Light-Enhanced Hypoxia-Responsive Gene Editing for Hypoxia-Resistant Photodynamic and Immunotherapy.

Hypoxia is a key hallmark of solid tumors and can cause resistance to various treatments such as photodynamics and immunotherapy. Microenvironment-responsive gene editing provides a powerful tool to overcome hypoxia resistance and remodel hypoxic microenvironments for enhanced tumor therapy. Here, a light-enhanced hypoxia-responsive multifunctional nanocarrier is developed to perform spatiotemporal specific dual gene editing for enhanced photodynamic and immunotherapy in breast cancer. As a gated molecule of nanocarrier, the degradation of azobenzene moieties under hypoxic conditions triggers controllable release of Cas9 ribonucleoprotein in hypoxic site of the tumor. Hyaluronic acid is conjugated with chloramine e6 to coat mesoporous silica nanoparticles for targeted delivery in tumors and generation of high levels of reactive oxygen species, which can result in increased hypoxia levels for effective cleavage of azobenzene bonds to improve gene editing efficiency and reduce toxic side effects with light irradiation. Moreover, dual targeting HIF-1α and PD-L1 in the anoxic microenvironments can overcome hypoxia resistance and remodel immune microenvironments, which reduces tumor plasticity and resistance to photodynamic and immunotherapy. In summary, a light-enhanced hypoxia responsive nanocomposite is developed for controllable gene editing which holds great promise for synergistic hypoxia-resistant photodynamic and immunotherapy.

Bioorthogonal Reaction-Mediated Tumor-Selective Delivery of CRISPR/Cas9 System for Dual-Targeted Cancer Immunotherapy.

CRISPR system-assisted immunotherapy is an attractive option in cancer therapy. However, its efficacy is still less than expected due to the limitations in delivering the CRISPR system to target cancer cells. Here, we report a new CRISPR/Cas9 tumor-targeting delivery strategy based on bioorthogonal reactions for dual-targeted cancer immunotherapy. First, selective in vivo metabolic labeling of cancer and activation of the cGAS-STING pathway was achieved simultaneously through tumor microenvironment (TME)-biodegradable hollow manganese dioxide (H-MnO2 ) nano-platform. Subsequently, CRISPR/Cas9 system-loaded liposome was accumulated within the modified tumor tissue through in vivo click chemistry, resulting in the loss of protein tyrosine phosphatase N2 (PTPN2) and further sensitizing tumors to immunotherapy. Overall, our strategy provides a modular platform for precise gene editing in vivo and exhibits potent antitumor response by boosting innate and adaptive antitumor immunity.

Cuproptosis-related gene CDKN2A as a molecular target for IPF diagnosis and therapeutics.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a progressive chronic interstitial lung disease with limited therapeutic options. Cuproptosis is a recently proposed novel form of programmed cell death, which has been strongly implicated in the development of various human diseases. However, the prognostic and therapeutic value of cuproptosis-related genes (CRGs) in IPF remains to be elucidated. METHODS: In the present study, weighted gene co-expression network analysis (WGCNA) was employed to identify the key CRGs associated with the development of IPF. The subsequent GSEA, immune cell correlation analysis, and single-cell RNA-Seq analysis were conducted to explore the potential role of the identified CRGs in IPF. In addition, ROC curves and survival analysis were used to assess the prognostic value of the key CRGs in IPF. Moreover, we explored the molecular mechanisms of participation of identified key CRGs in the development of pulmonary fibrogenesis through in vivo and in vitro experiments. RESULTS: The expression level of cyclin-dependent kinase inhibitor 2A (CDKN2A) is upregulated in the lung tissues of IPF patients and associated with disease severity. Notably, CDKN2A was constitutively expressed by fibrosis-promoting M2 macrophages. Decreased CDKN2A expression sensitizes M2 macrophages to elesclomol-induced cuproptosis in vitro. Inhibition of CDKN2A decreases the number of viable macrophages and attenuates bleomycin-induced pulmonary fibrosis in mice. CONCLUSION: These findings indicate that CDKN2A mediates the resistance of fibrosis-promoting M2 macrophages to cuproptosis and promotes pulmonary fibrosis in mice. Our work provides fresh insights into CRGs in IPF with potential value for research in the pathogenesis, diagnosis, and a new therapy strategy for IPF.

Biogenic materials for CRISPR delivery and therapeutics.

