Iminosugar Glucosidase Inhibitors Reduce Hepatic Inflammation in Hepatitis A Virus-Infected Ifnar1-/- Mice
Iminosugar Glucosidase Inhibitors Reduce Hepatic Inflammation in Hepatitis A Virus-Infected Ifnar1-/- Mice
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DOI：
10.1128/jvi.00058-21
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发表时间：
2021-06-01
影响因子：
5.4
通讯作者：
Lemon, Stanley M.
中科院分区：
文献类型：
--
作者：
Misumi, Ichiro;Li, Zhucui;Lemon, Stanley M.
亚氨基糖化合物是单糖模拟物，具有广泛但通常较弱的与糖生物学中涉及的酶的抑制相关的抗病毒活性。麦格司他（N-丁基-1-脱氧野尻霉素）获批用于治疗人类脂质沉积病，UV-4 [N-（9-甲氧基壬基）-1-脱氧野尻霉素]通过阻断HAV细胞进入所必需的神经节苷脂的合成，抑制细胞培养物中甲型肝炎病毒（HAV）的复制（50%抑制浓度[IC（50）分别为32.13 mM和8.05 mM）。我们使用甲型肝炎的小鼠模型和靶向质谱法来评估这些化合物消耗肝神经节苷脂和修改体内HAV感染过程的能力。通过灌胃给予Ifnar 1（-/-）小鼠麦格司他（4，800 mg/kg体重/天），肝神经节苷脂消耗了69%至75%，但引起了严重的胃肠道毒性，并且未能预防病毒感染。UV-4，类似地以高剂量（400 mg/kg/天）给药，耐受性良好，但14天后肝神经节苷脂仅消耗20%。神经节苷脂的UV-4消耗因类别而异。几种GM 2物种矛盾地增加，可能是由于抑制降解神经节苷脂的β-葡糖苷酶。这两种化合物增强，而不是减少，病毒复制。尽管如此，这两种亚胺糖都有令人惊讶的抗炎作用，阻止肝脏内炎症细胞的积累。UV-4治疗还导致与急性甲型肝炎相关的血清丙氨酸氨基转移酶（ALT）升高降低。这些抗炎作用可能是由于亚氨基糖抑制细胞α-葡萄糖苷酶，导致受损的成熟的聚糖部分的趋化因子和细胞因子受体，并指出在急性甲型肝炎的发病机制旁分泌信号的潜在重要性。重要性甲型肝炎病毒（HAV）是病毒性肝炎的常见原因。亚氨基糖化合物通过抑制HAV进入细胞所需的神经节苷脂的合成来阻断其在培养细胞中的复制，但尚未测试其在体内预防或治疗甲型肝炎的能力。我们发现，在缺乏关键干扰素受体的小鼠中，高剂量的亚胺糖麦格司他和UV-4未能充分消耗神经节苷脂以阻断HAV感染。尽管如此，这些化合物对HAV感染的肝脏具有显著的抗炎作用，降低了肝炎的严重程度，尽管增强了由肝细胞内在抗病毒应答引起的趋化因子和细胞因子表达。我们提出，亚氨基糖抑制细胞α-葡糖苷酶损害成熟的聚糖部分的趋化因子和细胞因子受体所需的有效信号。这些数据突出了旁分泌信号通路在HAV炎症反应中的潜在重要性，并增加了我们对小鼠HAV发病机制的理解。
Iminosugar compounds are monosaccharide mimetics with broad but generally weak antiviral activities related to inhibition of enzymes involved in glycobiology. Miglustat (N-butyl-1-deoxynojirimycin), which is approved for the treatment of lipid storage diseases in humans, and UV-4 [N-(9-methoxynonyl)-1-deoxynojirimy-cin] inhibit the replication of hepatitis A virus (HAV) in cell culture (50% inhibitory concentrations [IC(50)s] of 32.13mM and 8.05mM, respectively) by blocking the synthesis of gangliosides essential for HAV cell entry. We used a murine model of hepatitis A and targeted mass spectrometry to assess the capacity of these compounds to deplete hepatic gangliosides and modify the course of HAV infection in vivo. Miglustat, given by gavage to Ifnar1(-/-) mice (4,800 mg/kg of body weight/day) depleted hepatic gangliosides by 69 to 75% but caused substantial gastrointestinal toxicity and failed to prevent viral infection. UV-4, similarly administered in high doses (400 mg/kg/day), was well tolerated but depleted hepatic gangliosides by only 20% after 14 days. UV-4 depletion of gangliosides varied by class. Several GM2 species were paradoxically increased, likely due to inhibition of beta-glucosidases that degrade gangliosides. Both compounds enhanced, rather than reduced, virus replication. Nonetheless, both iminosugars had surprising anti-inflammatory effects, blocking the accumulation of inflammatory cells within the liver. UV-4 treatment also resulted in a decrease in serum alanine aminotransferase (ALT) elevations associated with acute hepatitis A. These anti-inflammatory effects may result from iminosugar inhibition of cellular alpha-glucosidases, leading to impaired maturation of glycan moieties of chemokine and cytokine receptors, and point to the potential importance of paracrine signaling in the pathogenesis of acute hepatitis A.IMPORTANCE Hepatitis A virus (HAV) is a common cause of viral hepatitis. Iminosugar compounds block its replication in cultured cells by inhibiting the synthesis of gangliosides required for HAV cell entry but have not been tested for their ability to prevent or treat hepatitis A in vivo. We show that high doses of the iminosugars miglustat and UV-4 fail to deplete gangliosides sufficiently to block HAV infection in mice lacking a key interferon receptor. These compounds nonetheless have striking anti-inflammatory effects on the HAV-infected liver, reducing the severity of hepatitis despite enhancing chemokine and cytokine expression resulting from hepatocyteintrinsic antiviral responses. We propose that iminosugar inhibition of cellular alpha-glucosidases impairs the maturation of glycan moieties of chemokine and cytokine receptors required for effective signaling. These data highlight the potential importance of paracrine signaling pathways in the inflammatory response to HAV and add to our understanding of HAV pathogenesis in mice.