CaN (calcineurin) promotes pathological cardiac remodeling but also cardioprotection in ischemia-reperfusion injury. CaN inhibitors are also immunosuppressants. This pleiotropy complicates targeting calcineurin in cardiovascular disease. CIP4/TRIP10 (Cdc42-interacting protein 4) is a scaffold protein that binds the CaNAβ (calcineurin Aβ) N-terminal polyproline domain and organizes a calcium and CaNAβ2 signaling compartment independent of contractile calcium. We showed that CIP4-CaNAβ2 signalosomes promote pathological cardiac hypertrophy induced by pressure overload. It is unknown whether CIP4-CaNAβ2 signalosomes contribute to cardioprotection and remodeling in ischemic disease. CIP4 conditional knockout mice were studied following ischemia-reperfusion and permanent left coronary artery ligation that induce myocardial infarction. C57BL/6NJ mice were transduced with cardiotropic adeno-associated virus expressing a CaNAβ2 small hairpin RNA to inhibit CaNAβ2 expression, a VIVIT peptide to inhibit CaN-NFAT (nuclear factor of activated T cells) signaling, or a CaNAβ polyproline peptide to block CIP4-CaNAβ2 binding and similarly studied by ischemia-reperfusion injury and left coronary artery ligation. CaNAβ polyproline-dependent signaling was also studied in T cells. CIP4 conditional knockout and cardiomyocyte-specific CaNAβ polyproline peptide expression improved cardiac function after ischemia-reperfusion injury and decreased infarct size and improved cardiac function after permanent left coronary artery ligation. In contrast, cardiomyocyte-specific CaNAβ2 depletion and VIVIT expression worsened outcome after myocardial infarction. The polyproline peptide had no effect on T-cell activation and cytokine expression invitro. CIP4-CaNAβ2 signalosomes promote adverse cardiac remodeling and are not cardioprotective. Proof of concept is provided for the treatment of ischemic cardiomyopathy by a polyproline peptide gene therapy. Targeting these complexes may be beneficial in cardiovascular diseases, including ischemic cardiomyopathy and acute myocardial infarction.

Mitochondrial dysfunction of mesenchymal stem cells (MSCs) has been implicated in impaired osteogenesis, resulting in bone loss following radiation therapy. However, the underlying mechanisms remain to be fully elucidated. This study reveals the critical role of Fis1 in regulating mitochondrial dynamics and MSC osteogenesis in radiation-induced bone injury. Specifically, radiation activates Fis1 expression, which induces excessive mitochondrial fission, leading to mitochondrial fragmentation, along with reduced capacities for oxidative phosphorylation, ATP synthesis, and antioxidant defense, that collectively impairs MSC osteogenesis and results in bone loss in radiation-induced bone injury. This process involves increased calcium (Ca2+) influx that stimulates calcineurin (CaN) to promote nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) dephosphorylation and nuclear translocation, which in turn, activates the transcriptional expression of Fis1. Consistent with the pivotal role of Fis1 in regulating mitochondrial fission and MSC osteogenesis, inhibition of Fis1 remarkably reduced mitochondrial fragmentation, enhanced MSC osteogenesis and reduced bone loss, highlighting the therapeutic potential of targeting Fis1 in radiation-induced bone injury. Our study provides new insights into the mechanisms and therapeutic strategies for radiation-induced bone injury.

Calcium dysregulation in Alzheimer's disease: unraveling the molecular nexus of neuronal dysfunction and therapeutic opportunities.

