Theranostics最新文献

Background: The switch to endothelial-to-mesenchymal transition (EndMT) in endothelial cells (ECs) induced by disturbed flow (d-flow) has been identified as the critical driver of the pathogenesis of inflammatory vascular disorders. We aimed to investigate the role of EndMT in abdominal aortic aneurysms (AAA) and the underlying mechanism. Methods: Immunoblotting, immunofluorescence and transmission electron microscope were used to assess d-flow-induced EndMT in human and mouse AAA models (Ang II/PPE). An Ibidi pump system was used to produce d-flow on human aortic endothelial cells (HAECs), and the expression of galectin-7 was enhanced and weakened using an adeno-associated virus. Furthermore, single-cell RNA sequencing was performed to explore the underlying mechanism of galectin-7-mediated EndMT. Results: EndMT induced by d-flow, which suppressed galectin-7 expression, was positively correlated with AAA. Enhanced galectin-7 expression inhibited d-flow-induced EndMT and AAA progression, whereas reduced galectin-7 expression resulted in the opposite effect. Mechanistically, we found a EndMT-related cluster in HAECs by single-cell RNA sequencing, and the SRGN gene in this cluster was considered the core gene. Galectin-7 bound competitively to the transcription factor CREB, resulting in the inhibition of SRGN transcription, which in turn prevented TGFβ/smad pathway activation, thereby restoring EndMT progression. Conclusions: EndMT transformation in ECs exposed to d-flow was the critical driver of AAA development. Furthermore, endothelium-enriched galectin-7 suppressed the EndMT process induced by d-flow and prevent AAA progression by transcriptionally inhibiting SRGN via competitive binding with CREB to restrict TGFβ/smad pathway.

Rationale: Current gastrointestinal endoscopy mainly depends on morphological changes for lesion diagnosis, thus often failing to detect early colorectal cancers (CRCs) with subtle morphological alterations. Optical molecular imaging via endoscopy may provide a unique means to identify early CRCs that precede the morphological changes observed via conventional endoscopy. In addition, optical imaging methods are utilized for intraoperative navigation when imaging tumors. However, the primary challenge in applying optical molecular imaging clinically is the restricted kinds of clinically endorsed targeted probes. Cerenkov luminescence (CL) can be observed with almost all clinically validated radiotracers. Therefore, Cerenkov luminescence imaging (CLI) does not require the development of new probes and can directly utilize clinically validated radiotracers. This study aimed to use Cerenkov luminescence endoscopy (CLE) for diagnosing early CRC effectively. Methods: In a prospective observational study, we use a self-produced CLE to diagnose colorectal lesions (mainly CRC). The CL images of the lesions were recorded and analyzed in comparison with PET/CT scans and histopathology. Results: A total of 20 colorectal lesions from 15 patients were included in the study. The agreement between CLE and PET/CT in diagnosing early CRC (stage Ⅰ CRC and advanced adenoma) was 100%. The level of agreement of CLE images with histopathology was 88.9% acceptable to high for early CRC. Compared with that of colorectal hyperplastic polyps, the signal-to-background ratio of CLE from early CRCs was significantly greater (1.33 ± 0.17 vs 0.99 ± 0.03, P < 0.001). In phantoms, tumor-bearing nude mice, and rectal pseudotumor model dogs, CLE detected CL at the corresponding lesion locations. Conclusions: This study demonstrated for the first time that CLE could utilize Cerenkov luminescence molecular imaging to diagnose early CRCs, overcoming the limitations of current endoscopic diagnosis based on morphological changes. (ClinicalTrials.gov, NCT05575765).

Neurodegenerative diseases (NDDs), including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), Huntington's disease (HD) and multiple sclerosis (MS), are characterized by progressive neuronal dysfunction and limited therapeutic options, largely due to the restrictive nature of the blood-brain barrier (BBB). Exosomes, naturally occurring extracellular vesicles (EVs), have gained attention as innovative drug delivery vehicles owing to their intrinsic ability to cross the BBB, minimal immunogenicity, high biocompatibility, and capability to carry diverse therapeutic cargos such as proteins, nucleic acids, and small molecules. Furthermore, exosomes can be bioengineered to enhance drug-loading efficiency and targeting specificity, positioning them as a versatile and effective platform for treating NDDs. In this review, we summarize recent advances in exosome biogenesis, secretion, and engineering, with an emphasis on innovative strategies for exosome isolation, drug loading, and surface modification. We further explore their roles in modulating neuroinflammation, promoting neural regeneration, and enabling precise therapeutic delivery. Critical challenges associated with large-scale production, quality control, and regulatory compliance under Good Manufacturing Practices (GMP) are also discussed. Collectively, these developments underscore the transformative potential of engineered exosomes in advancing precision therapies for neurodegenerative disorders and offer strategic insights into their clinical translation.

