Overactive lytic regulated cell death (RCD) causes excessive inflammation and impairs tissue repair. Current strategies rely on small-molecule inhibitors targeting the initiation of lytic RCD but these show off-target effects and reduced pro-repair factor release. During lytic RCD, a transitional agonal stage exists between initiation and terminal death, during which cells either proceed to terminal death or undergo resuscitation. Here, we found that agonal cells actively upregulated uptake of extracellular vesicles (EVs), and internalised EVs fused with the plasma membrane via SNARE complexes to enhance membrane repair. We demonstrated that enhancing the membrane repair capacity of agonal cells through artificially prepared EV-mimetic nano-platelet vesicles (NPVs) effectively promoted their resuscitation. Moreover, resuscitated cells secreted substantial amounts of prostaglandin E2 and N1-Acetylspermidine to further promote tissue repair. Our therapeutic strategy for lytic RCD-related delayed tissue repair is based on EV-mediated membrane repair and aims to establish a pro-regenerative niche using NPVs that can drive agonal cell resuscitation.

Neuro-immune crosstalk is increasingly recognized in Parkinson's disease (PD), and ATP13A2 is well known for its neuroprotective role. However, it remains unclear whether ATP13A2 mutations carried by PD patients contribute to immune dysfunction that exacerbates disease progression. Here, we systematically demonstrate that many ATP13A2 mutations result in a loss-of-expression phenotype. ATP13A2 is highly expressed in macrophages. Myeloid ATP13A2 deficiency causes uncontrolled NLRP3 inflammasome activation driven by lysosomal alkalization and subsequent disrupted mitochondrial homeostasis, rendering mice susceptible to a PD-like phenotype. PD-linked ATP13A2 loss-of-expression mutants fail to restore the ATP13A2 levels required to suppress NLRP3 hyperactivation in ATP13A2-depleted human THP-1 monocytes. Macrophages from a PD patient carrying the ATP13A2 loss-of-expression L927P mutation exhibit excessive NLRP3 activation due to lysosomal-mitochondrial dysfunction. Our findings provide insight into PD pathogenesis, emphasizing genetic factor-driven dysregulated macrophage NLRP3 activation, particularly in ATP13A2 loss-of-expression mutation cases.

Aging is driven in part by progressive collapse of cellular bioenergetics: mitochondrial dysfunction depletes adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide phosphate (NADPH), weakening energy metabolism, antioxidant defense, and regenerative capacity. However, no current intervention directly and simultaneously restores these two metabolites in senescent cells. Here, multi-omics analyses of aged skin identified ATP deficiency as a key upstream metabolic driver of skin aging. We engineered plant-derived nano-thylakoid units cloaked with fibroblast plasma membranes (NTU@MM) that enter senescent fibroblasts through membrane fusion and avoid lysosomal degradation. Under red-light irradiation, intracellular NTU@MM function as ectopic photosynthetic organelles, continuously generating ATP and NADPH. This dual supply of energy and reducing power restored mitochondrial membrane potential and respiratory function, alleviated oxidative stress and senescence phenotypes, and shifted the transcriptomic landscape of aged fibroblasts toward a youthful state. For local translation, NTU@MM were incorporated into dissolvable hyaluronic acid microneedle arrays for sustained intradermal delivery and localized photoactivation. Across UVB-induced photoaging and naturally aged mouse models, representing extrinsic and intrinsic aging, respectively, treatment improved skin architecture, collagen density, and mechanical properties. These findings establish a controllable cross-kingdom bioenergetic strategy that targets a shared metabolic driver across distinct aging etiologies and provides a minimally invasive approach to skin rejuvenation.

Oxygen is essential for cellular metabolism and bioenergy production, and insufficient oxygen supply is a hallmark of multiple pathological conditions, including diabetic foot ulcers and impaired wound healing. Persistent hypoxia severely compromises tissue regeneration, making these wounds particularly difficult to treat. Here, we developed platelet-membrane (PM)-coated nanothylakoid units (NTU@PMs) for targeted oxygen delivery to hypoxic cells, particularly macrophages involved in wound repair. Under light illumination, NTU@PMs efficiently generated oxygen, alleviated macrophage hypoxia, reprogrammed macrophage metabolism, and promoted macrophage migration and phagocytosis. In diabetic wound and flap mouse models, NTU@PMs alleviated wound hypoxia and accelerated tissue repair. These findings identify NTU@PMs as a promising strategy for the treatment of diabetic wounds and potentially other hypoxia-associated disorders.

