKit. A detailed literature search was conducted in search engines such as Google Scholar and Medline, to identify pleiotropic compounds with both GPCR and kinase affinity. Outcome: 34 compounds were identified to interact with proteins within both the kinase and GPCR protein families. Notable examples included the drugs loratadine, terfenadine, clozapine, thioridazine, aripiprazole, fluspirilene, sorafenib, dasatinib, and fasudil. Conclusions: Drug pleiotropy among GPCRs and kinases is a commonly occurring phenomenon. Structural factors that may contribute to pleiotropy include chemical similarity to endogenous ligands, lipophilicity, and planarity of chemical structure, which may guide the development of drugs with intentional multi-target effects. While pleiotropy presents challenges in ensuring specificity, it also creates opportunities for innovative therapeutic strategies if strategically applied.

The selection and optimization of drug leads is largely driven by equilibrium parameters such as IC50 values. However, this approach does not account for the time-dependence of drug-target interactions which is important given that drug and target concentrations fluctuate in the human body. A fundamental understanding of drug-target binding kinetics is particularly important when the formation and breakdown of the drug-target complex is slow compared to the rate of drug elimination and becomes critical when dealing with covalent drugs where the drug is irreversibly bound to the target. The parameters that define covalent inhibition, specifically kinact and KI, can be determined by quantifying target binding as a function of drug concentration and time. However, such assays are difficult to implement in early stages of drug discovery. Here, we review practical approaches to quantify irreversible inhibition including progress-curve kinetics, multi-timepoint IC50 fitting (EPIC) for scalable library triage, intact- and peptide-level mass spectrometry to confirm adducts and sites, label-free biophysics (SPR, NMR) for orthogonal validation, and washout/jump-dilution experiments to diagnose reversibility and measure residence time (τ). We then describe the translation of these microscopic rates to pharmacology through three system determinants: protein turnover, target vulnerability, and pharmacokinetics using case studies from BTK, JAK3, KRASG12C, and EGFR to illustrate how aligning chemistry with context yields durable efficacy. Finally, we propose reporting standards (IC50 with incubation time; kinact/KI; τ; turnover-driven occupancy) and a kinetics-first design paradigm to replace static potency with mechanism-anchored optimization of covalent inhibitors.

The Mediterranean diet (MedDiet) is among the most extensively studied dietary patterns and has been consistently associated with reduced risk of all-cause mortality, cardiovascular disease, metabolic disorders, cognitive decline, and several types of cancers. It is defined by a high intake of vegetables, fruits, legumes, nuts, (whole) grains, fish, seafood, and extra virgin olive oil, along with moderate consumption of red wine. Several physiological pathways may contribute to its beneficial effects, including favourable modulation of lipid profiles, enhanced insulin sensitivity, reduction of inflammatory and oxidative stress markers, improved endothelial function, and antithrombotic activity. These effects are largely attributed to bioactive compounds such as polyphenols, mono- and polyunsaturated fatty acids, and dietary fibre. This updated review summarizes the most recent evidence and discusses both established and emerging mechanisms of action related to individual dietary components within the MedDiet. It aims to provide a refined understanding of how this dietary pattern contributes to chronic disease prevention and overall health.

¿p#1 Background and Purpose: Calcitonin gene-related peptide (CGRP) is a potent, clinically relevant endogenous vasodilator, but the precise intracellular signalling pathways remain unclear, particularly in small coronary arteries. Experimental Approach: This study used ex vivo myography and intracellular recording techniques to investigate α-CGRP-induced vasorelaxation and vascular smooth muscle cell (VSMC) hyperpolarization in myogenically-active rat isolated coronary arteries. Key Results: CGRP-induced vasorelaxation was not dependent on the endothelium, but relied heavily on K + channel activation. Immunohistochemistry indicated Kv7 and BK Ca channel expression in VSMC and endothelial cells (ECs). A combination of the K V7 channel inhibitor, linopirdine, and the BK Ca channel inhibitor, paxilline, significantly attenuated CGRP-induced vasorelaxation in endothelium-intact or denuded arteries. Electrophysiology confirmed that CGRP induced hyperpolarization and showed this was prevented by linopirdine and paxilline, also demonstrating a role for K V7 and BK Ca channels in suppressing depolarizing smooth muscle spikes. These spikes were also suppressed by NO• and HNO donors, resulting in hyperpolarization. Gβγ subunit inhibition with gallein markedly right-shifted CGRP-induced vasorelaxation in both endothelium-intact and denuded coronary arteries. Conclusion and Implications: α-CGRP stimulates robust vasorelaxation in the coronary microvasculature that relies on Gβγ subunit activated VSMC hyperpolarization involving K V7 and BK Ca channels. These data enhance understanding of coronary microvascular physiology and inform the design of future therapeutic strategies targeting coronary vascular dysfunction.