CRISPR, as an emerging gene-editing technology, has been widely used in multidisciplinary fields, including genetic diseases and some cancers. However, it remains a challenge to efficiently deliver CRISPR for safe and efficient genome editing. Currently, biomimetic materials have become an attractive delivery strategy for CRISPR-mediated genome editing due to their low immunogenicity and application safety. The biomimetic materials delivery is involved in the improvement of cellular uptake of nanoparticle vectors, and the gene editing efficiency. In this review, we summarize the current delivery strategies of CRISPR/Cas systems based on biogenic materials such as viruses, bacteria, cells, bioactive substances, etc., focusing on the potential applications in disease research and therapy. Finally, the prospects and limitations of CRISPR-based systems in therapeutics are discussed.

Tumor Microenvironment-Responsive Nanoherb Delivery System for Synergistically Inhibition of Cancer Stem Cells.

Multidrug resistance in cancer stem cells (CSCs) is a major barrier to chemotherapy; hence, developing CSC-specific targeted nanocarriers for efficient drug delivery is critical. In this study, monodisperse hollow-structured MnO2 (H-MnO2) with a mesoporous shell was created for efficient targeted drug delivery. An effective therapeutic compound isoliquiritigenin (ISL) was confirmed to inhibit the lung cancer stem-cell phenotype by natural compound screening based on integrated microfluidic devices. The resultant H-MnO2 showed a high drug-loading content of the potent CSC-targeting compound ISL and near-infrared fluorescent dye indocyanine green (ICG). In addition, H-MnO2 was successively modified with hyaluronic acid (HA) to enhance targeting CSCs with high CD44 expression levels. The H-MnO2@(ICG + ISL)@HA nanocomposites displayed promising chemotherapeutic and photothermal treatment capabilities, as well as NIR-triggered drug release, which showed excellent CSC-killing effects and tumor inhibition efficacy. Meanwhile, the development of the tumor was effectively restrained by NIR-triggered phototherapy and prominent chemotherapy without obvious side effects after tail vein injection of the nanocomposites in vivo. In summary, the prepared nanocomposites accomplished synergistic cancer therapy that targets CSCs, offering a versatile platform for lung cancer diagnosis and treatment.

Na, K-ATPase α1 cooperates with its endogenous ligand to reprogram immune microenvironment of lung carcinoma and promotes immune escape.

Dysregulated endocrine hormones (EHs) contribute to tumorigenesis, but how EHs affect the tumor immune microenvironment (TIM) and the immunotherapy of non-small cell lung cancer (NSCLC) is still unclear. Here, endogenous ouabain (EO), an adrenergic hormone, is elevated in patients with NSCLC and closely related to tumor pathological stage, metastasis, and survival. EO promotes the suppression of TIM in vivo by modulating the expression of immune checkpoint proteins, in which programmed cell death protein ligand 1 (PD-L1) plays a major role. EO increases PD-L1 transcription; however, the EO receptor Na- and K-dependent adenosine triphosphatase (Na, K-ATPase) α1 interacts with PD-L1 to trigger the endocytic degradation of PD-L1. This seemingly contradictory result led us to discover the mechanism whereby EO cooperates with Na, K-ATPase α1 to finely control PD-L1 expression and dampen tumoral immunity. In conclusion, the Na, K-ATPase α1/EO signaling facilitates immune escape in lung cancer, and manipulation of this signaling shows great promise in improving immunotherapy for lung adenocarcinoma.

Customizable trajectory of trapped particle in quadruple-beam optical trap.

We have presented and demonstrated a customizable trajectory of a trapped particle in the Quadruple-beam optical trap. The orbital motion of the trapped microsphere was realized by modulating the trapping power. The motion trajectories could be designed by adjusting the modulation frequency, amplitude, and phase. By using this method, we have realized the triangle, bowknot, ellipse, straight line, and hooklike trajectories. The motion frequencies and circumferences were also modulated. The customizable trajectory in the optical trap may result in more possibilities for directional movement, microfluidic mixing, driven machines, and even painting freely.

Simulations and Experiments on the Vibrational Characteristics of Cylindrical Resonators with First Three Harmonic Errors.

