Theranostics最新文献

Rationale: Traumatic spinal cord injury (SCI) initiates a cascade of local and systemic inflammatory events that exacerbate tissue damage, hinder regeneration, and impair functional recovery. Interleukin-4 (IL-4) is an anti-inflammatory cytokine that promotes M2-macrophage polarization, but its functional benefit in SCI and the underlying mechanisms remain incompletely defined. We evaluated whether systemic IL-4 therapy can enhance recovery and modulate neuroinflammation in a rat model of SCI, and examined the translational relevance of key cytokine signatures in human SCI.

Methods: Female Wistar rats (n = 120) were randomized to sham surgery, SCI with vehicle, or SCI with IL-4 treatment. SCI was induced at T10 by clip contusion-compression; IL-4 (0.5 µg/kg) or vehicle was administered intraperitoneally twice daily for up to 7 days post-injury (dpi). Functional recovery was assessed with the Basso-Beattie-Bresnahan (BBB) scale, CatWalk XT gait analysis, and gridwalk testing. Spinal cords collected at 1, 3, 7, 14, and 28 dpi underwent immunohistochemistry, RNA sequencing, and proteomic profiling. Serum cytokines were quantified in rats by bead-based multiplex assays and compared with longitudinal cytokine profiles from SCI patients.

Results: IL-4-treated rats demonstrated significantly improved BBB scores and multiple CatWalk XT gait parameters by 14 dpi versus vehicle. RNA-seq and proteomics identified upregulation of pathways related to axonogenesis, tissue repair, and reduced TNF-α-mediated pro-inflammatory signaling. Immunohistochemistry confirmed increased IBA1⁺/ARG1⁺ and IBA1⁺/CD206⁺ M2-macrophages, reduced IBA1⁺/iNOS⁺ M1-macrophages, smaller cystic cavity area, and higher APC⁺ oligodendrocyte counts in IL-4-treated animals. Serum profiling showed suppression of acute/subacute pro-inflammatory cytokine surges (1-7 dpi) with IL-4. In SCI patients, lower circulating levels of these cytokines were associated with better neurological outcomes.

Conclusions: Repeated systemic IL-4 administration after SCI promotes functional recovery, shifts macrophage polarization toward a regenerative phenotype, reduces astrogliosis and oligodendrocyte loss, and suppresses systemic inflammation. Multi-omics integration together with patient data suggests IL-4 targets convergent pathways of neuroprotection and immune modulation, supporting its further development as a therapeutic candidate for SCI.

Background: Aortic dissection (AD) is a life-threatening vascular emergency with limited effective pharmacological treatments. Recent studies have identified Src homology 2 domain-containing transforming protein C1 (p66Shc) as a crucial mediator of oxidative stress, apoptosis, and inflammation in aortic cells, thereby contributing to cellular dysfunction and vascular remodeling implicated in AD development and progression. Despite its established role in promoting vascular dysfunction and remodeling, the protective potential of targeting p66Shc in AD remains unclear. Methods: We quantified activated protein C (aPC) levels in clinical plasma samples from control subjects and AD patients using enzyme-linked immunosorbent assay (ELISA). To evaluate changes in p66Shc expression, we analyzed aortic tissues by Western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) staining. An in vivo AD model was established in thrombomodulin (TM)-mutant ApoE^-/- mice, which display impaired TM-dependent PC activation, and exogenous PC was administered to evaluate its therapeutic effect. In parallel, mechanistic studies were performed in human endothelial cells using WB, co-immunoprecipitation (Co-IP), dual-label IF staining, chromatin immunoprecipitation (ChIP), luciferase reporter assays, and mitochondrial functional analyses. Results: In this study, we demonstrate that aPC, a coagulation protease with known cytoprotective properties, downregulates p66Shc expression through epigenetic modifications. Additionally, aPC can modulate the expression of a cold shock protein Y-box-binding protein 1 (YB1), which acts as a transcription factor, leading to elevated O-linked N-acetylglucosamine transferase (OGT) levels. This upregulation enhances the O-glycosylation of p66Shc on its 29th tyrosine residue, preventing its mitochondrial translocation, preserving mitochondrial membrane potential, and reducing reactive oxygen species (ROS) production. Consequently, these molecular mechanisms inhibit the onset and progression of AD. Conclusions: aPC epigenetically represses p66Shc transcription and promotes its O-glycosylation at Thr29 via the YB-1/OGT axis, thereby inhibiting mitochondrial ROS production and preventing vascular injury.

