Theranostics最新文献

Mitophagy, a selective autophagic pathway that clears damaged or dysfunctional mitochondria, has emerged as a promising therapeutic approach. Mitophagy maintains a delicate balance between cell survival and death, while mounting evidence suggests that it predominantly promotes tumor cell survival under stress, particularly in responses to cancer therapy. Moreover, aberrant regulation of mitophagy results in cancer pathology with characteristic hallmarks, including remodeling of metabolic plasticity, maintenance of cancer stem cell characteristics, and immune regulation of the tumor microenvironment. This review synthesizes multifaceted roles of mitophagy in cancer biology, from tumor initiation and progression to therapy responses. It also summarizes molecular mechanisms underlying mitophagy. How cancer cells exploit mitophagy to survive therapy has been harnessed to develop therapeutic strategies. We elaborate the evolution of mitophagic therapy from small-molecule modulators to nanotechnology-based targeted delivery systems. Finally, we highlight the promise of targeting mitophagy in overcoming treatment resistance and improving clinical outcomes for patients.

Rationale:​​ CAR-T cell therapy has demonstrated remarkable promise for managing specific autoimmune disorders. However, it remains unclear, whether long-term immunosuppressive therapy in autoimmune patients adversely affects the phenotype and function of patient-derived CAR-T products. This study aimed to compare the characteristics of T cells and manufactured CAR-T cells from patients with multiple myeloma (MM) and chronic inflammatory demyelinating polyneuropathy (CIDP). ​​Methods:​​ T cells isolated from MM and CIDP patients, as well as healthy volunteers (for baseline comparisons only), were analyzed. CAR-T cells were generated using an identical manufacturing process. A comprehensive analysis was conducted, including flow cytometry for phenotypic and functional assessment, transcriptomic profiling via RNA sequencing, and in vitro functional assays such as cytokine secretion and cytotoxicity tests. ​​Results:​​ T cells from CIDP patients showed phenotypes and functional profiles more comparable to those from healthy volunteers. In contrast, MM-derived T cells showed increased CD8⁺ T cell frequency, elevated exhaustion markers, reduced naïve and less-differentiated subsets, and enhanced effector molecule production upon non-specific stimulation. CAR-T manufacturing reduced these inherent differences, yielding similar differentiation states, transcriptomic profiles, and convergent cytotoxic capacities. However, distinct immunomodulatory features persisted, as CIDP-derived CAR-T cells displayed reduced activation markers and lower IFN-γ secretion upon antigen stimulation compared to MM-derived CAR-T cells. ​​Conclusions:​​ Our study reveals that CAR-T manufacturing process can reduce pre-existing T-cell heterogeneity across different patient populations. These findings support the feasibility of autologous CAR-T therapies in immunosuppressed autoimmune patients, demonstrating that critical cytolytic functions are preserved despite residual alterations in cytokine profiles.

Rationale: Bone defects pose a persistent challenge in orthopedic medicine due to their limited self-repair capacity. Although guided bone regeneration scaffolds have shown therapeutic potential, their clinical efficacy remains constrained by their suboptimal osteoinductive capability. Methods: Herein, we developed biodegradable piezoelectric polyhydroxybutyrate-barium titanate (PHB-BT) nanofiber scaffolds capable of generating synergistic piezoelectric stimulation for bone repair when integrated with low-intensity pulsed ultrasound (LIPUS). Results: Compared with conventional PHB scaffolds, ​PHB-BT nanofiber scaffolds​ ​showed enhanced piezoelectric properties​ and ​excellent biocompatibility, ​thereby facilitating​ sustained osteogenic activity. ​In vitro​ studies revealed that these scaffolds ​significantly promoted​ the osteogenic differentiation of bone marrow mesenchymal stem cells under LIPUS stimulation. ​Notably, ​in vivo​ evaluations ​demonstrated​ that these scaffolds ​substantially accelerated bone defect repair​, with complete scaffold degradation observed after eight weeks. Mechanistically, PHB-BT nanofibers improved osteogenesis via activating the Ca²⁺/calcineurin/nuclear factor of activated T-cells signaling pathway in response to ultrasound stimulation. Conclusions: These findings have significant implications for the design of next-generation, implantable electrical stimulators capable of providing sustained electromechanical cues for personalized bone tissue engineering applications.