A cylindrical resonator gyroscope is a kind of Coriolis gyroscope, which measures angular velocity or angle via processing of the standing wave. The symmetry of a cylindrical resonator is destroyed by different degrees of geometric nonuniformity and structural damage in the machining process. The uneven mass distribution caused by the asymmetry of the resonator can be expressed in the form of a Fourier series. The first three harmonics will reduce the anti-interference ability of the resonator to the external vibration, as well as increase the angular random walk and zero-bias drift of the gyroscope. In this paper, the frequency split of different modes caused by the first three harmonic errors and the displacement of the center of the cylindrical resonator bottom plate are obtained by simulation, and the relationship between them is explored. The experimental results on five fused silica cylindrical resonators are consistent with the simulation, confirming the linear relationship between the n = 1 frequency split and second harmonic error. A method for evaluating the first three harmonic errors of fused silica cylindrical resonators is provided.

A High-Precision Method of Stiffness Axes Identification for Axisymmetric Resonator Gyroscopes.

Axisymmetric resonators are key elements of Coriolis vibratory gyroscopes (CVGs). The performance of a CVG is closely related to the stiffness and damping symmetry of its resonator. The stiffness symmetry of a resonator can be effectively improved by electrostatic tuning or mechanical trimming, both of which need an accurate knowledge of the azimuth angles of the two stiffness axes of the resonator. Considering that the motion of a non-ideal axisymmetric resonator can be decomposed as two principal oscillations with two different natural frequencies along two orthogonal stiffness axes, this paper introduces a novel high-precision method of stiffness axes identification. The method is based on measurements of the phase difference between the signals detected at two orthogonal sensing electrodes when an axisymmetric resonator is released from all the control forces of the force-to-rebalance mode and from different initial pattern angles. Except for simplicity, our method works with the eight-electrodes configuration, in no need of additional electrodes or detectors. Furthermore, the method is insensitive to the variation of natural frequencies and operates properly in the cases of either large or small frequency splits. The introduced method is tested on a resonator gyroscope, and two stiffness axes azimuth angles are obtained with a resolution better than 0.1°. A comparison of the experimental results and theoretical model simulations confirmed the validity of our method.

The Rb-Xe coupling effect in optically pumped NMR gyroscopes.

Optically pumped nuclear magnetic resonance (NMR) gyroscope has been widely concerned as a future gyroscope with great development potential. In this study, the Rb-Xe coupling effect in this gyroscope between the alkali metal magnetometer and the noble gas NMR is investigated. The theoretical formulae about this effect are obtained based on Bloch equation and demonstrated by experiments. In order to see the influence of this effect on the gyro signal, the Xe NMR frequency changes with the carrier frequency and demodulation phase of the Rb magnetometer are studied with the deduced formulae. More importantly, theoretical analysis shows that this coupling effect can also significantly reduce the efficiency of the dual species scheme, which is used to suppress the influence of magnetic fluctuation. Therefore, to remove the Rb-Xe coupling effect, a simple and preliminary decoupling scheme operated by locking the phase difference to zero is proposed and demonstrated. All this work is of great significance to the improvement of the NMR gyro performance and future applications.

Broad ultra-potent neutralization of SARS-CoV-2 variants by monoclonal antibodies specific to the tip of RBD.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) continue to wreak havoc across the globe. Higher transmissibility and immunologic resistance of VOCs bring unprecedented challenges to epidemic extinguishment. Here we describe a monoclonal antibody, 2G1, that neutralizes all current VOCs and has surprising tolerance to mutations adjacent to or within its interaction epitope. Cryo-electron microscopy structure showed that 2G1 bound to the tip of receptor binding domain (RBD) of spike protein with small contact interface but strong hydrophobic effect, which resulted in nanomolar to sub-nanomolar affinities to spike proteins. The epitope of 2G1 on RBD partially overlaps with angiotensin converting enzyme 2 (ACE2) interface, which enables 2G1 to block interaction between RBD and ACE2. The narrow binding epitope but high affinity bestow outstanding therapeutic efficacy upon 2G1 that neutralized VOCs with sub-nanomolar half maximal inhibitory concentration in vitro. In SARS-CoV-2, Beta or Delta variant-challenged transgenic mice and rhesus macaque models, 2G1 protected animals from clinical illness and eliminated viral burden, without serious impact to animal safety. Mutagenesis experiments suggest that 2G1 is potentially capable of dealing with emerging SARS-CoV-2 variants in the future. This report characterized the therapeutic antibodies specific to the tip of spike against SARS-CoV-2 variants and highlights the potential clinical applications as well as for developing vaccine and cocktail therapy.

