Malaria remains one of the most devastating infectious diseases worldwide, predominantly caused by Plasmodium falciparum. Although the RTS,S and R21 vaccines have demonstrated partial efficacy, durable and broad protection across multiple parasite life stages remains an unmet need. Here, we report the development of a lipid-polyplex (LPP)-formulated mRNA vaccine platform targeting both the pre-erythrocytic and blood stages of P. falciparum. We designed mRNA constructs encoding the circumsporozoite protein (PfCSP) and the reticulocyte-binding protein homolog 5 (PfRH5), delivered either individually or as dual-antigen tandem forms. The optimized construct, RH5-T2A-CSP (Mal05), exhibited efficient antigen expression and elicited high-titer, durable antibodies capable of inhibiting sporozoite invasion and erythrocytic growth in vitro. In murine models, Mal05 induced robust Th1-biased CD4⁺ and CD8⁺ T-cell responses and conferred 100% sterile protection against challenge with transgenic Plasmodium berghei expressing PfCSP. Collectively, these findings demonstrate that dual-antigen mRNA-LPP vaccination can establish a potent, multistage immunological barrier by integrating neutralizing antibody activity with strong cellular immunity. Mal05 therefore represents a promising next-generation vaccine candidate with the potential to achieve both pre-erythrocytic and blood-stage protection. More broadly, this work highlights the potential of LPP-based mRNA technology for developing multistage vaccines against complex parasitic pathogens.
Chronic pain has emerged as the third leading health issue, following cardiovascular and cerebrovascular diseases and cancer. Clinically, patients with chronic pain often exhibit comorbid depression-like behaviors. However, the underlying mechanisms of chronic pain-related depression remain unclear. In this study, a chronic pain-related depression model was established in C57BL/6J, Vgat-Cre, and TH-Cre mice through sciatic nerve ligation (SNI) surgery. Viral-mediated DNA methyltransferase 1 (DNMT1) knockdown, optogenetic/chemogenetic modulation, electrophysiological recordings and molecular biology techniques were used to investigate the role of DNMT1 in regulating the projection from central amygdala GABAergic neurons (CeAGABA) to locus coeruleus noradrenergic neurons (LCNA) and its impact on chronic pain-related depression-like behaviors. Viral tracing identified LC as a major downstream target of the CeA. Immunohistochemical staining and electrophysiological recordings confirmed the functional synaptic connection between CeA-GABAergic neurons and LC-NAergic neurons. Behavioral assays showed the bidirectional modulation of pain-related depression phenotypes through LC-NAergic system: activation of LC-NAergic neurons induced depression-like behaviors, whereas their inhibition improved SNI-induced depression-like behaviors. Notably, CeA-GABAergic neurons were found to specifically innervate LC-NAergic neurons in mediating chronic pain-related depression-like behaviors. Furthermore, inhibition of the CeAGABA → LCNA circuit abolished the improvement of depressive behaviors in DNMT1 knockdown SNI mice, establishing DNMT1 as a key epigenetic regulator of this pathway. Taken together, our findings suggest that DNMT1 may suppress the GABAergic projections from the CeA to LC-NAergic neurons, thereby contributing to chronic pain-related depression-like behaviors. These findings provide novel insight into the circuit-related epigenetic basis of pain-depression comorbidity.
