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  2. Paraspeckle Promotes Hepatocellular Carcinoma Immune Escape by Sequestering IFNGR1 mRNA

Paraspeckle Promotes Hepatocellular Carcinoma Immune Escape by Sequestering IFNGR1 mRNA

  • Cell Mol Gastroenterol Hepatol. 2021;12(2):465-487. doi: 10.1016/j.jcmgh.2021.02.010.
Jie Zan 1 Xuya Zhao 2 Xiya Deng 3 Hongda Ding 4 Bi Wang 5 Minyi Lu 1 Zijing Wei 6 Zhi Huang 7 Shuai Wang 8
Affiliations

Affiliations

  • 1 School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou.
  • 2 Department of Interventional Radiology, Affiliated Cancer Hospital of Guizhou Medical University, Guiyang.
  • 3 School of Life Sciences, Westlake University, Hangzhou.
  • 4 Department of General Surgery, Shengjing Hospital of China Medical University, Shenyang, China.
  • 5 School of Basic Medical Science, Guizhou Medical University, Guiyang, China.
  • 6 Department of interventional radiology, the Affiliated Hosptial of Guizhou Medical University, Guiyang, China.
  • 7 Department of interventional radiology, the Affiliated Hosptial of Guizhou Medical University, Guiyang, China; School of Basic Medical Science, Guizhou Medical University, Guiyang, China. Electronic address: doctor@huangzhi.com.
  • 8 School of Life Sciences, Westlake University, Hangzhou. Electronic address: wangshuai@westlake.edu.cn.
Abstract

Background & aims: Hepatocellular carcinoma (HCC) is the most common type of hepatic malignancies, with poor prognosis and low survival rate. Paraspeckles, which are unique subnuclear structures, are recently found to be involved in the development of various tumors, including HCC, and are related to induction in chemoresistance of HCC. This study aimed to investigate the possibility of paraspeckle in HCC cells participating in immune escape and its underlying mechanism in vitro and in vivo.

Methods: Expression of NEAT1_2, the framework of paraspeckle, in HCC cells and tissues was detected by qRT-PCR and RNA-FISH. mRNAs interacted with NEAT1_2 were pull-downed and sequenced in C-terminal S1-aptamer-tagged NEAT1_2 endogenously expressed HCC cells constructed using CRISPR-CAS9 knock-in technology. The effects of paraspeckle on HCC sensitivity to T-cell-mediated cytolysis were detected by T-cell mediated tumor cell killing assay. The roles of NEAT1_2 or NONO on IFNGR1 expression and IFN-γ signaling by applying gene function loss analysis in HCC cells were detected by qRT-PCR, RNA immunoprecipitation, Western blotting, and ELISA. The role of paraspeckle during adoptive T-cell transfer therapy for HCC in vivo was performed with a subcutaneous xenograft mouse.

Results: Paraspeckle in HCC cells is negatively related to T-cell-mediated cytolysis. Destruction of paraspeckle in HCC cells by knockdown of NEAT1_2 or NONO significantly improved the sensibility of resistant HCC cells to T-cell killing effects. Furthermore, IFNGR1 mRNA, which is sequestered by NEAT1_2 and NONO, is abundant in paraspeckle of T-cell killing-resistant HCC cells. Incapable IFN-γ-IFNGR1 signaling accounts for paraspeckle mediated-adoptive T-cell therapy resistance. Moreover, NEAT1_2 expression negatively correlates with IFNGR1 expression in clinical HCC tissues.

Conclusions: Paraspeckle in HCC cells helps tumor cells escape from immunosurveillance through sequestering IFNGR1 mRNA to inhibiting IFN-γ-IFNGR1 signaling, thereby avoiding T-cell killing effects. Collectively, our results hint that NEAT1_2 highly expressed HCC patient is more resistant to T-cell therapy in clinic, and NEAT1_2 may be potential target for HCC immunotherapy.

Keywords

Hepatocellular Carcinoma; Immunotherapy; Interferon Gamma Receptor 1; Long Non-coding RNA Nuclear Enriched Abundant Transcript 1; Non-POU Domain-Containing Octamer-Binding Protein; Paraspeckle.

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