Calcineurin (CaN), a Ca2+-dependent phosphatase, plays key roles in eukaryotic cell signaling. We investigated whether Leishmania amazonensis’ two infective forms—promastigotes and amastigotes—exhibit differences in CaN expression, localization, and functional impact, using two canonical inhibitors (cyclosporin A, CsA; tracolimus, FK506). At high 40 µM CsA, promastigotes showed reduced viability, whereas amastigotes remained resistant. FK506 had no effect on either form. At a sub-lethal 25 µM CsA, parasite proliferation remained unaffected. In parasite–macrophage co-incubation assays, phosphorylation patterns differed: amastigotes—but not promastigotes—showed increased serine/threonine phosphorylation upon CaN inhibition. Western blotting and in silico data revealed higher CaN catalytic (CaNA2) and regulatory (CaNB) subunit expression in amastigotes than promastigotes. Immunofluorescence localized CaNA prominently in both cytoplasm and nucleus of promastigotes, but predominantly cytoplasmic in amastigotes; CaNB was largely cytoplasmic in both. In silico localization predictions suggested strong membrane associations for CaNA in Leishmania, contrasting with mammalian models. Subcellular fractionation confirmed CaNA enrichment in membrane fractions, with CaNB in cytoplasmic and nuclear fractions. Collectively, these findings reveal form-specific differences in expression, subcellular distribution, and inhibitor responses of CaN in L. amazonensis, highlighting its potential as a stage-specific therapeutic target in leishmaniasis.

Extracellular proteins from Perkinsus olseni: LC-MS/MS-based proteomic profiling and functional insights into host-parasite interactions.

Sublethal and lethal exposure to broflanilide disrupt sex pheromone biosynthesis signaling and reproductive success in the Spodoptera frugiperda.

Background: Immunosuppressive drugs are critical for managing organ transplantation and autoimmune diseases by targeting aberrant T lymphocyte activation and immune responses. However, existing therapies often cause significant side effects, driving the urgent need for safer alternatives. Natural products, particularly from marine-derived sources like mangrove endophytic fungi, have emerged as promising candidates due to their bioactive diversity and minimal toxicity profiles. Methods: To explore this potential, this study investigated pestalotiopyrone M (PyM), a new pyrone derivative obtained from the endophytic fungus Pestalotiopsis sp. HHL101, which was isolated from the Chinese mangrove plant Rhizophora stylosa. Its immunosuppressive activity targeting calcineurin (CN) was evaluated, and spleen cell viability was tested by CCK-8. The effects of PyM on T cell proliferation were assessed using Con A-induced splenocyte proliferation and unidirectional mixed lymphocyte response (MLR) models. Cell cycle and apoptosis analysis were performed to determine potential cytostatic effects. Additionally, the impact of PyM on NFAT1 dephosphorylation, translocation, and subsequent cytokines secretion (IL-2, IL-4, TNF-α, and IFN-γ) was examined in ConA-stimulated cells. Results: We verified that PyM obviously inhibited Con A-induced splenocyte proliferation and unidirectional MLR. Crucially, it demonstrated markedly reduced cytotoxicity, exhibiting approximately 38-fold lower cytotoxic effects compared to the clinically used drug CsA. It arrested the cell cycle from G1-phase to S-phase and G2-phase. Furthermore, we confirmed that PyM suppressed ConA-stimulated activation of NFAT1 dephosphorylation and blocked NFAT1 translocation in vitro, subsequently inhibiting the transcription and expression of IL-2, IL-4, TNF-α, and IFN-γ. Conclusions: These results indicate that PyM is an efficient inhibitor of the CN/NFAT signaling pathway. It shows promising immunosuppressive activity by targeting T cell activation and proliferation, indicating its potential as a candidate for development into an immunosuppressive agent for treating adverse immune responses.

Aim: Skeletal muscle unloading leads to the upregulation of proteolytic gene expression, downregulation of protein synthesis markers, and the development of muscle atrophy. These changes are accompanied by alterations in calcium signaling. We investigated the role of Inositol 1,4,5-triphosphate (IP3) receptors (IP3Rs) in regulating calcium signaling and controlling gene expression in skeletal muscles during unloading. Methods : Male Wistar rats were randomly assigned to 4 groups (n = 8 per group). C - vivarium control; C+2APB - vivarium control with daily intraperitoneal injections of the IP3 receptor inhibitor 2-aminoethoxydiphenyl borate (2-APB, 10 mg/ kg b.w.); HU - 3-day hind limb unloading; HU+2APB - 3-day hind limb unloading with daily 2-APB injections. After the intervention, soleus muscles were analyzed for markers of calcium metabolism and proteostasis. Results: Three days of unloading resulted in a significant increase in nuclear phosphorylated Ca2+/calmodulin-dependent protein kinase II (p-CaMK II) content and calcineurin (CaN) expression compared to the C group (P < 0.05); this effect was prevented in the HU+2APB group. In HU+2APB rats, the decline in soleus muscle weight-to-body weight ratio was partially prevented, and the downregulation of protein synthesis markers (as seen in the HU group) was also prevented. However, proteolytic signaling markers were equally upregulated in the HU+2APB and HU groups compared to C. Conclusion: IP3 receptor inhibition during 3-day hind limb suspension in rats partially prevented the decline in m. soleus weight index and protein synthesis markers. This effect may be attributed to alterations in the regulation of nuclear calcium signaling.

