Targeted PLK1 suppression through RNA interference mediated by high-fidelity Cas13d mitigates osteosarcoma progression via TGF-ß/Smad3 signalling.

Osteosarcoma is the most common primary bone malignancy in children and adolescents. Overexpression of polo-like kinase 1 (PLK1) is frequent in osteosarcoma and drives disease progression and metastasis, making it a promising therapeutic target. In this study, we explored PLK1 knockdown in osteosarcoma cells using RNA interference mediated by high-fidelity Cas13d (hfCas13d). PLK1 was found to be significantly upregulated in osteosarcoma tumour tissues compared to normal bone. sgRNA-mediated PLK1 suppression via hfCas13d transfection inhibited osteosarcoma cell proliferation, induced G2/M cell cycle arrest, promoted apoptosis, reduced cell invasion and increased expression of the epithelial marker E-cadherin. Proximity labelling by TurboID coupled with co-immunoprecipitation identified novel PLK1 interactions with Smad3, a key intracellular transducer of TGF-ß signalling. PLK1 knockdown impaired Smad2/3 phosphorylation and modulated TGF-ß/Smad3 pathway inactivation. Finally, in vivo delivery of hfCas13d vectors targeting PLK1 substantially attenuated osteosarcoma xenograft growth in nude mice. Taken together, this study highlights PLK1 as a potential therapeutic target and driver of disease progression in osteosarcoma. It also demonstrates the utility of hfCas13d-mediated gene knockdown as a strategy for targeted therapy. Further optimization of PLK1 suppression approaches may ultimately improve clinical outcomes for osteosarcoma patients.

Photonic Gauge Potential in One Cavity with Synthetic Frequency and Orbital Angular Momentum Dimensions.

We explore a single degenerate optical cavity supporting a synthetic two-dimensional space, which includes the frequency and the orbital angular momentum (OAM) axes of light. We create the effective gauge potential inside this synthetic space and show that the system exhibits topologically protected one-way edge states along the OAM axis at the boundaries of the frequency dimension. In this synthetic space, we present a robust generation and manipulation of entanglement between the frequency and OAM of photons. Our Letter shows that a higher-dimensional synthetic space involving multiple degrees of freedom of light can be achieved in a "zero-dimensional" spatial structure, pointing towards a unique platform to explore topological photonics and to realize potential applications in optical communications and quantum information processing.

Migration of gastric cancer is suppressed by recombinant Newcastle disease virus (rL-RVG) via regulating α7-nicotinic acetylcholine receptors/ERK- EMT.

BACKGROUND: Nicotinic acetylcholine receptors (nAChRs) have been reported to be overexpressed in malignancies in humans and is associated with tumorigenesis and cell migration. In previous studies of gastric cancer, alpha7 nicotinic acetylcholine receptor (α7-nAChR) overexpression leads to epithelial-mesenchymal transition (EMT) and promotes the migration of gastric cancer cells. Recombinant avirulent LaSota strain of Newcastle disease virus (NDV) expressing the rabies virus glycoprotein (rL-RVG) may promote apoptosis of gastric cancer cells and reduces the migration of lung cancer metastasis. However, whether rL-RVG inhibits migration of gastric cancer cells and what the underlying functional mechanism is remains unknown. METHODS: The gastric cancer cell lines BGC and SGC were randomly divided into 3 groups: rL-RVG, NDV and Phosphate Buffered Solution (PBS) control groups. Furthermore,we adopted ACB and MLA,α7nAChR-siRNA for the overexpression and silencing of α7-nAChR.Corynoxenine was used for inhibiting the MEK-ERK pathway. Western blot, Immunofluoresce,cell proliferation assays,cell migration analyses through wound-healing assays and Transwell assays were used to explore the underlying mechanisms. A mouse xenograft model was used to investigate the effects of rL-RVG,NDV on tumor growth. RESULTS: In this study, our findings demonstrate that rL-RVG suppressed the migration of gastric cancer cells and reduced EMT via α7-nAChR in vitro. Furthermore rL-RVG decreased the phosphorylation levels of the MEK/ERK signaling pathway such as down-regulating the expression of P-MEK and P-ERK. Additionally, rL-RVG also reduced the expression level of mesenchymal markers N-cadherin and Vimentin and enhanced the expression of the epithelial marker E-cadherin. Lastly, rL-RVG inhibited nicotinic acetylcholine receptors (nAChRs) to suppress cell migration and epithelial to mesenchymal transition (EMT) in gastric cell. We also found that rL-RVG suppresses the growth of gastric cancer subcutaneous tumor cells in vivo. CONCLUSION: rL-RVG inhibits α7-nAChR-MEK/ERK-EMT to suppress migration of gastric cancer cells.

