Agonists for toll-like receptors (TLRs) have shown promising activities against cancer. In the present study, a squalene-based nanoemulsion (NE) was loaded with resiquimod, a TLR7/8 agonist for therapeutic delivery. R848 NE was developed and characterized for long-term stability. In vitro and in vivo antitumor immunity of R848 NE were also evaluated in combination with SD-101, a CpG-containing TLR9 agonist. In vitro studies demonstrated strong long-term stability and immune responses to R848 NE. When combined with SD-101, strong antitumor activity was observed in MC38 murine colon carcinoma model with over 80% tumor growth inhibition. The combination treatment showed a 4-fold increase in systemic TNFa production and a 2.6-fold increase in Cd8a expression in tumor tissues, suggesting strong cell-mediated immune responses against the tumor. The treatment not only demonstrated a strong antitumor immunity by TLR7/8 and TLR9 activations but also induced PD-L1 upregulation in tumors, suggesting a potential therapeutic synergy with immune checkpoint inhibitors.

The ability of lipid nanoparticles (LNPs) to deliver nucleic acids have shown a great therapeutic potential to treat a variety of diseases. Here, an optimized formulation of QTsome lipid nanoparticles (QTPlus) is utilized to deliver an anti‐miR‐21 (AM21) against cancer. The miR‐21 downstream gene regulation and antitumor activity is evaluated using mouse and human cancer cells and macrophages. The antitumor activity of QTPlus encapsulating AM21 (QTPlus‐AM21) is further evaluated in combination with erlotinib and atezolizumab (ATZ). QTPlus‐AM21 demonstrates a superior miR‐21‐dependent gene regulation and eventually inhibits A549 non‐small cell lung cancer growth in vitro. QTPlus‐AM21 further induces chemo‐sensitization of A549 cells to erlotinib with a combination index of 0.6 in inhibiting A549 cell growth. When systemically administers to MC38 tumor‐bearing mouse model, QTPlus‐AM21 exhibits an antitumor immune response with over 80% tumor growth inhibition (TGI%) and over twofold and fourfold PD‐1 and PD‐L1 upregulation in tumors and spleens. The combination therapy of QTPlus‐AM21 and ATZ further shows a higher antitumor response (TGI% over 90%) and successfully increases M1 macrophages and CD8 T cells into TME. This study provides new insights into the antitumor mechanism of AM21 and shows great promise of QTPlus‐AM21 in combination with chemotherapies and immunotherapies.

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The strawberry (Fragaria × ananassa Duch.) is an important horticultural crop that is widely grown all over the world. Its sweetness, aroma, nutritional value and bright color make it popular. The woodland strawberry (Fragaria vesca) is a model plant for studying non-climacteric fruits because its respiration rate does not change significantly during fruit ripening. The B-box (BBX) protein family is made up of zinc-finger transcription factors important in plant growth and development. In this study, we identified 22 FveBBX genes from the newly released woodland strawberry genome database by comprehensive bioinformatics analysis. Phylogenetic analysis divided these FveBBX genes into five subfamilies. A promoter cis-acting element analysis detected 29 elements related to plant development, light response, abiotic stress and hormone response in the promoter of FveBBX genes. According to transcriptome data, relatively few BBX genes had tissue-specific expression, with examples including FveBBX12, which was expressed only in pre-fertilization cortex and pitch, and FveBBX19, which was specifically expressed in mature anthers. During fruit ripening, the expressions of eight FveBBX genes decreased by more than two-fold, and three FveBBX gene expressions increased more than two-fold both in “Ruegen” and “Yellow Wonder”. After cold and heat stresses, around half of the FveBBX genes displayed altered expression, especially FveBBX16 which showed an 8.3-fold increase in expression after heat treatment, while FveBBX14 showed at least an 11-fold decrease in expression after cold treatment. According to the result of quantitative real-time PCR (qRT-PCR), FveBBX genes’ expression differed depending on the photoperiod. Notably, FveBBX7 gene expression was the opposite during the first 16 h of the long-day (LD) and short-day (SD) conditions. This study provides helpful information for further research on BBX gene activity of the woodland strawberry in plant growth and development and adaptation to temperature and photoperiod environmental conditions.