Visualization of the process of a nanocarrier-mediated gene delivery: stabilization, endocytosis and endosomal escape of genes for intracellular spreading.

Nanoparticles have been widely applied as gene carrier for improving RNA interference (RNAi) efficiency in medical and agricultural fields. However, the mechanism and delivery process of nanoparticle-mediated RNAi is not directly visualized and elucidated. Here we synthesized a star polymer (SPc) consisted of a hydrophilic shell with positively-charged tertiary amine in the side chain, which was taken as an example to investigate the mechanism in gene delivery. The SPc could assemble with dsRNA spontaneously through electrostatic force, hydrogen bond and van der Waals force. Interestingly, the SPc could protect dsRNA from degradation by RNase A and insect hemolymph, thus remarkably increasing the stability of dsRNA. Meanwhile, the SPc could efficiently promote the cellular uptake and endosomal escape for intracellular spreading of dsRNA. Transcriptome analysis revealed that the SPc could up-regulate some key genes such as Chc, AP2S1 and Arf1 for activating clathrin-mediated endocytosis. Furthermore, the suppression of endocytosis hindered the cellular uptake of SPc-delivered dsRNA in vitro, and the subsequent RNAi effect was also disappeared in vivo. To our knowledge, our study is the first direct visualization of the detailed cellular delivery process and mechanism of nanocarrier-mediated gene delivery. Above mechanism supports the application of nanocarrier-based RNAi in gene therapy and pest management.

[Advances of CAR-T cell therapy in treating colorectal cancer].

Globally, colorectal cancer (CRC) ranks as the third most common cancer and the second leading cause of cancer-related fatalities. According to the World Health Organization, there are over 1.9 million annual cases of CRC diagnosed worldwide, resulting in more than 900 000 deaths. In recent years, chimeric antigen receptor T (CAR-T) cell therapy has shown clinical success in treating certain hematological malignancies and is now being explored for its potential in targeting solid tumors like CRC. Currently, CAR-T cell therapies targeting carcinoembryonic antigen (CEA), natural killer group 2, member D ligand (NKG2DL), and other markers have achieved remarkable results in clinical trials, albeit encountering significant challenges. This review summarizes the promising targets of CAR-T cell therapy for CRC and highlights progress made in clinical trials and preclinical studies. Additionally, the review discusses the challenges faced by CAR-T cell therapy in CRC treatment, including a shortage of tumor-specific antigens, cytokine release syndrome, adverse tumor microenvironment, and limited infiltration of CAR-T cells. In summary, this review provides an overview of the latest research progress and challenges in CAR-T cell therapy for CRC, aiming to contribute fresh insights for the clinical treatment of this disease.

Nanocarrier-delivered dsRNA suppresses wing development of green peach aphids.

RNA interference (RNAi) has developed rapidly as a potential "green" pest management strategy. At present, most studies have focused on the screening of aphid lethal genes, whereas only a few studies have been conducted on wing development, which is crucial for aphid migration and plant-virus dissemination. Here, the Myzus persicae genes vestigial (vg) and Ultrabithorax (Ubx) related to wing development, were cloned. These two genes were expressed in various tissues of 3rd-instar winged aphids. The mRNA level of vg was high in 3rd-instar nymphs, whereas the expression level of Ubx was high in adults. The nanocarrier-mediated delivery system delivered double-stranded RNAs for aphid RNAi using topical and root applications. The expression levels of vg and Ubx were downregulated by 44.0% and 36.5%, respectively, using the topical application. The simultaneous RNAi of the two target genes caused 63.3% and 32.2% wing aberration rates using topical and root applications, respectively. The current study provided a promising method for controlling aphid migration to alleviate the spread of insect transmitted plant diseases.