Search

Radiotherapy is a mainstay of glioblastoma (GBM) treatment; however, the development of therapeutic resistance has hampered the efficacy of radiotherapy, suggesting that additional treatment strategies are needed. Here, an in vivoloss-of-function genome-wide CRISPR screen was carried out in orthotopic tumors in mice subjected to radiation treatment to identify synthetic lethal genes associated with radiotherapy. Using functional screening and transcriptome analyses, glutathione synthetase (GSS) was found to be a potential regulator of radioresistance through ferroptosis. High GSS levels were closely related to poor prognosis and relapse in patients with glioma. Mechanistic studies demonstrated that GSS was associated with the suppression of radiotherapy-induced ferroptosis in glioma cells. The depletion of GSS resulted in the disruption of glutathione (GSH) synthesis, thereby causing the inactivation of GPX4 and iron accumulation, thus enhancing the induction of ferroptosis upon radiotherapy treatment. Moreover, to overcome the obstacles to broad therapeutic translation of CRISPR editing, we report a previously unidentified genome editing delivery system, in which Cas9 protein/sgRNA complex was loaded into Angiopep-2 (Ang) and the trans-activator of the transcription (TAT) peptide dual-modified extracellular vesicle (EV), which not only targeted the blood–brain barrier (BBB) and GBM but also permeated the BBB and penetrated the tumor. Our encapsulating EVs showed encouraging signs of GBM tissue targeting, which resulted in high GSS gene editing efficiency in GBM (up to 67.2%) with negligible off-target gene editing. These results demonstrate that a combination of unbiased genetic screens, and CRISPR-Cas9-based gene therapy is feasible for identifying potential synthetic lethal genes and, by extension, therapeutic targets.

As a kind of economic forest fruit tree with medicinal, edible, greening, and ornamental functions, Cornus officinalishas been introduced and cultivated in a large area in the People’s Republic of China. However, great differences exist in the content of active secondary metabolites of C. officinalisfrom different regions. We wonder whether local rhizosphere soil properties and soil microorganisms have a critical impact on the accumulation of active components in the fruit. Eight sampling sites were selected in the main C. officinalisproducing area to collect rhizosphere soil and mature fruits. The contents of soil pH, nitrate nitrogen, ammonia nitrogen, phosphate, and active secondary metabolites in C. officinalisfruits were measured, respectively, and the fungal community composition of rhizosphere soil were determined by Illumina sequencing. The content of secondary metabolites was more abundant in C. officinalisfruits A3, A4, A6, and A7 than A1, A2, and A5 (p< 0.05) (A represents fruit sample). Fungal community compositions were similar between production areas S3, S4, and S6, whereas S7 (S represents rhizosphere soil sample) was apparently different from them. Soil pH, nitrate nitrogen, and ammonia nitrogen content were significantly correlated with the content of gallic acid, morroniside, sweroside, and loganin (p< 0.05). The quality of C. officinalisfruits in introduction cultivation areas is not necessarily worse than those in traditional cultivation areas (for example, the content of monoside in A7 is higher than others). The contents of secondary metabolites of C. officinalisfruits seem to have a positive correlation with the abundance of a certain fungus of the Cladosporiaceae family Cladosporium genus in the rhizosphere soil. Such findings had important implications for the cultivation, development, and utilization of C. officinalis.

Corni Fructus (CF) has great potential as a food product in the treatment of T2DM (thirst-quenching). As the main active ingredient of CF, loganin (LOG) possesses multiple pharmacological propertiesincluding hypoglycemic, hypolipidemic, anti-inflammatory and antibacterial. In this study, we firstly verified the efficacy of LOG in improving the disordered glycolipid metabolism and liver damages in a murine insulin resistance (IR) model, and then determined the ameliorative effect of LOG on IR-HepG2 cell model. To clarify the mechanism of LOG on IR, a series of differentially expressed metabolites (DEM) and genes (DEG) were screened out through non-targeted metabolomics and transcriptomics techniques. In addition, integrative metabolomics and transcriptomics analyses revealed a few key pathways (foxO signaling pathway, glycine serine and threonine metabolism, glutamatergic synapse, cocaine addiction, amphetamine addiction, and valine leucine and isoleucine biosynthesis), important DEM (l-Glutamate), and DEGs (Sgk2, Grin3b, Acadsb, Grik1) were reversely regulated by LOG as compared with the IR model. It is hypothesized that LOG may exert the anti-IR effect via finely tuning amino acid metabolism. Taken together, our data may shine light on the screening of new intervention targets and developing of new LOG-related agents for IR treatment.

The recently reported inhibitory effects of angiotensin 1–7 (Ang-(1–7)) on various cancers indicate its potential use as a therapeutic agent for primary and metastatic cancers. However, its extremely short half-life in the circulation greatly compromises its potential applications. Here, we reported an Ang-(1–7) analogue peptide with the amino and carboxy termini protected by acetylation and amination. The in vitro and in vivo degradation of the resulting analogue, Ang-AA, were determined using high-performance liquid chromatography (HPLC). At the same time, small RNA interference and competition studies were performed to evaluate the specific capacity of Ang-AA to bind to the cell surface Mas receptor. Cell Counting Kit-8 (CCK8), wound-healing, and Boyden chamber assays were performed to investigate the inhibitory effects of Ang-AA on A549 cells. Finally, the synergistic inhibitory effects of Ang-AA and paclitaxel (PTX) on A549 xenografts in mice were observed using animal imaging systems and survival observations. The toxicity of Ang-AA in mice was evaluated. Our results showed that acetylation and amination significantly inhibited the hydrolyzation of Ang-(1–7) in vitro and in vivo. The half-life of Ang-(1–7) in rats was prolonged from 2.4 ± 0.6 min to 238.7 ± 61.3 min (p< 0.001). The specific binding of Ang-AA to the Mas receptor was well preserved, and Ang-AA exerted significantly greater inhibitory effects on the proliferation, migration, and invasion of A549 cells than Ang-(1–7). The combination of Ang-AA and PTX exhibited a significantly greater synergistic inhibitory effect on A549 xenografts than the combination of Ang-(1–7) and PTX. Ang-AA did not display obvious toxicity in mice. Our findings indicate acetylation and amination is a simple and effective method for producing Ang-(1–7) as a bioactive peptide.

CD20, a B-cell-specific protein, is a primary target for immunotherapy of B-cell lymphomas. We used a mimotopes of CD20 to construct vaccines for B-cell-related disorders. The immunogenicity of the vaccines was tested in BALB/c mice. Results of this study suggest that the mimotope may be a useful tool for the construction of a functional vaccine to treat B cell-related disorders.

Plants are a promising expression system for the production of recombinant proteins. However, low protein productivity remains a major obstacle that limits extensive commercialization of whole plant and plant cell bioproduction platform. Plastid genetic engineering offers several advantages, including high levels of transgenic expression, transgenic containment via maternal inheritance, and multigene expression in a single transformation event. In recent years, the development of optimized expression strategies has given a huge boost to the exploitation of plastids in molecular farming. The driving forces behind the high expression level of plastid bioreactors include codon optimization, promoters and UTRs, genotypic modifications, endogenous enhancer and regulatory elements, posttranslational modification, and proteolysis. Exciting progress of the high expression level has been made with the plastid-based production of two particularly important classes of pharmaceuticals: vaccine antigens, therapeutic proteins, and antibiotics and enzymes. Approaches to overcome and solve the associated challenges of this culture system that include low transformation frequencies, the formation of inclusion bodies, and purification of recombinant proteins will also be discussed.