Biological & pharmaceutical bulletin最新文献

Duchenne muscular dystrophy (DMD) is a hereditary disease that requires gene or nucleic acid therapy, which involves muscle-targeted delivery of therapeutic material. We previously developed liposomes targeting muscle tissue in DMD model mice (mdx) using an A2G80 peptide, which has an affinity for α-dystroglycan abundantly expressed on the muscle cell membrane. However, these liposomes did not carry gene or nucleic acids. In this study, we aimed to develop muscle-targeting lipid nanoparticles (LNPs) encapsulating luciferase mRNA and evaluate gene expression levels after systemic administration of these LNPs. We first evaluated the efficiency of mRNA delivery based on luciferase activity using polyethylene glycol (PEG)-dimyristoyl glycerol (DMG) and PEG-distearoyl glycerol (DSG) in mdx systemic administration. PEG-DSG-LNPs showed lower luciferase expression in the liver and spleen and higher expression in mdx muscle tissue than PEG-DMG-LNPs. The addition of the A2G80 peptide to LNPs using PEG-DSG (A2G80-DSG-LNPs) significantly increased their activity in mdx but not in normal mice. These results suggest that A2G80-DSG-LNPs allow for muscle-targeted mRNA delivery and are useful tools for DMD treatment.

One of the most significant conceptual changes brought about by the discovery of clock genes and development of circadian-clock mutant mice is the recognition that impaired circadian rhythmicity extends its impact far beyond sleep, driving pathogenesis of a wide variety of disorders such as cancer, obesity, and hypertension. However, despite this growing clinical evidence, chronobiology still lacks a coherent answer to the converse question: can restoration of circadian rhythms ameliorate-or even reverse-such diseases? In this review, three complementary pharmacological strategies-each still in preclinical development-are explored. First, direct modulation of the transcription-translation feedback loop (TTFL)-the core gene-regulatory circuit that generates 24-h rhythms in almost all nucleated cells-is reviewed as an approach to manipulation of cellular circadian biology. Second, the suprachiasmatic nucleus (SCN)-enriched G-protein-coupled receptor Gpr176 is highlighted as a central-clock target, given its ligand-independent, G_z-mediated control of cAMP signaling and demonstrated ability to reset the master pacemaker. Third, the concept of rhythmic enhancement of output function is introduced and exemplified by describing re-activation of circadian oxidized form of nicotinamide adenine dinucleotide (NAD⁺)-dependent 3β-hydroxy-steroid dehydrogenase (3β-HSD) activity in the meibomian gland-using nicotinamide mononucleotide (NMN)-to restore peripheral clock-driven steroidogenesis in this tissue, which leads to amelioration of meibomian gland dysfunction, a leading cause of dry eye disease. This review aims to highlight the molecular logic of each strategy; both mechanistic insights and safety/efficacy considerations are discussed.

Bone resorption inhibitors, such as bisphosphonates, infrequently cause refractory medication-related osteonecrosis of the jaw (MRONJ). Oral care, including maintaining oral hygiene and preventing oral infections, is important for preventing MRONJ. However, the extent of oral care in patients taking bisphosphonates is unclear. We recommended dental visits to 790 outpatients taking bisphosphonates who visited the pharmacy at Ehime University Hospital. The effect of this recommendation was determined by evaluating changes in the rate of dental visits within 6 months among patients with and without a family dentist. Following recommendations by pharmacists, the dental visit rate increased from 46.2 to 67.6%, and the proportion of patients with a family dentist increased from 55.9 to 76.5%. Furthermore, patients with a family dentist had a significantly higher rate of dental visits than those without a family dentist (79.9 vs. 6.8%). The findings in this study suggest that pharmacists' recommendations for oral health care may lead to early detection and avoidance of MRONJ.

Parthanatos is a noncanonical form of regulated cell death mediated by the overactivation of poly(ADP-ribose) polymerase-1, yet its regulatory mechanisms are not fully understood. To fully elucidate its regulatory mechanisms, it is necessary to establish useful research tools to investigate parthanatos. We have previously identified the human fibrosarcoma HT1080 cells as highly sensitive cells to parthanatos, and cefotaxime, a 3rd-generation cephem antibiotic, as the parthanatos inducer. In this study, we comprehensively characterized the ability of various cephem antibiotics, including cephalothin (also called cefalotin) (CET), cefoxitin, ceftriaxone, cefoperazone, ceftezole, and cefalexin, to induce parthanatos. Among them, CET exhibited the lowest LD₅₀. Therefore, our results show that CET works as the more potent and useful parthanatos inducer.

The high plasma glucose induced in glucose metabolism disorders leads to secondary pathologies, including bone disease. Fish scales, similar to mammalian bone, are composed of osteoblasts, osteoclasts, and calcified bone matrix and have been used as a system to analyze hyperglycemia-induced bone abnormalities. Here, we developed a hyperglycemia model in fish to study abnormalities in bone metabolism linked to increased plasma glucose and to analyze the function of calcitonin, the suppressor of osteoclastic activity, while maintaining high glucose levels. Following a 1-d fast and exposure to 5% glucose, plasma glucose concentrations increased significantly. We then examined plasma calcium and osteoclast activity of scales related to bone metabolism in goldfish treated with glucose for 5 d after a 1-d fast. The results showed that glucose treatment significantly increased plasma calcium levels at 3 and 5 d with a decrease in calcium content in the scales of goldfish. Hyperglycemia in glucose-exposed goldfish induced osteoclastic activation in scales, as indicated by the ratio of the osteoclastic activating factor (rankl) to the osteoclast inhibiting factor (osteoprotegerin, opg). Plasma calcitonin was found to be increased in glucose-exposed goldfish, which appears to suppress bone resorption by regulating the rankl/opg ratio. This hyperglycemia model, capable of examining both glucose and bone metabolism, would be valuable for analyzing the mechanism underlying abnormal bone metabolism caused by hyperglycemia.