Osteoporosis is a systemic bone disease in which excessive differentiation of osteoclasts leads to reduced bone mass, weakened bone microstructure, and increased fragility. These overactive osteoclasts require low lysosomal pH to function. Therefore, alkalinizing lysosome represents a potential therapeutic strategy for osteoporotic bone defects. Recently, platelet-derived nanovesicles have shown promise in tissue repair and drug delivery. Here, we developed nanosized platelet vesicles (NPVs) loaded with sodium bicarbonate (NaHCO3@NPVs) for the repair of osteoporotic bone defects. These vesicles were encapsulated in an extracellular matrix hydrogel (NaHCO3@NPVs-EH) made from fibrinogen and thrombin. When applied to bone defects, the hydrogel releases NaHCO3@NPVs at the defect site. After being internalized by bone marrow-derived macrophages (BMDMs), NaHCO3@NPVs can inhibit osteoclast differentiation and activity through a “biochemical dual-action” mechanism. On the one hand, NaHCO3 can effectively alkalinize lysosomes, promote lysosomal calcium depletion, eliminate receptor activator of nuclear factor kappa-B ligand (RANKL)-induced intracellular calcium oscillations. On the other hand, the bioactive factors contained in NPVs can directly or indirectly exert an inhibitory effect on osteoclasts. In vivo experiments in a rat osteoporotic bone defect model revealed that the NaHCO3@NPVs-EH reduced the number of overactive osteoclasts and enhanced bone repair. These findings suggest that NaHCO3@NPVs-EH could be a promising tool for the treatment of osteoporotic bone defects. This study also provides a new perspective on acid-base neutralization for the treatment of osteoporosis.

The maintenance of homeostasis in hematopoietic stem and progenitor cells (HSPCs) is essential for the proper development of the entire hematopoietic system. However, the mechanisms underlying this regulatory equilibrium remain elusive. Here, we report that Prdm15 deficiency in HSPCs induces the accumulation of immature hematopoietic stem cells in mice. A series of transplantation assays shows that these cells display impaired reconstitution capacity and competitive fitness, which are associated with abnormal differentiation trajectories and transcriptional alterations identified by single-cell RNA sequencing. Mechanistically, integrated multi-omics analyses including ATAC-seq and CUT&Tag sequencing of HSPCs indicate that Prdm15 deficiency induces significant transcriptional and epigenetic alterations, particularly affecting the methyltransferase KMT2C and altering H3K4me1 and H3K27ac modifications at the promoters of hematopoietic developmental genes. Collectively, our findings establish PRDM15 as a critical epigenetic regulator of HSPCs, offering valuable insights into the molecular mechanisms underlying hematopoietic homeostasis.

Alcohol-associated liver disease (ALD) is a leading cause of morbidity and premature mortality worldwide, characterized by hepatic steatosis and inflammatory cell infiltration. CD73, predominantly expressed in hepatocytes, has been implicated in the regulation of hepatocyte metabolism and liver homeostasis. However, whether and how hepatocyte CD73 contributes to ALD pathogenesis remains poorly understood and warrants systematic investigation. In this study, we observed that CD73 expression was significantly upregulated in ethanol (EtOH)-exposed hepatocytes and in liver tissues from mice with alcohol-associated steatohepatitis (ASH). Liver-specific CD73 knockout (CD73-LKO) altered circadian locomotor activity, exacerbated EtOH-induced liver injury and inflammatory infiltration, and promoted hepatic fatty acid uptake and de novo lipogenesis in vivo. In vitro, CD73 knockdown in hepatocytes aggravated cellular injury, enhanced fatty acid synthesis, and increased lipid deposition, whereas CD73 overexpression conferred protective effects. Furthermore, we found that CD73 modulates the expression of adenosine receptors and circadian clock components. In primary hepatocytes from EtOH-fed mice, CD73-LKO upregulated A1R and A2bR protein levels, downregulated A2aR expression, and concurrently suppressed the core clock gene BMAL1. Notably, adenosine receptor signaling regulated BMAL1 expression in EtOH-stimulated AML-12 cells. Furthermore, liver-specific knockout of BMAL1 (BMAL1-LKO) aggravated liver injury and steatosis in EtOH-fed mice. In vitro, BMAL1 knockdown promoted lipid accumulation and cellular damage, while BMAL1 overexpression alleviated these pathological phenotypes. Collectively, these findings demonstrated that CD73-LKO could affect mice circadian rhythm, regulate the expression of liver adenosine receptors and BMAL1, which provides new insights into the molecular mechanisms underlying the role of CD73 in ALD.

