Theranostics最新文献

Background: Radiotherapy resistance in breast cancer remains a major clinical challenge. The key molecular determinants and cellular populations driving this resistance are not fully understood. Methods: A radiotherapy resistance (RR) gene panel was identified from TCGA-BRCA and GSE120798 cohorts. Single-cell and spatial transcriptomics characterized RRhigh epithelial cells (RRhighepi). A prognostic model, named SuperPC and StepCox-based Radiotherapy Resistance model (SSRR), was built via machine learning and Mendelian randomization. Functional roles of Prolyl 4-Hydroxylase Subunit Alpha 2 (P4HA2) were validated in vitro. Results: The RR gene panel was upregulated in tumors and enriched for cell cycle pathways. RRhighepi cells exhibited elevated stemness, activated cell cycle and metabolic programs, and enhanced DNA damage repair. RRhighepi represented a developmental origin and communicated with endothelial cells. The SSRR model stratified patients into high-risk groups with poorer survival and distinct therapeutic responses. P4HA2, a key model gene, was upregulated in multiple cancers. P4HA2 knockdown suppressed proliferation, invasion, and colony formation, and synergized with radiotherapy to reduce stemness and enhance DNA damage. WGCNA confirmed co-module membership of P4HA2 and the RR panel. Conclusions: This study, through multi-omics analysis, proposes a potential mechanistic model associated with radiotherapy resistance in breast cancer. P4HA2 is a potential therapeutic target that sensitizes breast cancer to radiotherapy. The RR gene panel and SSRR model provide insights into resistance mechanisms and prognostic stratification.

The impaired healing of diabetic wounds is caused by complex multifactorial pathologies and conventional therapeutic approaches often show limited efficacy. In recent years, stimulus-responsive hydrogels based on cascade reactions have become a promising approach in the management of diabetic wounds. These hydrogels are designed to react to particular characteristics of the wound microenvironment, such as glucose concentration, pH, reactive oxygen species (ROS) and enzyme activity, allowing spatiotemporally controlled drug release and synergistic multi-target control. This review focuses on the recent development in understanding of the pathophysiology of diabetic wounds, immune microenvironment modulation, and the development of stimuli-responsive cascade hydrogels, as well as the challenges. By integrating responsive moieties, these hydrogels dynamically control the polarization of immune cells and scavenging of ROS. Furthermore, cascade systems, from single-step to multistep design, enable precise spatiotemporal activation and coordinate antibacterial, antioxidant and pro-regenerative effects. Additionally, emerging technologies such as AI-assisted modeling, biosensing-guided feedback, and organ-on-a-chip platforms have great potential to improve the rational design and predictive validation of cascade hydrogel systems, paving the way for intelligent and personalized diabetic wound therapies.

Background: Splicing factors play pivotal roles in mRNA processing and are implicated in tumor progression. The aberrant expression of splicing factors is closely associated with the invasiveness and secretion profiles of pituitary neuroendocrine tumors (PitNETs). In this study, we explored the involvement of splicing factors in PIT1-lineage PitNET progression and assessed the feasibility of targeting the splicing process as a therapeutic approach. Methods: Statistical data on PitNET subtypes were obtained from the National Brain Tumor Registry of China (NBTRC), and gene expression analysis was conducted on 40 clinical samples collected for this study. Transcriptome analysis and RNA immunoprecipitation sequencing (RIP-seq) were utilized to examine FUS-mediated alternative splicing and to identify mRNA binding sites in PitNET cells. Minigene splicing assays were employed to confirm the specific exonic and intronic regions. Additionally, Annexin V/PI assays and JC-1 staining were conducted to evaluate apoptosis. Results: The expression of the splicing factor FUS was elevated in PIT1-lineage PitNETs and was correlated with increased proliferative capacity and reduced apoptosis levels. Transcriptome sequencing revealed that the knockdown of FUS led to extensive exon skipping and activated the p53 pathway. In addition to RIP-seq analysis, these findings suggest that FUS contributes to the inclusion of exon 3 to generate full-length MDM2, a well-established negative regulator of p53. Antisense oligonucleotides (ASOs) specifically designed to target binding sequences on pre-mRNAs effectively disrupted the FUS-mediated splicing process, consequently impeding the progression of PitNETs. Conclusions: Our study elucidated the critical function of FUS as a splicing factor in PitNETs. Furthermore, we illustrated that targeting the splicing mechanism associated with MDM2 could restore p53 levels, thereby impeding the progression of PitNETs. This discovery presents a potentially novel strategy for the clinical management of PIT1-lineage PitNETs.