The aim of this study was to investigate whether ginsenoside Rb1 attenuates cockroach extract (CRE)-induced asthma by interfering with mitochondrial dysfunction. After induction with CRE, mice were administered different doses of Rb1. Hematoxylin-eosin (H&E) staining, enzyme-linked immunosorbent assay (ELISA), and flow cytometry analysis revealed that inflammatory cell infiltration, total immunoglobulin E (IgE) and CRE-specific IgE in serum, and inflammatory cytokines in bronchoalveolar lavage fluid were effectively inhibited by Rb1. Through Western blot, TUNEL, and immunofluorescence colocalization assays, we observed Rb1 also inhibited endogenous reactive oxygen species (ROS), tightly associated with increased superoxide dismutase, catalase levels, and decreased malondialdehyde levels. Subsequently, the silent information regulator sirtuin 1 (SIRT1)/peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) pathway was activated, whereas the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway was inhibited. Additionally, Rb1 could rescue mitochondrial dysfunction by promoting the mitochondrial fusion protein mitofusion 1 (MFN1) and inhibiting dynamin-related protein 1 (DRP1) expression and apoptosis in the lungs. In BEAS-2B cells, Rb1 plays a role similar to that of a SIRT1 agonist (SRT1720), including enhancing mitochondrial membrane potential and decreasing mitochondrial ROS and DRP1 translocation to mitochondria. Our findings suggest that Rb1 maintains mitochondrial integrity by activating SIRT1/PGC-1α and inhibiting PI3K/AKT, thereby ameliorating asthmatic airway inflammation.

Retinoic acid receptor-related orphan receptor gamma (RORγ) is a key transcriptional factor that plays a crucial role in the differentiation and activation of Type 17 cells, such as interleukin-17 (IL-17)-producing CD4⁺ T (Th17) cells and CD8⁺ T (Tc17) cells, which are known to boost antitumor responses. Although a RORγ agonist (LYC-55716) has been under clinical evaluation, the precise effects of RORγ agonists on immune cells within tumor environments remain unclear. In our study, we investigated the role of tumor-infiltrating immune cells in the MC38 syngeneic mouse model of colorectal cancer using Compound-34, a novel orally available RORγ-selective agonist we discovered. Our findings revealed that Compound-34 exerts its antitumor efficacy by modulating immune cell activity rather than directly targeting tumor cells. Specifically, Compound-34 increased the infiltration of effector T cells, including Th17 and Tc1 (interferon [IFN]-γ⁺ CD8⁺ T) cells, as well as innate immune cells like natural killer (NK) and natural killer T (NKT) cells, within the MC38 tumor tissue. Following the administration of Compound-34, there was an increase in IFNγ and Granzyme B within the MC38 tumor tissue, accompanied by an increase in the infiltration of cytotoxic immune cells. Moreover, the addition of Th17-derived cytokines to MC38 cells stimulated the release of CXCL10, a chemokine crucial for immune cell recruitment. These results offer valuable insights into the immunomodulatory and therapeutic potential of RORγ agonists in cancer immunotherapy, highlighting their role in enhancing immune cell infiltration and activity within tumors.

Exposure to ionizing radiation during early pregnancy poses serious risks to fetal development, particularly during organogenesis. In this study, we evaluated the protective effects of AHCC^®, a standardized extract of Lentinula edodes mycelia with known immunomodulatory and metabolic properties, using an ICR mouse model. Intraperitoneal administration of AHCC prior to 1.4-Gy γ-irradiation on gestational day 9 significantly reduced the incidence of tail malformations and diaphragmatic hernias, without affecting implantation or fetal survival. Additionally, lifelong AHCC ingestion across three generations did not alter reproductive parameters, namely, gestation length and litter size, supporting its safety during pregnancy. These findings suggest that AHCC may mitigate radiation-induced teratogenic effects through maternal immune modulation, without compromising reproductive outcomes.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by selective motor neuron cell death. A GGGGCC hexanucleotide repeat expansion (HRE) within the chromosome 9 open reading frame 72 (C9orf72) gene is a major causative factor in ALS. This abnormal HRE triggers five types of dipeptide repeat protein (DPR), each composed of two alternating amino acid expressions. Among the DPRs, arginine-rich Poly-PR localizes predominantly to the nucleus, exerting particularly strong toxicity on motor and cortical neurons. Several mechanisms have been proposed for poly-PR-induced neurotoxicity. In this study, poly-PR-expressing NSC34 motor neuron-like cells showed an increase in oxidative stress. Fibroblast growth factor receptor 1 (FGFR1) is known to promote neurogenesis and inhibit apoptosis in neurons. However, its neuroprotective effects against DPR-induced toxicity have not been previously reported. Here, we demonstrated that FGFR1 activation reduced oxidative stress by upregulating nuclear factor erythroid 2-related factor 2 (NRF2) expression. Furthermore, we propose that the increase in NRF2 through FGFR1 activation may result from the alleviation of protein translation impairment. Overall, these findings suggest that FGFR1 activation provides neuroprotection against poly-PR toxicity and may represent a potential therapeutic strategy for ALS.