Molecular Pharmacology最新文献

{"title":"Trimethylamine N-oxide induces macrophage M1 polarization via the Piezo1/Yes-associated protein pathway.","authors":"Haiyan Xiang, Congcong Li, Yanhua Tang, Juesheng Yang","doi":"10.1016/j.molpha.2026.100120","DOIUrl":"10.1016/j.molpha.2026.100120","url":null,"abstract":"<p><p>Trimethylamine N-oxide (TMAO), a gut microbiota-derived metabolite, has been implicated in cardiometabolic inflammation; however, the intracellular signaling mechanisms linking TMAO to macrophage polarization remain incompletely defined. This study aimed to investigate whether TMAO promotes macrophage M1 polarization through a Piezo1-Yes-associated protein (YAP) signaling axis. Gain- and loss-of-function approaches were performed in RAW264.7 cells and primary bone marrow-derived macrophages, including Piezo1 overexpression, small interfering RNA-mediated knockdown of Piezo1 or YAP, and pharmacological inhibition of YAP. Macrophage polarization markers were evaluated by immunofluorescence, flow cytometry, real-time quantitative polymerase chain reaction, and Western blot analysis. Intracellular Ca²⁺ levels were assessed using Fluo-4 acetoxymethyl ester fluorescence imaging. TMAO increased inducible nitric oxide synthase expression and reduced arginase-1 levels (P < .01), accompanied by elevated IL-1β and IL-6 and decreased IL-4 and IL-10 (P < .01). Piezo1 overexpression promoted M1 polarization (P < .01), whereas Piezo1 knockdown under TMAO exposure attenuated inflammatory cytokine induction (P < .001). TMAO enhanced intracellular Ca²⁺ influx (P < .001) and reduced inhibitory phosphorylation of YAP (Ser127) (P < .01). Inhibition or silencing of YAP mitigated TMAO-induced M1 marker expression and downstream kappa B/extracellular signal-regulated kinase activation (P < .05). Similar results were confirmed in primary bone marrow-derived macrophages. These findings suggest that TMAO promotes macrophage M1 polarization in association with Piezo1-dependent Ca²⁺ influx and YAP activation, identifying the Piezo1-YAP axis as a potential mechanistic interface linking microbiota-derived metabolites to inflammatory remodeling. SIGNIFICANCE STATEMENT: This study identifies the Piezo1-Yes-associated protein signaling axis as a potential drug target in trimethylamine N-oxide-induced macrophage polarization toward a proinflammatory phenotype. Inhibition of this pathway attenuates inflammatory responses, suggesting a potential pharmacological strategy for microbiota-associated cardiovascular diseases.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 5-6","pages":"100120"},"PeriodicalIF":3.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147840185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural and molecular determinants of glutamate transporter allosteric modulators.","authors":"Katelyn L Reeb, Satyaki Saha, Xiaowei Bogetti, Adi N R Poli, Joseph M Salvino, Mary Hongying Cheng, Ole V Mortensen, Ivet Bahar, Andréia C K Fontana","doi":"10.1016/j.molpha.2026.100122","DOIUrl":"10.1016/j.molpha.2026.100122","url":null,"abstract":"<p><p>Excitatory amino acid transporters (EAATs) are critical regulators of synaptic glutamate levels in the central nervous system. Dysregulated central nervous system glutamatergic homeostasis is implicated in many neurological diseases, highlighting the key role of EAATs in neurological health. We previously identified a library of small compounds that function as either positive allosteric modulators (PAMs) or negative allosteric modulators of EAATs, with diverse selectivity for subtypes EAAT1, EAAT2, and EAAT3, including astrocytic EAAT1 and EAAT2 and neuronal EAAT3. In this work, we characterized compounds from our library using molecular modeling, mutagenesis, and pharmacologic approaches. We focused on 3 representative compounds: NA-014, an EAAT2-selective PAM, DA-038, an EAAT1-3 PAM, and NA-010, an EAAT2-selective negative allosteric modulators. Binding studies demonstrated that these compounds do not interact with the orthosteric glutamate-binding site, confirming an allosteric action. Docking studies suggested several potential binding poses of NA-014 between the scaffold and transport domains of EAAT2, which we then studied with mutagenesis approaches. We identified potential binding sites of representative compounds in transmembrane domains 1, 5, 8, and hairpin 2 and demonstrated that these are necessary for their activity. Ten key amino acid residues within a subdomain of EAAT2 substituted into EAAT1 conferred EAAT2-selective PAM activity, demonstrating these residues are required and sufficient to enable selective PAM function. Collectively, these studies identified crucial subdomains and key amino acids linked to PAM activity, advancing our understanding of how to modulate EAAT activity. This knowledge can be integrated into future studies to develop EAAT allosteric modulators for neurological disorders. SIGNIFICANCE STATEMENT: We identified modulators of glutamate transporters, key regulators of central nervous system excitability and neuronal health. Using molecular modeling, mutagenesis, and pharmacology, we mapped their allosteric binding sites and identified ten residues that confer selective transport enhancement. This mechanism of transporter activation may guide development of therapies for disorders involving glutamatergic dysregulation, including stroke, neuropathic pain, and substance use disorders.