Synergistic photothermal cancer immunotherapy by Cas9 ribonucleoprotein-based copper sulfide nanotherapeutic platform targeting PTPN2.

Photothermal therapy (PTT) is a promising strategy for the treatment of advanced malignant neoplasm. However, the anti-tumor efficacy by PTT alone is insufficient to control tumor growth and metastasis. Here, we report a multifunctional nanotherapeutic system exerting a combined PTT and immunotherapy to synergistically enhance the therapeutic effect on melanoma. In particular, we selected the semiconductor nanomaterial copper sulfide (CuS), which served not only as a near-infrared (NIR) light-triggered photothermal converter for tumor hyperthermia but as a basic carrier to modify Cas9 ribonucleoprotein targeting PTPN2 on its surface. Efficient PTPN2 depletion was observed after the treatment of CuS-RNP@PEI nanoparticles, which caused the accumulation of intratumoral infiltrating CD8 T lymphocytes in tumor-bearing mice and upregulated the expression levels of IFN-ᵧ and TNF-α in tumor tissue, thus sensitizing tumors to immunotherapy. In addition, the effect worked synergistically with tumor ablation and immunogenic cell death (ICD) induced by PTT to amplify anti-tumor efficacy. Taken together, this exogenously controlled method provides a simple and effective treatment option for advanced malignant neoplasm.

Dual-beam intracavity optical tweezers with all-optical independent axial and radial self-feedback control schemes.

The feedback control to optical tweezers is an obvious approach to improve the optical confinement. However, the electronic-based feedback controlling system in optical tweezers usually consists of complex software and hardware, and its performance is limited by the inevitable noise and time-delay from detecting and controlling devices. Here, we present and demonstrate the dual-beam intracavity optical tweezers enabling all-optical independent radial and axial self-feedback control of the trapped particle's radial and axial motions. We have achieved the highest optical confinement per unit intensity to date, to the best of our knowledge. Moreover, both the axial and radial confinements are adjustable in real-time, through tuning the foci offset of the clockwise and counter-clockwise beams. As a result, we realized three-dimensional self-feedback control of the trapped particle's motions with an equivalent level in the experiment. The dual-beam intracavity optical tweezers will significantly expand the range of optical manipulation in further studies of biology, physics and precise measurement, especially for the sample that is extremely sensitive to heat.

Experimental Study on Variation of Surface Roughness and Q Factors of Fused Silica Cylindrical Resonators with Different Grinding Speeds.

For the axisymmetric shell resonator gyroscopes, the quality factor (Q factor) of the resonator is one of the core parameters limiting their performances. Surface loss is one of the dominating losses, which is related to the subsurface damage (SSD) that is influenced by the grinding parameters. This paper experimentally studies the surface roughness and Q factor variation of six resonators ground by three different grinding speeds. The results suggest that the removal of the SSD cannot improve the Q factor continuously, and the variation of surface roughness is not the dominant reason to affect the Q factor. The measurement results indicate that an appropriate increase in the grinding speed can significantly improve the surface quality and Q factor. This study also demonstrates that a 20 million Q factor for fused silica cylindrical resonators is achievable using appropriate manufacturing processes combined with post-processing etching, which offers possibilities for developing high-precision and low-cost cylindrical resonator gyroscopes.

Hypoxia-Responsive Gene Editing to Reduce Tumor Thermal Tolerance for Mild-Photothermal Therapy.

Near-infrared (NIR)-light-triggered photothermal therapy (PTT) is usually associated with undesirable damage to healthy organs nearby due to the high temperatures (>50 °C) available for tumor ablation. Low-temperature PTT would therefore have tremendous value for clinical application. Here, we construct a hypoxia-responsive gold nanorods (AuNRs)-based nanocomposite of CRISPR-Cas9 for mild-photothermal therapy via tumor-targeted gene editing. AuNRs are modified with azobenzene-4,4'-dicarboxylic acid (p-AZO) to achieve on-demand release of CRISPR-Cas9 using hypoxia-responsive azo bonds. In the hypoxic tumor microenvironment, the azo groups of the hypoxia-activated CRISPR-Cas9 nanosystem based on gold nanorods (APACPs) are selectively reduced by the overexpression of reductases, leading to the release of Cas9 and subsequent gene editing. Owing to the knockout of HSP90α for reducing the thermal resistance of cancer cells, highly effective tumor ablation both in vitro and in vivo was achieved with APACPs under mild PTT.