Exosomes exert a protective effect on motor functional recovery after spinal cord injury (SCI) but are limited by the lack of effective preservation in situ and the indeterminate underlying mechanism. Firstly, we fabricated a versatile and optimized controlled release system for mesenchymal stromal cells-derived exosomes, precise concentration of Laponite dispersion combines with exosomes via electrostatic incorporation to form a 3D porous structure, exhibiting excellent biocompatibility and a sustained release of exosomes. Secondly, we assess multiple therapeutic benefits of the Lap@exo gel, including motor recovery, axonal rehabilitation, scar growth, inflammatory response and remyelination. The comprehensive demonstration a unique synergistic effect of Laponite gel and exosomes. Last, we analysis the functional underlying mechanism of the exosomes. We used ExoRNA sequencing and bioinformatic analysis to find that microtubule polymerization and cytoskeleton organization pathways were related to the axon regeneration. Additionally, the microtubule dynamic regulated by Lap@exo was studied for the axonal regeneration as well as the distribution of mitochondria. The study demonstrated that Lap@exo released exosomes for a sustained period of time and protected its bioactivities, and the combination of exosomes with bioactive multifunctional gel could lead to satisfactory recovery of locomotor function for patients with diseases of the central nervous system.

164 Background: Aberrant activation of fibroblast growth factor receptor (FGFR) pathways contributes to tumor progression in colorectal cancer (CRC), often through ligand-mediated stimulation of extracellular domains. To address this, we evaluated the preclinical activity of OM-RCA-01, a humanized monoclonal antibody selectively targeting FGFR1, and compared its efficacy with bevacizumab and chemotherapy in preclinical models. Methods: For in vitro study, SW620 and HCT116 CRC cells with confirmed FGFR1 expression were exposed to OM-RCA-01, bevacizumab, or their combination at concentrations ranging from 2×10⁻⁴ to 10⁻⁷ g/ml (prepared by serial 1:2 dilutions). Following antibody treatment, cells were stimulated with FGF2 (5 ng/ml), or VEGF (10 ng/ml), or both ligands, and proliferation was quantified. For in vivo study, SW620 cells were implanted subcutaneously into the thighs of female NU–A/A Tyrc/Tyrc Foxn1nu/Foxn1nu mice. When tumors reached ~150 mm³, animals were randomized to receive: 1) OM-RCA-01 (30 mg/kg, i.p., twice weekly for 3 weeks); 2) bevacizumab (5 mg/kg, i.p., twice weekly for 3 weeks); 3) OM-RCA-01 + bevacizumab combination; 4) 5-fluorouracil (30 mg/kg, i.p., on Days 1–3 of each week for 3 weeks); 5) triple combination, or 6) saline control. Tumor dimensions were recorded every 3 days for 4 weeks. Results: In vitro, FGF2 stimulation markedly increased proliferation of both cell lines, which was dose-dependently inhibited by OM-RCA-(IC₅₀ < 25 μg/ml). Bevacizumab had no measurable effect on either FGF2- or VEGF-driven proliferation (IC₅₀ > 100 μg/ml). However, combining bevacizumab with OM-RCA-01 under dual FGF/VEGF stimulation produced a more pronounced inhibitory effect (IC₅₀ 11.2–20 μg/ml). In vivo, all treatment regimens significantly reduced tumor burden compared with saline control (Table). OM-RCA-01 monotherapy showed robust tumor suppression (P < 0.0001), with a median tumor volume of 956.7 mm³ versus 2129.9 mm³ in controls at study endpoint. The dual-antibody approach yielded tumor/control ratios (T/C) similar to either monotherapy (47.1% vs. 52.5% vs. 44.9%). Notably, the triple combination of OM-RCA-01, bevacizumab, and 5-FU demonstrated the greatest antitumor activity, achieving the smallest median tumor size (633.7 mm³; T/C = 29.8%), consistent with a synergistic interaction. Conclusions: Targeting FGFR1 effectively inhibited CRC cell proliferation and tumor progression in preclinical models. While dual FGFR1/VEGF antibody therapy did not enhance antitumor efficacy compared with single agents, the addition of chemotherapy produced a marked improvement in therapeutic activity. Tumor growth inhibition. Vehicle OM-RCA-01 Bevacizumab OM-RCA-01 + Bevacizumab 5-FU OM-RCA-01 + Bevacizumab + 5-FU Tumor volume, median, mm 3 2129.9 956.7 1117.1 1003.8 895.6 633.7 T/C% – 44.9 52.5 47.1 42.1 29.8 TGI% – 55.1 47.6 52.9 58.0 70.3

