Journal of Pharmacology and Experimental Therapeutics最新文献

Diabetes mellitus is an escalating global health challenge, with current therapies primarily focused on symptom management rather than targeting the root causes of the disease. Emerging mRNA and gene therapies offer a new frontier by targeting disease mechanisms at the molecular level. This review explores the mechanistic landscape, delivery systems, and therapeutic outcomes of mRNA and gene-based interventions in diabetes. mRNA therapies, notably delivered via lipid nanoparticles, have demonstrated potential in restoring insulin production, enhancing angiogenesis, and modulating immune responses. mRNA therapy for both type 1 and type 2 diabetes using mRNAs of vascular endothelial growth factor-A, fibroblast growth factor 21, and glucagon-like peptide-1 linked to the IgG4 Fc region has shown immense potential in preclinical models of diabetes. Gene therapy for long-term correction of insulin signaling and pancreatic cell reprogramming is widely administered using viral (adeno-associated virus, lentivirus) vectors; however nonviral methods such as lipid nanoparticles and chitosan-based nanoparticles are being investigated in preclinical stages. CRISPR/Cas9 based genome-editing tools are another emerging therapeutic option for diabetes that correct metabolic defects and protect β cells. Despite these advances, delivery efficiency, immunogenicity, and off-target effects remain key translational hurdles for these therapies. Ongoing clinical trials for diabetes, including vascular endothelial growth factor A mRNA therapy for diabetic wound healing and fibroblast growth factor 21 gene therapy for type 1 diabetes, highlight the promising applications of these technologies. With careful attention to safety and pharmacokinetics, mRNA and gene therapies hold transformative promise as next-generation treatments for diabetes and its complications. SIGNIFICANCE STATEMENT: This review highlights emerging mRNA and gene therapies targeting the root causes of diabetes. By focusing on molecular mechanisms, delivery systems, and clinical advances, the review outlines next-generation strategies for treating diabetes and its complications.

Multidrug resistance of cancer cells is attributed to drug-induced alteration of numerous intracellular processes. Using clinically relevant models of triple-negative breast and non-small lung cancer cells we previously showed that these cells respond to repeated paclitaxel exposure by inter alia lysosome enrichment in ABCC3, ABCC5, and ABCC10, which contribute to drug sequestration in these organelles and reduced drug cytotoxicity. In this study, we provide experimental evidence that transcription of the above-mentioned ABCC genes is enabled by BRG1-based SWI/SNF chromatin remodeling complex. Pharmacological inhibition of SWI/SNF with PFI3 or ACBI1, the PROTAC degrader of SMARCA2/4, substantially reduced transcription of ABCC3, ABCC5, and ABCC10. A similar effect was caused by transient silencing of SMARCA4 (BRG1), but not SMARCA2 (BRM). The deficiency of BRG1 led to extralysosomal distribution of anticancer drugs, their deeper penetration of spheroids, and substantial increase in drug cytotoxicity. Interestingly, in BRG1-deficient cell line paclitaxel triggered mutations, which reverted BRG1 truncating deletion in SMARCA4, thereby restoring SWI/SNF ATPase expression in paclitaxel-resistant cells and increasing transcription of ABCC. Acquisition of drug resistance was associated with BRG1 redistribution in the genome, de novo occurrence at the promoters of genes functionally linked to endolysosomal system, and stronger co-occurrence with EP300. Our study indicates possible target--SWI/SNF complex for anticancer combinatorial interventions in paclitaxel-induced multidrug resistant phenotypes. SIGNIFICANCE STATEMENT: This study provides evidence that BRG1 inhibition with PFI3 and degradation of SMARCA4 mRNA substantially declines lysosomal drug sequestration and potentiate drug toxicity. Therefore, BRG1 targeting can be considered as candidate for combinatorial anticancer therapy with some standard chemotherapy drugs.

Cardiovascular disease (CVD) remains the leading cause of death worldwide despite decades of therapeutic advances. Emerging insights into its etiology have revealed previously unappreciated cellular and molecular drivers beyond traditional risk factors, prompting the development of treatments that target newly identified culprit proteins and cells within cardiovascular tissues. Protein-based biologics-particularly monoclonal antibodies and multispecific proteins-are known for their strength and specificity in targeting and their established use as treatments for other diseases. However, extending biologics to new indications faces challenges: achieving durable effects in diseased tissues and minimizing side effects in healthy tissue. Addressing these long-standing challenges requires fine-tuning biologics' pharmacokinetic properties and pharmacodynamic effects according to target- and disease-specific requirements. In this review, we examine foundational pharmacokinetic and pharmacodynamic principles in the context of cardiovascular-targeted biologics, highlighting the role of protein design in controlling distribution, efficacy, and safety. Additionally, we discuss emerging preclinical and clinical biologics specifically designed for CVDs, as well as emerging opportunities in this landscape. These advances point toward a future where pharmacokinetics guide the rational design of next-generation protein therapeutics for CVD. SIGNIFICANCE STATEMENT: Protein-based biologics hold promise for treating cardiovascular diseases (CVD); however, their successful translation requires understanding how proteins' properties and cardiovascular physiology shape pharmacokinetic and pharmacodynamic behavior. This minireview connects foundational pharmacology principles with strategies in protein engineering suitable for CVD applications. Pharmacokinetic-guided design will accelerate the development of protein therapies that can transform CVD treatment.