We read with considerable interest the comprehensive review by Geng et al., which synthesizes the complex role of N6-methyladenosine (m6A) modification in autoimmune diseases and cancer [1]. The authors successfully chart the expansive ”epitranscriptomic landscape,” illustrating how dysregulation of writers, erasers, and readers contributes to pathogenesis. While this map is undeniably valuable, the journey from this mechanistic cartography to genuine clinical therapeutics is far more treacherous than the review might suggest. We offer a critical appraisal to highlight the substantial gaps and challenges that must be addressed to realize the proposed ”medicinal value.”

Background/Purpose The G protein-coupled estrogen receptor (GPER) participates in breast cancer and nociception. The GPER inverse agonist PLMI was studied on pain-like symptoms, in murine models of chemotherapy-induced peripheral neuropathy. Experimental Approach Genetic and pharmacological experiments were performed in mice. The functional localization of GPER was explored by using nociceptors and dorsal horn GPER knockouts. We used the PLMI and the GPER antagonist G15 to study the role of GPER in mechanical allodynia in the model of paclitaxel-induced peripheral neuropathy. The effect of PLMI and/or G15 was assessed in dorsal root ganglia primary cultures to explore the role of GPER in neuronal calcium flux. The anti-nociceptive and neuroprotective effects resulting from a chronic administration of PLMI were investigated in the paclitaxel-induced neuropathy model. Cognitive alterations, addiction and acute effects in bortezomib- and oxaliplatin-induced pain were evaluated. NMR and CD spectroscopy were used to determine the conformation of the peptide PLMI, in solution. Key Results In our paclitaxel-induced pain model, peripheral, spinal and supraspinal GPER participates in pain. The peptide PLMI decreases nociception by lowering intraneuronal free calcium flux. Chronic PLMI treatment reduces pain-like behaviours and protects against nerve conduction velocity deficits, without causing cognitive impairments or addiction. PLMI alleviates oxaliplatin- and bortezomib-induced neuropathic pain. The peptide PLMI adopts a turn conformation. Conclusion/Implications Our results suggest that GPER inverse agonists could be used to alleviate pain and to protect against paclitaxel-induced peripheral neuropathy. The turn conformation of the PLMI peptide in solution is in favour of a bioactive GPCR-interacting peptide.

Cannabinoid receptor 2 (CBS) is a G-protein coupled receptor that controls immune responses and has recently emerged as a regulators of renal injury and repair. Over the past 15 years, numerous pharmacological and genetics studies have explored the role of CB2 in acute kidney injury (AKI) and chronic kidney disease (CKD). Using a wide array of chemical, metabolic, ischemia and obstructive mouse models, CB2 agonism has consistently preserved tubular epithelial cell integrity, reduced inflammation, and limited tubulointerstitial fibrosis. These protective effects were supported by the opposing outcomes in Cnr2 knockout mice, which showed worsened injury. More selective CB2 ligands with defined pharmacokinetic and pharmacodynamic profiles further reinforced the renoprotective role of CB2 agonism. In recent years, however, a small number of studies have suggested that CB2 may contribute to tubular damage, though there are issues of ligand selectivity, off-target activity, and methodological differences that complicate accurate interpretation. With this in mind, the prevailing evidence supports CB2 activation as beneficial in AKI and CKD and warrants continued progress to develop CB2 agonists for therapeutic applications for the kidney.

Background and Purpose: Sepsis induces immunosuppression, contributing to high mortality. Plasmablasts, a regulatory B cell subset that expands during sepsis, are known to suppress immune function, but the consequences of their specific removal on sepsis outcome remain unclear. This study aimed to determine whether depleting plasmablasts could improve sepsis prognosis and to investigate the underlying mechanisms, with a focus on neutrophil function. Experimental Approach: We utilized a genetic approach to selectively deplete plasmablasts in vivo by using CD138-diphtheria toxin receptor (DTR) transgenic mice subjected to cecal ligation and puncture (CLP)-induced sepsis. The effects of plasmablast depletion on survival, bacterial load, inflammatory mediators, and neutrophil number and function were assessed. Furthermore, the proteasome inhibitor bortezomib was evaluated for its ability to suppress plasmablasts and its therapeutic efficacy in wild-type mice with sepsis. Key Results: Depletion of plasmablasts in CD138-DTR mice significantly improved survival and reduced bacterial load in the peritoneal cavity. This was associated with a marked increase in the number and antibacterial function of neutrophils, including enhanced reactive oxygen species production and phagocytic capacity. In vitro co-culture experiments demonstrated that plasmablasts directly suppressed neutrophil activity, an effect mediated via adenosine and IL-10 signaling. Treatment of wild-type septic mice with bortezomib effectively reduced plasmablast abundance and CD39 expression, and a specific dosing regimen (0.05 mg/kg) mirrored the protective effects of genetic ablation, improving survival, reducing bacterial burden and inflammation, and enhancing neutrophil-mediated bacterial clearance. Conclusion and Implications: Our findings establish plasmablasts as a critical contributor to sepsis immunosuppression by directly impairing neutrophil function. The clinically available drug bortezomib can target this population, positioning plasmablast inhibition as a novel and translatable therapeutic strategy to improve bacterial clearance and survival in sepsis.