UTX (ubiquitously transcribed X chromosome tetratricopeptide repeat protein), a histone H3K27 demethylase, has been implicated in diverse cancer-related processes. However, its role and regulatory mechanisms in lung adenocarcinoma (LUAD) remain unclear. This study investigated the tumor-suppressive function of UTX and its underlying epigenetic and transcriptional mechanisms in LUAD. UTX expression and prognostic significance were assessed using The Cancer Genome Atlas (TCGA), Gene Expression Profiling Interactive Analysis (GEPIA), and Kaplan-Meier Plotter databases. LUAD cells with UTX overexpression or dual specificity phosphatase 8 (DUSP8)/p53 knockdown were established. Functional assays included MTT proliferation assays, Transwell assays, reactive oxygen species (ROS) detection. Transcriptome sequencing identified downstream targets of UTX. Luciferase reporter, chromatin immunoprecipitation (ChIP), and co-immunoprecipitation (Co-IP) assays were performed to investigate the regulatory mechanism of UTX. MAPK and PI3K/AKT signaling were evaluated by western blotting and immunohistochemistry, while xenograft models were used for in vivo validation. UTX expression was reduced in tumor tissues and significantly associated with poorer clinical outcomes and unfavorable pathological features. UTX overexpression suppressed LUAD cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), accompanied by inhibition of MAPK and PI3K/AKT signaling and an increase in intracellular ROS levels. Integrated transcriptomic analyses identified DUSP8 as a key downstream effector positively correlated with UTX. Mechanistically, UTX promoted DUSP8 transcription through p53-dependent H3K27me3 demethylation at the DUSP8 promoter. p53 facilitated the recruitment of UTX to the DUSP8 promoter, and disruption of the UTX-p53 interaction impaired DUSP8 transcriptional activation. DUSP8 knockdown in UTX-overexpressing cells reactivated the MAPK pathway, diminished ROS buildup, and largely negated the tumor-suppressive phenotype induced by UTX both in vitro and in vivo. UTX acts as a tumor suppressor in LUAD by epigenetically activating DUSP8 via p53-dependent H3K27me3 demethylation, thereby dampening MAPK-driven oncogenic signaling. These findings reveal a previously unrecognized UTX/p53-DUSP8 regulatory axis with potential therapeutic relevance for LUAD.

Treatment strategies focusing on the induction of multiple programmed cell death (PCD) have gained attention in non-small-cell lung cancer (NSCLC). PCD represented by apoptosis and ferroptosis is closely linked to oxidative stress. This study aimed to investigate whether icariside II (ICS II) induces dual PCD and elucidate the underlying mechanisms. The current study showed that ICS II inhibited cell progression and metastasis and was more selective towards NSCLC cells. Transcriptomic analysis revealed that ICS II induced ferroptosis, characterized by excessive levels of reactive oxygen species (ROS), depletion of reductive compounds, and mitochondrial impairment. Functional analysis revealed that ICS II suppressed Nrf2-mediated SLC7A11/GPX4/HO-1 transcription and activated ACSL4-mediated lipid peroxidation, which were rescued by ferrostatin-1 and Nrf2 overexpression. Simultaneously, ICS II activated caspase family proteins and reduced PARP activity, thereby triggering apoptosis. N-acetyl-L-cysteine abolished the effects of ICS II on NSCLC cells and restored cell viability, confirming the functional contribution of ROS in ICS II-induced PCD. Finally, a mouse subcutaneous tumor model was established to assess the drug efficacy and toxicity. As expected, ICS II inhibited tumor development in a mouse subcutaneous tumor model with minimal toxicity. Taken together, ICS II exerts anti-NSCLC efficacy by weakening cellular antioxidative capacity (inhibition of Nrf2 activity) and promoting the accumulation of substrates for lipid peroxidation (ACSL4 activation), triggering oxidative stress, and exhibiting dual PCD induction. Our findings demonstrate the potential of ICS II for use in clinical NSCLC therapy.