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 5-6","pages":"100122"},"PeriodicalIF":3.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13339737/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148239559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and characterization of selective inhibitors of Nav1.9 voltage dependent sodium channels.","authors":"Franck Potet, Sonia Santos, Allison Gallucci, Jonathan Theile, Katrina Yoger, Zhixin Lin, Sylvain Lebreton, Robert Volkmann, Robert Dow, Mark Elban, Chongyang Han, Jared Spletstoser, Jay Spampanato, Gareth Young, Neil A Castle","doi":"10.1016/j.molpha.2026.100125","DOIUrl":"10.1016/j.molpha.2026.100125","url":null,"abstract":"<p><p>Voltage-gated sodium channels play fundamental roles in physiological and pathophysiological electrical excitability of the nervous and muscular systems. This involves several sodium channel subtypes, including Nav1.9, which has been genetically linked to altered pain sensation in humans. This association makes Nav1.9 a promising target for new pain therapies. However, progress has been limited by the historical challenges of expressing functional recombinant Nav1.9 as well as the absence of selective pharmacological tools. The current study expands on previous success of: (1) functionally expressing recombinant Nav1.9; (2) developing electrophysiological assays which enabled a more detailed assessment of the sensitivity of Nav1.9 to existing sodium channel modulators and; (3) screening for, identifying and characterizing novel potent Nav1.9 subtype selective sodium channel inhibitors. This study shows that previously described sodium channels inhibitors exhibit similar sensitivity and gating dependence at Nav1.9 as to other Nav1.x family members. This study also describes the identification of several novel Nav1.9 subtype selective inhibitors including ICA604025, which inhibits human Nav1.9 in a state dependent manner, with EC50s of 9 nM and 1.1 μM for the activated/inactivated state versus resting closed state. ICA604025 exhibits a similar potency for inhibition of cynomolgus monkey Nav1.9 but ∼50-fold lower potency for inhibition of rodent Nav1.9. It is a 300- to 1800-fold more potent inhibitor of human Nav1.9 compared with other human Nav1.x family members. In conclusion, this work demonstrates that subtype selective inhibition of Nav1.9 is achievable and offers valuable tools for further exploration of its physiological role and therapeutic potential. SIGNIFICANCE STATEMENT: Nav1.9 is a voltage-gated sodium channel genetically associated with human pain disorders and thus is a key target for new analgesics. Several novel potent and highly subtype selective Nav1.9 inhibitors have been identified that may enable further advances in pain research and therapeutic development.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 5-6","pages":"100125"},"PeriodicalIF":3.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147988328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Maraviroc alleviates neuropathic pain symptoms in a mouse model of spared nerve injury.","authors":"Wei Zhao, Cigdem Sevim Bayrak, Fiona Chen, Hsiao-Yun Lin, Bin Zhang, Jun Wang","doi":"10.1016/j.molpha.2026.100126","DOIUrl":"10.1016/j.molpha.2026.100126","url":null,"abstract":"<p><p>Chronic pain represents a major health problem in the health care system. According to the CDC data brief in 2020, 20.4% of adults have chronic pain. There has been no promising therapy for chronic pain. Currently available treatments include medications such as nonsteroidal anti-inflammatory drugs, antiepileptic drugs, tricyclic antidepressants, corticosteroids, opioids, and cannabinoids, all of which may cause various negative side effects. Thus, there is an urgent need to develop novel, efficacious, and safe interventions for treating pain. Studies have shown that proinflammatory cytokines and chemokines make important contributions to the initiation and persistence of pain. We have found that C-C motif chemokine ligand 5 levels increased at day 14 post-spared nerve injury (SNI). This study was designed to investigate the effect of maraviroc (MVC), an FDA-approved CCR5 antagonist, on neuropathic pain in a mouse model of SNI. We