Background and Purpose: The pathogenesis of type 2 diabetes mellitus (T2DM)-induced cardiomyopathy involves cardiac fibrosis that leads to diastolic dysfunction. We established that replacement of lost substance P (SP) that occurs in T2DM reduces cardiac fibrosis and decreases inflammation in T2DM mice and non-human primates. This study aimed to identify the specific anti-fibrotic SP receptor. Experimental Approach: Age-matched male wild type (WT) and Lepr db/db mice at 12 weeks of age were treated with either saline or the neurokinin 1 receptor (NK-1R) agonist, GR73632 (300 μg/kg/day) for 4 weeks. The hearts were assessed for cardiac function, fibrosis, mast cells and macrophage phenotype. Mouse cardiac fibroblast cultures were treated with high glucose and GR73632 to assess collagen release and signaling pathways. Proteomics analysis was conducted to assess the cardiac proteomic profile between WT and Lepr db/db mice, and the effects of GR73632. Key Results: NK-1R activation decreased cardiac fibrosis, improved diastolic function, decreased mast cell numbers and promoted an anti-inflammatory macrophage phenotype in Lepr db/db mice. NK-1R activation reduced collagen I production in high glucose treated mouse cardiac fibroblasts. NK-1R activation increased Smad7, whilst decreasing P65 phosphorylation (NF-κB) and CCL2 chemokine release. Proteomic analysis revealed a distinct proteome profile between WT and Lepr db/db mouse hearts. Conclusion and Implications: The NK-1R is the anti-fibrotic SP receptor and improves diastolic function in the diabetic heart. This likely involves direct effects on cardiac fibroblasts. This study provides a potential target for treatment of diabetic cardiomyopathy.

Background and Purpose: The study evaluated human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as a non-animal new approach methodology (NAM) for assessing cardiovascular repolarization risk in drug development. The research compared hiPSC-CM data with traditional methods (hERG assays and animal studies), to determine if hiPSC-CMs could potentially replace or reduce animal testing in regulatory decision-making, aligning with FDA Modernization Act 2.0 goals. Experimental Approach: The study analyzed 29 Investigational New Drug (IND) applications submitted to FDA/CDER between 2020 and 2023, comparing hiPSC-CM studies using multielectrode array (MEA) and voltage sensing optical (VSO) platforms with hERG assays, multi-channel assessments, and in vivo non-rodent QT studies. The analysis focused on concordance between these methods and their ability to predict clinical QT prolongation risk. Key Results: hiPSC-CM studies demonstrated higher overall accuracy in predicting clinical QT prolongation risk compared to standard hERG and in vivo non-rodent QT studies alone. Notably, to achieve comparable accuracy to hiPSC-CMs, additional animal studies beyond the standard in vivo QT assessment were needed. These additional studies included testing at higher exposures, using different animal species, or conducting repeated dose studies. The standard in vivo QT study had lower sensitivity, likely due to limitations in achieving high drug exposures in animals. hiPSC-CMs could evaluate effects at much higher exposure multiples relative to clinical concentrations. Conclusion and Implications: hiPSC-CMs offer a promising alternative to extensive animal testing for cardiovascular risk assessment. However, limitations include small sample size, early-phase INDs, variability in hiPSC-CM study rigor, and challenges in establishing exposure-response relationships. Despite these limitations, this research provides evidence supporting hiPSC-CMs as potential alternatives to animal studies in regulatory assessments. Further research is needed to establish robust protocols for integrating hiPSC-CM data into regulatory workflows, balancing the promise of reduced animal testing with the need for comprehensive safety assessment.

