Advanced biology最新文献

Type I interferons （IFN-I） are key proteins in antiviral response and immunomodulation. Negative regulators avoid abnormal activation of the interferon pathway or overactivation of interferon-activating proteins through multiple mechanisms. Loss-of-function mutations in negative regulator genes lead to the development of a variety of type I interferonopathy. It is of great significance in theory and clinic to discover new negative regulators and study their functions, but there is no effective screening system at present. Here, a screening system is established through a resistance reporter, which is designed to be expressed only in cells with interferon pathway activation. In conjunction with CRISPR knockout library, cells are screened for interferon pathway activation due to knockout of negatively regulators, which led to the identification of negatively regulator candidate genes such as PCGF3/5, UCK2, and ITPKA. The expression products of these genes functioned by targeting MAVS and promoted EMCV (encephalomyocarditis virus) infection by inhibiting interferon pathway activation. This study deepens the understanding of the regulatory network of the interferon pathway and provides a new theoretical basis for the study of the pathogenesis of autoimmune diseases.

The invasive pest Halyomorpha halys (Hemiptera: Pentatomidae) poses a significant threat to agriculture and requires control methods beyond chemical pesticides. This study investigates RNA interference (RNAi) as a targeted gene silencing approach to manage H. halys populations. However, RNAi efficacy varies between insect orders, including hemipterans, due to factors such as the rapid degradation of double-stranded RNA (dsRNA) by a DNA/RNA non-specific nuclease (HhNSE) present in the saliva of H. halys. Notably, this study proves that double-stranded DNA (dsDNA) can stabilise dsRNA in saliva, probably by competitively inhibiting HhNSE, which is highly expressed in salivary glands. In vivo tests targeting the clathrin heavy chain gene (HhCHC) demonstrate that a mixture of dsRNA-CHC and dsDNA result in enhanced gene silencing when fed to H. halys, compared to dsRNA alone. While dsRNA-CHC injection causes almost complete mortality, the dsDNA formulation do not significantly increase mortality when fed together with dsRNA-CHC. These findings highlight the need to further investigate factors beyond nucleases such as dsRNA uptake and release mechanisms in the insect gut. Nevertheless, this study provides promising insights for improving RNAi delivery in H. halys, and perhaps other pests with such nucleases, in support of sustainable pest management solutions.

Oesophageal squamous cell carcinoma (ESCC) is the most prevalent type of oesophageal cancer. It is an aggressive malignancy with a poor prognosis, and recent studies have revealed the critical role of the microbiota in its occurrence and development. In this review, the current understanding of the impact of microbiota is summarized on the tumour immune microenvironment (TIME) in ESCC, focusing on intratumoural microbes and the oral-gut microbiota axis as potential therapeutic targets. The mechanisms are discussed by which specific bacterial species, such as Fusobacterium nucleatum (F. nucleatum), Porphyromonas gingivalis (P. gingivalis), Streptococcus spp., and Lactobacillus spp., influence immune responses and contribute to the progression of ESCC. Additionally, the potential of the microbiota is highlighted as a biomarker for early detection, prognosis, and prediction of treatment responses, and explore emerging strategies in microbiota-based immunotherapy that exploit the tumour-targeting properties of bacteria to improve cancer treatment outcomes. Despite these promising developments, the complex interactions between the microbiota and the immune system remain unclarified, and translating research findings into clinical practice is a significant challenge. In this review, the current advancements and emphasise the need for further investigation is summarized into the mechanisms of microbiota-mediated immunotherapy, while outlining future directions for developing personalised treatments for ESCC

