The CXC chemokine receptor type 4 (CXCR4) receptor and its ligand, CXCL12, are overexpressed in various cancers and mediate tumor progression and hypoxia-mediated resistance to cancer therapy. While CXCR4 antagonists have potential anticancer effects when combined with conventional anticancer drugs, their poor potency against CXCL12/CXCR4 downstream signaling pathways and systemic toxicity had precluded clinical application. Herein, BPRCX807, known as a safe, selective, and potent CXCR4 antagonist, has been designed and experimentally realized. In in vitro and in vivo hepatocellular carcinoma mouse models it can significantly suppress primary tumor growth, prevent distant metastasis/cell migration, reduce angiogenesis, and normalize the immunosuppressive tumor microenvironment by reducing tumor-associated macrophages (TAMs) infiltration, reprogramming TAMs toward an immunostimulatory phenotype and promoting cytotoxic T cell infiltration into tumor. Although BPRCX807 treatment alone prolongs overall survival as effectively as both marketed sorafenib and anti–PD-1, it could synergize with either of them in combination therapy to further extend life expectancy and suppress distant metastasis more significantly.

Successful siRNA therapy requires suitable delivery systems with targeting moieties such as small molecules, peptides, antibodies, or aptamers. Galactose (Gal) residues recognized by the asialoglycoprotein receptor (ASGPR) can serve as potent targeting moieties for hepatocellular carcinoma (HCC) cells. However, efficient targeting to HCC via galactose moieties rather than normal liver tissues in HCC patients remains a challenge. To achieve more efficient siRNA delivery in HCC, we synthesized various galactoside derivatives and investigated the siRNA delivery capability of nanoparticles modified with those galactoside derivatives. In this study, we assembled lipid/calcium/phosphate nanoparticles (LCP NPs) conjugated with eight types of galactoside derivatives and demonstrated that phenyl β-d-galactoside-decorated LCP NPs (L4-LCP NPs) exhibited a superior siRNA delivery into HCC cells compared to normal hepatocytes. VEGF siRNAs delivered by L4-LCP NPs downregulated VEGF expression in HCC in vitro and in vivo and led to a potent antiangiogenic effect in the tumor microenvironment of a murine orthotopic HCC model. The efficient delivery of VEGF siRNA by L4-LCP NPs that resulted in significant tumor regression indicates that phenyl galactoside could be a promising HCC-targeting ligand for therapeutic siRNA delivery to treat liver cancer.

Background Metastasis and chemoresistance are major culprits of cancer mortality, but factors contributing to these processes are incompletely understood. Methods Bioinformatics methods were used to identify the relations of Smyca expression to clinicopathological features of human cancers. RNA-sequencing analysis was used to reveal Smyca-regulated transcriptome. RNA pull-down and RNA immunoprecipitation were used to examine the binding of Smyca to Smad3/4 and c-Myc/Max. Chromatin immunoprecipitation and chromatin isolation by RNA purification were used to determine the binding of transcription factors and Smyca to various gene loci, respectively. Real-time RT-PCR and luciferase assay were used to examine gene expression levels and promoter activities, respectively. Xenograft mouse models were performed to evaluate the effects of Smyca on metastasis and chemoresistance. Nanoparticle-assisted gapmer antisense oligonucleotides delivery was used to target Smyca in vivo. Results We identify lncRNA Smyca for its association with poor prognosis of many cancer types. Smyca potentiates metabolic reprogramming, migration, invasion, cancer stemness, metastasis and chemoresistance. Mechanistically, Smyca enhances TGF-β/Smad signaling by acting as a scaffold for promoting Smad3/Smad4 association and further serves as a Smad target to amplify/prolong TGF-β signaling. Additionally, Smyca potentiates c-Myc-mediated transcription by enhancing the recruitment of c-Myc/Max complex to a set of target promoters and c-Myc binding to TRRAP. Through potentiating TGF-β and c-Myc pathways, Smyca synergizes the Warburg effect elicited by both pathways but evades the anti-proliferative effect of TGF-β. Targeting Smyca prevents metastasis and overcomes chemoresistance. Conclusions This study uncovers a lncRNA that coordinates tumor-relevant pathways to orchestra a pro-tumor program and establishes the clinical values of Smyca in cancer prognosis and therapy.

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Figure 2. Comparison of size and detection limit between PalmGRET-and dye-labeled bEVs and sEVs. a) NTA of 4T1-bEVs and -sEVs demonstrating distinct size distributions (left; representative data of three independent experiments) and mean sizes (right; determined from three independent experiments). n.s., p > 0.05; *p < 0.05 with 2-tailed Student's t-test. b) Schematic of inner EV membrane labeling using PalmGRET (top) and outer EV membrane labeling using lipophilic fluorescent dyes (bottom). c) Dot blot analysis showing that PalmGRET labeled the inner membrane of bEVs and sEVs. The positive control was whole cell lysates (WCL). d) Representative TEM images of 4T1-PalmGRET-bEVs and -sEVs. The same imaging parameters were used to acquire bEV and sEV TEM images. Images on the right of each row depict the enlarged images of the boxed regions (red). Black scale bar: 200 nm; red scale bar: 100 nm. e) Mean particle size (left) and particle size distribution (right) of 4T1-PalmGRET-bEVs and -sEVs determined from TEM image analysis. The bEV and sEV particles (N = 400 per group) in the captured TEM images were analyzed using Fiji software (ImageJ, NIH) to measure individual particle sizes (left). Size distribution analysis of bEVs and sEVs (right) demonstrating that bEVs are largely composed of 201-400 nm particles, while sEVs mainly consist of 1-100 and 101-200 nm particles. ****p < 0.0001 with 2-tailed Student's t-test. f-h) NTA of PalmGRET-and dye-labeled 4T1-EVs. f) Size distribution and g) mean size of bEVs and sEVs labeled with PalmGRET or colabeled with PKH26, DiD, and DiR are shown. h) Analysis of particle size composition of the bEVs and sEVs showed that PalmGRET did not change the size distribution in all size divisions as compared to the WT EV controls. The 101-200 nm particles were decreased, while the 201-300 nm and 301-500 nm particles were increased among the lipophilic-dye-labeled bEVs. Similarly, the 101-200 nm particles were decreased and 201-300 nm particles were increased among the lipophilic-dye-labeled sEVs. Data are from three independent experiments. n.s., p > 0.05; *p < 0.05; **p < 0.01 with 1-way ANOVA followed by Dunnett's post hoc test versus the control.