Rationale: Parkinson's disease (PD) is a progressive neurodegenerative disorder that affects movement, muscle control, and balance. Effective therapeutic options for this condition are limited. Natural therapies, including lifestyle changes, probiotics, and muscle relaxants, have received attention for symptomatic relief. Colostrum, particularly its extracellular vesicles (C-EVs), has emerged as a promising nutraceutical with the potential to improve therapeutic outcomes in divergent diseases. Methods: We purified and characterized (C-EVs) as a putative cell-based therapy. Theranostic (biodistribution, diagnostic, and therapeutic) efficacy studies were performed in C-EV-treated mice intoxicated with methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The C-EV tissue distribution, anti-inflammatory and neurorestorative activities were examined. These include transcriptomic, immune, and neuroprotective profiles linked to disease outcomes. Results: C-EV-treated MPTP mice showed reduced microglial activation and restored neuronal responses. RNA sequencing and transcriptomic analyses have demonstrated reduced immune cell recruitment and activation in the disease-affected brain subregions. The activation of canonical inflammasomes, pro-inflammatory cytokines, and chemokine expression was controlled. C-EV treatment reduced the levels of disease-associated immune-regulatory transcription factors. Simultaneously, Treg-associated adaptive immune responses increased. Multiple C-EV-miR-isolated immune regulatory cargos are linked to neuroinflammation and nigral preservation. C-EVs loaded with miR-20a-5p, miR-23b-3p, let-7a-5p, miR-22-3p, and miR-30a-3p mimics attenuated pro-inflammatory cytokines in activated microglia. Conclusions: C-EVs elicit neuroprotective responses in MPTP-intoxicated mice. These responses control microglial activation and facilitate neuroprotective responses.

Maintaining telomere integrity is essential for cellular survival, and reactivation of telomerase or alternative lengthening of telomeres (ALT) represents a hallmark of cancer, ensuring replicative immortality. Osteosarcoma (OS), a malignancy in which many tumors rely on ALT for telomere maintenance, lacks effective therapeutic strategies targeting this pathway. This study aimed to identify and characterize novel molecular regulators of ALT activity and explore their potential as therapeutic targets in OS. Methods: Immunohistochemistry was performed to evaluate the expression of phosphorylated NPM1 (pT199-NPM1) in OS tissues. Functional experiments including NPM1 knockdown and rescue assays were conducted to assess the impact of NPM1 on break-induced telomere replication (BITR) and cell viability in ALT-positive cells. Mechanistic studies involving phosphorylation analysis, ubiquitination assays, and co-immunoprecipitation were used to determine how ATR-mediated phosphorylation of NPM1 regulates POLD3 stability and its interaction with the CST complex. Pharmacological screening was performed to identify compounds that inhibit ALT activity, followed by in vitro proliferation assays and in vivo mouse xenograft experiments to evaluate therapeutic efficacy and synergy with doxorubicin. Results: We identified pT199-NPM1 as a novel, highly expressed protein factor in ALT-positive OS tissues. NPM1 depletion impaired break-induced telomere replication and significantly reduced the viability of ALT-positive cells. ATR signaling phosphorylated NPM1 at Thr199, which stabilized POLD3 by preventing its ubiquitin-mediated degradation. Recruitment and function of pT199-NPM1 at telomeric damage sites required STN1, defining a CST/pT199-NPM1/POLD3 regulatory axis essential for ALT activity. Clinically, elevated Thr199 phosphorylation correlated with poor survival in OS patients, while expression of a phosphorylation-deficient T199A mutant failed to sustain ALT telomere maintenance. Pharmacological screening identified EPZ-6438, an EZH2 inhibitor, as a potent ALT suppressor that reduced NPM1 transcription, inhibited homologous recombination-mediated telomere synthesis, and suppressed OS cell proliferation. In mouse xenografts, EPZ-6438 enhanced OS cell sensitivity to doxorubicin, suggesting therapeutic synergy. Conclusions: This study uncovers a novel CST/pT199-NPM1/POLD3 regulatory module that is critical for ALT telomere maintenance in OS. Targeting NPM1 or its downstream effectors effectively suppresses ALT activity and enhances chemotherapy response. These findings provide new mechanistic insights into telomere regulation in ALT-positive tumors and highlight the therapeutic potential of NPM1-centered pathways in OS.