Rationale: Although reperfusion has been established as the main strategy for stroke treatment, it frequently causes irreversible secondary injury with dynamic pathological changes. The ischemic microenvironment in the brain continues to deteriorate after reperfusion, resulting in progressive expansion of the injured area and subsequent neuronal death. Therefore, it is urgent to find a therapeutic strategy for long-term management of ischemic stroke at post-reperfusion stages. Methods: In this study, we designed an inflammation cascade-directed therapy using a biomimetic polydopamine nanosystem (MPP-B@MM). The synthesis of the nanosystem was confirmed by transmission electron microscopy, scanning electron microscopy, dynamic laser scattering, proton nuclear magnetic resonance, and so forth. The capability of blood-brain barrier crossing was investigated by cellular study with fluorescence imaging. Meanwhile, reactive oxygen species assay, apoptosis detection, and flow cytometry were employed to evaluate intracellular anti-apoptotic effects of the nanosystem. For in vivo evaluation, a middle cerebral artery occlusion model was established. Anti-stroke efficacy and mechanism of action of the nanosystem were assessed through multiple analytical methods, including behavioral tests, immunohistochemical staining, mRNA sequencing, and blood biochemical analysis. Results: In vitro experiments demonstrated that MPP-B@MM exhibited superior cellular uptake and blood-brain barrier crossing, significantly attenuated mitochondrial damage, and rescued injured neurons. Comprehensive in vivo studies, spanning both acute and chronic phases, confirmed the superior long-term therapeutic performance of the nanosystem. Importantly, mRNA sequencing and pharmacological intervention experiments demonstrated that Homx1 served as the predominant molecular target for acidosis-responsive drug release. Conclusions: This tailored nanosystem demonstrated acute neuroprotective effects during the initial phase of ischemic stroke. As inflammation progressed, the acidosis-responsive motif in the nanosystem could serve as an on-demand therapeutic method, spatiotemporally increasing neurotrophic factor expression at the stroke lesion, which significantly contributed to brain recovery.

Rationale: Myocardial ischemia-reperfusion (MI/R) injury induces apoptosis, metabolic dysregulation, and ventricular remodeling through complex pathological mechanisms. Although nanozyme engineering has the potential for antioxidation, reoxygenation, and pro-vascularization, achieving responsive modulation of the pathological microenvironment remains significantly challenging. Methods: A layered double hydroxide (LDH)-based nanozyme (ZnSrMo-LDH/Cu) was synthesized via a low-temperature hydrothermal/isomorphic substitution method for MI/R treatment. The reactive oxygen species (ROS) scavenging ability and responsive ion release performance of ZnSrMo-LDH/Cu were evaluated through various spectroscopic characterization methods. The biosafety and therapeutic efficiency of ZnSrMo-LDH/Cu-BSA nanozymes were assessed by in vitro and in vivo experiments. Results: ZnSrMo-LDH/Cu demonstrated cascade superoxide dismutase (SOD) and catalase (CAT) activities, effectively overcoming acidic microenvironment limitations to maintain CAT activity rather than peroxidase (POD) activity while scavenging ROS to generate oxygen, with a ROS scavenging capacity 2.97 times that of Fe₃O₄. Moreover, the acid-triggered Sr²⁺ release promoted vascular regeneration and synergistically improved the ischemic-hypoxic microenvironment. Consequently, after bovine serum albumin (BSA) modification, ZnSrMo-LDH/Cu-BSA demonstrated excellent cytoprotective effects, reducing the cardiomyocyte apoptosis rates to 9.4% (in vitro) and 20.7% (in vivo) of the levels in the MI/R group. In vivo studies further validated that ZnSrMo-LDH/Cu-BSA enhanced cardiac function and attenuated ventricular remodeling by inhibiting oxidative stress and promoting angiogenesis. Mechanistically, ZnSrMo-LDH/Cu-BSA provided a cardioprotective effect by inhibiting the TGF-β signaling pathway, thereby alleviating cell damage caused by MI/R. Conclusions: The pathologically responsive LDH-based nanozyme represents a promising avenue for MI/R treatment.