Each year, ≈1.6 million women are diagnosed with breast cancer worldwide. Of these cases, 0.5 million result in death, with over 90% of these deaths resulting from metastasis. Since it is one of the top 5 cancers with the highest mortality rates, the development of breast cancer models that are physiologically relevant to the human body is severely needed. This includes models of the breast tissue microenvironment, the microenvironment of metastatic sites (e.g., brain, lungs, bone, liver), and models specifically mimicking each individual step of the cancer metastatic cascade. This review focuses on models of the primary tumor environment for studying cell invasion and spread within the mammary tissue, prior to metastatic outgrowth. Using a combination of methods such as tumor spheroids, 3D printed biomaterials, and microfluidic designs, a variety of 3D in vitro modeling methods exist to recapitulate specific aspects of the tumor microenvironment and understand sources of tumor heterogeneity. An effective model can be specified for each patient, eliminating the need for human trials. Subsequently, as the mechanisms involved in breast cancer metastasis are studied utilizing more effective in vitro models, targeted therapeutics can be discovered, thus advancing clinical treatment strategies.

Solasonine, extracted from Solanum nigrum, has been proven to exert anti-tumor effects in various tumors. However, the role of solasonine in gastric cancer remains unclear. This study aims to elucidate the therapeutic effects of solasonine in suppressing gastric cancer progression. This study explores the regulatory mechanism of solasonine in vitro cells and xenograft tumor mouse models. Methylation-specific PCR (MSP) is used for DNA methylation analysis; immunohistochemical and flow cytometry are performed for MHC class I polypeptide-related sequence A (MICA) expression profiling. Solasonine inhibits the proliferation and migration of gastric cancer cells. MICA is identified as a regulatory target for solasonine in gastric cancer cells. Mechanistically, DNA methylation of MICA is suppressed by solasonine. DNA methyltransferases (DNMT) family members, DNMT1, DNMT3A, and DNMT3B, are downregulated in solasonine-treated tumor tissues. Importantly, solasonine restores the sensitivity of HGC-27 cells to natural killer (NK) cells through upregulating MICA, suggesting the potential value of solasonine as an immunotherapy drug in gastric cancer. Solasonine inhibits gastric cancer progression and restores sensitivity of gastric cancer to NK cells through inducing DNA demethylation of the MICA promoter in vitro and in vivo. This study provides application prospects for solasonine anti-tumor therapy.

Adipose mesenchymal stem cell-derived exosomes and the PDE4 inhibitor roflumilast (ROF) are investigated as potential treatments for chronic kidney disease (CKD). The exosomes are extracted and analyzed using electron microscopy and flow cytometry, then employed with ROF for in vivo implantation in a CKD animal model. Animals aredivided into seven groups. Group (I) Control; (II) exosomes; (III) ROF; (IV) Adriamycin (ADR); (V) ADR + exosomes, (VI) ADR + ROF, and (VII) ADR + Exosomes+ ROF. Biochemical serum indicators (creatinine, BUN), antioxidant status (GSH, MDA), and the mRNA expressions of TGF-β1, Smad3, IL-6, BAX, Wnt-7, FN, and miRNA145-5p are determined using qRT-PCR. Histology assessment using H&E staining, ultrastructural observation using TEM, and protein expression in kidney tissue (FN1 and BAX) are assessed. The isolated exosomes showed cup-shaped morphologyand expressed CD81, CD9, and CD63. Exosomes and ROF increased glutathione (GSH) levels while decreasing malondialdehyde (MDA) levels. Further, ROF and exosomes treatment lowered the expression of the apoptotic indicators BAX, the fibrotic markers TGFβ1, Smad3, Wnt7a, and FN1, and the inflammatory marker IL6, and increased the expression of miRNA-145. Moreover, ROF and exosomes improved histological and ultrastructural examination. In conclusion, exosomes and ROF can protect against CKD by reducing apoptosis and fibrosis.