LaSota Strain Expressing The Rabies Virus Glycoprotein (rL-RVG) Suppresses Gastric Cancer by Inhibiting the Alpha 7 Nicotinic Acetylcholine Receptor (α7 nAChR)/Phosphoinositide 3-Kinase (PI3K)/AKT Pathway.

BACKGROUND The recombinant avirulent Newcastle disease virus (NDV) LaSota strain expressing the rabies virus glycoprotein (rL-RVG) can induce much greater apoptosis than can NDV in gastric carcinoma cells, but the mechanisms involved remains unclear. MATERIAL AND METHODS The 2 gastric carcinoma cell lines were divided into the rL-RVG group, the NDV group, and the PBS group. MTT assay was used to detect and analyze cell viability. siRNA for alpha7-nAChR, alpha7-nAChR antagonist, or alpha7-nAChR agonist, AKT antagonist, and p-AKT agonist were used for pretreatment. The protein expressions of RVG, NDV, alpha7-nAChR, cleaved caspase-3, p-AKT, PI3K, Bcl-2, and Bax proteins were detected by Western blot assay. Immunofluorescence was used to detect expressions of alpha7-nAChR proteins. Light microscopy, flow cytometry, and TUNEL assay were used to assess apoptosis. RESULTS The results showed that 2 virus concentrations over 10³ dilution caused greater cell proliferation inhibition. rL-RVG treatment increased the expression of alpha7-nAChR, cleaved caspase-3, and Bax protein but decreased the expression of p-AKT, PI3K, and Bcl-2 protein. When the groups were pretreated with alpha7-nAChR antagonist, the alpha7-nAChR, cleaved caspase-3, and Bax protein expression increased, but the expression of p-AKT, PI3K, and Bcl-2 protein was clearly decreased. However, the results in the alpha7-nAChR agonist group were the opposite. When treated with the AKT antagonist, the result was the same as in the rL-RVG treatment group. The result in the AKT agonist group was the opposite of that in the AKT antagonist group. Compared with the NDV group, the results of light microscopy, FCM, and TUNEL assay showed that alpha7-nAChR antagonist significantly affected the apoptosis of gastric cancer cells in the rL-RVG group. CONCLUSIONS rL-RVG leads to much greater apoptosis through the alpha7-nAChR/PI3K/AKT pathway.

Frozen Condition of Quantum Coherence for Atoms on a Stationary Trajectory.

The quantum coherence (QC) of two comoving atoms on a stationary trajectory is investigated. We develop a formalism to characterize the properties of atoms on a stationary trajectory. We give a criterion under which QC is frozen to a nonzero value. The frozen condition that vanishing super- or subradiant decay rate is not so sensitive to the initial condition of state. We show that enhanced QC and a subradiant state can be gained from the initial state.

Small Ceria Nanoclusters with High ROS Scavenging Activity and Favorable Pharmacokinetic Parameters for the Amelioration of Chronic Kidney Disease.

The over production of reactive oxygen species (ROS) plays a critical role in the progression of chronic kidney disease (CKD). Organic ROS scavengers currently used for CKD treatment do not satisfy low dosage and high efficiency requirements. Ceria nanomaterials featured with renewable ROS scavenging activity are potential candidates for CKD treatment. Herein, a method for the synthesis of ceria nanoclusters (NCs) featured with small size of ≈1.2 nm is reported. The synthesized NCs are modified by three hydrophilic ligands with different molecular weights, including succinic acid (SA), polyethylene glycol diacid 600 (PEG600), and polyethylene glycol diacid 2000 (PEG2000). The surface modified NCs exhibit excellent ROS scavenging activity due to the high Ce3+ /Ce4+ ratio in their crystal structures. Compared with bigger-sized ceria nanoparticles (NPs) (≈45 nm), NCs demonstrate smoother blood concentration-time curve, lower organ accumulation, and faster metabolic rate superiorities. The administration of NCs to CKD mice, especially PEG600 and PEG2000 modified NCs, can effectively inhibit oxidative stress, inflammation, renal fibrosis, and apoptosis in their kidneys. Due to these benefits, the constructed NCs can ameliorate the progression of CKD. These findings suggest that NCs is a potential redox nanomedicine for future clinical treatment of CKD.