Cell survival depends upon the ability to adapt to changing environments. Environmental stressors trigger an acute stress response program that rewires cell physiology, downregulates proliferation genes and pauses the cell cycle until the cell adapts. After the acute response is resolved, cells resume cycling but at a reduced rate. The importance of cell cycle changes for survival in chronic stress is not clear. Here, we show that dynamic phosphorylation of the yeast cell cycle-regulatory transcription factor Hcm1 is required to maintain fitness in chronic stress. Hcm1 is activated by cyclin dependent kinase (CDK) during S-phase and is inactivated by the phosphatase calcineurin (CN) in response to stressors that signal through increases in cytosolic Ca2+. Cells expressing a constitutively active, phosphomimetic Hcm1 mutant exhibit a reduction in fitness in stress, suggesting Hcm1 inactivation promotes survival. However, a comprehensive analysis of Hcm1 phosphomutants revealed that Hcm1 activity is also important to survive stress, and that all mutants with fixed phosphorylation states are less fit in stress. Moreover, our data suggests that pulses of Hcm1 activity are necessary to maximize target gene expression in stress. These findings demonstrate that expression levels of Hcm1 target genes influence fitness in stress and suggest that the dynamic phosphorylation of cell cycle regulators plays a crucial role in promoting survival in stressful environments.

Binge-drinking behavior is a prevalent and costly burden that many face. To expand potential treatments for alcohol use disorder and improve treatment efficacy, calcineurin (CN) inhibiting cyclosporine A (CsA) is investigated as a potential treatment for this disorder based on a neuroinflammation-driven approach to addictive behaviors and stress maladaptation. CsA has previously been shown to reduce binge-drinking behaviors, and this study aims to provide a connection between binge-drinking reduction and stress response to the expression of the neuroinflammasome. CamKIIa and CRF neuronal CN knockout mice were characterized for knockout of CN expression through immunohistochemistry and RNA scope imaging of selected brain regions. Others from this cohort were then subjected to 6-week binge-drinking behavioral experiments in the format of "Drinking-in-the-Dark" (DID). Experimental mice were tracked for ethanol intake overtime and following injection of either vehicle only intraperitoneal injection or CsA and vehicle injection for any changes. These transgenic mice were separately subjected to 1-hour restraint stress experiments with exposure to either CsA or vehicle only. These cohorts were sacrificed with their brain tissue harvested and microdissected for rtPCR characterization of inflammatory gene products. Expression of CD45, COX- 2, CYC, Iba-1, IL-1b, IL-6, TNF-a, ACTB, CCL2, and CCR2 was quantified. The expression for CsA exposed mice was then compared to vehicle-only mice through the log2 fold change analysis for final comparison. Immunohistochemistry with RNA scope imaging for calcineurin expression revealed widespread CN knockout in the experimental lineage. CN knockout revealed no effect on ethanol consumption in DID models for both CamkIIa and CRF neuronal CN knockout compared to wild type. CsA did still induce a large reduction in ethanol intake for both lines of CN knockout mice compared to baseline. In the restraint stress study, rtPCR log2 fold analysis revealed that CsA reduced a wide-range of stress-induced neuroinflammatory markers. Specifically, the authors observed a generalized reduction in inflammatory gene expression with IL-1b seeing a nearly 6-fold decrease in both the central nucleus of the amygdala as well as the paraventricular nucleus. The findings of normal baseline drinking behaviors and improved parameters following treatment with CsA in wild type and neuronal CN knockout lineages indicate that CN at the level of neurons is not responsible for CsA-induced reduction in binge-drinking behavior. These findings, in conjunction with the reduction of neuroinflammatory gene product expression, implicate glial cells as possibly responsible for these changes. Further investigation into glia cell specific CN knockout is warranted under similar conditions. Ultimately, these findings improve the characterization of the mechanism of CsA-induced reduction in binge drinking behaviors and aid in discovering new treatments for alcohol use disorder.