Early-life sevoflurane exposure is associated with long-term cognitive deficits. Given that hippocampal development relies on precise critical windows, disruption of developmental processes during these periods is likely the origin of these impairments. However, the molecular mechanisms underlying sevoflurane-induced perturbations during critical development periods and their progression to lasting cognitive dysfunction remain elusive. Here, we determined that the first three postnatal weeks are a critical window of vulnerability to early-life sevoflurane exposure. Mechanistically, sevoflurane exposure suppressed the physiological upregulation of activity-regulated cytoskeleton-associated protein (Arc; also known as Arg3.1) during the critical period of hippocampal development, a process driven by glycogen synthase kinase-3 beta (GSK3β)-mediated protein degradation. We demonstrated that transient suppression of Arc via hippocampal injection of Arc antisense oligonucleotide (ASO) during the third postnatal week was sufficient to recapitulate the sevoflurane-induced phenotype, impairing microglial synaptic pruning and causing initial synaptic redundancy. This early pathology subsequently evolved into aberrant microglial phagocytic activation in adolescence and adulthood, resulting in excessive synaptic loss and cognitive deficits. Crucially, restoration of Arc expression specifically during this critical period using a doxycycline (Dox)-inducible Tet-On system successfully reversed this pathological pruning trajectory and prevented long-term cognitive impairment. Our findings highlight that Arc upregulation during this critical period is essential for microglial function and synaptic homeostasis, establishing Arc as a time-sensitive therapeutic target for preventing the developmental neurotoxicity associated with pediatric anesthesia. This diagram illustrates the role of the Arc protein in microglial synaptic pruning and cognitive development under physiological conditions (upper panel, blue; Arc is highly expressed during the critical period) and sevoflurane exposure (lower panel, red; Arc expression during the critical period is downregulated). Under normal conditions, Arc is expressed during the critical period and is localized to synapses, where it facilitates the tagging of redundant synapses for elimination. Synaptic pruning peaks during the critical period, followed by further refinement during adolescence, leading to mature cognitive function in adulthood. Neonatal sevoflurane exposure leads to aberrant activation of GSK3β during the critical period. This promotes Arc degradation, thereby disrupting its synaptic localization. This leads to microglial dysfunction in terms of synaptic engulfment. In adolescence, this disruption leads to excessive microglial phagocytosis and significant synaptic loss, ultimately resulting in cognitive deficits. Graphical elements: GSK3β, green circles; Arc, orange circles; phosphorylation site, pale yellow circle containing the letter "P"; C1q, yellow umbrella shapes.
Insufficient tumor-specific immunogenicity and cytotoxic T lymphocytes infiltration limit the therapeutic efficacy of combined chemotherapy and immune checkpoint blockade for triple-negative breast cancer (TNBC). To address this limitation, we developed a paclitaxel prodrug-loaded synthetic high-density lipoprotein nanoparticle (P-sHDL) that both targets tumors and induces immunogenic cell death (ICD) to potentiate immune checkpoint blockade. Leveraging scavenger receptor class B type 1-mediated uptake, P-sHDL efficiently accumulated at tumor sites and triggered ICD in tumor cells, thereby enhancing antitumor immunogenicity. Building on this immunogenicity enhancement, P-sHDL was combined with an anti-programmed death-ligand 1 antibody (αPD-L1) to enhance intratumoral CD8+ T cell activity and alleviate immunosuppression. As a result, this regimen achieved marked therapeutic efficacy in tumor-bearing mice. Collectively, this study highlights that P-sHDL-based immunogenicity enhancement combined with immune checkpoint blockade represents a promising strategy for TNBC chemoimmunotherapy.
Alzheimer's disease (AD) is a neurodegenerative disorder that is characterized by the accumulation of amyloid-beta (Aβ) aggregates, in the form of fibrils and plaques. While it has largely been stated that Aβ oligomers are the main toxic species, significant evidence indicates that fibrils may also be relevant to AD pathogenesis. Notably, evidence indicates that while fibrils, through direct interaction with neuronal membranes, contribute to synaptic dysfunction and cellular damage, no direct evidence between fibrils and their impact on neuronal functions, including plasticity, was investigated. This study inquired into the impact of Aβ fibrils complex on neuronal function and membrane integrity, shedding light on their contribution to synaptic defects. Aβ fibrils were generated from Aβ1-42 oligomers, and their effects were evaluated on synaptic plasticity in ex vivo hippocampal slices from both female and male mice. Compared to Aβ1-42 oligomers, fibrils induced more severe damage in synaptic plasticity, emphasizing their potent neurotoxicity effect underlying cognitive decline. Additionally, using a liposomal model to examine fibril-membrane interactions, it was observed that Aβ fibrils are able to affect membrane fluidity compared to the Aβ1-42 oligomeric species, indicating that the size and aggregation state of Aβ fibrils are crucial for their toxicity. These findings challenge the view that oligomers are the primary toxic species in AD, showing that Aβ fibrils also play an active role in cellular dysfunction, promoting a synaptic dysfunction correlated to the cognitive impairment observed in AD.
