设计有效的长效Wnt抑制剂DKK2变体

Richelle Sopko, Joshua W. Mugford, A. Lehmann, R. Shapiro, M. Rushe, Abhishek Kulkarni, Joe Worrall, Joseph Amatucci, Dingyi Wen, N. Pederson, Brenda K. Minesinger, J. Arndt, B. Pepinsky
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引用次数: 2

摘要

从胚胎发育到组织修复和再生,各种各样的生物过程都需要Wnt信号通路。Dickkopf-2 (DKK2)通常被定义为典型的Wnt抑制剂,尽管它可能在急性损伤后内皮网络形成的背景下激活非典型Wnt通路。在这里,我们报道了DKK2多肽的融合伙伴的发现,该融合伙伴显著改善了DKK2多肽的表达、生化特性和药代动力学(PK)。具体来说,人血清白蛋白(HSA)被认为是一个非常有效的融合伙伴。通过替换DKK2中选定的氨基酸残基来减少HSA-DKK2变体对硫酸肝素的结合,进一步改善了DKK2分子在啮齿动物体内的PK特性。HSA-DKK2变体是单体的,与野生型一样热稳定,并且通过其结合和阻止Wnt协受体LRP6磷酸化的能力来测量其活性。我们的工程努力导致了典型Wnt抑制剂DKK2的有效长寿命变体,适用于通过系统给药或在组织损伤部位集中给药来操纵Wnt通路。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Engineering potent long-acting variants of the Wnt inhibitor DKK2
Abstract Wnt signaling pathways are required for a wide variety of biological processes ranging from embryonic development to tissue repair and regeneration. Dickkopf-2 (DKK2) is classically defined as a canonical Wnt inhibitor, though it may play a role in activating non-canonical Wnt pathways in the context of endothelial network formation after acute injury. Here we report the discovery of a fusion partner for a DKK2 polypeptide that significantly improves the expression, biochemical properties and pharmacokinetics (PK) of the DKK2 polypeptide. Specifically, human serum albumin (HSA) was identified as a highly effective fusion partner. Substitution of selected amino acid residues in DKK2 designed to decrease heparan sulfate binding by HSA-DKK2 variants, further improved the PK properties of the molecule in rodents. The HSA-DKK2 variants were monomeric, as thermally stable as wild type, and active as measured by their ability to bind to and prevent phosphorylation of the Wnt coreceptor LRP6. Our engineering efforts resulted in potent long-lived variants of the canonical Wnt inhibitor DKK2, applicable for Wnt pathway manipulation either by systematic delivery or focused administration at sites of tissue injury.
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