Low shear stress exacerbates atherosclerosis by inducing the generation of neutrophil extracellular traps via Piezo1-mediated mechanosensation.

BACKGROUND AND AIMS: Atherosclerosis is a chronic lipid-driven inflammatory disease largely influenced by hemodynamics. Neutrophil extracellular trap (NET)-mediated inflammation plays an important role in atherosclerosis. However, little is known about the relationship between low shear stress (LSS) and NET generation, as well as the underlying mechanism. METHODS: We induced LSS by partial ligation of the left carotid artery in high-fat diet-fed male ApoE-/- mice. To further validate the direct relationship between LSS and NET formation invitro, differentiated human promyelocytic leukemia HL-60 cells and bone marrow-derived neutrophils were suspended in fluid flow under normal or low shear stress using a parallel-plate flow chamber system. RESULTS: Four weeks after surgery, ligated carotid arteries had more lipid deposition, larger plaque area, and increased NET formation than unligated arteries. Inhibition of NETosis could significantly reduce plaque formation in ApoE-/- mice. Invitro, LSS could promote NET generation directly through downregulation of Piezo1, a mechanosensitive ion channel. Downregulation of Piezol could activate neutrophils and promote NETosis in static conditions. Conversely, Yoda1-evoked activation of Piezo1 attenuated LSS-induced NETosis. Mechanistically, downregulation of Piezo1 resulted in decreased Ca2+ influx and increased histone deacetylase 2 (HDAC2), which increased reactive oxygen species levels and led to NETosis. LSS-induced NET generation also promoted apoptosis and adherence of endothelial cells. CONCLUSION: LSS directly promotes NETosis through the Piezo1-HDAC2 axis in atherosclerosis progression. This study uncovers the essential role of Piezo1-mediated mechanical signaling in NET generation and plaque formation, which provides a promising therapeutic strategy for atherosclerosis.

Ultra-efficient radio-immunotherapy for reprogramming the hypoxic and immunosuppressive tumor microenvironment with durable innate immune memory.

Radiosensitization efficacy of conventional tumor radiosensitizers has been frequently limited by insufficient competence for tumor microenvironment (TME) regulation and unfavorable cellular uptake at biological barriers. Here, we reported an ultra-efficient radiotherapy (RT) strategy by synthesizing an extracellular vesicles (EVs)-encapsulated hollow MnO2 to load metformin (Met@HMnER). It demonstrated significant RT enhancement by morphological control of catalyst and cellular respiratory depression against conventional solid MnO2. Furthermore, the target-modified EVs clothing retains outstanding metformin loading capacity while endowing enhanced biological barrier penetration. A noticeably durable innate immune activation of NK cells was triggered with this nanoplatform via the cGAS-STING pathway. The enhanced immunocompetence was verified on distal metastasis and in-situ recurrence model in vivo, This work paved a new path for synergistic and robust innate immunity in clinical cancer treatment.

A smart TESTER for reliable discrimination of cancer-derived small extracellular vesicles.

Cancer-derived small extracellular vesicles (csEVs) are crucial liquid biopsy indicators that reflect the presence and progression of many malignancies. However, reliable discrimination of csEVs remains a great challenge owing to the interference from normal sEVs (nsEVs) and low abundance in the early stages of cancer. In this work, we developed a Two-Elements Selectively Triggered csEVs Recognization (TESTER) strategy for selective identification of csEVs from the complex clinical body fluid samples. This method was based on the MNAzyme-controlled synchronous recognition to EpCAM and CD63 proteins on the membrane of csEVs. Efficient recognition to csEVs via EpCAM aptamer and CD63 aptamer prompted the release of Partzyme A and Partzyme B probes to induce a MNAzyme structure formation, resulting in the cyclic cleavage of substrate chain to produce cascade fluorescence signal amplification. The detection threshold of the developed TESTER approach for csEVs in complicated biological samples was 72 particles µL-1, accomplishing the highly sensitive and selective quantification of csEVs. At the same time, we successfully constructed a new platform for bimolecular simultaneous recognition, which provides a good idea for the construction of bimolecular-activated detection switch in the future.

Liposome fusogenic enzyme-free circuit enables high-fidelity determination of single exosomal RNA.