The 3' untranslated regions (3'UTRs) have been known to regulate mRNA location, stability, and translation. 3'UTR length regulation is involved in the pathogenesis of cardiac dysfunction; however, more about the roles of 3'UTRs in cardiac remodeling remains elusive. In this study, we found slit guidance ligand 1 (SLIT1) 3'UTR with 3074 nt in length, which was 10-fold higher than SLIT1 coding sequence (CDS), was significantly decreased in the myocardium of patients with heart failure (HF) (n = 40) in comparison with healthy organ donors (n=17). We revealed that SLIT1 3'UTR and the 1526 nt fragment of SLIT1 3'UTR (FS1UTR) mainly and specifically combined miR-34a-5p, and improved cardiac remodeling through the miR-34a-5p/SIRT1 axis independently of Slit1 expression. Furthermore, a 260 nt restructured RNA derived from FS1UTR, S1UTRSP5, which contains 5 binding sites of miR-34a-5p seed sequence, alleviated cardiac remodeling in vitro and in vivo. We demonstrated that S1UTRSP5 blocked the function of miR-34a-5p and activated the SIRT1-PGC-1α-Nrf2 axis in cardiomyocytes, and promoted the SIRT1/Smad3 signal in cardiac fibroblasts and the SIRT1-eNOS-VEGFA axis in endothelial cells, collectively contributing to the amelioration of cardiac remodeling. These results provide new insights into the development of S1UTRSP5 as a novel inhibitor of miR-34a-5p for cardiac remodeling and HF. The human SLIT1 3'UTR or FS1UTR combines miR-34a-5p to increase SIRT1 level in CMs, CFs and ECs. Notably, S1UTRSP5, a 260-nt stable RNA derived from SLIT1 3'UTR, efficiently sponged miR-34a-5p to activate SIRT1-PGC-1α-Nrf2 pathway in CMs, and to promote SIRT1/Smad3 signal in CFs and the SIRT1-eNOS-VEGFA pathway in ECs, collectively contributing to amelioration of cardiac remodeling.
The intestinal epithelium maintains host-microbiota homeostasis, while inflammatory conditions, such as inflammatory bowel disease (IBD), induce pathological shifts in intestinal epithelial cell (IEC) subtypes. We unveil METTL3, an RNA m6A methyltransferase, as a pivotal regulator of this balance. METTL3 is enriched in intestinal stem cells and transit-amplifying cells (TACs), and upregulated in patients with IBD and a mouse model of IBD. DSS-challenged, intestine-specific Mettl3 knockout mice exhibited exacerbated colitis as exemplified by more weight loss, elevated disease activity index (DAI), and higher extent of colon shortening. Single-cell transcriptomics of colonic tissues from DSS-challenged intestine-specific Mettl3 knockout mice revealed that Mettl3 ablation depleted epithelial lineages (TACs, goblet cells, enterocytes) but amplified immune infiltration (macrophages, neutrophils, T cells) within the intestinal mucosa. Crucially, METTL3 loss impaired TAC multipotency and increased epithelial-neutrophil crosstalk mediated by the TNF pathway. Mechanistically, METTL3-mediated m6A modification increases Slc39a8 expression, whose knockdown in colon organoids phenocopied METTL3 deficiency in impairing self-renewal. Our work establishes METTL3 as a dual guardian of intestinal homeostasis-preserving epithelial regeneration and restraining inflammation by calibrating epithelial-immune dialogue. The former is mediated at least in part by regulating Slc39a8 expression through m6A modification.
