A nuclear function for an oncogenic microRNA as a modulator of snRNA and splicing

Background miRNAs are regulatory transcripts established as repressors of mRNA stability and translation that have been functionally implicated in carcinogenesis. miR-10b is one of the key onco-miRs associated with multiple forms of cancer. Malignant gliomas exhibit particularly striking dependence on miR-10b. However, despite the therapeutic potential of miR-10b targeting, this miRNA’s poorly investigated and largely unconventional properties hamper the clinical translation. Methods We utilized Covalent Ligation of Endogenous Argonaute-bound RNAs and their high-throughput RNA sequencing to identify miR-10b interactome and a combination of biochemical and imaging approaches for target validation. They included Crosslinking and RNA immunoprecipitation with spliceosomal proteins, a combination of miRNA FISH with protein immunofluorescence in glioma cells and patient-derived tumors, native Northern blotting, and the transcriptome-wide analysis of alternative splicing. Results We demonstrate that miR-10b binds to U6 snRNA, a core component of the spliceosomal machinery. We provide evidence of the direct binding between miR-10b and U6, in situ imaging of miR-10b and U6 co-localization in glioma cells and tumors, and biochemical co-isolation of miR-10b with the components of the spliceosome. We further demonstrate that miR-10b modulates U6 N-6-adenosine methylation and pseudouridylation, U6 binding to splicing factors SART3 and PRPF8, and regulates U6 stability, conformation, and levels. These effects on U6 result in global splicing alterations, exemplified by the altered ratio of the isoforms of a small GTPase CDC42, reduced overall CDC42 levels, and downstream CDC42 -mediated effects on cell viability. Conclusions We identified U6 snRNA, the key RNA component of the spliceosome, as the top miR-10b target in glioblastoma. We, therefore, present an unexpected intersection of the miRNA and splicing machineries and a new nuclear function for a major cancer-associated miRNA.

Direct Cytotoxic and Indirect, Immune-Mediated Effects of Local Anesthetics Against Cancer

Local anesthetics are frequently employed during surgery in order to control peri- and postoperative pain. Retrospective studies have revealed an unexpected correlation between increased long-term survival and the use of local anesthetics during oncological surgery. This effect of local anesthetics might rely on direct cytotoxic effects on malignant cells or on indirect, immune-mediated effects. It is tempting to speculate, yet needs to be formally proven, that the combination of local anesthetics with oncological surgery and conventional anticancer therapy would offer an opportunity to control residual cancer cells. This review summarizes findings from fundamental research together with clinical data on the use of local anesthetics as anticancer standalone drugs or their combination with conventional treatments. We suggest that a better comprehension of the anticancer effects of local anesthetics at the preclinical and clinical levels may broadly improve the surgical treatment of cancer.

Silicon fertilisation affects morphological and immune defences of an insect pest and enhances plant compensatory growth

Herbivorous insects have evolved various anti-predator defences, including morphological, behavioural, and immune defences, which can make biocontrol of herbivorous pests challenging. Silicon (Si) accumulation in plants is a potent physical defence against mandibulate insects. However, it remains uncertain how Si affects the anti-predator defences of insect herbivores and plant defences following herbivory. We grew the model grass, Brachypodium distachyon, hydroponically with (+Si) or without (–Si) Si and investigated the plant-mediated effects of Si on the anti-predator defences of the cotton bollworm, Helicoverpa armigera, integrating morphological (i.e. integument resistance and thickness), behavioural, and immune defences. We also examined the effects of Si on plant compensatory growth and leaf trichome production. Larval growth, leaf consumption, and integument resistance were lower when feeding on +Si plants compared to when feeding on –Si plants. Larval integument thickness, defensive behaviours, haemocyte density, and lysozyme-like activity in the haemolymph were unaffected by Si. Larvae fed on +Si plants had higher haemolymph phenoloxidase (PO) and total-PO activities than larvae fed on –Si plants, although this did not enhance the melanisation response of larvae. Furthermore, Si supplies increased plant compensation for herbivory and constitutive trichome production, whereas herbivory induced trichome production only on –Si plants. We provide the first evidence for plant-mediated effects of Si on anti-predator defences of an insect herbivore. We suggest that the lower integument resistance of larvae when feeding on Si-supplemented plants could contribute to their vulnerability to natural enemies and that high PO activity may impose fitness costs (e.g. delayed development).

The Relationship between Wellness and Stress of Home Training Mediating Effect

The purpose of this study was to analyze the mediated effects of home training between wellness and stress in three areas. Recently, the number of people who do home training has increased rapidly due to lack of outdoor activities in the aftermath of COVID-19. Thus, in this study, we understand the effect of three types of wellness on home training and stress and conducted an analysis focusing on the mediated effects of home training between wellness and stress. In order to obtain the above results, this study surveyed a total of 389 students and officials related to physical education at S University from July 5 to July 23, 2021. First, among the three types of wellness, physical and emotional wellness have a positive effect on home training, but social wellness has no significant effect on home training. Second, all three types of wellness have been shown to have a positive effect on stress. Third, home training has been shown to serve as a partial mediator between the other two types (physical wellness, emotional wellness) and stress, excluding social wellness. The implications of this study are to determine the role of the mediated effects of home training in the relationship between wellness and stress. Through this, it was found that higher physical and emotional wellness has a positive effect on trying home training, and more home training has a positive effect on stress. It also found that social wellness has a positive effect on stress, but not on trying to train flaws.

Immune-Mediated Effects of Microplanar Radiotherapy with a Small Animal Irradiator

Spatially fractionated radiotherapy has been shown to have effects on the immune system that differ from conventional radiotherapy (CRT). We compared several aspects of the immune response to CRT relative to a model of spatially fractionated radiotherapy (RT), termed microplanar radiotherapy (MRT). MRT delivers hundreds of grays of radiation in submillimeter beams (peak), separated by non-radiated volumes (valley). We have developed a preclinical method to apply MRT by a commercial small animal irradiator. Using a B16-F10 murine melanoma model, we first evaluated the in vitro and in vivo effect of MRT, which demonstrated significant treatment superiority relative to CRT. Interestingly, we observed insignificant treatment responses when MRT was applied to Rag−/− and CD8-depleted mice. An immuno-histological analysis showed that MRT recruited cytotoxic lymphocytes (CD8), while suppressing the number of regulatory T cells (Tregs). Using RT-qPCR, we observed that, compared to CRT, MRT, up to the dose that we applied, significantly increased and did not saturate CXCL9 expression, a cytokine that plays a crucial role in the attraction of activated T cells. Finally, MRT combined with anti-CTLA-4 ablated the tumor in half of the cases, and induced prolonged systemic antitumor immunity.