Rationale: Hemodynamic shear stress critically influences atherosclerosis progression, yet the molecular mechanisms linking biomechanical stimuli to endothelial activation and vascular pathology remain poorly understood. While circular RNAs (circRNAs) participate in endothelial mechanotransduction, the role of mechanosensitive small nucleolar RNA (snoRNA)-like circRNA-a unique subclass harboring snoRNA sequences-in atherosclerosis is unexplored. Methods: We characterized sno-circCNOT1 using high-throughput RNA sequencing, RNA interference, immunofluorescence, and co-immunoprecipitation. Functional studies were performed in endothelial cells and ApoE⁻/⁻ mice to assess its role in pyroptosis and atherogenesis. Mechanistic investigations included RNA pull-down, mass spectrometry, and gain- and loss-of-function assays to identify sno-circCNOT1-interacting proteins and downstream signaling. Results: We identified sno-circCNOT1, a circular RNA derived from CNOT1 exon 17 and intron 17, which incorporates snoRNA SNORA50A. Its expression was upregulated by pro-atherogenic interleukin-1β and pathological oscillatory shear stress, but downregulated by laminar shear stress. Functionally, sno-circCNOT1 mediated shear stress-dependent regulation of endothelial pyroptosis and inflammation. Endothelial-specific overexpression of sno-circCNOT1 aggravated atherosclerotic lesion formation in ApoE⁻/⁻ mice. Mechanistically, its snoRNA-like motif was essential for nuclear localization and function. sno-circCNOT1 bound the IF-ROD domain of lamin A/C (LMNA), stabilizing LMNA and facilitating its interaction with the N-terminal domain of methyltransferase-like 14 (METTL14-N), thereby enhancing METTL14 stability. This axis activated NOD-like receptor protein 3 (NLRP3) and amplified endothelial inflammation. Conversely, overexpression of METTL14-N to disrupt this signaling axis attenuates endothelial dysfunction and atherosclerosis progression. Conclusions: sno-circCNOT1 is a mechanosensitive snoRNA-like circRNA that promotes endothelial pyroptosis and atherogenesis via the LMNA/METTL14/NLRP3 axis. METTL14-N offers a protein-based therapeutic approach, positioning this regulatory pathway as a druggable target for atherosclerosis.

Rationale: Macrophage phagocytosis is essential for pathogen clearance during sepsis. We previously demonstrated that the glycolytic enzyme 6-phosphofructokinase, muscle type (PFKM), modulates macrophage functions and its deficiency alleviates sepsis in mice. However, the function of PFKM in regulating macrophage phagocytosis remains unclear. Methods: CD14⁺ monocytes were sorted by flow cytometry from healthy volunteers and septic patients, and the subcellular localization of PFKM was assessed by immunofluorescence. Nuclear translocation mechanisms and PFKM-p53 interaction were identified by Co-immunoprecipitation coupled with mass spectrometry (Co-IP/MS) and validated by Co-IP. Transcriptomic sequencing was used to identify the downstream target of the PFKM-p53 complex. Inflammatory cytokine levels were detected by ELISA and real-time RT-PCR, and the phagocytosis of macrophages was assessed by flow cytometry. Dual-luciferase reporter assays and ChIP were employed to investigate whether PFKM acts as a co-regulator of p53 in mediating Pdcd1 transcription. Nanobodies targeting PFKM-p53 were screened and subsequently synthesized according to the sequences. The effect of nuclear PFKM and the therapeutic effect of nanobodies were evaluated on the well-established sepsis mouse models induced by Escherichia coli or cecal ligation and puncture. Results: PFKM translocated to the macrophage nucleus during sepsis. Nuclear accumulation of PFKM impaired phagocytosis through a non-glycolytic "moonlighting" function and exacerbated sepsis. Mechanistically, PFKM interacts with p53, which facilitates its nuclear translocation. Subsequently, PFKM promotes p53 acetylation at K120, enhancing p53 binding to the Pdcd1 promoter and driving its transcription, thereby suppressing macrophage phagocytosis. Blocking the PFKM-p53 interaction with a nanobody, Nb07, restored phagocytosis of macrophages and alleviated sepsis in mice. Conclusion: Our data reveal the PFKM-p53-PD-1 axis that suppresses macrophage phagocytosis in sepsis and highlight the therapeutic potential of targeting this pathway with nanobody-based strategies.