A Novel Method for Estimating and Balancing the Second Harmonic Error of Cylindrical Fused Silica Resonators.

The cylindrical resonator gyroscope (CRG) is a type of Coriolis vibratory gyroscope which measures the angular velocity or angle through the precession of the elastic wave of the cylindrical resonator. The cylindrical fused silica resonator is an essential component of the CRG, the symmetry of which determines the bias drift and vibration stability of the gyroscope. The manufacturing errors breaking the symmetry of the resonator are usually described by Fourier series, and most studies are only focusing on analyzing and reducing the fourth harmonic error, the main error source of bias drift. The second harmonic error also is one of the obstacles for CRG towards high precision. Therefore, this paper provides a chemical method to evaluate and balance the second harmonic error of cylindrical fused silica resonators. The relation between the frequency split of the n = 1 mode and the second harmonic error of the resonator is obtained. Simulations are performed to analyze the effects of the first three harmonic errors on the frequency splits. The relation between the location of the low-frequency axis of n = 1 mode and the heavy axis of the second harmonic error is also analyzed by simulation. Chemical balancing experiments on two fused silica resonators demonstrate the feasibility of this balancing procedure, and show good consistency with theoretical and simulation analysis. The second harmonic error of the two resonators is reduced by 86.6% and 79.8%, respectively.

Optical confinement efficiency in the single beam intracavity optical tweezers.

Single beam intracavity optical tweezers characterizes a novel optical trapping scheme where the laser operation is nonlinearly coupled to the motion of the trapped particle. Here, we first present and establish a physical model from a completely new perspective to describe this coupling mechanism, using transfer matrices to calculate the loss of the free-space optical path and then extracting the scattering loss that caused by the 3D motions of the particle. Based on this model, we discuss the equilibrium position in the single beam intracavity optical tweezers. The influences of the numerical aperture, pumping power, particle radius and refractive index on the optical confinement efficiency are fully investigated, compared with standard optical tweezers. Our work is highly relevant for guiding the experiments on the single beam intracavity optical tweezers to achieve higher optical confinement efficiency.

A 5.86 Million Quality Factor Cylindrical Resonator with Improved Structural Design Based on Thermoelastic Dissipation Analysis.

The cylindrical resonator is the core component of cylindrical resonator gyroscopes (CRGs). The quality factor (Q factor) of the resonator is one crucial parameter that determines the performance of the gyroscope. In this paper, the finite element method is used to theoretically investigate the influence of the thermoelastic dissipation (TED) of the cylindrical resonator. The improved structure of a fused silica cylindrical resonator is then demonstrated. Compared with the traditional structure, the thermoelastic Q (QTED) of the resonator is increased by 122%. In addition, the Q factor of the improved cylindrical resonator is measured, and results illustrate that, after annealing and chemical etching, the Q factor of the resonator is significantly higher than that of the cylindrical resonators reported previously. The Q factor of the cylindrical resonator in this paper reaches 5.86 million, which is the highest value for a cylindrical resonator to date.

Angiotensin II contributes to intratumoral immunosuppressionvia induction of PD-L1 expression in non-small cell lung carcinoma.

The formation of an immunosuppressive microenvironment and up-regulation of PD-L1 protein are the main causes of tumor immune escape. Previous reports suggest that Angiotensin II (Ang II) can modulate the immune status of tumor microenvironment in non-small cell lung cancer (NSCLC), but the underlying mechanism remains not fully understood. Here we demonstrated that AngII treatment causes the reduction of intratumoral infiltrating CD4 T lymphocytes in tumor-bearing mice, increases the accumulation of immunosuppressive granulocytes and TAMs in tumor tissue, and upregulates the expression levels of immunosuppressive marker genes. In addition, AngII/AGTR1 axis triggers cell PD-L1 expression through a mechanism involving increases in PD-L1 mRNA stability by human antigen R (HuR), an AU-rich element (ARE)-binding protein. Collectively, AngII/AGTR1 signaling promotes the tumor immunosuppressive microenvironment by upregulating PD-L1 in NSCLC, the mechanism of which is largely accounted by HuR-mediated PD-L1 mRNA stabilization.