Rationale: Glycosylase-derived base editors enable transversion base substitutions, expanding the scope of genome engineering for both basic research and clinical applications. However, the variable outcomes and low efficiency of B (C/G/T)-to-A editing in mammalian cells hinder their broader utility, likely due to inefficient thymine translesion synthesis (TLS) across apurinic/apyrimidinic (AP) sites.

Methods and results: We developed a nucleotide metabolism-based strategy to enhance B-to-A editing by leveraging endogenous nucleotide metabolism. We showed that elevating intracellular deoxythymidine triphosphate (dTTP) levels via exogenous thymidine (dT) supplementation, which activates the thymidine kinase 1 (TK1)-dependent salvage pathway for the production of dTTP, increased C-to-A, G-to-A, and T-to-A editing efficiencies by up to 4-fold, 1.8-fold, and 1.8-fold, respectively, and improved A-product purity by up to 2.7-fold. Moreover, supplementation with dA increased T outcomes, albeit at a relatively modest level. In a disease-relevant single nucleotide variation (SNV) model, dT treatment enabled efficient generation of pathogenic mutations otherwise inaccessible to base editing.

Conclusion: Our findings establish metabolic modulation as a powerful means to control base editing outcomes and expand the functional capabilities of glycosylase-derived editors.

Rationale: Glioblastoma (GBM), an aggressive malignant brain tumour associated with a dismal prognosis, is characterized by metabolic reprogramming that drives tumour progression, with the Warburg effect being a central contributor. This effect not only causes significant lactate buildup but also fuels lactylation, a novel post-translational modification implicated in the development of gliomas and various other cancers. Nevertheless, the exact molecular mechanisms by which lactylation promotes GBM progression remain largely elusive. Methods: Lactylation levels in normal brain and GBM tissues were analysed using immunohistochemistry, immunofluorescence, and Western blotting. Glycolysis inhibitors and LDHA/LDHB knockdown were used to modulate histone lactylation in subsequent in vitro and in vivo experiments assessing GBM cell proliferation, invasion, and migration. CUT&Tag and RNA sequencing were used to identify H4K8la target genes, and NUPR1 expression was validated via ChIP‒qPCR and Western blotting. Autophagic flux was examined using transmission electron microscopy, EGFP-mCherry-LC3B probes, and LysoTracker staining. The therapeutic effects of NUPR1 inhibitor ZZW-115 were evaluated in both cellular and animal models. Results: Histone lactylation, notably that of H4K8la, was markedly increased in GBM cells. Targeting lactate metabolism and lactylation levels attenuated GBM malignancy in vitro and in vivo. Genome-wide analysis revealed H4K8la enrichment at promoter regions, where it transcriptionally activated the autophagy regulator NUPR1. Functionally, NUPR1 enhanced protective autophagy via autophagosome‒lysosome fusion. Pharmacological inhibition of NUPR1 with ZZW-115 suppressed GBM growth by impairing autophagic flux, demonstrating therapeutic potential. Conclusion: In summary, this study defines the functional and prognostic significance of histone lactylation in the progression of GBM. We identified the H4K8la-NUPR1 axis as a key regulatory pathway that mediates protective autophagy and developed targeted therapeutic strategies to disrupt this pathway. These findings provide novel insights into epigenetic regulation and targeted therapy for GBM.

