Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons.
Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons.
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DOI：
10.3390/ijms232416013
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发表时间：
2022-12-16
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
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作者：
ALS相关突变在SOD1蛋白内诱导异常构象，这被认为是SOD1介导的ALS发病机制的基础。虽然SOD1基因沉默的临床试验正在进行中，但这些方法可以减少SOD1的野生型和突变形式。在这里，我们试图开发抗SOD1纳米体，对人类SOD1突变和错误折叠形式具有选择性，而不是野生型SOD1。两种抗SOD1纳米体的表征表明，这些生物制剂在体外稳定了SOD1突变体。此外，通过在永生化细胞中共表达抗SOD1纳米体，SOD1的表达水平得到增强，突变体SOD1的生理亚细胞定位得以恢复。在携带SOD1 A4V突变的人类运动神经元中，抗SOD1纳米体的表达促进了神经突的生长，证明了抗SOD1纳米体对其他不健康细胞的保护作用。体外实验显示，抗SOD1纳米体对人SOD1突变体的选择性优于内源性小鼠SOD1，从而支持抗SOD1纳米体在ALS动物模型中的临床前应用。总之，本文开发和提出的抗sod1纳米体代表了进一步在人类和小鼠ALS模型中进行临床前测试的可行生物制剂。
ALS-linked mutations induce aberrant conformations within the SOD1 protein that are thought to underlie the pathogenic mechanism of SOD1-mediated ALS. Although clinical trials are underway for gene silencing of SOD1, these approaches reduce both wild-type and mutated forms of SOD1. Here, we sought to develop anti-SOD1 nanobodies with selectivity for mutant and misfolded forms of human SOD1 over wild-type SOD1. Characterization of two anti-SOD1 nanobodies revealed that these biologics stabilize mutant SOD1 in vitro. Further, SOD1 expression levels were enhanced and the physiological subcellular localization of mutant SOD1 was restored upon co-expression of anti-SOD1 nanobodies in immortalized cells. In human motor neurons harboring the SOD1 A4V mutation, anti-SOD1 nanobody expression promoted neurite outgrowth, demonstrating a protective effect of anti-SOD1 nanobodies in otherwise unhealthy cells. In vitro assays revealed that an anti-SOD1 nanobody exhibited selectivity for human mutant SOD1 over endogenous murine SOD1, thus supporting the preclinical utility of anti-SOD1 nanobodies for testing in animal models of ALS. In sum, the anti-SOD1 nanobodies developed and presented herein represent viable biologics for further preclinical testing in human and mouse models of ALS.
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影响因子：
15.1
作者：
Baron DM;Kaushansky LJ;Ward CL;Sama RR;Chian RJ;Boggio KJ;Quaresma AJ;Nickerson JA;Bosco DA
通讯作者：
Bosco DA
影响因子：
3.9
作者：
Abel, Olubunmi;Powell, John F.;Al-Chalabi, Ammar
通讯作者：
Al-Chalabi, Ammar
影响因子：
6.1
作者：
Ghadge, Ghanashyam D.;Kay, Brian K.;Roos, Raymond P.
通讯作者：
Roos, Raymond P.
影响因子：
4.6
作者：
Hilton JB;Mercer SW;Lim NK;Faux NG;Buncic G;Beckman JS;Roberts BR;Donnelly PS;White AR;Crouch PJ
通讯作者：
Crouch PJ
影响因子：
25
作者：
Bosco, Daryl A.;Morfini, Gerardo;Karabacak, N. Murat;Song, Yuyu;Gros-Louis, Francois;Pasinelli, Piera;Goolsby, Holly;Fontaine, Benjamin A.;Lemay, Nathan;McKenna-Yasek, Diane;Frosch, Matthew P.;Agar, Jeffrey N.;Julien, Jean-Pierre;Brady, Scott T.;Brown, Robert H., Jr.
通讯作者：
Brown, Robert H., Jr.