Endothelial and macrophage inflammation and sterol transport play an important role in atherogenesis. Serum/glucocorticoid-regulated kinase 1 (SGK1) is a member of the serine/threonine kinase family sharing approximately 54% identity with Akt. It has been implicated in smooth muscle cell calcification and macrophage inflammation during atherosclerosis; however, the role of SGK1 in endothelial dysfunction and endothelial or macrophage lipid metabolism is less characterized. In this study, we intraperitoneally injected high-cholesterol diet-fed male ApoE-knockout mice with the SGK1 inhibitor EMD638683 (10 mg/kg) every other day for 2 weeks, followed by histopathological and transcriptome analysis of the atherosclerotic lesion. To study the SGK1-associated mechanism in vascular inflammation in vitro, SGK1 silencing was performed in primary human umbilical vein endothelial cells (HUVECs), followed by treatment with pro-inflammatory cytokines. In THP-1-differentiated macrophages, SGK1 silencing followed by treatment with lipopolysaccharides was used. We showed that ApoE-knockout mice treated with EMD638683 show reduced atherosclerotic plaque area and attenuated endothelial and macrophage inflammation. Further transcriptome analysis of thoracic aortae showed that SGK1 inhibition downregulated inflammation and lipid metabolism-associated genes. In vitro, interleukin-1β treatment induces SGK1 phosphorylation. SGK1 inhibitor or siRNA reduced endothelial inflammation induced by pro-inflammatory cytokine treatments in HUVECs through nuclear factor κ light chain enhancer of activated B cell (NF-κB) signaling. In THP-1-differentiated macrophages, SGK1 inhibition or knockdown by siRNA is associated with reduced levels of pro-inflammatory cytokines, NF-κB, and sterol regulatory element binding protein 1 (SREBP1) pathway activation, following lipopolysaccharide treatment. SREBP1 inhibition by fatostatin or siRNA reduced p65 phosphorylation. In conclusion, the inhibition of SGK1 has been shown to reduce atherosclerotic plaque area and attenuate endothelial and macrophage inflammation in ApoE-knockout mice via NF-κB and SREBP1 signaling. These results not only contribute to our understanding of the complex interplay between vascular inflammation and lipid metabolism but also hold promise for the development of novel therapeutic strategies for atherosclerosis.
Metabolic hijacking of tryptophan (Trp) via the IL4I1-AHR axis is a pivotal immune evasion mechanism in cancers, yet therapeutic strategies to disrupt this pathway remain unexplored. Here, we report the identification of ZY-MY-111, a selective small-molecule inhibitor of interleukin-4-induced-1 (IL4I1), through an in-house compound library screening and structural optimization. ZY-MY-111 exhibits potency (IC50 = 1.86 ± 0.13 μM) in blocking IL4I1-mediated oxidative deamination. Mechanistically, ZY-MY-111 acts as a mixed-type inhibitor, competitively occupying the catalytic pocket of IL4I1 and disrupting Trp-AHR signaling in cells. Functionally, ZY-MY-111 promotes T cell proliferation, enhancing immune responses against the tumor cells. In syngeneic tumor models, ZY-MY-111 achieved 49% tumor growth inhibition in CT26 colon carcinoma (P < 0.001) and 56% tumor growth inhibition in A20 lymphoma (P < 0.001) by remodeling the immunosuppressive microenvironment: increasing CD8+/CD4+ T cell ratios, reducing myeloid-derived suppressor cells (MDSCs, 59% decrease), and enhancing effector memory T cell infiltration. Our findings position IL4I1 inhibition as a potential strategy to restore anti-tumor immunity.
Palbociclib, ribociclib, and abemaciclib, which are FDA-approved CDK4/6 inhibitors, constitute the standard first-line treatment for advanced hormone receptor-positive (HR+) breast cancers. Although these drugs share nominal targets and demonstrate comparable first-line efficacy, significant divergences exist in their cross-line therapeutic responses upon first-line progression, indicating distinct mechanisms of action. To systematically elucidate their similarities and differences, we employed quantitative proteomics and phosphoproteomics to investigate and compare the molecular expression characteristics induced by these drugs. As a result, the three CDK4/6 inhibitors exhibited distinct anti-proliferative potencies in HR+ breast cancer cells. Dynamic proteomic and phosphoproteomic profiling demonstrated cell cycle arrest and mTORC1 pathway downregulation by all inhibitors, with abemaciclib exerting the most potent suppression. Subsequent kinase activity analysis indicated an upregulation of AKT1 kinase activity following CDK4/6 inhibitor stimulation. Furthermore, acridine orange staining and flow cytometry indicated that abemaciclib elevated lysosomal proteins and acidification while increasing reactive oxygen species (ROS). Collectively, this study deepens the understanding of shared and drug-specific molecular characteristics and mechanisms of CDK4/6 inhibitors at multi-omic levels. Moreover, it provides an experimental basis and potential directions for customizing combination therapies based on pathway vulnerabilities, as well as exploration of novel therapeutic modes and drugs.
