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用 VHH 内抗体阻断磷蛋白增强心力衰竭时的收缩和舒张功能。

Blocking phospholamban with VHH intrabodies enhances contractility and relaxation in heart failure.

机构信息

Discovery Sciences, R&D, AstraZeneca, Cambridge, UK.

Karolinska Institutet/AstraZeneca Integrated Cardio Metabolic Centre (KI/AZ ICMC), Department of Medicine, Karolinska Institutet, Huddinge, Sweden.

出版信息

Nat Commun. 2022 May 31;13(1):3018. doi: 10.1038/s41467-022-29703-9.

DOI:10.1038/s41467-022-29703-9
PMID:35641497
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9156741/
Abstract

The dysregulated physical interaction between two intracellular membrane proteins, the sarco/endoplasmic reticulum Ca ATPase and its reversible inhibitor phospholamban, induces heart failure by inhibiting calcium cycling. While phospholamban is a bona-fide therapeutic target, approaches to selectively inhibit this protein remain elusive. Here, we report the in vivo application of intracellular acting antibodies (intrabodies), derived from the variable domain of camelid heavy-chain antibodies, to modulate the function of phospholamban. Using a synthetic VHH phage-display library, we identify intrabodies with high affinity and specificity for different conformational states of phospholamban. Rapid phenotypic screening, via modified mRNA transfection of primary cells and tissue, efficiently identifies the intrabody with most desirable features. Adeno-associated virus mediated delivery of this intrabody results in improvement of cardiac performance in a murine heart failure model. Our strategy for generating intrabodies to investigate cardiac disease combined with modified mRNA and adeno-associated virus screening could reveal unique future therapeutic opportunities.

摘要

两种细胞内膜蛋白（肌浆/内质网 Ca2+-ATP 酶及其可逆抑制剂磷酸化肌球蛋白轻链）之间失调的物理相互作用通过抑制钙循环诱导心力衰竭。虽然磷酸化肌球蛋白轻链是一个真正的治疗靶点，但选择性抑制该蛋白的方法仍难以捉摸。在这里，我们报告了源自骆驼重链抗体可变域的细胞内作用抗体（内抗体）在调节磷酸化肌球蛋白轻链功能方面的体内应用。使用合成的 VHH 噬菌体展示文库，我们鉴定出对磷酸化肌球蛋白轻链不同构象状态具有高亲和力和特异性的内抗体。通过对原代细胞和组织的修饰 mRNA 转染进行快速表型筛选，有效地鉴定出具有最理想特征的内抗体。腺相关病毒介导的这种内抗体的递送至心力衰竭的小鼠模型中改善了心脏功能。我们用于研究心脏疾病的产生内抗体的策略结合了修饰的 mRNA 和腺相关病毒筛选，可以揭示独特的未来治疗机会。

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