Rationale: Metabolic dysfunction-associated steatohepatitis (MASH) is a severe liver disease with limited therapeutic options. This study aimed to investigate the protective effects of elemicin (Ele) against MASH and its underlying mechanisms, focusing on the interaction between AMP-activated protein kinase (AMPK) and hyaluronan synthase 1 (Has1). Methods: HFHC diet-induced MASH mouse models and palmitic acid/oleic acid (PO)-treated primary hepatocytes were used. Transcriptomic and lipidomic analyses, immunohistochemistry, western blotting, and molecular docking were employed to assess gene expression, lipid metabolism, inflammation, and fibrosis. Interactions between Ele, AMPK, and Has1 were validated via SPR, Co-IP, and CETSA. Results: Ele significantly ameliorated hepatic steatosis, inflammation, and fibrosis in MASH mice. Systematic profiling of transcriptomic and lipidomic landscapes reveals that Has1-mediated lipid metabolism is strongly correlated with MASH severity in dietary mouse models. Using loss-of-function studies, liver-specific inhibition of Has1 ameliorates hepatic steatosis, inflammation and fibrosis in vivo and in vitro. The anti-MASH effects of Ele are largely dependent on interrupting the formation of AMPK/Has1 complex. Furthermore, Ele normalized hepatic phospholipid profiles, particularly increasing phosphatidylethanolamine to improve mitochondrial function. Conclusions: Ele protects against MASH by interrupting AMPK/Has1 interaction, regulating lipid metabolism, and restoring mitochondrial function. Collectively, these findings highlight Ele as a potential therapeutic agent and Has1 as a novel target for MASH treatment.

Rationale: Inflammation and myocardial remodeling are major contributors to the progression of cardiac diseases. mRNA-based therapeutics have emerged as a promising modality for cardiovascular intervention; however, their clinical translation remains constrained by challenges in achieving efficient and spatially precise delivery to diseased cardiac tissue, particularly following myocardial injury. To address this unmet need, a dual-active magnetic nanocarrier was engineered for targeted mRNA delivery to damaged cardiovascular tissue. Methods: The interleukin-10 anti-inflammatory cytokine mRNA (IL-10 mRNA) was encapsulated in lipid nanoparticles, which were fused with nanovesicles derived from mesenchymal stem cells (NVs) and functionalized with cardiac-targeting peptides (T peptides) to form IL-10 mRNA-loaded T-NVs (m10@T-NVs). Magnetic nanoparticles (MNPs) were conjugated with azide-modified antibodies against CD63 and myosin light chain 3 (MLC3), which are overexpressed in damaged myocardial tissue via click chemistry, to enable targeted delivery to injured cardiac tissue. Subsequently, the m10@T-NVs were combined with functionalized MNPs via CD63 interactions to form m10@T-MNVs. Results: m10@T-MNVs were developed and characterized, confirming the functionalization of NVs and MNPs. Under guided of an external magnetic field, m10@T-MNVs exhibited a 4.5-fold increase in accumulation in H₂O₂-induced injured cardiomyocytes and damaged cardiac regions, achieving significantly higher delivery efficiency. In a mouse model of myocardial infarction (MI), administration of m10@T-MNVs enhanced intramyocardial IL-10 mRNA expression and cytokine production. This led to the polarization of macrophages toward an M2 anti-inflammatory phenotype, mitigation of tissue injury, reduced apoptosis, attenuation of fibrosis, and suppression of pathological myocardial remodeling. Conclusions: Dual-active targeting of injured cardiac tissue using magnetic nanocarriers constitutes a promising therapeutic strategy for cardiovascular diseases by addressing key challenges associated with tissue-selective mRNA delivery in the injured myocardium.

Rationale: Glioblastoma multiforme (GBM) is an aggressive brain tumor marked by diffuse infiltration, a complex microenvironment, and poor drug delivery due to the blood-brain barrier. Fibroblast activation protein (FAP), widely expressed by cancer-associated fibroblasts (CAFs), emerges as a promising yet underexploited target for drug delivery. Methods: Here, a modular ferritin-based drug carrier (FDC) functionalized with an optimized FAP-targeting ligand using site-specific sortase A-mediated ligation was developed. This approach ensures precise surface modification while preserving ferritin's structure and drug-loading capacity. Monomethyl auristatin E (MMAE), a potent cytotoxin, is stably encapsulated to create a dual-targeting nanocarrier aimed at FAP and transferrin receptor 1. Results: In orthotopic glioma mouse models, the resulting FDC enables pH-responsive MMAE release, enhances tumor targeting and cellular uptake, reduces tumor burden, prolongs survival, and minimizes systemic toxicity compared to free MMAE. Furthermore, spatial transcriptomic analyses and immunohistochemistry data reveal that this therapeutic approach reshapes the tumor microenvironment by enhancing the spatial proximity between CAFs and cytotoxic immune cells and activating multiple immune pathways. Conclusions: This study presents a precision-engineered nanoplatform for FAP-targeted GBM therapy, provides novel insights into the stromal-immune dynamics of GBM under therapeutic pressure and supports the rationale for combining CAF modulation with immunotherapy to achieve durable tumor control.