Uterine leiomyosarcoma (ULMS) is a rare yet aggressive uterine malignancy with high recurrence and poor survival, prompting an urgent search for better treatments. We investigated whether atorvastatin, an HMG-CoA reductase inhibitor, could suppress ULMS growth by targeting mevalonate pathway-dependent prenylation. Human ULMS cell lines received atorvastatin ± isoprenoids to assess proliferation, cell-cycle distribution, and smooth muscle contractility, whereas proteomic profiling (LC-MS/MS) and in vivo xenografts were used to evaluate molecular pathways and antitumor efficacy. Atorvastatin inhibited ULMS proliferation in a dose-dependent manner, induced G₀/G₁ cell-cycle arrest, and diminished the contractile phenotype. Geranylgeranyl pyrophosphate rescued these effects, implicating geranylgeranylation as the key dependency, and Rap1A/Rap1B Western blotting confirmed functional GGPP depletion. In xenografts, atorvastatin suppressed ULMS tumor growth by ~50% with minimal toxicity, as evidenced by normal serum ALT and creatinine levels and preserved organ histology. These findings identify protein geranylgeranylation as a novel therapeutic vulnerability in ULMS and support statin repurposing as a promising treatment strategy.
Epilepsy, a neurological condition, is characterized by frequent, spontaneous seizures. Both apolipoprotein E (ApoE) and TAM (Tyro3, Axl and MerTK) receptors, the key efferocytosis receptors, have been associated with the pathological mechanisms underlying epilepsy. Nevertheless, little is known about how they interact or the molecular mechanisms. Here, we explored how ApoE and microglial efferocytosis interact in a mouse model of acute seizures caused by kainic acid (KA). KA exposure reduced microglial efferocytosis that is dependent on growth arrest-specific protein 6 (Gas6)/MER proto-oncogene tyrosine kinase (MerTK). Additionally, KA treatment increased microglial ApoE expression, whereas ApoE knockout restored Gas6/MerTK-mediated microglial efferocytosis and alleviated KA-induced seizures and neuronal injury. Furthermore, KA exposure promoted ApoE immunoprecipitation with nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3). ApoE knockout suppressed KA-induced NLRP3 inflammasome activation. Interestingly, KA-induced seizures and neuronal injury were lessened by hippocampal NLRP3 knockdown, which further preserved Gas6/MerTK-dependent microglial efferocytosis. Furthermore, exogenous delivery of recombinant Gas6 had anti-seizure and neuroprotective benefits in addition to restoring microglial efferocytosis function. Collectively, ApoE stimulates NLRP3 inflammasome activation and targets Gas6/MerTK in seizure-prone mice, thereby impairing microglial efferocytosis. For the treatment of seizures, the ApoE-NLRP3-Gas6/MerTK axis may represent a potential therapeutic target.
Triple-negative breast cancer (TNBC) represents an aggressive breast cancer subtype with unfavorable clinical outcomes. Because actionable therapeutic targets remain limited, current treatment options are suboptimal and patient prognosis is generally poor. Our study aims to address the dual challenges of proliferation control and metastasis inhibition in TNBC treatment. Based upon previous findings, although Unc-51 like kinase 1 (ULK1) activators can inhibit proliferation, their anti-metastatic effects may be antagonized by the abnormally activated Extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. Thus, we propose an innovative therapeutic strategy that synergistically targets ULK1 (activation) and ERK1/2 (inhibition). Accordingly, we successfully developed the first ULK1/ERK1/2 dual-target small-molecule modulator, UE01. Kinase activity analysis showed that UE01 can effectively activate ULK1 and inhibit ERK1. Its dual-target binding properties were validated through molecular docking, kinetic simulations, and cellular thermal shift assays (CETSA). The concentration-dependent inhibition of TNBC cell proliferation, colony formation, and migration by UE01 in vitro was superior to or equivalent to single-target positive drugs. In animal models, UE01 significantly inhibits tumor growth and lung metastasis, without showing a remarkable toxicity. The anti-tumor effect of UE01 was achieved on the one hand by inducing autophagic cell death and apoptosis, and on the other hand, its excellent anti-metastatic ability stems from key regulation of the Exo70/Cav-1/MMPs axis. By synergistically activating ULK1 and inhibiting ERK1/2, UE01 could disrupt the functional coupling of Exo70/Cav-1, thereby inhibiting the secretion of MMP-2/MMP-9 and extracellular matrix degradation. Taken together, these results demonstrate that a dual-target strategy against ERK1/2 attenuates TNBC growth and metastatic dissemination by disrupting the Exo70/Cav-1/MMPs axis, highlighting UE01 as a potential lead compound for future targeted therapy.