Environmental cues that have been associated with drug-taking can evoke drug-craving and drug-seeking and drive relapse. Using the New Response Acquisition procedure, we evaluated the extent to which activation of delta opioid receptors (DORs) changes responding for cocaine-associated stimuli. We hypothesized that activation of DORs, either directly via agonists or indirectly via protected concentrations of endogenous enkephalin peptides, would increase the conditioned reinforcing effects of cues. First, animals undergo Pavlovian conditioning during which rats received 5 infusions of cocaine (0.32 mg/kg/inf) and either paired or unpaired presentations of a stimulus (light + tone) per day for 10 days. Next, nosepokes were added to the operant chamber and rats were allowed to respond for presentations of cocaine-associated stimuli (acquisition). Consistent with previous findings, animals assigned to paired Pavlovian conditioning emitted more responses for cue presentations than animals assigned to the unpaired control. Interestingly, acute administration of SNC80 (DOR agonist; 3.2 mg/kg s.c.) on acquisition session 4 led to robust increases in responding for the cocaine-paired cues in the paired, but also increased responding for cues in rats assigned to unpaired and saline control groups. Further, the enkephalinase inhibitor RB101 (10 mg/kg intravenous), which maintains extracellular concentrations of enkephalins, increased active responding in a DOR-dependent manner. These data suggest that activation at DORs influences behaviors maintained by cues and sheds light on the neurobiology underlying the conditioned reinforcing effects of drug-associated stimuli. SIGNIFICANCE STATEMENT: We used a more rigorous test of conditioned reinforcement to show that activation of delta opioid receptors increases the reinforcing effects of cocaine-paired cues, depending on conditioning history, which implicates the delta opioid receptor system as a target to reduce relapse.

Efficient delivery of small interfering RNA (siRNA) remains a major challenge in gene therapy, particularly due to poor cellular uptake, enzymatic degradation, and endosomal entrapment of the cargo. Cell penetrating peptides (CPPs) offer a promising strategy for intracellular delivery of nucleic acids; however, peptide-based gene delivery carriers possess limited nucleic acid condensation capability, demonstrating reduced transfection efficacies in mammalian cells. This study aimed to enhance siRNA delivery efficacies of CPP-based siRNA therapeutics by combining nucleic acid condensation efficacies of methylated protamine with the cell permeation capabilities of cyclic peptides to improve physiological stability, cellular uptake, and gene silencing efficacies in HER2⁺ breast cancer cells. Cyclic CPPs (cCPPs) of different cationic and amphipathic characters (namely, cTAT, cKALA, and cC105Y) were synthesized by solid phase peptide synthesis. Methylated protamine/siRNA complexes of net neutral nanoparticles (NNPs) obtained at nitrogen-to-phosphate ratio of 1 were functionalized with cCPPs at various cCPP/siRNA w/w ratios, and the sizes and net charges of the formulated nanoparticles were analyzed using dynamic light scattering and a zeta potential instrument, respectively. cCPP-functionalized NNPs demonstrated improved nucleic acid condensation efficacy and protection against enzymatic degradation compared with bare NNPs. cCPP-NNPs formulated using various types of peptides overall demonstrated superior siRNA transfection efficacies with ∼60% HER2 gene silencing efficacies in HER2⁺ breast cancer cells. cC105Y-NNPs particularly showed improved physiological stability and cellular uptake compared with the other cCPP-functionalized nanoparticles. Endosomal escape remained a limiting factor in the gene expression efficacies of cCPP-NNPs, and incorporation of an endosome-disrupting agent improved HER2 gene knockdown efficacies (∼80%) to levels that were comparable to the Lipofectamine control. Furthermore, the potential of cCPP-NNPs for simultaneous knockdown of HER2 and HER3 was demonstrated to improve the anticancer efficacies of the nanoparticle for HER2⁺ breast cancer treatment. SIGNIFICANCE STATEMENT: Cyclic cell penetrating peptide-functionalized nanoparticles with improved physiological stability and optimized nucleic acid condensation and release efficacies formulated by simple deposition of cationic peptides on protamine small interfering RNA complexes demonstrate superior gene knockdown and anticancer efficacies in breast cancer cells.