found that MVC alleviated SNI-induced mechanical allodynia at 3, 7, and 14 days postinjury. MVC treatment also prevented SNI-mediated thermal hypersensitivity at 7 and 14 days postinjury in both male and female cohorts. SNI resulted in weight-bearing deficits, which were corrected by MVC administration in male mice. RNA sequencing analysis revealed that MVC rescued SNI-induced dysregulation of sex-specific canonical pathways in the spinal cord. Collectively, our findings showed that MVC could reduce neuropathic pain following peripheral nerve injury, providing a base for the repurposing of this FDA-approved human immunodeficiency virus drug as a pain reducer in clinical applications. SIGNIFICANCE STATEMENT: Spared nerve injury-induced neuropathic pain is associated with upregulation of the C-C motif chemokine ligand 5. Targeting the C-C motif chemokine ligand 5-CCR5 axis with FDA-approved maraviroc alleviated pain phenotype through modulating different pathways in male and female mice.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 5-6","pages":"100126"},"PeriodicalIF":3.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148055626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The CaMKII D135N mutation blocks kinase activity and reduces GluN2B binding.","authors":"Matthew E Larsen, C Madison Barker, Raul Satoshi Vargas, Steven J Coultrap, K Ulrich Bayer","doi":"10.1016/j.molpha.2026.100127","DOIUrl":"10.1016/j.molpha.2026.100127","url":null,"abstract":"<p><p>Three recent studies claimed that induction of long-term potentiation of synaptic strength requires structural rather than enzymatic functions of the Ca²⁺/calmodulin-dependent protein kinase II (CaMKII). One study used the CaMKII D135N mutation, which was claimed to abolish enzymatic activity without affecting the structural function, that is, binding to GluN2B. We found here that the D135N mutant indeed abolished enzymatic kinase activity and autophosphorylation at T286. However, D135N mutation additionally reduced binding to GluN2B and prevented persistence of co-condensation with GluN2B. Similar to the T286A mutant, GluN2B binding of the D135N mutant could be partially rescued by AS283, an inhibitor that directly enhances GluN2B binding. This partial effect on GluN2B binding has to be taken into account when using D135N to distinguish between enzymatic versus structural functions of CaMKII. Nonetheless, as discussed here, the D135N mutant indeed supports a structural rather than enzymatic CaMKII function in long-term potentiation induction. SIGNIFICANCE STATEMENT: This study characterized the Ca²⁺/calmodulin-dependent protein kinase II D135N mutant and found abolished kinase activity and reduced GluN2B binding. These findings support the nonenzymatic role of Ca²⁺/calmodulin-dependent protein kinase II in long-term potentiation induction but provide important information for the interpretation of results obtained with this tool mutant.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 5-6","pages":"100127"},"PeriodicalIF":3.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148101692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Loss of α-cell GRK2 modulates glucagon response and supports cardiac function.","authors":"Jonathan Snyder, Lilly Underwood, Chun-Sun Jiang, Victoria Kaml, Snekha Rajasekaran, Lily Xie, C Reid Dotson, Priscila Y Sato","doi":"10.1016/j.molpha.2026.100115","DOIUrl":"10.1016/j.molpha.2026.100115","url":null,"abstract":"<p><p>Pancreatic α-cells secrete glucagon to maintain glucose homeostasis, yet the molecular mechanisms regulating hormone release are understudied. G-protein coupled receptor kinases (GRKs) regulate receptor desensitization; however, their role in α-cells remains unknown. Here, we generated an inducible α-cell-specific GRK2 knockout (αGRK2KO) mouse model to investigate the role of GRK2 in islet physiology, systemic metabolism, and cardiac function. Loss of GRK2 in α-cells reduced islet GRK2 protein by ∼20%, consistent with α-cell islet abundance, and produced negligible alterations in glucose tolerance without affecting insulin secretion. Notably, αGRK2KO mice had altered fast/fed glucagon responses, reduced adiposity, and lower body weight. Despite minimal effects on systemic glucose handling, αGRK2KO animals exhibited improved cardiac function, characterized by enhanced ejection fraction and fractional shortening, without signs of hypertrophy. High-fat, high-sucrose diet feeding abated these changes, underscoring the diet-dependent impact of α-cell GRK2. Together, these findings identify GRK2 as a previously unrecognized regulator of α-cell glucagon secretion that influences systemic energy balance and cardiac performance through endocrine crosstalk. This work establishes a framework for α-cell G protein-coupled receptor regulation and highlights GRK2 as a potential therapeutic node linking islet function, metabolism, and the heart. SIGNIFICANCE STATEMENT: This study explores the role of GRK2 in α-cell biology as a regulator of glucagon dynamics and cardiac function, thereby establishing a new endocrine link between islet signaling and cardiac biology.