Accurate determination of single exosomal inclusions in situ presents a significant challenge due to their extremely low abundance as well sub-100 nm vesicle dimensions. Here, we created a Liposome Fusogenic Enzyme-free circuit (LIFE) approach for the high-fidelity identification of exosome-encapsulated cargoes without destroying the vesicle integrity. The probe-loaded cationic fusogenic liposome could capture and fuse with a single target exosome, enabling probes delivery and target biomolecule-initiated cascaded signal amplification in situ. Then the DNAzyme probe encountered conformal change upon exosomal microRNA activation, and generated a convex DNAzyme structure to cleave the RNA site of substrate probe. After that, the target microRNA could be released to introduce a cleavage cycle to yield amplified fluorescence readout. Therefore, trace cargoes in a single exosome could be accurately determined by elaborately controlling the ratio of introduced LIFE probe, paving the way toward the exploration of a universal sensing platform for the assessment of exosomal cargoes to facilitate early disease diagnosis and personalized treatment.

Intelligent Dual-Lock Deoxyribonucleic Acid Automatons Boosting Precise Tumor Imaging.

An early and accurate cancer diagnosis holds the potential to improve treatment and prognosis. Nevertheless, the complexity of the biological system limits the selectivity of existing approaches and makes tumor imaging in vivo particularly challenging. In this study, tumor-specific fluorescence imaging was achieved by building intelligent dual-lock deoxyribonucleic acid automatons (IDEAs) that employed a DNA walking system standing on ZrMOF@MnO2 multifunctional nanocomposites for controllable molecular recognition. The IDEAs exhibited significantly enhanced fluorescence signals only in the coexistence of both miRNA and GSH of tumor cells, enabling accurate distinguishing of tumor cells from healthy ones. Furthermore, the feasibility and specificity of IDEAs were also validated in vivo with tumor bearing mice successfully. This work highlights the potential of the proposed IDEA strategy for tumor-specific imaging, paving the way for successful precision diagnosis and treatment.

Cancer-Derived Small Extracellular Vesicles PICKER.

Cancer-derived small extracellular vesicles (csEVs) play critical roles in the genesis and development of various cancers. However, accurate detection of low-abundance csEVs remains particularly challenging due to the complex clinical sample composition. In the present study, we constructed a Programmable Isothermal Cascade Keen Enzyme-free Reporter (PICKER) for the reliable detection and acquisition of the relative abundance of csEVs in total sEVs (tsEVs) by integrating dual-aptamer recognition (cancer-specific protein EpCAM and tetraspanin protein CD63) with a catalytic hairpin assembly (CHA) amplification. By employing this strategy, we were able to achieve a detection limit of 420 particles/µL csEVs. Particularly, we proposed a novel particle ratio index of csEV against tsEV (PRcsEV/tsEV) to greatly eliminate errors from inconsistent centrifugation, which was calculated from the fluorescence ratio produced by csEVs and tsEVs. The PICKER showed a 1/10,000 discrimination capability by successfully picking out 1.0 × 103 csEV from 1.0 × 107 tsEV per microliter. We also found that the PRcsEV/tsEV value increased proportional to the stages of breast cancer by analyzing EVs from clinical patients' plasma. Taken together, we established a PICKER strategy capable of accurately discriminating csEVs, and the proposed PRcsEV/tsEV had been proven a potential indicator of breast cancer staging, paving the way toward facilitating cancer diagnosis and precision therapeutics.

Ultracentrifugation-Free Enrichment and Quantification of Small Extracellular Vesicles.

Cancer is a malignant tumor with the highest mortality of human diseases. The early diagnosis of cancer can greatly reduce its mortality. Ultracentrifugation is the most commonly employed technique to separate small extracellular vesicles (sEVs) due to their small size and rare abundance, but the low separation efficiency is a major concern. Herein, we proposed a DNAzyme-triggered assembly and disassembly system that converted single nano-sized sEVs into clusters that could be conveniently enriched by ordinary centrifugation and then be broken into single sEVs in the presence of magnesium ions. The simultaneous quantification of sEVs was realized by recording the increase in fluorescence upon nucleic acid cleavage, and a detection limit as low as 54 particles/µL was achieved. The whole analytical procedure could be completed in 1.5 h without the assistance of ultracentrifugation. Efficient enrichment and accurate quantification of sEVs are enabled through the proposed approach, broadening the potentials of sEVs in biological science, biomedical engineering, and personalized medicine.