Rationale: The death of chondrocytes triggered by extracellular acidification represents a critical factor in the degradation of cartilage tissue and bone, thereby exacerbating the progression of rheumatoid arthritis (RA). Our previous research demonstrated that acid-sensing ion channel 1a (ASIC1a) serves as a key acid sensor mediating the destruction of articular cartilage in RA, which is closely associated with mitochondrial damage of chondrocytes. However, its regulatory mechanism remains unclear.Methods: Cartilage samples from RA patients and collagen-induced arthritis (CIA) rat models were examined to determine the levels of mitophagy and PANoptosis. In parallel, primary rat articular chondrocytes were cultured and subjected to either ASIC1a activation or silencing. Mitochondrial function, mitophagy, and PANoptotic markers were evaluated using immunoblotting, immunofluorescence, and transmission electron microscopy. Additionally, the subcellular distribution of SIRT3 to clarify its role in maintaining mitochondrial homeostasis.Results: We observed a significant increase in the levels of mitophagy and PANoptosis within the cartilage tissue of both RA patients and collagen-induced arthritis (CIA) rat models. Activation of ASIC1a by extracellular acidification triggered mitophagy, ultimately resulting in PANoptosis of chondrocytes. The loss of ASIC1a protected chondrocytes from PANoptosis, thereby alleviating disease progression in CIA rats. Mechanistically, we demonstrated that the transport of SIRT3 from cytoplasm to mitochondria was inhibited upon ASIC1a activation. ASIC1a upregulated calcineurin (CaN) expression, which competitively bound to HSP70, disrupting the SIRT3-HSP70 complex and thereby impairing SIRT3 mitochondrial translocation. The reduced levels of SIRT3 in mitochondria induced mitochondrial dysfunction and excessive mitophagy in primary rat articular chondrocytes, ultimately leading to PANoptosis of chondrocytes. Restoration of SIRT3 improved mitochondrial dysfunction and inhibited excessive mitophagy in the process of ASIC1a-induced PANoptosis of chondrocytes.Conclusion: Our study demonstrated that ASIC1a induces the destruction of articular cartilage through the disruption of the equilibrium between mitochondrial quality control and cell fate. This suggests that ASIC1a is a promising therapeutic target to improve the clinical treatment of RA.

BACKGROUNDDiabetic cognitive dysfunction (DCD) is one of the chronic complications of diabetes, but its mechanism is currently unknown. Studies have shown that mitochondrial fission mediated by calcium overload is an important mechanism of DCD. Blocking calcium overload and restoring calcium homeostasis are key steps in treatment. Transient receptor potential melastatin 7 (TRPM7) is a novel player in causing calcium overload. Our previous studies have shown that genetic silencing of TRPM7 in type 1 diabetic rats leads to significant improvements in cognitive function, but the specific mechanism remains unclear. Troxerutin, extracted from the flowers of Sophora japonica, is one of the derivatives of rutin and has been shown to have neuroprotective effects. However, its association with TRPM7 remains unclear.AIMTo use animal and cellular models, we investigated whether TRPM7 mediated mitochondrial fission by upregulation of calcineurin (CaN)/dynamin-related protein 1 (Drp1)ser637 in DCD, and whether Troxerutin improved DCD by inhibiting TRPM7-mediated mitochondrial division.METHODSIn this study, we used db/db mice and hippocampal neuronal cell lines (HT22) treated with high-concentration glucose as our study subjects. We evaluated cognitive function using Morris water maze, novel object recognition tasks, and Nesting tests. We observed mitochondrial morphology using transmission electron microscopy and measured mitochondrial energy metabolism indicators using a spectrophotometer. We also detected mRNA and protein expression of TRPM7, CaN, p-Drp1ser637, caspase-3, B-cell lymphoma 2 associated X protein, and B-cell lymphoma 2 using quantitative real-time polymerase chain reaction, western blotting, and immunofluorescence.RESULTSIn the db/db diabetic mice with cognitive dysfunction, as well as in hippocampal neurons exposed to high-concentration glucose, TRPM7 and CaN expression were upregulated, phosphorylated Drp1ser637 expression was downregulated, and mitochondrial fission was increased. By modulating (inhibiting or overexpressing) TRPM7, it was further validated that TRPM7 activates the CaN/Drp1ser637 pathway, resulting in an increase in mitochondrial fission and neuronal cell apoptosis. Troxerutin downregulated TRPM7/CaN/Drp1ser637, reduced mitochondrial fission, and improved DCD.CONCLUSIONTRPM7 promotes mitochondrial fission via the CaN/Drp1ser637 pathway. Troxerutin improves mitochondrial function and reduces neuronal damage by inhibiting this pathway, suggesting TRPM7 as a potential therapeutic target for DCD.