Rationale: Sustained hypertension induces adverse cardiac remodeling. Platelet activation is instrumental in exacerbating inflammation by engaging with macrophages. C-C chemokine motif ligand 5 (CCL5) is contained within platelet α-granules, and released following platelet activation. This work delineated the specific contributions of CCL5 to platelet function, platelet-induced macrophage polarization, and hypertensive cardiac remodeling. Methods: CCL5 knockout (KO) mice infused with Angiotensin II (Ang II) were used to identify the role of CCL5 in vivo. CCL5 absence on platelet activation were evaluated on washed platelets. Two distinct models of platelet depletion and reconstitution were utilized to investigate the impact of platelets lacking CCL5. An in vitro co-culture system was established to explore the roles of CCL5-mediated platelet activation in M2 macrophage polarization. Results: CCL5 KO attenuated the adverse cardiac effects induced by Ang II, including fibrosis, hypertrophy, and functional impairment, accompanied by reduced platelet activation and M2 macrophage polarization in cardiac tissue. Platelet inhibitor administration and platelet depletion/reconstitution experiments revealed that the suppression of platelet activation by CCL5 KO contributed to the amelioration of Ang II-promoted cardiac M2 macrophage polarization and cardiac remodeling. CCL5 KO markedly suppressed the activation of TGF-β1 and NF-κB signaling, an effect observed both in cardiac tissue from Ang II-infused mice and in platelets following ADP stimulation in vitro. In in vitro co-culture systems, rmTGF-β1 reversed CCL5 KO platelet-impaired M2 macrophage polarization. NF-κB inhibition abolished recombinant CCL5 (rmCCL5)-induced platelet activation, while blocking antibodies against CCR1 and CCR3 inhibited rmCCL5-induced NF-κB signaling and platelet activation. Conclusions: These findings underscore the detrimental role of CCL5-mediated platelet activation in promoting M2 macrophage polarization during hypertensive cardiac remodeling and elucidate the molecular mechanism that CCL5 facilitates platelet-derived TGF-β1 signaling by promoting NF-κB activation via CCR1 and CCR3 receptors. These findings support CCL5 inhibition as a promising strategy against inflammation and cardiac damage.

Rationale: Extensive leukocyte diapedesis is a defining step in inflammation and contributes critically to myocardial ischemia/reperfusion injury (MI/RI). Infiltrating leukocytes amplify local inflammation and exacerbate myocardial damage. However, the upstream control of the trans-endothelial migration step remains incompletely understood. Methods: Peripheral blood myeloid cells were isolated from MI/RI patients and healthy donors to examine MAP3K3 expression and its correlation with cardiac markers. Mouse MI/RI models were established to investigate MAP3K3 expression of myeloid cells in the heart. Myeloid-specific Map3k3 deficiency mice were used to evaluate the impact of MAP3K3 depletion on MI/RI severity and on myeloid cell diapedesis from the bone marrow. RNA sequencing and various manipulations of the MAP3K3/TAL1/JAM-A axis were used to elucidate its role in diapedesis. Finally, the therapeutic potential of pazopanib, a MAP3K3 inhibitor, was evaluated in the mouse MI/RI model. Results: MAP3K3 expression was upregulated in both monocytes and neutrophils from MI/RI patients and was positively correlated with the severity of MI/RI. In mice, MAP3K3 in cardiac myeloid cells peaked at day 3 post-MI/RI. Myeloid cell-specific depletion of MAP3K3 alleviated MI/RI by reducing the infiltration of myeloid cells into cardiac tissue. Functionally, MAP3K3 facilitated myeloid cell de-adhesion and transmigration across endothelial barriers. Further mechanistic studies identified the MAP3K3/TAL1/JAM-A signaling pathway as a key regulator of myeloid cell diapedesis. MAP3K3 phosphorylates TAL1 at Ser-122, leading to its ubiquitination and attenuating its transcriptional repression of F11r (encoding JAM-A). Through JAM-A, MAP3K3 promotes integrin internalization, thereby enhancing de-adhesion and myeloid cell transmigration. Treatment with pazopanib, a MAP3K3 inhibitor, ameliorated MI/RI injury and reduced myeloid cell diapedesis into the heart by blocking MAP3K3 phosphorylation activity. Conclusions: MAP3K3 orchestrates myeloid cell diapedesis via a TAL1/JAM-A dependent program during MI/RI. Targeting MAP3K3, exemplified by pazopanib, may offer a therapeutic strategy for MI/RI and related inflammatory conditions.