Ischemic stroke (IS) is accompanied by high disability and mortality. Thrombolysis and neuroprotection are the predominant therapeutic strategies for IS. However, thrombolytic drugs commonly suffer from hemorrhagic risks and unsatisfactory thrombus-targeting delivery. Additionally, the blood-brain barrier (BBB) presents a significant challenge for the effective delivery of neuroprotective drugs. In recent years, nanodrug delivery systems (nano-DDS) have garnered significant attention for their ability to improve drug efficacy in vivo and facilitate BBB penetration. Specifically, thrombus- and cerebral ischemic lesion-targeted nano-DDS have emerged as a versatile toolbox for the precise treatment of IS. Herein, this paper provides an overview on the latest advancements in nano-DDS for IS therapy, covering conventional nanomedicines, cell membrane-camouflaged biomimetic nano-DDS, and exosome-involved nanotherapeutics, with a particular focus on the potential and application of cutting-edge nano-drug delivery techniques. Finally, we discuss the future perspectives and challenges of nano-DDS in the context of IS treatment.

Rationale: Neuroblastoma (NB) is a predominant extra-cranial malignancy in childhood, while molecular drivers of its progression and effective treatment strategies have yet to be clarified. Methods: RNA sequencing was performed to identify transcriptional regulators and corresponding target genes. To explore the biological effects and underlying mechanisms of these regulators, a comprehensive methodology was utilized, encompassing chromatin immunoprecipitation, dual-luciferase reporter assay, qRT-PCR, western blot, alongside gene over-expression and silencing techniques, co-immunoprecipitation, and mass spectrometry. The MTT assay, soft agar colony formation, Matrigel invasion, and nude mouse xenograft models were applied to assess oncogenic properties. Patient survival was analyzed using the log-rank test. Results: Armadillo repeat containing 12 (ARMC12) was identified as a MYC-interacting modulator within liquid condensates to up-regulate critical nucleoporin-encoding targets (NUP62/NUP93/NUP98), which promoted nuclear pore complex (NPC) biogenesis to facilitate nuclear trafficking of oncogenic effectors, thereby enhancing invasion and metastasis of NB. As a protein within extracellular vesicles of Malassezia globosa colonizing NB tissues, MGL_0381 also facilitated MYC transactivation via physical interaction to accelerate NPC biogenesis and NB progression. Tioconazole (TCZ) and UU-T02 were identified as efficient inhibitors blocking ARMC12-MYC and MGL_0381-MYC interaction, and synergistically reduced NPC number and aggressive features of NB. High ARMC12, MYC, NUP62, NUP93, or NUP98 levels served as markers of unfavorable patient outcomes in clinical cohorts. Conclusions: These findings collectively demonstrate that dual targeting of AMRC12 and Malassezia globosa disrupts MYC liquid condensates-driven NPC biogenesis during NB progression.

