Intramuscular delivery of formulated RNA encoding six linked nanobodies is highly protective for exposures to three Botulinum neurotoxin serotypes.
Intramuscular delivery of formulated RNA encoding six linked nanobodies is highly protective for exposures to three Botulinum neurotoxin serotypes.
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DOI：
10.1038/s41598-022-15876-2
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发表时间：
2022-07-08
影响因子：
4.6
通讯作者：
Shoemaker, Charles B.
中科院分区：
文献类型：
--
作者：
Mukherjee, Jean;Ondeck, Celinia A.;Tremblay, Jacqueline M.;Archer, Jacob;Debatis, Michelle;Foss, Alexa;Awata, Junya;Erasmus, Jesse H.;McNutt, Patrick M.;Shoemaker, Charles B.
单结构域抗体（sdAb），也称为纳米抗体，相对于常规抗体具有显著的生物物理优势，并且越来越多地被用作免疫抑制剂的组分。一个特别有利的特性是将不同sdAb连接成异多聚体的能力。该特征允许产生能够同时靶向多于一种抗原的单分子。此外，多个连接的sdAb与相同靶标上的非重叠表位的协同结合可以产生靶标亲和力、变体特异性和体内效力的协同改善。在此，我们试图通过测试不同的sdAb异源六聚体来测试这些异源多聚体中增加组分sdAb的选择，其中六种骆驼sdAb组分（VHH）中的每一种都可以中和三种不同的肉毒神经毒素（BoNT）血清型A、B或E中的一种。每种异源六聚体结合所有三种靶向BoNT血清型，并保护小鼠免受每种血清型的至少100个MIPLD 50。为了测试编码长sdAb异多聚体的mRNA治疗剂的潜力，将一种异六聚体编码为复制RNA（repRNA），与阳离子纳米载体一起配制，并通过肌内注射递送至小鼠。通过处理后8小时容易地检测到异源六聚体抗毒素血清表达水平，在约两天达到5-10 nM的峰值，并且持续超过三天。治疗后一天用配制的repRNA治疗的小鼠在用100 MIPLD 50的每种毒素血清型的攻击中存活，证明了所有六种组分VHH的功能。作为蛋白质或repRNA施用的长sdAb多聚体的使用在基于抗体的治疗剂的开发中提供了显著改善的多功能性的潜力。
Single domain antibodies (sdAbs), also called nanobodies, have substantial biophysical advantages over conventional antibodies and are increasingly being employed as components of immunotherapeutic agents. One particularly favorable property is the ability to link different sdAbs into heteromultimers. This feature allows production of single molecules capable of simultaneously targeting more than one antigen. In addition, cooperative binding of multiple linked sdAbs to non-overlapping epitopes on the same target can produce synergistic improvements in target affinity, variant specificity, and in vivo potencies. Here we seek to test the option of increased component sdAbs in these heteromultimers by testing different sdAb heterohexamers in which each of the six camelid sdAb components (VHHs) can neutralize one of three different Botulinum neurotoxin (BoNT) serotypes, A, B or E. Each heterohexamer bound all three targeted BoNT serotypes and protected mice from at least 100 MIPLD50 of each serotype. To test the potential of mRNA therapeutics encoding long sdAb heteromultimers, one heterohexamer was encoded as replicating RNA (repRNA), formulated with a cationic nanocarrier, and delivered to mice via intramuscular injection. Heterohexamer antitoxin serum expression levels were easily detected by 8 h post-treatment, peaked at 5–10 nM around two days, and persisted for more than three days. Mice treated with the formulated repRNA one day post-treatment survived challenge with 100 MIPLD50 of each toxin serotype, demonstrating the function of all six component VHHs. Use of long sdAb multimers, administered as proteins or repRNA, offer the potential for substantially improved versatility in the development of antibody-based therapeutics.
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影响因子：
4.2
作者：
Lou J;Wen W;Conrad F;Meng Q;Dong J;Sun Z;Garcia-Rodriguez C;Farr-Jones S;Cheng LW;Henderson TD;Brown JL;Smith TJ;Smith LA;Cormier A;Marks JD
通讯作者：
Marks JD
影响因子：
12.4
作者：
Erasmus, Jesse H.;Khandhar, Amit P.;Van Hoeven, Neal
通讯作者：
Van Hoeven, Neal
影响因子：
4.8
作者：
Vance, David J.;Tremblay, Jacqueline M.;Shoemaker, Charles B.
通讯作者：
Shoemaker, Charles B.
影响因子：
8
作者：
Vazquez-Cintron, Edwin;Machamer, James;McNutt, Patrick
通讯作者：
McNutt, Patrick
影响因子：
8
作者：
Dumoulin, M;Conrath, K;Matagne, A
通讯作者：
Matagne, A