The cysteine residue 674 (C674) of sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2 (SERCA2) is pivotal in maintaining SERCA2 activity. The C674S mutation leads to SERCA2 dysfunction and exacerbates atherosclerosis by inducing endoplasmic reticulum stress and inflammation in bone marrow-derived macrophages (BMDMs) and endothelial cells (ECs). This study aimed to explore if SERCA2 dysfunction aggravates atherosclerosis, by disrupting fatty acid metabolism and promoting the formation of macrophage foam cells. Heterozygous SERCA2 C674S gene mutation knock-in (SKI) mice were used to simulate SERCA2 dysfunction under pathological conditions. Serum from SKI mice and their littermate wild-type mice were taken for metabolomic testing. The entire aorta and aortic root were isolated for histological analysis. BMDMs were used for protein expression, lipid uptake and accumulation analysis. In SKI BMDMs, SERCA2 dysfunction induced the expression of calcineurin (CaN), which promoted nuclear translocation of forkhead box O1 (FoxO1) and transcription of its downstream target fatty acid-binding protein 4 (FABP4), leading to increased fatty acid synthesis and foam cell formation. Inhibition of the CaN/FoxO1/FABP4 pathway corrects aberrant lipid metabolism and inhibits the formation of foam cells in SKI BMDMs. Pharmacological interventions targeting either FoxO1 or FABP4, or FABP4 partial deficiency, significantly ameliorated atherosclerosis progression. SERCA2 dysfunction accelerates the progression of atherosclerotic lesions by stimulating the CaN/FoxO1/FABP4 pathway and promoting the formation of foam cells. Our findings highlight the importance of SERCA2 function in the context of atherosclerosis and reveal a novel therapeutic strategy to combat lipid accumulation and atherosclerosis.

Hyperhomocysteinemia (HHcy)-inducing diets recapitulate cerebral small vessel disease phenotypes in mice including cerebrovascular pathology/dysfunction, neuroinflammation, synaptic deficits, and cognitive decline. We recently showed that astrocyte signaling through calcineurin(CN)/nuclear factor of activated T cells (NFATs) plays a causative role in these phenotypes. Here, we assessed the impact of astrocytic signaling on microglia, which set the inflammatory tone in brain. Seven-to-eight-week-old male and female C57BL/6 J mice received intrahippocampal injections of adeno-associated virus (AAV) expressing EGFP (AAV2/5-Gfa2-EGFP) or AAV expressing the NFAT inhibitor VIVIT (i.e., AAV2/5-Gfa2-VIVIT-EGFP). Mice were then fed with control chow (CT) or B-vitamin-deficient chow for 12 weeks to induce HHcy. Immunohistochemistry and Western blot analyses suggested that expression of the homeostatic microglial marker, P2RY12, responded differently to AAV treatments depending on diet and sex. We next conducted single-cell RNA sequencing (scRNA-seq) to determine if microglial genes and/or clustering patterns were differentially sensitive to diet and AAV, depending on sex. In males, disease-associated microglial genes and subclusters were overrepresented in HHcy-treated mice, while VIVIT promoted the appearance of homeostatic microglial genes and clusters. In contrast, microglial genes in females were less sensitive to diet and AAV treatments, though disease-like patterns were also observed in the HHcy condition. Very few of the HHcy-sensitive microglial genes in females were affected by VIVIT. Though based on small sample sizes, the results suggest a sexually dimorphic influence of astrocyte signaling on microglial transcriptional phenotypes in the context of HHcy and small cerebral vessel disease. However, these interpretations will need to be bolstered with additional biological replicates and more stringent statistical analyses.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12974-025-03523-2.

Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling

Numerous investigations indicate that cyclosporine A (CsA) significantly influences the prevention or progression of Alzheimer's disease (AD) through various mechanisms. The precise pathways by which CsA, an inhibitor of calcineurin (CN) and peptidyl-prolyl isomerase A (PPIA), operates remain incompletely elucidated. In this study, we explored the effects of CsA, a widely utilized immunosuppressive agent, on the gene expression of various factors related to inflammation, cell adhesion, intercellular communication, and apoptosis in SH-SY5Y cells. We employed the semi-quantitative real-time PCR (RT-qPCR) method for gene expression analysis to assess these effects. Furthermore, network pharmacology was utilized to identify the potential targets of CsA and to further analyze. Our results demonstrated that CsA enhances the mRNA levels of most factors examined, including IL1B, S100A9, CD147, TREM2, LRP1, MAPK1, MAPK14, GSK3B, Dynamin 1, Dynamin 3, NLRP3, NLRP1, CASPASE 3, CASPASE 1, AIFM1, and STAT3. At the same time, it suppresses the mRNA levels of PPIA, TLR2, TLR4, PYCARD, RAGE, and BCL2. The network pharmacology analysis revealed seven target genes overlapping our experimental findings (PPIA, S100A9, TLR4, STAT3, MAPK1, MAPK14, and BAX). Our results suggest that CsA modulates gene expression in SH-SY5Y cells, with the potential for either beneficial/ harmful effects on cellular function, depending on the specific roles of the assessed genes.

Calcineurin, the Ca 2+ /calmodulin-activated protein phosphatase, recognizes substrates and regulators via short linear motifs, PxIxIT and LxVP, which dock to distinct sites on calcineurin to determine enzyme distribution and catalysis, respectively. Calcimembrin/C16orf74 (CLMB), an intrinsically disordered microprotein whose expression correlates with poor cancer outcomes, targets calcineurin to membranes where it may promote oncogenesis by shaping calcineurin signaling. We show that CLMB associates with membranes via lipidation, i.e. N-myristoylation and reversible S-acylation. Furthermore, CLMB contains an unusual composite 'LxVPxIxIT' motif, that binds the PxIxIT-docking site on calcineurin with extraordinarily high affinity when phosphorylated, 33 LDVPDIIITPP(p)T 44 . Calcineurin dephosphorylates CLMB to decrease this affinity, but Thr44 is protected from dephosphorylation when PxIxIT-bound. We propose that CLMB is dephosphorylated in multimeric complexes, where one PxIxIT-bound CLMB recruits calcineurin to membranes, allowing a second CLMB to engage via its LxVP motif to be dephosphorylated. In vivo and in vitro data, including nuclear magnetic resonance (NMR) analyses of CLMB-calcineurin complexes, support this model. Thus, CLMB with its composite motif imposes unique properties to calcineurin signaling at membranes including sensitivity to CLMB:calcineurin ratios, CLMB phosphorylation and dynamic S-acylation.