Dysfunction of the autonomic nervous system (ANS) is strongly linked to the pathophysiological mechanisms of depression. Unfortunately, the correlation between depressive-like behaviors and the activation or inhibition of the sympathetic/parasympathetic nervous system has not been systematically explored. This study demonstrated that the pharmacological activation of sympathetic α-/β-adrenergic receptors increased stress susceptibility, whereas their antagonists enhanced stress resistance, thus suggesting a close correlation between sympathetic activity and depressive-like behaviors. Furthermore, arecoline, an agonist of the peripheral parasympathetic muscarinic (M) receptors, has exhibited significant antidepressant effects in multiple murine depression models, and its antidepressant effects could be blocked by the M receptor antagonist known as atropine, thus indicating that parasympathetic excitability regulates depressive-like behaviors. Moreover, arecoline acted on peripheral M receptors to activate glutamatergic neurons in the anterior cingulate cortex (ACCGlu) via the vagus nerve and nucleus tractus solitarius (NTS) in sequence. Chemogenetic manipulation confirmed that the antidepressant effect of arecoline is dependent on the activation of ACCGlu neurons, and direct activation of these ACCGlu neurons exerts an antidepressant effect. Notably, pharmacological activation of peripheral parasympathetic M receptors can regulate the activation of the nodose ganglion (NG)-NTS-ACCGlu neural axis and modulate depressive-like behaviors. For the first time, this study indicates that peripheral M receptors are promising targets for antidepressants development and proposes a potential antidepressant strategy involving the pharmacological modulation of peripheral autonomic balance.
Heart failure (HF) following myocardial infarction (MI) remains a major threat to health worldwide. While transcriptomics has revealed numerous genes whose expression is altered in HF, distinguishing therapeutic targets remains challenging. In this study, we aimed to identify novel therapeutic targets for HF and explore potential pharmacological interventions. We integrated human HF datasets with weighted gene coexpression network analysis (WGCNA) and machine learning (LASSO/SVM-RFE) to screen for candidate genes and applied Mendelian randomization (MR) to assess causality. SLCO5A1 emerged as a prioritized candidate, as it showed a genetically supported protective association with HF and was consistently downregulated in the ischemic failing myocardium. In mice, cardiomyocyte-targeted SLCO5A1 overexpression attenuated post-MI systolic dysfunction and pathological remodeling. Using drug-gene signature mining followed by biophysical and cellular validation, we identified 3-iodothyronamine (T1AM) as a small molecule that directly binds to SLCO5A1 and increases SLCO5A1 protein levels. Pharmacological administration of T1AM increased post-MI survival, improved cardiac function and reduced fibrosis; these benefits were markedly weakened by cardiomyocyte-specific SLCO5A1 knockdown, supporting a functional requirement for SLCO5A1. Mechanistically, SLCO5A1 reduced cardiomyocyte transforming growth factor beta 1 (TGF‑β1) secretion, thereby limiting fibroblast Smad3 activation and myofibroblast marker expression in conditioned-medium assays. In conclusion, our findings demonstrate that SLCO5A1 is a cardioprotective regulator of cardiomyocyte-fibroblast communication in post-MI HF and support a pharmacological increase in SLCO5A1 levels as a potential therapeutic strategy. SLCO5A1 serves as a novel therapeutic target for heart failure. Enhancing SLCO5A1 expression protects against MI-induced HF by inhibiting TGF-β1/Smad3-mediated cardiomyocyte-fibroblast crosstalk.