Ocular drug delivery faces significant challenges because of the eye's complexity and anatomical barriers, such as the cornea, conjunctiva, and blood-retinal barrier, which limit drug penetration and bioavailability. Recent advances in nanotechnology-based drug delivery have led to the development of innovative delivery platforms, enabling targeted, sustained, and minimally invasive delivery for ocular diseases and injuries. This review outlines the recent advances in nanosystems-based ocular drug delivery and highlights the latest progress in targeting technologies based mainly on preclinical studies and selected clinical trial data. It covers a variety of nanosystems, including organic nanoparticles (NPs) such as liposomes, nanomicelles, nanosuspensions, nanoemulsions, dendrimers, and nanofibers. Additionally, it addresses inorganic NPs, which include gold NPs, silver NPs, silica NPs, and carbon nanotubes. Besides, we summarize the clinical challenges and regulatory aspects in nanotechnology-based ocular drug delivery. Finally, inspired by current advances and therapeutic strategies, we provide an insight into clinical applications and future research directions on nanosystems-based drug delivery. We highlight the need to overcome the challenges of using nanosystems in ocular drug delivery and fill the knowledge gap on their nanotoxicity and future development. SIGNIFICANCE STATEMENT: This review highlights recent advances in nanosystem applications for ocular drug delivery, summarizes up-to-date clinical trials utilizing nanosystems for ocular drug delivery, and discusses clinical challenges and directions for future development.

The clinical effectiveness of peptide and protein therapeutics is often limited by their short circulating half-life, necessitating frequent injections. Therefore, half-life extension strategies have emerged as centerpieces in biopharmaceutical development to enhance therapeutic efficacy, reduce dosing frequency, and improve patient outcomes. A variety of approaches have been being investigated and used to prolong systemic exposure of protein drugs. Increasing hydrodynamic volume and enabling neonatal Fc receptor recycling are 2 mechanisms that are being applied to extend the half-life of many therapeutic proteins. In this review, we discuss elimination mechanisms of protein therapeutics, underlying mechanisms and strategies for their half-life extension, impact of half-life extension on primary pharmacokinetic parameters, and critical factors to be considered to choose effective half-life extension strategy. Despite these advances, challenges remain in balancing half-life extension with biological activity, stability, and manufacturability. Ongoing research aims to optimize these technologies to meet the growing demand for long-acting biologics in chronic disease management. As half-life extension continues to evolve, it holds promise not only in improving therapeutic performance but also in expanding the applicability of protein drugs across a broader range of clinical indications. SIGNIFICANCE STATEMENT: The clinical utility of many protein and peptide therapeutics is limited by insufficient circulation time. Half-life extension strategies such as PEGylation, albumin fusion, and Fc fusion have demonstrated benefits clinically and permit extended dosing windows.

Adiponectin receptor (AdipoR) agonists protect against glomerular inflammation and injury in obesity-related glomerulopathy (ORG), but their molecular mechanisms remain unclear. Given the implication of the ceramide signaling pathway in the pathogenesis of ORG, the present study tested whether AdipoR agonists target acid ceramidase (AC) to inhibit NLRP3 inflammasome activation in podocytes, thereby blocking glomerular inflammation and injury during obesity. Confocal microscopy showed that adiponectin attenuated visfatin-induced NLRP3 inflammasome activation and IL-1β-containing multivesicular body (MVB) formation in podocytes. Nanoparticle tracking analysis revealed that adiponectin suppressed visfatin-induced extracellular vesicle release, an effect dependent on AC activity. Structured illumination microscopy demonstrated that visfatin reduced lysosome-MVB interaction in podocytes, which was restored by adiponectin via enhancement of TRPML1 channel-mediated Ca²⁺ release. The rescue of lysosome-MVB interaction and TRPML1 channel activity by adiponectin was mimicked by the AC enhancer but interfered with by the AC inhibitor. In vivo, high-fat diet (HFD) treatment induced NLRP3 inflammasome activation and T cell infiltration in glomeruli and increased urinary extracellular vesicle excretion in mice, which were exaggerated by podocyte-specific Smpd1 gene (gene code of acid sphingomyelinase) overexpression in Smpd1^trg/Podo^cre mice compared with WT/WT mice. AdipoRon, a synthetic AdipoR agonist, reduced HFD-induced glomerular inflammation in both WT/WT and Smpd1^trg/Podo^cre mice, but its effect was blocked by AC inhibition. Moreover, podocyte-specific Smpd1 gene overexpression aggravated HFD-induced podocyte injury, proteinuria, and glomerular sclerosis, which were mitigated by AdipoRon in an AC-dependent manner. Additionally, we found that the protective actions of AdipoRon may be mainly attributed to the activation of AdipoR1, but not AdipoR2. Taken together, our findings suggest that AC activation mediates the protective effects of AdipoR agonists against glomerular inflammation and injury in ORG, highlighting AC as a potential therapeutic target. SIGNIFICANCE STATEMENT: This study identifies acid ceramidase as a key mediator of adiponectin receptor agonist action in podocytes, linking its activation to suppression of NLRP3 inflammasome and extracellular vesicle release, and highlighting a novel therapeutic target in obesity-related kidney disease.