Rationale: Immunotherapy has emerged as a crucial component in cancer treatment, particularly for the long-term reduction of cancer metastasis and recurrence. However, its development is hindered by limited activation of cellular immune response and suboptimal delivery of vaccine to antigen-presenting cells. Methods: The vaccine was encapsulated within mesoporous silica nanoparticles, followed by functionalization by mannose and phenylboronate ester (MSN-NH-DPM), which facilitates targeting antigen-presenting cells via mannose receptors and enables intracellular delivery through endosomal escape, thereby activating cellular immunity. The nanoparticles were then integrated into chitosan microneedle patches (MNs), which are engineered to deliver the nanoparticles into the skin that is abundant in immune cells, and improve the immune response through the adjuvant properties of chitosan. Results: The chitosan MNs incorporating MSN-NH-DPM (CTS-MN@MSN-NH-DPM) significantly activated the cellular immune response through the MHC-I pathway. The antigen-presenting cells that uptake the vaccine migrated to nearby lymph nodes, inducing systemic immunity to eliminate cancer cells. Compared with subcutaneous injection, the application of CTS-MN@MSN-NH-DPM significantly inhibited the growth of B16/OVA melanoma tumors and extended the survival time of the melanoma mouse model. Conclusions: The MNs with targeted and intracellular delivery represent a promising platform for various vaccines to improve the cellular immune response, thus providing a potential solution for cancer treatment.

Background: Acute kidney injury (AKI) is a severe and prevalent nephrotic syndrome which lack of definitive therapies. Alpha-amino-β-carboxymuconic acid-ε-semialdehyde decarboxylase (ACMSD) is a metabolic enzyme mainly expressed in the kidney which exacerbated AKI injury by promoting TCA cycle and inhibiting nicotinamide adenine dinucleotide (NAD⁺) production, whereas lack of effective intervention strategies for ACMSD-targeted therapy. Methods: Herein, we knocked out ACMSD in vitro through CRISPR-Cas9 method, and developed a reactive oxygen species (ROS)-responsive neutrophil-derived cellular vesicles (CVs) drugs (RNAi@ROS-CVs), which efficiently mediated ACMSD knockdown in vivo, exploring the mechanism of ACMSD-induced ferroptosis process in AKI. Results: ACMSD knockout effectively alleviated cisplatin (CP)-induced mitochondrial damage, suppressed TCA cycle progression, promoted NAD⁺ synthesis, and inhibited ferroptosis in HK2 cells. In mice AKI model, RNAi@ROS-CVs effectively targeted the injured kidneys, downregulated ACMSD expression in renal tubular epithelial cells, reduced ROS production and lipid peroxidation, and alleviated CP or ischemia/reperfusion (I/R)-induced ferroptosis. Conclusion: These findings highlight the therapeutic potential of ACMSD-targeted knockout in AKI intervention and introduce a versatile and efficient controlled-release drug delivery platform for AKI-targeted therapy, with potential applicability to other acute renal diseases.

Bioimaging technologies visually resolve spatiotemporal dynamics of biomolecules, cells, and tissues, enabling essential insights into gene regulation, disease mechanisms, and drug metabolism. CRISPR/Cas-based fluorescent probes transform CRISPR from "genetic scissors" into "molecular microscopes," providing an indispensable tool for in situ decoding of molecular events in living systems. Their high nucleic acid specificity establishes CRISPR/Cas as a pivotal technology for dynamically monitoring genomic and transcriptomic events at live-cell and in vivo levels. This work systematically outlines design strategies and functional mechanisms of mainstream CRISPR/Cas fluorescent probes for bioimaging, encompassing five categories: fluorescent proteins, synthetic dyes, smart gated probes, nanomaterials, and multimodal integrated probes. Recent advances and persistent challenges in achieving high-sensitivity targeted imaging, effective signal amplification, and precise delivery control are comprehensively examined, including analysis of their advantages, limitations, and adaptability in complex biological environments. Building on breakthroughs in in vivo delivery systems, diverse carriers demonstrate significant potential for enhancing CRISPR/Cas transport efficiency, improving tissue penetration, and enabling spatiotemporal controlled release. Continued innovation drives CRISPR/Cas imaging platforms toward higher sensitivity, enhanced biocompatibility, and multifunctional integration, thereby fostering the convergence and broad application of gene editing and molecular diagnostics.