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

Yichi Chen; Haitao Shang; Chunyue Wang; Jiaqi Zeng; Shentao Zhang; Bolin Wu; Wen Cheng

doi:10.2147/IJN.S343361

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

Int J Nanomedicine. 2022 Jan 7:17:105-123. doi: 10.2147/IJN.S343361. eCollection 2022.

Authors

Yichi Chen¹, Haitao Shang¹, Chunyue Wang¹, Jiaqi Zeng², Shentao Zhang¹, Bolin Wu^{1

3}, Wen Cheng^{1

3}

Affiliations

¹ Department of Ultrasound, Harbin Medical University Cancer Hospital, Harbin, People's Republic of China.
² School of Life Science and Technology, Computational Biology Research Center, Harbin Institute of Technology, Harbin, People's Republic of China.
³ Department of Interventional Ultrasound, Harbin Medical University Cancer Hospital, Harbin, People's Republic of China.

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.

Keywords: HCC; RSL3; ferroptosis; sonodynamic therapy; synergistic therapy.

MeSH terms

Carcinoma, Hepatocellular* / therapy
Ferroptosis*
Humans
Liver Neoplasms* / therapy
Nanomedicine / methods*
Oxygen
RNA-Seq
Tumor Microenvironment
Ultrasonics

Substances

Oxygen

Grants and funding

This research was supported by the National Natural Science Foundation of China (grant number 81873900 and 82171947), Natural Science Foundation of Heilongjiang Province (grant number YQ2020H037 and ZD2021H005), and HAI YAN Science Foundation of Harbin Medical University Cancer Hospital (grant number JJMS2021-30).