Homodimeric Minimal Factor H: In Vivo Tracking and Extended Dosing Studies in Factor H Deficient Mice.
Homodimeric Minimal Factor H: In Vivo Tracking and Extended Dosing Studies in Factor H Deficient Mice.
复制标题
DOI：
10.3389/fimmu.2021.752916
复制
发表时间：
2021
影响因子：
7.3
通讯作者：
Marchbank KJ
中科院分区：
文献类型：
--
作者：
Kamala O;Malik TH;Hallam TM;Cox TE;Yang Y;Vyas F;Luli S;Connelly C;Gibson B;Smith-Jackson K;Denton H;Pappworth IY;Huang L;Kavanagh D;Pickering MC;Marchbank KJ
C3肾小球病变(C3G)与补体的替代途径(AP)调节失调有关，治疗方案仍然不足。因子H(FH)是AP的有力调节因子。最近发表了一项对FH相关蛋白二聚化最小(Mini)-FH结构的深入分析。这项分析表明，在mini-FH中添加二聚模块不仅延长了血清半衰期，还改善了补体调节功能，从而为C3G提供了一种潜在的治疗选择。在这里，我们描述了一种小鼠版本的同源二聚体微型FH的生产[mHDM-FH(mFH1-5^18-20^R1-2)]，开发用于在C3G和其他补体驱动的小鼠模型的长期实验中降低抗药抗体形成的风险。我们对mHDM-FH的分析表明，与小鼠(M)FH(mHDM-FH Kd=505 nM；MFH Kd=1370 nM)相比，它与WT mC3b具有更高的亲和力和亲和力，类似于我们对各自人类蛋白质的观察。结合亲和力的提高导致了溶血试验中补体调节功能的增强。在CFH-/-小鼠中的延长间隔给药研究(每72小时5 mg/kg)是部分有效的，通过体内成像技术在CFH-/-小鼠中的生物分布分析表明，mHDM-FH优先沉积并在肾脏(和肝脏)保持固定长达4天。使用AAV-Human HDM-FH(hHDM-FH)构建的延长剂量可使CFH-/-小鼠的C3水平在3个月内完全正常化，并与肾小球C3染色显著减少有关。我们的数据证明了微小FH构建体的基因治疗能够增强体内补体调节，并支持将这种方法作为一种新的治疗策略应用于C3G等疾病。
C3 glomerulopathy (C3G) is associated with dysregulation of the alternative pathway (AP) of complement and treatment options remain inadequate. Factor H (FH) is a potent regulator of the AP. An in-depth analysis of FH-related protein dimerised minimal (mini)-FH constructs has recently been published. This analysis showed that addition of a dimerisation module to mini-FH not only increased serum half-life but also improved complement regulatory function, thus providing a potential treatment option for C3G. Herein, we describe the production of a murine version of homodimeric mini-FH [mHDM-FH (mFH1–5^18–20^R1–2)], developed to reduce the risk of anti-drug antibody formation during long-term experiments in murine models of C3G and other complement-driven pathologies. Our analysis of mHDM-FH indicates that it binds with higher affinity and avidity to WT mC3b when compared to mouse (m)FH (mHDM-FH KD=505 nM; mFH KD=1370 nM) analogous to what we observed with the respective human proteins. The improved binding avidity resulted in enhanced complement regulatory function in haemolytic assays. Extended interval dosing studies in CFH-/- mice (5mg/kg every 72hrs) were partially effective and bio-distribution analysis in CFH-/- mice, through in vivo imaging technologies, demonstrates that mHDM-FH is preferentially deposited and remains fixed in the kidneys (and liver) for up to 4 days. Extended dosing using an AAV- human HDM-FH (hHDM-FH) construct achieved complete normalisation of C3 levels in CFH-/- mice for 3 months and was associated with a significant reduction in glomerular C3 staining. Our data demonstrate the ability of gene therapy delivery of mini-FH constructs to enhance complement regulation in vivo and support the application of this approach as a novel treatment strategy in diseases such as C3G.