Circadian rhythmicity in seizure susceptibility has been documented in patients with epilepsy. We established a chemogenetic epilepsy model induced using Gq-coupled designer receptors exclusively activated by designer drugs to examine the mechanisms underlying the circadian rhythmicity of seizure susceptibility. Intraperitoneal administration of clozapine N-oxide (CNO) induced dose-dependent seizure behaviors characteristic of mild (Racine scale III or less) and severe (Racine scale IV-V) seizures. CNO at a dose of 0.5 mg/kg elicited significantly more severe seizures during the light phase (Zeitgeber time 3 [ZT 3]) compared with the dark phase (ZT 15). Chemogenetic activation of the Gi instead of the Gq pathway in hippocampal astrocytes suppressed the number of mild seizures exclusively during the light phase. Astrocyte-specific knockdown of adrenergic α2 but not α1 or β receptors exacerbated seizure susceptibility specifically in the light phase, an effect partially reversed by administration of 0.2 mg/kg of dexmedetomidine, an adrenergic α2 agonist. The administration of atipamezole, an α2 antagonist, significantly upregulated norepinephrine-induced lactate production in both intra- and extra-cellularly cultured astrocytes. Supplementation with medium-chain fatty acids in epileptic mice rescued astrocytic α2 receptor knockdown-induced seizure exacerbation, which produce blood-brain-barrier-permeable ketone bodies that likely balance the pro-epileptic lactate burden. Taken together, these findings suggest that astrocytic adrenergic α2 signaling at least partly mediates circadian rhythmicity in seizure susceptibility.
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disorder of the central nervous system characterized by an imbalance between effector T cells (Th1/Th17) and regulatory T-cell (Treg) function. Current disease-modifying therapies (DMTs) reduce relapse frequency mainly through broad immunomodulation, but their non-specific mechanisms often lead to adverse effects and incomplete disease control, especially in progressive MS. These limitations have driven efforts to develop next-generation T-cell-targeted approaches, such as antigen-specific tolerance induction and adoptive Treg therapy. However, the highly oxidative inflammatory microenvironment within the central nervous system (CNS) of MS limits the development of these advanced T-cell-targeted therapies. Excessive reactive oxygen and nitrogen species (ROS/RNS) destabilize FoxP3 expression, interfere with IL-2 signaling, and promote Th17 differentiation, thereby reducing the stability and function of therapeutic T cells. This review summarizes the development of existing DMTs and emerging T-cell-targeted immunotherapies and reviews the current understanding of the underlying mechanisms. We further summarize evidence on bioactive compounds derived from Traditional Chinese Medicine (TCM) that can regulate ROS/RNS levels, reduce oxidative and inflammatory stress, and potentially serve as adjunct agents to improve the performance of cell-targeted therapies. In conclusion, we propose that future therapeutic strategies for MS may require the simultaneous targeting of autoreactive T-cell pathways and modulation of the CNS redox environment to maintain immune tolerance. Taken together, integrating T-cell immunotherapy with redox microenvironment modulation may enhance the durability of therapeutic responses and provide a conceptual rationale for combining TCM-based redox strategies with emerging cellular immunotherapies in the management of autoimmune neurological disorders.