Rationale: Traumatic brain injury (TBI) poses a significant global health concern, necessitating a comprehensive understanding of its pathophysiology to devise effective treatments. The correlation between the intensity of head impact during injury and resultant neuropathology and behavioral changes in TBI remains unclear. Methods: The Quantitatively Controlled and Measured-TBI (QCM-TBI) system, a novel closed-head injury model, enables precise control and measurement of impact during collision. The QCM-TBI system is designed with a unique gravity-compensating animal support system that replicates natural head motion in human TBI. Using QCM-TBI in conjunction with a multimodal sensor fusion technique, we measured instantaneous force over the time of collision, while compensating distortion led by extreme acceleration of the force sensor. To address whether TBI affects neuropathology and behaviors of mice in a force-dependent manner, we conducted transcriptome analysis, electron microscopy, and confocal microscopy in QCM-TBI model. We further compared molecular and pathological features of QCM-TBI mice with chronic traumatic encephalopathy (CTE) patients. Results: Transcriptome analysis of QCM-TBI mice showed a significant downregulation of neuronal genes associated with synaptic function and neurogenesis, particularly in the hippocampus, which correlated with the severity of neuropathological features. Molecular and neuropathological characteristics of QCM-TBI mice partially resemble those of chronic traumatic encephalopathy (CTE) patients. Levels of phosphorylated Tau (p-Tau) and amyloid precursor protein (APP) correlate with impact magnitude, while neurofilament levels diminish in QCM-TBI mice. Neurons exhibit ultrastructural axonal damage in an impact-dependent manner. Conclusions: Overall, this study suggests head impact intensity leads to decreased adult hippocampal neurogenesis, increased levels of phosphorylated Tau (p-Tau), and axonal damage, reflecting key neuropathological signatures of traumatic brain injury. Consequently, therapeutic strategies for TBI should account for the impact's severity in determining neuropathological outcomes.

Rationale: Osteoarthritis (OA) lacks disease-modifying therapies. Although systemic denosumab delays OA progression, it causes uneven drug distribution and off-target effects, whereas intra-articular injections are invasive and risk joint infection. We aimed to develop a minimally invasive microneedle platform that delivers denosumab locally to achieve therapeutic efficacy comparable to intra-articular injection while avoiding systemic exposure. Methods: A dissolvable denosumab-loaded microneedle array (MNs@De) was fabricated for transcutaneous intra-articular delivery. OA was induced in rodents and Beagle dogs; animals were treated with MNs@De, systemic denosumab, intra-articular denosumab, or vehicle. Synovial inflammation, cartilage erosion, and pain were evaluated histologically and behaviorally. Single-cell RNA sequencing and immunofluorescence were performed to assess macrophage senescence and chondrocyte metabolism. Secretion of pro-inflammatory and catabolic factors was quantified in vitro using senescent macrophage-chondrocyte co-cultures. Results: MNs@De delivered denosumab effectively into joints, significantly reducing synovial inflammation, cartilage erosion, and pain compared with systemic administration and achieving outcomes comparable to intra-articular injection. Single-cell profiling revealed that denosumab markedly decreased senescent macrophage abundance within synovial tissue. Mechanistically, denosumab inhibited senescent macrophage-derived pro-inflammatory and catabolic factor release, thereby shifting chondrocytes from catabolic to anabolic states. Conclusions: Targeting senescent macrophages via MNs@De attenuates OA progression without requiring intra-articular injections or increasing systemic drug exposure. Microneedle-mediated denosumab delivery offers a minimally invasive, localized therapeutic strategy for OA.

Mitochondria are involved in energy production, signal conduction, and cellular differentiation in the human body, and they determine the direction of tumorigenesis and development. Mitochondria-targeted therapy in cancer cells has been reported since researchers discovered the relationship between mitochondria and cancer. However, the complexity of the tumor microenvironment (TME) can impair the therapeutic effect. Understanding the mechanisms of mitochondrial function in various cells of TME (e.g., tumor-associated macrophages (TAMs), cancer-associated fibroblasts (CAFs), cancer stem cells (CSCs), T cells, natural killer (NK) cells, tumor-associated neutrophils (TANs)), as well as mediated crosstalk with cancer cells, would be beneficial for accelerating these therapeutic strategies into clinical practice and leading to more effective disease treatment. Subsequently, we summarized representative small-molecule drugs targeting mitochondrial homeostasis, energy metabolism, and mitochondrial DNA (mtDNA) and evaluated their limitations. Building on this foundation, we reviewed the latest multifunctional nanomedicines. These agents leverage TME responsiveness, surface-targeting engineering, and multimodal synergy (combining chemotherapy, photodynamic therapy (PDT), sonodynamic therapy (SDT), radiodynamic therapy (RDT), and immunotherapy) to precisely deliver drugs, ions, genetic material, and even whole mitochondria to target organelles. This approach simultaneously remodels the immunosuppressive microenvironment and induces immunogenic cell death (ICD).