Rationale: Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are a standard therapy for non-small cell lung cancer (NSCLC). Despite their clinical efficacy, dose-limiting systemic toxicity and the eventual development of acquired resistance limit their long-term benefit. Therefore, innovative drug delivery strategies are highly demanded to optimize the therapeutic window, minimizing toxicity of EGFR-TKIs at lower doses without compromising their efficacy. Methods: We developed a red blood cell (RBC)-based biomimetic platform for the systemic delivery of EGFR-TKIs. Osimertinib-loaded poly(lactic-co-glycolic acid) nanoparticles were camouflaged with a biotinylated RBC membrane (Osi-RNPs). They were then conjugated to the surface of RBCs via high-affinity biotin-streptavidin interactions to form a stable construct (Osi-RNP-SA-RBC). The physicochemical characteristics, cellular uptake, and in vitro antitumor activity of Osi-RNPs were characterized. We further assessed the pharmacokinetics, biodistribution, therapeutic efficacy, and safety profile of Osi-RNP-SA-RBC in subcutaneous and orthotopic NSCLC mouse models. Results: The Osi-RNP-SA-RBC platform demonstrated stable attachment, favorable hematocompatibility and excellent biosafety. Compared to the Osi-RNP-RBC (nonspecific adsorption), Osi-RNP-SA-RBC presented prolonged blood circulation (1.6-fold) and enhanced tumor accumulation (2.2-fold). Upon intravenous injection of Osi-RNP-SA-RBC at a reduced dose and frequency, superior tumor suppression was observed in both subcutaneous (16.8-fold increase) and orthotopic (4.2-fold increase) NSCLC mouse models compared to the free osimertinib at same administration dosage. Conclusion: This study demonstrates the potential of RBC-conjugated biomimetic nanomedicine as a promising strategy for enhancing the treatment efficiency of EGFR-TKI against NSCLC in vivo.

Background: Compared to the lymphodepleting chemotherapy and radiotherapy, photodynamic therapy (PDT) is an oncotherapeutic modality inherently stimulating immune responses by inducing immunogenic cell death (ICD). However, the immunosuppressive tumor microenvironment (TME) frequently attenuates PDT-elicited immune responses, limiting its efficacy in eradicating distant and metastatic tumor cells. Methods: To maximize the immunotherapeutic efficacy of PDT, we developed a photodynamic immunotherapeutic liposomal nanoplatform (PDIT-liposome) integrating components targeting sequential stages of the antitumor immune response: 1) a phthalocyanine photosensitizer to induce ICD, 2) a factor Xa inhibitor (rivaroxaban) to promote T-cell priming, 3) and a program death-ligand 1 inhibitor to augment cytotoxic T lymphocyte (CTL) attack. To enable light-controlled drug release at tumor sites, the liposome was constructed with reactive oxygen species-sensitive phospholipids in response to the PDT effect. Results: PDIT-liposomes were characterized via multiple physicochemical and optical evaluations. Comprehensive in vitro and in vivo investigations confirmed that PDIT-liposomes significantly enhanced antitumor efficacy compared to monotherapies and dual combinations. In a subcutaneous implantation tumor model, PDIT-liposome achieved a 91.7% antitumor rate compared to 21.83% (P-liposome), 46.78% (PD-liposome), and 51.08% (PR-liposome) (p < 0.001). Mechanistic analysis revealed enhanced dendritic cell maturation (8-fold increase in CD11c⁺ cells) and T-cell priming (2.3-fold increase in CD8⁺ T cells) in tumor-draining lymph nodes (TDLNs), and CTL-mediated cytotoxicity (5.4-fold increase in CD107a⁺ activated CTLs) in TME. Notably, PDIT therapy induced long-term immunological memory, which suppressed 90.68% tumor reoccurrence and metastasis. Conclusion: This study presents a strategy to amplify PDT-elicited immunotherapeutic efficacy by synergizing agents targeting distinct stages of the immune response. It also theoretically validates the synergy of PDT, anticoagulation therapy, and immune checkpoint inhibition in cancer treatment.