Rationale: Pathological cardiac hypertrophy, triggered by persistent neurohumoral or hemodynamic stress, is a key precursor of ventricular dysfunction and heart failure. Deubiquitinating enzymes (DUBs) have emerged as critical regulators of cardiovascular biology. This study examined the function of a DUB, ovarian tumor domain-containing 7B (OTUD7B), in cardiac hypertrophy. Methods: Cardiomyocyte-specific OTUD7B knockout and overexpression mouse models were generated to evaluate myocardial hypertrophy and cardiac dysfunction in response to angiotensin II (Ang II) infusion or transverse aortic constriction (TAC). Quantitative ubiquitinome analysis, site-directed mutagenesis, and co-immunoprecipitation assays were performed to explore the substrate and mechanism of OTUD7B. Results: Transcriptomic and experimental validation demonstrated that cardiomyocyte OTUD7B was increased in hypertrophic hearts of both humans and mice. Cardiomyocyte-specific deletion of OTUD7B significantly mitigated angiotensin II (Ang II)- and transverse aortic constriction (TAC)-induced cardiac hypertrophy and dysfunction in mice. Mechanistically, quantitative ubiquitinome analysis identified sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase 2a (SERCA2a) as a direct substrate of OTUD7B. OTUD7B bound to SERCA2a and removed K63-linked ubiquitin at K628 through its catalytic site C194. This deubiquitination promoted SERCA2a-phospholamban (PLN) interaction, thereby restricting SERCA2a activity in Ca²⁺ handling and driving hypertrophic response in cardiomyocytes. Moreover, cardiomyocyte-specific OTUD7B overexpression exacerbated TAC-induced cardiac hypertrophy and dysfunction by deubiquitinating SERCA2a at K628. Conclusions: This study defines a novel OTUD7B-SERCA2a regulatory axis and identifies OTUD7B as a promising therapeutic target for cardiac hypertrophy and dysfunction.

Rationale: Mucins are epithelial transmembrane glycoproteins involved in inflammation and kidney dysfunction, yet the role of the transmembrane mucin MUC20 in renal injury and fibrosis remains unclear. This study aimed to define the functional significance and underlying mechanisms of MUC20 in kidney fibrosis. Methods: Muc20-deficient mice and tubular epithelial cell models were used to evaluate renal fibrosis and pyroptosis in induced kidney injury. Molecular and biochemical approaches were applied to assess protein interactions, RAS activation, 2'3'-cGAMP production, cGAS-STING signaling, lysosomal integrity, potassium efflux, and NLRP3 inflammasome activation. Results: Loss of MUC20 significantly exacerbated renal fibrosis and increased pyroptosis in tubular epithelial cells. Mechanistically, MUC20 interacted with MET to promote RAS activation. MUC20 deficiency decreased GTP-bound RAS levels, leading to increased 2'3'-cGAMP production and activation of the cGAS-STING pathway. STING activation induced lysosomal membrane permeabilization, potassium efflux, and subsequent NLRP3 inflammasome-mediated pyroptosis. Conclusions: MUC20 acts as a key protective regulator in kidney by restraining RAS-cGAS-STING-NLRP3-driven pyroptosis and fibrotic progression. Targeting MUC20-related signaling pathways may offer therapeutic potential for kidney fibrosis and chronic kidney disease.

Background: The diagnosis of metastatic clear cell renal cell carcinoma (mccRCC) remains challenging due to the tumor's molecular heterogeneity, often resulting in low sensitivity and a high false-positive rate. In this study, we introduce and validate a new imaging modality for mccRCC based on the first radioligand targeting the type 2 vasopressin receptor (V2R) suitable for positron emission tomography (PET). V2R is ectopically expressed in mccRCC. This imaging approach utilizes [¹⁸F]F-MQ232, a radiolabeled peptide derived from snake venom which exhibits high in vivo selectivity for V2R.

Methods: The V2R-selective peptide MQ232 conjugated with either a cyanine 5 (Cy5) or a fluorine 18 (¹⁸F) group were chemically synthetized. V2R mRNA was quantified and protein expression assessed by flow cytometry using Cy5-MQ232. The selectivity and tumor targeting ability of the modified MQ232 peptides were assessed using in vivo fluorescence imagery in tumor-bearing mice using CHO-V2R tumors with graded expression. Metabolic stability and PET pharmacokinetics of [¹⁸F]F-MQ232 were assessed in rodents. Specific tumor targeting and imaging contrast were validated in vivo using V2R-expressing tumors.

Results: [¹⁸F]F-MQ232 is a highly relevant radioligand whose tumor uptake directly correlates with V2R expression levels in tissues, demonstrating its specificity to V2R-expressing tumors. Replacing the peptide moiety by an isoform unable to interact with V2R leads to a drastic decrease in the radioligand's tumor uptake, highlighting its origin in a specific, ligand/receptor type interaction between the MQ232 moiety and V2R. PET/CT imaging of Caki-1 xenografted mice demonstrated the ability of [¹⁸F]F-MQ232 to allow specific detection of the tumor compartment associated with high tumor-to-background contrast. RT-qPCR screening of metastatic and non-metastatic ccRCC biopsies from patients confirms V2R expression.