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 4","pages":"100115"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147530586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression of concern: \"The Cyclopentenone Prostaglandin 15-Deoxy-delta^12,14-Prostaglandin J₂ Attenuates the Development of Acute and Chronic Inflammation\" [Mol. Pharmacol. 2002; 61: 997-1007] by Cuzzocrea S, Wayman N, Mazzon E, et al.","authors":"","doi":"10.1016/j.molpha.2026.100107","DOIUrl":"https://doi.org/10.1016/j.molpha.2026.100107","url":null,"abstract":"","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 4","pages":"100107"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular domain-dependent modulation of class B1 G-protein-coupled receptor signaling.","authors":"Alex D White, Leah R Padgett, Francis S Willard","doi":"10.1016/j.molpha.2026.100111","DOIUrl":"10.1016/j.molpha.2026.100111","url":null,"abstract":"<p><p>The class B1 subfamily of G-protein-coupled receptors (GPCRs) comprises 15 members that respond to endogenous peptides to regulate an array of important physiological processes. Activation by peptide ligands is considered to follow a 2-step binding mechanism that involves an initial interaction with the receptor ectodomain (ECD) followed by engagement with transmembrane regions that collectively result in effector coupling. This model suggests that the ECD acts as an affinity trap that generally does not regulate downstream signaling events. Here, we show that members of the glucagon receptor subfamily of class B1 GPCRs contain a conserved 6-amino acid motif within their ECDs and that individual residues regulate cAMP and/or βarr pathways by either allosterically promoting effector coupling or as determinants required for proper receptor expression and function via modulation of posttranslational receptor glycosylation. We also highlight that this conserved motif surprisingly alters observed signaling phenomena in both a ligand- and receptor-dependent manner. Our study unveils an unexpected role of the ECD in regulating class B1 GPCR signaling properties independently of ligand binding affinity and suggests that the well-established 2-step binding paradigm requires further investigation to understand the nuances of activation mechanisms for this receptor class. SIGNIFICANCE STATEMENT: Class B1 G-protein-coupled receptors respond to endogenous peptide ligands to fine-tune biological processes. These receptors contain the hallmark structural feature of a large ectodomain at their N-termini that has been shown to be critical for ligand binding affinity, but whether this region regulates downstream signaling remains unclear. We identify a conserved site within the glucagon-like peptide-1 receptor, glucagon receptor, and glucose-dependent insulinotropic polypeptide receptor localized to the ectodomain that modulates both effector coupling as well as proper receptor expression and functionality in ligand- and receptor-dependent manners. Given the benefit of targeting these receptors in the treatment of type 2 diabetes and obesity, our findings may provide insights toward a better understanding of their activation mechanisms and facilitate rational drug discovery approaches.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 4","pages":"100111"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147372955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression of concern: \"Adelmidrol, a Palmitoylethanolamide Analogue, as a New PharmacologicalTreatment for the Management of Inflammatory Bowel Disease\" [Mol. Pharmacol. 2016; 90: 549-561] by Cordaro M, Impellizzeri D, Gugliandolo E, et al.","authors":"","doi":"10.1016/j.molpha.2026.100109","DOIUrl":"https://doi.org/10.1016/j.molpha.2026.100109","url":null,"abstract":"","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 4","pages":"100109"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression of concern: \"Glucocorticoid-Induced Tumor Necrosis Factor Receptor-Related (GITR)-Fc Fusion Protein Inhibits GITR Triggering and Protects from the Inflammatory Response after Spinal Cord Injury\" [Mol. Pharmacol. 2008; 73: 1610-1621] by Nocentini G, Cuzzocrea S, Genovese T, et al.","authors":"","doi":"10.1016/j.molpha.2026.100108","DOIUrl":"https://doi.org/10.1016/j.molpha.2026.100108","url":null,"abstract":"","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"108 4","pages":"100108"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817629","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}