Local anesthetics (LAs), commonly used for regional and general anesthesia, have gained attention in recent years for their potential role during cancer curative surgery, as they may reduce cancer recurrence and progression. Studies in both laboratory and animal models have shown that LAs can inhibit tumor growth and cell proliferation, trigger apoptosis, and reduce metastasis by limiting cancer cell invasion and migration. In addition, LAs impact the tumor microenvironment by modulating inflammation, enhancing the immune response, blocking angiogenesis, and interfering with tumor innervation. The mechanisms behind these effects involve both voltage-gated sodium channel-dependent and independent pathways, such as AKT/mTOR, RAS/ERK, and SRC/STAT3, as well as regulating microRNAs, circular RNAs, and apoptosis-related proteins, among others. Furthermore, LAs may enhance the efficacy of chemotherapy and counteract chemoresistance. The aim of this review is to provide a comprehensive summary of the current literature on the various mechanisms through which LAs influence tumorigenesis, alter metastasis processes, modulate immune responses, and affect angiogenesis within the tumor microenvironment.

Spheroids are being widely studied as potential building blocks for complex organ engineering, tools for drug screening and cancer study. However, formation time has become the bottleneck of applications due to the need for large-scale high-quality spheroids production. Formation time is often dominated by ECM construction and not cell aggregation. Therefore, this study focuses on the influence of ultrasound detachment replacing conventional enzyme detachment on spheroid formation processes. Thanks to cell surface protein preservation in ultrasound detachment, cell aggregation time is reduced while decreasing the formation variabilities. Moreover, it is confirmed that cells are intrinsically more capable of aggregating through enzyme-free detachment. On top of that, transplantations into rats showed equally successful engraftment properties for enzyme-free detached cells. Finally, the impact on the real co-cultured spheroid application was shown to be beneficial through more localized cell groups inside of the spheroids, possibly improving therapeutic effects and vascularization. Through this study, it is proved that ultrasound detachment can replace enzyme detachment without degrading the final spheroid properties but reducing the formation time, and variability and improving robustness and cell distribution. This opens up a new range of applications for better and faster spheroid formation in numerous bioengineering applications.

Molecular optogenetics allows the control of molecular signaling pathways in response to light. This enables the analysis of the kinetics of signal activation and propagation in a spatially and temporally resolved manner. A key strategy for such control is the light-inducible clustering of signaling molecules, which leads to their activation and subsequent downstream signaling. In this work, an optogenetic approach is developed for inducing graded clustering of different proteins that are fused to eGFP, a widely used protein tag. To this aim, an eGFP-specific nanobody is fused to Cryptochrome 2 variants engineered for different orders of cluster formation. This is exemplified by clustering eGFP-IKKα and eGFP-IKKβ, thereby achieving potent and reversible activation of NF-κB signaling. It is demonstrated that this approach can activate downstream signaling via the endogenous NF-κB pathway and is thereby capable of activating both an NF-κB-responsive reporter construct as well as endogenous NF-κB-responsive target genes as analyzed by RNA sequencing. The generic design of this system is likely transferable to other signaling pathways to analyze the kinetics of signal activation and propagation.

Degenerative disc disease is strongly associated with low back pain, making it a leading cause of disability. With injury and age, cellular remodeling of the disc tissue leads to compositional changes, stiffening, and loss of stress relaxation, particularly in the central gelatinous nucleus pulposus (NP) region of the disc. As part of this extracellular matrix (ECM) remodeling, there is an increase in the deposition of fibronectin, a strongly adhesive integrin ligand that is known to regulate inflammatory signaling. However, it is unclear how these pathological changes in cellular adhesion regulate cell phenotype, and which domains of fibronectin are specifically involved. Here, a dextran vinyl sulfone (DexVS) hydrogel system is employed for presentation of specific fibronectin domains. Fibronectin peptides are found to enhance YAP signaling, inflammatory NF-κB signaling, cellular adhesion, and cellular contractility in NP cells, which leads to a decrease in aggrecan gene expression. Covalent modification of these DexVS hydrogels with bioactive peptides allows for targeted interactions with specific integrin receptors that are involved in healthy or degenerative signaling. In doing so, the integrin binding peptides from fibronectin are identified to activate a contractile phenotype in NP cells.