Background and Purpose: New alarming trends show that vaping synthetic cannabinoid receptor agonists, such as JWH-018, by e-cigarettes is increasing among youths. However, the sex-related consequences of these trends are unclear. We therefore characterized the neuropharmacological effects of adolescent JWH-018 inhalation in male and female rats. Experimental Approach: Adolescent rats inhaled passively JWH-018 vapor (0.3 or 0.6 mg/ml qd) for 21 days. During vapor exposure, JWH-018 and main metabolites plasma levels, body weight, and locomotion were measured at different time points. During drug-free period, behavioural (withdrawal signs, anxiety, repetitive-like behaviour) and microdialysis (NAc shell/mPFC dopamine responsiveness to intraoral chocolate, taste reactivity) studies were performed 24 hours and 7 days after last JWH-018 inhalation, respectively. Key Results: Repeated adolescent JWH-018 inhalation induced sex-dependent effects with (i) higher plasma levels in males; (ii) increased body weight gain and withdrawal signs in females; (iii) transient hypolocomotion in female and dose-dependent biphasic locomotion in males; (iv) higher taste aversion in male; (v) sex- and dose-related adaptive changes of NAc shell and mPFC dopamine to single/repeated chocolate exposure in early adulthood, as follows: in the NAc shell, either low or high dose decreased dopamine sensitivity to chocolate in males, low dose abolished habituation whereas high dose blunted dopamine responsiveness in females; in the mPFC, the low dose blunted responsiveness in male and induced habituation in females while the high dose induced habituation only in males. Conclusion and Implications: Using a highly translational model, we showed that the impact of adolescent JWH-018 inhalation differs between sexes.

The purinergic receptor family is primarily activated by nucleotides and contains members of both G protein coupled-receptor (GPCR) superfamily (P1, P2Y) and ligand-gated ion channel (P2X). The P2Y receptors are widely expressed in the human body, and given the ubiquitous nature of nucleotides, purinergic signalling is involved with a plethora of molecular physiological functions. The widespread nature of P2Y receptors make them a viable therapeutic target, but with the negative risk of on-target side effects. Some of the side effects associated with P1 and P2Y receptors arise due to the pleotropic nature of many GPCRs, as a singular GPCR can activate several G proteins, as well as recruit β-arrestins. The utilization of ligands that exhibit downstream pathway-specific activation, also known as biased signalling, at the P2Y receptors could circumvent these issues. This review will cover the signalling nature and impact of the P2Y family, with an emphasis on individual activity patterns of the P2Y receptors. This review will also discuss current literature on the development of biased ligands for these receptors, with an aim to highlight the most beneficial targets and outcomes.

Background and Purpose: Mesenchymal stem cells (MSCs) are widely utilized in regenerative medicine due to their multipotency and immunomodulatory properties. Compared to conventional two-dimensional (2D) monolayer cultures, three-dimensional (3D) spheroid cultures better mimic the in vivo microenvironment, influencing the metabolic activity and therapeutic efficacy of MSCs. This study aims to evaluate how 2D and 3D culture conditions affect the behavior, proliferation, and functional properties of MSCs. Experimental Approach: Metabolomic and transcriptomic analyses were conducted on MSCs cultured under both 2D and 3D conditions. To assess metabolic differences between 2D and 3D cultured MSCs, polar metabolites were extracted and analyzed using ¹H-NMR spectroscopy. The data was processed with Chenomx and subjected to multivariate statistical analysis. For transcriptomic analysis, RNA sequencing was performed, followed by differential gene expression and gene set enrichment analysis. Key Results: The findings reveal that MSCs in 3D spheroids exhibit reduced proliferation, enhanced stemness, and distinct metabolic adaptations, including increased glycolysis and altered nutrient metabolism. Additionally, genes associated with ribosome biogenesis and cell cycle progression were downregulated in 3D MSCs. These changes promote a more quiescent state, favoring its applications on tissue repair and immune modulation. Conclusion and Implications: Understanding these metabolic adaptations offers valuable insights for optimizing culture conditions, improving MSC-based therapies, and identifying novel therapeutic targets and biomarkers.

Background: In some individuals, opioid use leads to a decreased interest in socially relevant rewards. Recent studies showed that after extended-access heroin self-administration rats strongly prefer social interaction over single unit-dose heroin infusions. We hypothesized that this strong social preference results from access to a suboptimal heroin dose during testing, and individual differences in heroin versus social choice would emerge if rats were given access to their ‘preferred’ heroin dose. Methods: In Experiment 1, we trained male rats to lever-press for social interaction, followed by heroin self-administration under continuous-access, no-timeout schedule, which promotes burst-patterned (or binge-like) heroin intake. We then tested the rats for choice between single-unit heroin dose and 1-minute full-contact social interaction, or 5-minute heroin-access (sufficient for binge-like intake) and 5-minute social interaction. In Experiment 2, we extended the 5-minute access procedure to female rats and tested heroin versus limited-contact (screen-based) social interaction. We also manipulated response requirements (effort) for heroin. Results: When rats were given a single-unit heroin dose during choice testing, they strongly preferred social interaction. In contrast, when given 5-minute heroin-access, large individual differences in heroin preference emerged. These differences were independent of sex, social-interaction conditions, and effort manipulations. High heroin intake and binge-like intake during self-administration, and high heroin seeking during abstinence predicted individual differences in heroin preference. Conclusions: Access to ‘preferred’ heroin doses during the choice tests leads to stable and effort-independent individual differences in heroin preference. This procedure provides a platform to study mechanisms of resilience and vulnerability to opioid addiction.