AASLD2019[691]一种新的肝定向锁定核 酸靶向PD-L1表达可调节HBV

StephenW

A NOVEL LIVER-DIRECTED LOCKED NUCLEIC
ACID TARGETING PD-L1 EXPRESSION REVERTS HBV-SPECIFIC
IMMUNE TOLERANCE AND INDUCES SUSTAINED CLEARANCE
OF HBV INFECTION
Jie Zhao1, Lykke Pedersen2, Julie Blaising3, Malika Ait-
Goughoulte3, Fang Shen1, Youjun Yu1, Johanna Walther3,
Anna-Maria Geretti3, Timothy Tellinghuisen3, John Young3,
Soren Ottosen2 and Souphalone Luangsay3, (1)Roche Pharma
Research and Early Development, Roche Innovation Center
Shanghai, Shanghai, China, (2)Roche Pharma Research and
Early Development, Roche Innovation Center Copenhagen,
Hørsholm, Denmark, (3)Roche Pharma Research and
Early Development, Roche Innovation Center Basel, Basel,
Switzerland
Background: Functional impairment of HBV-specific T cell
responses is the hallmark of chronic HBV infection. The
PD-1/ PD-L1 pathway is a key immune checkpoint regulator
responsible for delivering a strong immune-inhibitory signal
in the liver of chronic hepatitis B patients. We present a new
technology that uses a liver-targeted locked nucleic acid (LNA)
single stranded antisense oligonucleotide (ASO) to suppress
PD-L1 expression in hepatocytes (PD-L1 LNA) and disrupt the
PD-1/ PD-L1 pathway in the liver microenvironment, thereby
restoring local HBV-specific T cell effector function. Methods:
We generated a GalNAc-conjugated LNA ASO directed
against mouse PD-L1 and evaluated several subcutaneous
doses ranging from 3 to 15 mg/kg, once weekly (QW), in
the AAV-HBV mouse model. Circulating HBV parameters
(HBsAg, HBeAg, HBV DNA) were monitored weekly during
4 weeks of dosing and for 8 weeks after the end of dosing
Intrahepatic expression of PD-L1 mRNA (by RT-qPCR), HBVspecific
immune response (by IFN-γ Elispot) and liver-resident
and circulating immune cell subsets (by flow cytometry) were
evaluated in mouse liver at the end of the study Results:
Mice receiving 5 QW doses of PD-L1 LNA showed a marked
inhibition of PD-L1 mRNA expression at all tested doses (e g ,
~70% reduction after 2 doses at 5 mg/kg) Inhibition of PD-L1
expression was maintained (~50%) for at least 8 weeks postdosing,
and was associated with a significant and sustained
reduction of HBsAg, HBeAg and HBV DNA levels (2.4, 2.1 and
1 6 log10 drop, respectively), with no evidence of post-dosing
rebound After 2 to 5 doses, concomitant with the decline of
viral parameters, and depending on the dose, we observed a
transient serum ALT increase, without changes in markers of
liver function Interestingly, mice treated with PD-L1 LNA (5
QW doses at 5 mg/kg) showed a marked increase (2.8 fold)
in intrahepatic immune cell numbers, consisting of mainly
T cell subsets and antigen presenting cells, together with
a significant increase in HBV specific IFN-γ secreting cells
(~40 fold) compared to the vehicle control group Treatment
was well tolerated with no significant changes in body weight.
Conclusion: The findings support the hypothesis that a livertargeted
PD-L1 LNA ASO can result in effective suppression
of PD-L1 expression in the liver This new immunotherapy
offers the potential to overcome the immune tolerance that
characterises HBV infection, and achieve sustainable control
of viral replication.

StephenW

一种新的肝定向锁定核
酸靶向PD-L1表达可调节HBV
免疫耐受性并保持持续的间隙
HBV感染
赵捷1，Lykke Pedersen2，Julie Blaising3，Malika Ait-
Goughoulte3，Fang Shen1，Youjun Yu1，Johanna Walther3，
Anna-Maria Geretti3，Timothy Tellinghuisen3，John Young3，
Soren Ottosen2和Souphalone Luangsay3，（1）Roche Pharma
罗氏创新中心研究与早期开发
中国上海，上海，（2）罗氏制药研究与开发
哥本哈根罗氏创新中心早期开发
丹麦，霍尔斯霍姆（3）Roche Pharma Research and
巴塞尔罗氏创新中心早期开发
瑞士
背景：HBV特异性T细胞功能受损
反应是慢性HBV感染的标志。的
PD-1 / PD-L1途径是关键的免疫检查点调节剂
负责传递强烈的免疫抑制信号
在慢性乙型肝炎患者的肝脏中。我们提出一个新的
使用肝脏靶向锁定核酸（LNA）的技术
单链反义寡核苷酸（ASO）可抑制
PD-L1在肝细胞中的表达（PD-L1 LNA）并破坏肝细胞
肝脏微环境中的PD-1 / PD-L1途径，
恢复局部HBV特异性T细胞效应子功能。方法：
我们生成了针对GalNAc的LNA ASO
针对PD-L1并评估了几种皮下注射
每周一次（QW），剂量范围为3至15 mg / kg
AAV-HBV鼠标模型。循环HBV参数
（HBsAg，HBeAg，HBV DNA）每周监测一次
给药4周，给药结束后持续8周
PD-L1 mRNA的肝内表达（通过RT-qPCR），HBV特异性
免疫反应（通过IFN-γElispot）和肝脏驻留
循环免疫细胞亚群（通过流式细胞仪）
研究结束时在小鼠肝脏中进行了评估：
接受5 QW剂量的PD-L1 LNA的小鼠表现出明显的
在所有测试剂量下，PD-L1 mRNA表达的抑制作用（例如，
5毫克/千克的2剂给药后减少约70％）抑制PD-L1
给药后至少8周保持表达（〜50％），
并与重大和持续
降低HBsAg，HBeAg和HBV DNA水平（2.4、2.1和
1 6 log10下降），无给药后证据
在2到5剂后反弹，伴随着
病毒参数，并根据剂量，我们观察到
暂时性血清ALT增加，而
肝功能有趣的是，用PD-L1 LNA处理的显微镜（5
5 mg / kg的QW剂量显示明显增加（2.8倍）
在肝内免疫细胞数量中，主要由
T细胞亚群和抗原呈递细胞
HBV特异性IFN-γ分泌细胞显着增加
（〜40倍）与车辆对照组比较
耐受性良好，体重无明显变化。
结论：这些发现支持以下假设：
PD-L1 LNA ASO可导致有效抑制
PD-L1在肝脏中的表达这种新的免疫疗法
提供克服免疫耐受的潜力
设计HBV感染并实现可持续控制
病毒复制。