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

Abstract

Background The combination of sonodynamic therapy and oxygenation strategy is widely used in cancer treatment. However, due to the complexity, heterogeneity and irreversible hypoxic environment produced by hepatocellular carcinoma (HCC) tissues, oxygen-enhancing sonodynamic therapy (SDT) has failed to achieve the desired results. With the emergence of ferroptosis with reactive oxygen species (ROS) cytotoxicity, this novel cell death method has attracted widespread attention. Methods In this study, nanobubbles (NBs) were connected with the sonosensitizer Indocyanine green (ICG) to construct a 2-in-1 nanoplatform loaded with RAS-selective lethal (RSL3, ferroptosis promoter) (RSL3@O2-ICG NBs), combined with oxygen-enhanced SDT and potent ferroptosis. In addition, nanobubbles (NBs) combined with low-frequency ultrasound (LFUS) are called ultrasound-targeted nanobubble destruction (UTND) to ensure specific drug release and improve safety. Results MDA/GSH and other related experimental results show that RSL3@O2-ICG NBs can enhance SDT and ferroptosis. Through RNA sequencing (RNA-seq), the differential expression of LncRNA and mRNA before and after synergistic treatment was identified, and then GO and KEGG pathways were used to enrich and analyze target genes and pathways related ferroptosis sensitivity. We found that they were significantly enriched in the ferroptosis-related pathway MAPK cascade and cell proliferation. Then, we searched for the expression of differentially expressed genes in the TCGA Hepatocellular carcinoma cohort. At the same time, we evaluated the proportion of immune cell infiltration and the identification of co-expression network modules and related prognostic analysis. We found that it was significantly related to the tumor microenvironment of hepatocellular carcinoma. The prognostic risk genes “SLC37A2” and “ITGB7” may represent new hepatocellular carcinoma ferroptosis-inducing markers and have guiding significance for treating hepatocellular carcinoma. Conclusion The therapeutic effect of the in vitro synergistic treatment has been proven to be significant, revealing the prospect of 2-in-1 nanobubbles combined with SDT and ferroptosis in treating HCC.

Recent Progress Toward Imaging Application of Multifunction Sonosensitizers in Sonodynamic Therapy

“… 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. 162 Because the reporter gene is inserted directly into the cell’s DNA, the imaging is generated only in the living cell and can retain accurate imaging for a long time as the cell divides and is passed on to daughter cells.…”

“…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.…”

“…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. 162 Because the reporter…”

Ferroptosis: From Basic Research to Clinical Therapeutics in Hepatocellular Carcinoma

“…It provided a novel clinical antitumor therapeutic strategy to induce ferroptosis via the consumption of GSH, disrupt redox balance by the Fenton reaction and doxorubicin-supplied hydrogen peroxide, synergistic cytotoxicity of doxorubicin inhibition of recurrence and metastasis of HCC, and reversing drug resistance in translational therapy. Chen et al 116 assessed the prospect of nanobubbles combined with SDT and ferroptosis for treating HCC. Because of their low immunogenicity, low cytotoxicity, and high biocompatibility, nanosized vesicle exosomes can be used as drug delivery systems.…”

Synergistic therapy for orthotopic gliomas via biomimetic nanosonosensitizer-mediated sonodynamic therapy and ferroptosis

“…In recent years, the combined treatment strategy of SDT and ferroptosis has been used in research on breast cancer and liver cancer. 17,23–26 However, its application in orthotopic brain tumors has not been studied.…”