Overexpression of miR-338-5p in exosomes derived from mesenchymal stromal cells provides neuroprotective effects by the Cnr1/Rap1/Akt pathway after spinal cord injury in rats.

Growing evidence has shown that microRNAs (miRNAs) play crucial roles in the physiopathology of spinal cord injury (SCI). Recent studies have confirmed that miR-338-5p regulates myelination, suggesting a potential role in the treatment of SCI. However, the molecular mechanism of miR-338-5p on SCI is still unknown. Recently, exosomes have emerged as an ideal vector to deliver therapeutic molecules such as miRNAs. Here, we explored the effects of miR-338-5p-overexpressing exosomes derived from bone marrow-derived mesenchymal stromal cells (BMSCs) on SCI. In vivo, a model of contusion SCI in rats was established, and we observed that overexpression of miR-338-5p in exosomes profoundly increased the expression levels of neurofilament protein-M and growth-associated protein-43 and decreased those of myelin-associated glycoprotein and glial fibrillary acidic protein, which provided neuroprotective effects after acute SCI. In an in vitro study, we found that overexpression of miR-338-5p in exosomes repressed cell apoptosis following H2O2-induced oxidative stress injury in PC12 cells. Additionally, we confirmed that cannabinoid receptor 1 (Cnr1) was the target gene of miR-338-5p by dual-luciferase reporter assays and that Rap1 was the downstream gene by the KEGG pathway analysis. We found that miR-338-5p increased cAMP accumulation as a consequence of downregulated expression of the target gene Cnr1, and then, Rap1 was activated by cAMP. Eventually, the activation of the PI3K/Akt pathway attenuated cell apoptosis and promoted neuronal survival by cAMP-mediated Rap1 activation. In brief, these findings showed that exosomes overexpressing miR-338-5p were a promising treatment strategy for SCI.

Circular RNA circAFF2 accelerates gastric cancer development by activating miR-6894-5p and regulating ANTXR 1 expression.

BACKGROUND: Circular RNAs (circRNAs) contain a new class of non-coding RNAs that play an important role in adjusting biological function and gene expression. But the function of circRNAs in gastric cancer remains unclear. In the present research, we explored the functions of circular RNA AFF2(circAFF2, hsa_circ_0001947) in gastric cancer cells and an animal model of gastric cancer. METHODS: The expression of circAFF2, microRNA-6894-5p (miR-6894-5p), and Anthrax toxin receptor 1 (ANTXR 1) were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell counting kit 8 (CCK-8) and transwell assays were used to analyze the knockdown effects of circAFF2, miR-6894-5p, and overexpression of ANTXR 1 on cell proliferation, migration, and invasion abilities. Binding interactions between, circAFF2 and miR-6894-5p and between, miR-6894-5p and ANTXR 1 were detected by Dual-luciferase reporter assays. Levels of protein expression were analyzed by Western blotting. Tumor models were established by subcutaneous injection of tumor cells in nude mice. RESULT: The result showed that circAFF2 expression was significantly increased in gastric cancer cell lines and tissues. The knockdown of circAFF2 dramatically suppressed the cell migration, invasion and proliferation of gastric cancer cells. In vivo studies showed that knockdown of circAFF2 delayed tumor growth. Furthermore, we revealed that circAFF2 functioned as a sponge to absorb miR-6984-5p and elevated the expression of ANTXR 1. CONCLUSION: CircAFF2 acts as an oncogene in gastric cancer and exerts its effects via miR-6894-5p/ANTXR 1 signaling.

Recombinant Newcastle disease virus expressing human IFN-λ1 (rL-hIFN-λ1) inhibits lung cancer migration through repolarizating macrophage from M2 to M1 phenotype.

