Single-cell RNA sequencing reveals the mechanism of sonodynamic therapy combined with a RAS inhibitor in the setting of hepatocellular carcinoma

Abstract

Background Ras activation is a frequent event in hepatocellular carcinoma (HCC). Combining a RAS inhibitor with traditional clinical therapeutics might be hampered by a variety of side effects, thus hindering further clinical translation. Herein, we report on integrating an IR820 nanocapsule-augmented sonodynamic therapy (SDT) with the RAS inhibitor farnesyl-thiosalicylic acid (FTS). Using cellular and tumor models, we demonstrate that combined nanocapsule-augmented SDT with FTS induces an anti-tumor effect, which not only inhibits tumor progression, and enables fluorescence imaging. To dissect the mechanism of a combined tumoricidal therapeutic strategy, we investigated the scRNA-seq transcriptional profiles of an HCC xenograft following treatment. Results Integrative single-cell analysis identified several clusters that defined many corresponding differentially expressed genes, which provided a global view of cellular heterogeneity in HCC after combined SDT/FTS treatment. We conclude that the combination treatment suppressed HCC, and did so by inhibiting endothelial cells and a modulated immunity. Moreover, hepatic stellate secretes hepatocyte growth factor, which plays a key role in treating SDT combined FTS. By contrast, enrichment analysis estimated the functional roles of differentially expressed genes. The Gene Ontology terms “cadherin binding” and “cell adhesion molecule binding” and KEGG pathway “pathway in cancer” were significantly enriched by differentially expressed genes after combined SDT/FTS therapy. Conclusions Thus, some undefined mechanisms were revealed by scRNA-seq analysis. This report provides a novel proof-of-concept for combinatorial HCC-targeted therapeutics that is based on a non-invasive anti-tumor therapeutic strategy and a RAS inhibitor.

RNA-Seq Explores the Mechanism of Oxygen-Boosted Sonodynamic Therapy Based on All-in-One Nanobubbles to Enhance Ferroptosis for the Treatment of HCC

“…As reported in previous studies, we have prepared RSL3@O 2 -ICG NBs using the thin-film hydration ultrasound method. 44 First, mix DSPC and DSPE-PEG-2000 at a mass ratio of 9:1, and then add 10ul of 5 mM RSL3 in methylene chloride and methanol (2:1, vol/vol). After entirely dissolving and mixing, transfer the above solution to a beaker and evaporate to form a lipid film.…”

Single-Cell Sequencing: Current Applications in Precision Onco-Genomics and Cancer Therapeutics

“…Identifying new combinations helps tremendously with immunotherapy strategies where most patients do not respond to monotherapy as a first-line treatment [45]. In a recent study, single-cell RNA sequencing was performed to show that a combinational strategy using both an RAS inhibitor and IR820 nanocapsuleaugmented sonodynamic therapy suppressed hepatocellular carcinoma and modulated immunity through differentially expressed genes [46]. In a similar study, single-cell sequencing was performed on CD45 + tumor-infiltrating lymphocytes (untreated controls or PDL1 and AB680 CD73 inhibitor treated) to find that CD73 inhibition is distinct from PDL1 inhibition [47].…”

Recent Progress Toward Imaging Application of Multifunction Sonosensitizers in Sonodynamic Therapy

“…Multiple studies have supported that multifunctional sonosensitizers could alter gene expression in cancer and immune-related cells and revealed therapeutic target genes in various signaling pathways by the data analysis. [158][159][160][161] For example, Wu et al designed IR820@NCP using RAS inhibitor Farnesyl-Thiosalicylic acid (FTS) to demonstrate that SDT combined with FTS predominantly achieved anti-tumour effects by inhibiting endothelial cells and simulating host immunity. 160 In Chen et al's research on the treatment of liver cancer by RSL3@O2-ICG NBs, essential genes were identified to affect the sensitivity of hepatocellular carcinoma to iron death.…”

“…[158][159][160][161] For example, Wu et al designed IR820@NCP using RAS inhibitor Farnesyl-Thiosalicylic acid (FTS) to demonstrate that SDT combined with FTS predominantly achieved anti-tumour effects by inhibiting endothelial cells and simulating host immunity. 160 In Chen et al's research on the treatment of liver cancer by RSL3@O2-ICG NBs, essential genes were identified to affect the sensitivity of hepatocellular carcinoma to iron death. 158 In addition to molecular imaging techniques, such gene and protein level monitoring could be achieved by reporter imaging techniques that combined reporter proteins with corresponding imaging probes.…”