抗乙肝病毒和抗PD1 gRNA / cas9聯合治療的抗病毒作用增強，可

StephenW

Enhanced antiviral benefit of combination therapy with anti-HBV and anti-PD1 gRNA/cas9 produces a synergistic antiviral effect in HBV infection
Shuai Zhen  1 , Rong Qiang  2 , Jiaojiao Lu  3 , Xiaoqian Tuo  3 , Xiling Yang  3 , Xu Li  3
Affiliations
Affiliations

    1
    Medical Heredity Research Center, Northwest Women's and Children's Hospital, Shaanxi, PR China. Electronic address: [email protected].
    2
    Medical Heredity Research Center, Northwest Women's and Children's Hospital, Shaanxi, PR China.
    3
    Center for Translational Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, PR China.

    PMID: 33340931 DOI: 10.1016/j.molimm.2020.12.004

Abstract

Targeted therapy for patients with hepatitis B virus (HBV) infection can lead to objective responses, although response times may be short. At the same time, the response rate to programmed cell death-1 (PD-1) treatment was more durable. It is speculated that HBV targeted therapy can synergistically enhance the antitumor activity with PD-1 blockade. To test this hypothesis, we evaluated the effect of crispr-cas9 on HBV and PD-1 in vitro and in vivo. We found that HBV targeting gRNA/cas9 induced a decrease in the expression of HBsAg, while the PD-1 gene could be knocked out by electroporation targeting gRNA / cas9 by polymerase chain reaction. In HBV transgenic mice, the immunophenotype and cytokine expression of human dendritic cells (DCS) were detected by crispr-cas9 system stimulation, flow cytometry and polymerase chain reaction. These results indicate that gRNA/cas9 treatment upregulates the expression of CD80, CD83 and CD86, and significantly increases the mRNA levels of IL-6, IL-12, IL-23 and tumor necrosis factor alpha. The combination of anti HBV and anti PD-1 therapy can inhibit HBV expression and significantly improve the survival of HBV transgenic mice. In addition, the combination therapy increased the production of interferon by T cells, and then enhanced the expression of Th1 related immunostimulatory genes, thereby reducing the transcription of regulatory / inhibitory immune genes. In general, this response can reshape the tumor microenvironment from immunosuppression to immune stimulation. Finally, anti HBV therapy can induce the expression of interferon dependent programmed cell death ligand-1 in HBV transgenic mice in vivo. To sum up, these results demonstrate that the combination of HBV targeted therapy and PD-1 immune checkpoint block has a strong synergistic effect, thus supporting the transformation potential of this combined therapy strategy in clinical treatment of HBV infection.

Keywords: CRISPR/Cas9; HBV; PD1.

Copyright © 2020 Elsevier Ltd. All rights reserved.

StephenW

抗乙肝病毒和抗PD1 gRNA / cas9聯合治療的抗病毒作用增強，可在乙肝病毒感染中產生協同抗病毒作用
帥真1，榮強2，郊教路3，小倩陀3，楊西靈3，徐力3
隸屬關係
隸屬關係

    1個
    陝西省西北婦女兒童醫院醫學遺傳研究中心電子地址：[email protected]。
    2
    陝西省西北婦女兒童醫院醫學遺傳研究中心
    3
    西安交通大學附屬第一醫院轉化醫學中心，陝西西安710061

    PMID：33340931 DOI：10.1016 / j.molimm.2020.12.004

抽象

儘管反應時間可能很短，但針對乙型肝炎病毒（HBV）感染的患者進行的靶向治療可導致客觀反應。同時，對程序性細胞死亡1（PD-1）治療的反應率更持久。推測HBV靶向治療可以通過PD-1阻滯協同增強抗腫瘤活性。為了驗證該假設，我們在體外和體內評估了crisp-cas9對HBV和PD-1的作用。我們發現靶向gRNA / cas9的HBV誘導了HBsAg表達的降低，而PD-1基因可通過聚合酶鏈反應靶向gRNA / cas9的電穿孔而被敲除。在HBV轉基因小鼠中，通過crisp-cas9系統刺激，流式細胞儀和聚合酶鏈反應檢測人樹突狀細胞（DCS）的免疫表型和細胞因子表達。這些結果表明，gRNA / cas9處理可上調CD80，CD83和CD86的表達，並顯著增加IL-6，IL-12，IL-23和腫瘤壞死因子α的mRNA水平。抗HBV和抗PD-1療法的組合可以抑制HBV表達，並顯著提高HBV轉基因小鼠的存活率。另外，聯合療法增加了T細胞產生的干擾素，然後增強了Th1相關免疫刺激基因的表達，從而減少了調節/抑制性免疫基因的轉錄。通常，這種反應可以使腫瘤的微環境從免疫抑制重塑為免疫刺激。最後，抗HBV療法可以在體內誘導HBV轉基因小鼠中乾擾素依賴性程序性細胞死亡配體-1的表達。綜上所述，這些結果表明，針對HBV的靶向療法與PD-1免疫檢查點阻滯劑的聯合具有強大的協同作用，從而支持了該聯合療法在HBV感染臨床治療中的轉化潛力。

關鍵詞：CRISPR / Cas9；乙肝病毒PD1。

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