Conclusions: This work validates the V2R-targeting strategy in mccRCC using [¹⁸F]F-MQ232 and demonstrates that human mccRCC tissues express V2R, confirming the suitability of this specific imaging technique for metastasis extension assessment.

[This corrects the article DOI: 10.7150/thno.20912.].

Background: Osteoarthritis (OA) characterized by progressive cartilage degeneration and chronic pain is hindered by the vicious inflammation-pain cycle. Nonsteroidal anti-inflammatory drugs (NSAIDs) can only alleviate clinical symptoms. Although electrical stimulation of the vagus nerve has achieved some results, the toxicity of the electrodes and the secondary damage caused by dismantling limited its clinical application. Methods: A "sono-piezoelectric-bioelectricity-neuroimmune" cascade modulation strategy based on ultrasound-driven piezoelectric ZnO nanoparticles was established to attenuate osteoarthritic neurogenic inflammation and pain with inhibited cartilage degradation. Results: The "sensory neuron-cholinergic anti-inflammatory pathway" reflex arc was activated by the dynamically and spatially-temporally programmed sono-piezoelectric field (0.7 V/36 μA peak) within deep joint tissues by targeting α7nAChR-P2RX7 neuroimmune axis. Consequently, our sono-piezoelectric neuroimmune modulation strategy significantly up-regulated α7nAChR expression, and synchronously inhibited pain mediator CX3CL1, opposed macrophage infiltration, inhibited P2RX7-mediated IL-1β/IL-6 inflammatory storm, restored IL-1β-injured chondrocyte activity and migration capacity, activated stromal genes (Col2a1) for matrix synthesis, and inhibited cartilage degradation-related MMP13 expression. All these actions re-established the balance of glycosaminoglycan (GAG)/deoxyribonucleic acid (DNA) metabolism to remodel joint immune homeostasis, and activated the cholinergic pathway to break the vicious cycle of "inflammation-pain", which restored mechanical pain threshold (Von Frey) and weight-bearing capacity to near-normal levels and reconstructed tidal structures in a rat OA model. Conclusions: Our pioneering "sono-piezoelectrical signal-neural reflex-immunomodulation" cascade strategy for regulating neuroinflammatory reflex-arc-mediated α7nAChR-P2RX7 axis provide deep insights into OA-represented neuroinflammatory diseases.

Rationale: Psoriasis features persistent activation of the mononuclear phagocyte system (MPS), yet the subset-specific pathogenic roles of colony-stimulating factor 1 receptor (CSF1R) remain undefined. We aimed to identify pathogenic CSF1R^high MPS subsets, characterize their ligand-receptor circuits, and define the CSF1R-PPARα axis in disease pathogenesis. Methods: By integrating human single-cell and spatial transcriptomics with murine imiquimod (IMQ)-induced psoriasis models, we employed genetic and pharmacologic interventions to achieve our aims. Results: We found a pathologic CSF1R^high MPS population was selectively expanded, forming localized cytokine hubs enriched for TNF-α, IL-1β, and IL-23. Ligand mapping showed CSF1 upregulation amplified MPS activation via autocrine loops. Systemic CSF1R targeting dismantled skin-blood MPS circuits and depleted pathogenic hubs, suppressing pro-inflammatory cytokines more effectively than local blockade. Mechanistically, CSF1R activation directly suppressed PPARα. Critically, the anti-inflammatory effect of CSF1R inhibition was abrogated by PPARα antagonism, demonstrating a non-redundant, downstream role for PPARα. Consequently, CSF1R suppression releases PPARα-mediated resolution programs. Pathogenic CSF1R^high MPS hubs sustain inflammation through ligand-driven expansion and PPARα suppression. Conclusions: Our work delineates a unidirectional CSF1R-PPARα pathogenic axis and demonstrates that systemic CSF1R targeting is required to disrupt this circuit, providing a mechanistic foundation for a novel treatment strategy.