Roy Arpita, Byrne Sylvester, Sarangi Nirod Kumar, Murphy Paul V, Keyes Tia E
School of Chemical Sciences and National Centre for Sensor Research, Dublin City University, Dublin, Ireland.
School of Biological and Chemical Sciences, University of Galway, Galway, Ireland.
Front Mol Biosci. 2022 Oct 13;9:1017338. doi: 10.3389/fmolb.2022.1017338. eCollection 2022.
Seasonal periodic pandemics and epidemics caused by Influenza A viruses (IAVs) are associated with high morbidity and mortality worldwide. They are frequent and unpredictable in severity so there is a need for biophysical platforms that can be used to provide both mechanistic insights into influenza virulence and its potential treatment by anti-IAV agents. Host membrane viral association through the glycoprotein hemagglutinin (HA) of IAVs is one of the primary steps in infection. HA is thus a potential target for drug discovery and development against influenza. Deconvolution of the multivalent interactions of HA at the interfaces of the host cell membrane can help unravel therapeutic targets. In this contribution, we reported the effect of a multivalent HA glycoprotein association on various glycosphingolipid receptors (GD1a, GM3, GM1) doped asymmetrically into an artificial host membrane spanned across an aqueous filled microcavity array. The extent of HA association and its impact on membrane resistance, capacitance, and diffusivity was measured using highly sensitive electrochemical impedance spectroscopy (EIS) and fluorescence lifetime correlation spectroscopy (FLCS). Furthermore, we investigated the inhibition of the influenza HA glycoprotein association with the host mimetic surface by natural and synthetic sialic acid-based inhibitors (sialic acid, Siaα2,3-GalOMe, FB127, 3-sialyl lactose) using electrochemical impedance spectroscopy and observe that while all inhibit, they do not prevent host binding. Overall, the work demonstrates the platform provides a label-free screening platform for the biophysical evaluation of new inhibitors in the development of potential therapeutics for IAV infection prevention and treatment.
甲型流感病毒（IAV）引发的季节性周期性大流行和流行病在全球范围内都与高发病率和高死亡率相关。它们频繁发生且严重程度不可预测，因此需要生物物理平台来深入了解流感病毒的毒力机制以及抗IAV药物对其的潜在治疗作用。IAV通过糖蛋白血凝素（HA）与宿主细胞膜结合是感染的主要步骤之一。因此，HA是开发抗流感药物的潜在靶点。解析HA在宿主细胞膜界面的多价相互作用有助于揭示治疗靶点。在本论文中，我们报道了多价HA糖蛋白与不对称掺杂到跨越充满水的微腔阵列的人工宿主膜中的各种糖鞘脂受体（GD1a、GM3、GM1）结合的影响。使用高灵敏度电化学阻抗谱（EIS）和荧光寿命相关光谱（FLCS）测量了HA结合的程度及其对膜电阻、电容和扩散率的影响。此外，我们使用电化学阻抗谱研究了天然和合成的基于唾液酸的抑制剂（唾液酸、Siaα2,3-GalOMe、FB127、3-唾液酸乳糖）对流感HA糖蛋白与宿主模拟表面结合的抑制作用，并观察到虽然所有抑制剂都有抑制作用，但它们并不能阻止宿主结合。总体而言，这项工作表明该平台为IAV感染预防和治疗潜在疗法开发中新型抑制剂的生物物理评估提供了一个无标记筛选平台。