BACKGROUND: Tumor-associated macrophages (TAMs) are frequently infiltrated in tumor microenvironment and promote tumor progression. Lung cancer development largely depends upon the essential contributions from the TAMs which generally polarize into M2 TAMs and produce abundant anti-inflammatory factors and facilitate tumor development. The recombinant Newcastle disease virus expressing human IFN-λ1 (rL-hIFN-λ1) could regulate Th1/Th2 immune response to produce anti-tumor microenvironment. However, the interaction between rL-hIFN-λ1 and macrophages polarization remains unclear. METHODS: The THP-1 cells were used to construct the THP-1-M0, THP-1-M1, THP-1-M2 and THP-1-rL-hIFN-λ1 macrophage models. qRT-PCR and Immunofluorescence were used to detect the polarization phenotype of macrophage polarized by rL-hIFN-λ1. The inhibitory properties of THP-rL-hIFN-λ1 on A549 cells and H446 cells were determined by a Clonogenic assay, as well as scratch migration assays and Transwell were used to explore the capability of migration. Furthermore, the M1/M2 infiltration density in different clinical stages of lung cancer tissues were examined. RESULTS: It was showed that rL-hIFN-λ1 could induce normal macrophages to differentiate into THP-1-M1 macrophages. Meanwhile, rL-hIFN-λ1 could also direct THP-1-M2 macrophages polarization into THP-1-M1 macrophages. Supernatants from rL-hIFN-λl induced macrophages inhibited colony formation, migration and invasion of lung cancer cells in vitro which was similar to THP-1-M1 macrophages. Moreover, analysis of clinical tumor tissues indicated that M1-type macrophages decreased gradually with the development of the clinical stage of lung cancer. CONCLUSIONS: Therefore, rL-hIFN-λl induced significant suppression of primary lung tumor growth and spontaneous lung metastases through regulating macrophages function, and it was expected to become a new biological therapy for lung cancer.

Recombinant virus expressing hIFN-λ1 (rL-hIFN-λ1) has important effects on endoplasmic reticulum stress, autophagy and apoptosis in small cell lung cancer.

BACKGROUND: Small cell lung cancer (SCLC) is an aggressive tumor with a poor prognosis. Human IFN-λ1 (IL-29), belonging to the type III IFN family, captured increasing attention recently due to its crucial role in developing tumors. Recent studies have revealed that the recombinant Newcastle Disease Virus (NDV) expressing human IFN-λ1 (rL-hIFN-λ1) plays a critical role in the development of tumors. However, the role of rL-hIFN-λ1 in SCLC is still unknown. METHODS: We determined the concentration of the virus intervention, followed by successfully infection in virus. We also investigated the effects of rL-hIFN-λ1 on endoplasmic reticulum stress (ERS), apoptosis and autophagy in H446 cells, and explored the interaction among the three. RESULTS: We found that the ERS, autophagy and apoptosis related proteins were significantly upregulated after infected with rL-hIFN-λ1 or NDV. In addition, both 4-phenylbutyric acid (4-PBA) or 3-Methyladenine (3-MA) could downregulate the expression of related proteins which increased by rL-hIFN-λ1. Furthermore, we found that both B-cell lymphoma-2 (BCL-2) knockdown or Rapamycin (Rapa) could increase ERS, autophagy and apoptosis. CONCLUSIONS: Our findings suggest that rL-hIFN-λ1 can induce ERS, autophagy and apoptosis in SCLC H446 cells, particularly, autophagy plays an important role during this process. Furthermore, rL-hIFN-λ1 might provide a potential biological treatment target for lung cancer treatment.

Molecular modeling and rational design of hydrocarbon-stapled/halogenated helical peptides targeting CETP self-binding site: Therapeutic implication for atherosclerosis.