ABSTRACT Background Alzheimer’s disease (AD), a chronic and progressive neurodegenerative disease, is the most common cause of dementia. An important pathological basis for AD lesions is the excessive generation and deposition of β-amyloid (Aβ) caused by increased expression of the β-secretase, known as the β-site amyloid precursor protein cleaving enzyme 1 (BACE1). Effective suppression of the BACE1 overexpression has become a key AD treatment. Nuclear factor of activated T cells (NFAT) is a key transcription factor that regulates the expression of BACE1 in AD lesions, while Calcineurin (CaN) is a key regulatory protein that affects the transcription function of NFAT. Several lines of evidence have indicated that FK506 may promote the Aβ degradation via upregulation of the matrix metalloproteinase-9 (MMP-9) expression, which is associated with reduction of Aβ plaque deposition in the cerebral cortex and hippocampus. Methods In this study, behavioral, histological, and biochemical methods were used to investigate the key role and molecular mechanisms of CaN inhibitor FK506 in cognitive dysfunction, regulation of BACE1 expression, and Aβ production in APPswe/PS1dE9 transgenic mice. Results The results indicate that FK506 inhibits NFAT1 levels in the cerebral cortex and hippocampus, thereby reducing the expression of BACE1 and mediating APP processing towards non-amyloidosis pathways, significantly reducing Aβ overproduction, which in turn saved cognitive deficits in APPswe/PS1dE9 transgenic mice. In addition, FK506 treatment had no significant effect on the expression of a disintegrin and metalloprotease (ADAM10) in α - secretase. Conclusions FK506 rescues cognitive deficits in APPswe/PS1dE9 mice by reducing Aβ production and deposition in the brain.

Immune checkpoint inhibitors have revolutionized cancer therapy; however, many patients exhibit suboptimal responses, which is due to inadequate T cell priming by the innate immune response. Metal ions play a critical role in modulating the innate immune response. However, the mechanisms by which metal ions facilitate dendritic cell maturation through the activation of interferon remain poorly understood. This research identifies a nanomaterial Calcium phosphate-containing liposome (NanoCa), characterized by an atypical crystal structure and pH-responsive profile. NanoCa promotes bone marrow-derived dendritic cell maturation and exhibits antiviral effects and anti-tumor properties in different tumor models. Also, NanoCa acts as an immunostimulant by fostering antibody production. Furthermore, when combined with programmed cell death 1 receptor (PD-1) blocking antibodies, NanoCa synergistically enhances anti-tumor efficacy in CT26 models. Mechanistically, NanoCa rapidly releases Ca2+ via the lysosome pathway post-endocytosis, subsequently triggering interferon through the Ca2+-calcineurin (CaN) - nuclear factor of activated T cells 2 (NFATc2) - protein kinase C beta (PKCβ) - interferon regulatory factor 3 (IRF3) signal pathway. Single-cell RNA sequencing (scRNA-seq) shows NanoCa increases the population of tumoral infiltrating dendritic cell (DC), C1qc+ TAM, and CD8T_eff cells and decreases the CD8T_ex and immunosuppressive SPP1+ TAM population in tumor-draining lymph nodes. Overall, NanoCa shows translational potential for anti-tumor immune therapeutics.

BACKGROUNDHepatic stellate cell (HSC) activation is key to liver fibrosis. Targeting DNA methylation shows promise. Zebularine, a methylation inhibitor, may suppress HSC activation via the calcineurin (CaN)/NFAT3 pathway. Magnetic resonance imaging (MRI) is a noninvasive tool for evaluating liver fibrosis evaluation tool, but multiparametric MRI for zebularine’s effects in liver fibrosis mouse models has not been studied.AIMTo clarify the anti-fibrosis mechanism and MRI-evaluated efficacy of zebularine.METHODSIn vitro, transforming growth factor (TGF)-β1-stimulated human HSCs (LX-2) were treated with zebularine. α-smooth muscle actin, fibrotic and anti-fibrotic gene levels, and regulator of calcineurin1 (RCAN1) regulation were measured. In vivo, carbon tetrachloride (CCl4)-induced liver fibrosis in mice was treated with zebularine, and fibrosis was evaluated using various biochemical, histopathological, and MRI methods.RESULTSZebularine upregulated RCAN1.4 protein (P < 0.01) and inhibited the CaN/NFAT3 pathway (P < 0.05). In HSCs, TGF-β1 reduced anti-fibrotic gene massage RNA (mRNA) and increased fibrotic mRNA (P < 0.05), whereas zebularine had the opposite effects (P < 0.01, P < 0.05). CCl4-treated mice exhibited increases in various fibrosis-related indices, all of which were reversed by zebularine treatment (P < 0.05).CONCLUSIONZebularine may reduce LX-2 activation and extracellular matrix deposition via RCAN1.4 and CaN/NFAT3 pathways. Multiparametric MRI can assess its efficacy, suggesting zebularine’s potential as a liver fibrosis treatment.