Colorectal cancer (CRC) is a major cause of cancer mortality worldwide. Here, we identified the antipsychotic drug penfluridol as a potent anticancer agent that induces cuproptosis in CRC through a newly defined signaling mechanism. Using real-world clinical datasets, we demonstrated that PDPK1 is significantly upregulated in CRC and further increases in its expression level are correlated with a worse outcome, whereas CTR1 is downregulated in CRC and further decreases in its expression level are correlated with favourable outcomes, directly establishing the clinical relevance of these two proteins to CRC. Mechanistically, drug affinity responsive target stability assays revealed PDPK1 as a direct binding target of penfluridol. Penfluridol inhibited PDPK1 kinase activity and reduced AKT1 phosphorylation, which in turn decreased CTR1 ubiquitination and stabilized CTR1 on the plasma membrane. Enhanced CTR1 expression promoted intracellular copper influx, leading to copper overload and cuproptosis. Functionally, penfluridol suppressed CRC growth in cell lines, patient-derived organoids, and PDX models and was well-tolerated with limited systemic toxicity. Genetic and pharmacologic modulation confirmed that the PDPK1-p-AKT1-CTR1 axis governs copper homeostasis and mediates penfluridol-induced cell death. Collectively, our findings revealed a previously unrecognized link between oncogenic kinase signaling and copper metabolism, established PDPK1 and CTR1 as clinically relevant biomarkers, and provided a strong rationale for repurposing penfluridol as a dual-function therapeutic that induces cuproptosis and enhances chemosensitivity in colorectal cancer.
Oncolytic virus M1 is a promising anticancer agent; however, its therapeutic efficacy is often limited by insufficient intratumoral viral replication and host antiviral immunity. Sirolimus, an mTOR inhibitor widely used in transplantation immunosuppression, has demonstrated potential to modulate antiviral responses. This study investigates whether sirolimus potentiates the efficacy of M1 virotherapy and elucidates the underlying mechanisms. Sirolimus significantly enhanced the antitumor efficacy of M1 virus in murine prostate cancer and liver cancer models, leading to reduced tumor growth. This synergistic effect remained evident in CD8⁺ T cell-depleted mice, indicating that the therapeutic benefit is independent of adaptive cytotoxic immunity. Mechanistically, sirolimus markedly increased M1 viral replication in tumor tissues, accompanied by enhanced tumor cell-cycle arrest and apoptosis. Notably, sirolimus selectively amplified viral load within tumors but not in normal organs, demonstrating tumor-specific viral enrichment and safety. Further analyses revealed that this increase in intratumoral virus was driven by mTOR pathway inhibition rather than alterations in macrophage or NK cell populations. Transcriptomic profiling and molecular validation indicated that sirolimus-mediated mTOR suppression downregulated key type I interferon-stimulated genes (Ifitm1, Stat1, Ifit3), thereby attenuating intrinsic antiviral defenses and facilitating viral amplification. In summary, sirolimus potentiates M1 oncolytic virotherapy by selectively enhancing viral replication in tumors via mTOR inhibition and suppression of type I interferon signaling, independent of CD8⁺ T cell-mediated immunity. These findings establish a mechanistic rationale for combining mTOR inhibitors with oncolytic viruses to achieve dual benefits of enhanced viral oncolysis and controlled immunosuppression, with translational relevance for cancer patients requiring long-term immunosuppressive therapy.
Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of foam cells within the arterial wall. Despite substantial progress, the molecular mechanisms governing foam cell formation remain incompletely understood. Here, we identify fibronectin leucine-rich transmembrane protein 2 (FLRT2) as a previously unrecognized regulator of macrophage-driven atherogenesis through the facilitation of foam cell formation. FLRT2 expression was markedly elevated in macrophages within both murine and human atherosclerotic plaques. Functionally, adenovirus-mediated FLRT2 overexpression aggravated atherosclerotic lesion development in Apoe-/- mice, whereas macrophage-specific Flrt2 deletion significantly reduced the plaque burden in both AAV8-PCSK9-injected and Apoe-/- atherosclerotic models. Mechanistically, the loss of Flrt2 attenuated local inflammatory responses, as indicated by lower proportions of proinflammatory Ly6Chi monocytes in the peritoneal cavity and bone marrow and fewer M1 macrophages in the spleen. Furthermore, FLRT2 promoted foam cell formation by suppressing the cholesterol efflux transporter ABCA1 at the post-transcriptional level. We demonstrate that FLRT2 enhances ABCA1 ubiquitin‒proteasome degradation by inhibiting USP22-mediated deubiquitination. Collectively, our findings identify FLRT2 as a key regulator of macrophage lipid metabolism and inflammation, revealing a novel FLRT2-USP22-ABCA1 axis that promotes foam cell formation and accelerates atherosclerosis. Therefore, targeting FLRT2 may represent a promising therapeutic strategy for atherosclerotic cardiovascular disease.