Tumor-associated macrophages (TAMs) are key regulators of the tumor microenvironment (TME), significantly influencing cancer progression and therapeutic responses. TAMs polarize into M1 or M2 phenotypes, exerting distinct functional roles. M1-type macrophages promote inflammation and tumor cell destruction, whereas M2-type macrophages facilitate immune suppression, angiogenesis, and metastasis. However, inconsistencies and mischaracterizations in the literature regarding TAM classification have led to confusion in the field, potentially impeding the development of effective macrophage-targeted immunotherapies. This commentary highlights the need for clear and standardized nomenclature, clarifies the functional distinctions between M1- and M2- type TAMs, and explores the signaling pathways and environmental factors driving their polarization. We also discuss emerging TAM subtypes and the therapeutic significance of accurate classification, including macrophage reprogramming strategies. Standardizing terminology and addressing misconceptions will be critical to advancing macrophage-based immunotherapies and improving clinical outcomes in cancer treatment.

BackgroundCirculating platelets are increasingly recognized for their critical involvement in thromboinflammatory complications during respiratory infections, underscoring the importance of comprehending their dual role in immunomodulation and hemostasis. The multifaceted roles of platelets are attributed to their inherent heterogeneity, where the molecular diversity enables their functional versatility. However, research remains deficient in dissecting platelet heterogeneity within respiratory infection contexts, capturing the platelet subpopulations exhibiting specialized functional roles, and elucidating the therapeutic modulation by dexamethasone and tocilizumab.MethodsLeveraging the adequate platelet population detected through single-cell RNA sequencing, this study established stratified study cohorts central to: (1) COVID-19 disease severity, (2) therapeutic responsiveness, and (3) non-COVID-19 respiratory infections. An integrative annotation framework was implemented in this study, which encompassed co-expressed gene module identification, cell-cell communication network mapping, and cell trajectory assessment, to hierarchically dissect the functional dynamics of platelet heterogeneity in disease pathologies and their modulation by therapeutic interventions.ResultsThis study unfolds six severity-associated platelet subpopulations, explores their modulation by corticosteroids and immune antibody treatments, and extends the findings to non-COVID-19 severe conditions. In severe COVID-19, platelet subpopulations with elevated innate immune responsiveness and high coagulation potential are intensively enriched, whereas following dexamethasone administration, this pro-coagulable subpopulation is remarkably repressed. Post-tocilizumab treatment, the instructive interaction from platelets to IFN-activated CD8+ T cells is reduced, while communications from memory B cells, CD4+ T cells, plasmacytoid dendritic cells, and natural killer cells to platelets are enhanced. A platelet trajectory, which participates in neutrophil chemotaxis and B cell-mediated humoral responses, emerges specifically in COVID-19 conditions and is absent in non-COVID-19.ConclusionsA systematic investigation of platelet transcriptomic heterogeneity in respiratory infections, combined with the delineation of its therapeutic modulation by dexamethasone and tocilizumab, advances the understanding of platelet involvement in immune-thrombotic dysregulation and provides a foundation for exploring potential therapeutic targets.

Bone regeneration is a complex and coordinated physiological process, and the different stages of this process have corresponding microenvironments to support cell development and physiological activities. However, biological scaffolds that provide different three-dimensional environments during different stages of bone regeneration are lacking. In this study, we report a novel composite scaffold (NPE@DCBM) inspired by the stages of bone regeneration; this scaffold was composed of a fibrin hydrogel loaded with nanoplatelet vesicles (NPVs), designated as NPE, and decellularized cancellous bone matrix (DCBM) microparticles. Initially, the NPE rapidly established a temporary microenvironment conducive to cell migration and angiogenesis. Subsequently, the DCBM simulated the molecular structure of bone and promoted new bone formation. In vitro, the NPVs regulated lipid metabolism in bone marrow mesenchymal stem cells (BMSCs), reprogramed the fate of BMSCs by activating the PI3K/AKT and MAPK/ERK positive feedback pathways, and increased BMSC functions, including proliferation, migration and proangiogenic potential. In vivo, NPV@DCBM accelerated bone tissue regeneration and repair. Initially, the NPE rapidly induced angiogenesis between DCBM microparticles, and subsequently, BMSCs differentiated into osteoblasts with DCBM microparticles at their core. In summary, the design of this composite scaffold that sequentially mimics different bone regeneration microenvironments may provide a promising strategy for bone regeneration, with clinical translational potential.