The human plasma cholesteryl ester transfer protein (CETP) collects triglycerides from very-/low-density lipoproteins (V/LDL) and exchanges them for cholesteryl esters from high-density lipoproteins (HDL), which has recognized as an important therapeutic target for atherosclerosis. The protein has a C-terminal amphipathic α-helix that serves as self-binding peptide to fulfill biological function by dynamically binding to/unbinding from its cognate site (termed self-binding site) in the same protein. Previously, we successfully derived and halogenated the helical peptide to competitively disrupt the self-binding behavior of CETP C-terminal tail. However, the halogenated peptides have only a limited affinity increase as compared to native helical peptide (∼3-fold), thus exhibiting only a moderate competitive potency. Here, instead of optimizing the direct intermolecular interaction of peptide with CETP self-binding site we attempt to further improve the peptide competitive potency by reducing its conformational flexibility with hydrocarbon-stapling technique. Computational analysis reveals that the helical peptide has large intrinsic disorder in unbound free state, which would incur a considerable entropy penalty upon rebinding to the self-binding site. All-hydrocarbon bridge is designed and optimized on native and halogenated peptides in terms of the helical pattern and binding mode of self-binding peptide. Dynamics simulation and circular dichroism indicate that the stapling can considerably reduce peptide disorder in free state. Energetics calculation and fluorescence assay conform that the binding affinity of stapled/halogenated peptides is improved substantially (by > 5-fold), thus exhibiting an effective competition potency with native peptide for the self-binding site. Structural examination suggests that the binding modes and nonbonded interactions of native and halogenated peptides are not influenced essentially due to the stapling.

Next-generation sequencing reveals hsa_circ_0058092 being a potential oncogene candidate involved in gastric cancer.

Gastric cancer is a serious problem for human health. As part of noncoding RNA, circular RNA (circRNA) plays a key role in the occurrence and development of malignant tumor. We used next generation sequencing technology to detect circRNA expression profiles in 5 paired human gastric cancer tissues. Then, bioinformatics analysis was carried out to analyze the function of dysregulated circRNAs. Hsa_circ_0058092 was selected as the object of follow-up analysis. After using the Cistrome DB dataset the data was used to predict specific transcription factors of hsa_circ_0058092. The relationship between hsa_circ_0058092 and PODXL was further validated using RT-PCR and immunohistochemical techniques. Survival data were collected using a Kaplan-Meier analysis of hsa_circ_0058092. We identified 319 aberrantly expressed circRNAs, Hsa_circ_0058092 was selected for our studies. Functional analysis of hsa_circ_0058092 revealed that it was related to metabolic processes. The prediction results suggested that hsa_circ_0058092 has a relationship with hsa-miR-4269 which could specifically bind to the PODXL sequence. Transcription factor CEBPB may regulate the transcription process of hsa_circ_0058092. The expression of hsa_circ_0058092 was positively correlated with PODXL expression. Immunohistochemical analysis of PODXL showed that the expression of PODXL protein in cancer tissues is higher than that in adjacent tissues. Kaplan-Meier analysis suggested that hsa_circ_0058092 was associated with survival of gastric cancer patients. All of these results showed that hsa_circ_0058092 was a potential oncogene.

RNA-Seq profiling of circular RNAs and potential function of hsa_circ_0002360 in human lung adenocarcinom.

CircRNAs have been identified play a key role in various different types of cancer. However, their role in lung adenocarcinoma remains unclear. In this study, we explored the specific circular transcriptome and characterized the circRNA expression profiles of five paired lung adenocarcinoma (LAC) tissues relative to adjacent normal tissues from LAC patients using next-generation sequencing (NGS). To illuminate circRNAs function, their gene targets were initially predicted before using, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses to further analyse the associated significant cell signalling pathways and functions. The potential interactions between circRNA-miRNA-mRNA were also investigated. Additionally, qRT-PCR assay, Western blot and Immunohistochemistry were performed to validate the differential expression of circRNA, microRNA and mRNA in the LAC group in comparison to the control group. Two-hundred-eighty-five dysregulated circular transcripts were found in LAC tissues, among which 102 and 183 were either up or down regulated, respectively. Our biological analysis suggested that the host genes of differentially expressed circRNAs targeted to cancer-related processes and mechanisms. The interaction maps of the circRNA-miRNA-target gene were constructed using Cytoscope. In further study, hsa_circ_0002360 was found to be the most significantly overexpressed circRNA in LAC tissues by interacting with miRNA and its corresponding mRNA. Our results showed that hsa_circ_0002360 was aberrantly and abundantly expressed and implicated in the development of LAC, suggesting a valuable therapeutic target for LAC treatment.