The intestinal mucus layer produced by goblet cells is a critical component of innate immunity. The key host factors and regulatory mechanisms controlling goblet cell function in mucus layer formation remain poorly understood. This study identifies a function for the microprotein FXYD domain-containing transport regulator 3 (FXYD3) in goblet cells in regulating mucus layer formation to maintain intestinal homeostasis. Deficiency of FXYD3 in mouse intestinal epithelial cells results in a damaged mucus barrier, leading to dysbiosis and increased susceptibility to colitis. Mechanistically, FXYD3 interacts with endoplasmic reticulum Ca2+-ATPase SERCA2 to enhance its pump activity. FXYD3 deficiency causes defects in ER Ca2+ homeostasis and mucin glycosylation, impairing mucus layer integrity. Furthermore, metabolites of gut microbiota, propionate, and butyrate promote FXYD3 expression. In ulcerative colitis patients, FXYD3 expression is significantly downregulated and correlates with disease severity. These findings indicate FXYD3 is a key mediator of host-microbiota interactions for intestinal health.

Dysfunction of anti-tumor immune responses is crucial for cancer progression. Immune checkpoint blockade (ICB), which can potentiate T cell responses, is an effective strategy for the normalization of host anti-tumor immunity. In recent years, immune checkpoints, expressed on both tumor cells and immune cells, have been identified; some of them have exhibited potential druggability and have been approved by the US Food and Drug Administration (FDA) for clinical treatment. However, limited responses and immune-related adverse events (irAEs) cannot be ignored. This review outlines the development and applications of ICBs, potential strategies for overcoming resistance, and future directions for ICB-based cancer immunotherapy.

Highly conserved homeobox genes are closely related to bone formation during embryogenesis, while their role in adult bone resorption remains unclear. In this study, we found that the homeobox gene MSX2 actively participates bone metabolism. Myeloid-specific Msx2 deficiency safeguards bone mass under physiological and pathological conditions. Loss of Msx2 acts as a "brake" in the fusion fate of osteoclasts, resulting in a larger population of pre-osteoclasts. Pre-osteoclasts secrete platelet-derived growth factor-BB (PDGF-BB), which promotes angiogenesis-mediated bone formation. Mechanistically, MSX2 directly binds to the vital osteoclastogenic transcription factor PU.1 and protects it from FBXW7-mediated ubiquitination degradation. Msx2 and Fbxw7 double knockout mitigated the protective effect of MSX2 deficiency on bone mass. Finally, we identified a natural compound, morusinol, that specifically destroys the combination of MSX2 and PU.1, promoting PU.1 degradation and attenuating ovariectomy-induced bone loss. Overall, our results demonstrate that targeting Msx2 is a promising anabolic therapy for osteoporosis.

Neuro-immune crosstalk has become increasingly recognized in Parkinson’s disease (PD). ATP13A2, a key genetic factor associated with PD, is well-known for its critical neuroprotective role. However, it remains unclear whether mutations in the ATP13A2 gene, found in PD patients, contribute to immune dysfunction that exacerbates disease progression. Here, we systematically demonstrate that a significant proportion of ATP13A2 mutations display a loss-of-expression phenotype. ATP13A2 is highly expressed in macrophages. Myeloid-specific ATP13A2 deficiency leads to uncontrolled NLRP3 inflammasome activation, driven by lysosomal alkalization and subsequent disrupted mitochondrial homeostasis, rendering mice more susceptible to a PD-like phenotype. PD-linked ATP13A2 loss-of-expression mutants fail to restore the ATP13A2 levels to suppress NLRP3 hyperactivation in stable ATP13A2-depleted human THP-1 monocytes. Notably, macrophages derived from a PD patient carrying the ATP13A2 loss-of-expression L927P mutation exhibit excessive NLRP3 activation due to lysosomal and mitochondrial dysfunction. Our findings provide insights into the PD pathogenesis, emphasizing genetic factor-driven dysregulated macrophage NLRP3 activation, particularly in cases involving ATP13A2 loss-of-expression mutations. The work was funded by the National Natural Science Foundation of China (82171719, 32100696, 81930041, U22A20307), the Natural Science Foundation of Zhejiang Province (Y23H100011), and the National Key Research and Development Program of China (STI2030-Major Projects- 2021ZD0202400, and 2022YFA1103800). Neuroimmunology (NEUR)