Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons

被引:3
|
作者
Kumar, Meenakshi Sundaram [1 ,2 ]
Fowler-Magaw, Megan E. [1 ,3 ]
Kulick, Daniel [4 ]
Boopathy, Sivakumar [1 ,2 ]
Gadd, Del Hayden [1 ]
Rotunno, Melissa [1 ,2 ]
Douthwright, Catherine [1 ,2 ]
Golebiowski, Diane [1 ,5 ]
Yusuf, Issa [6 ]
Xu, Zuoshang [6 ]
Brown Jr, Robert H. H. [1 ]
Sena-Esteves, Miguel [1 ,5 ]
O'Neil, Alison L. [7 ]
Bosco, Daryl A. [1 ,6 ]
机构
[1] Univ Massachusetts, Dept Neurol, Chan Med Sch, Worcester, MA 01605 USA
[2] Univ Massachusetts, Morningside Grad Sch Biomed Sci, Biochem & Mol Biotechnol Program, Chan Med Sch, Worcester, MA 01605 USA
[3] Univ Massachusetts, Morningside Grad Sch Biomed Sci, Neurosci Program, Chan Med Sch, Worcester, MA 01605 USA
[4] Wesleyan Univ, Dept Biol, Neurosci & Behav Program, Middletown, CT 06459 USA
[5] Univ Massachusetts, Horae Gene Therapy Ctr, Chan Med Sch, Worcester, MA 01605 USA
[6] Univ Massachusetts, Dept Biochem & Mol Biotechnol, Chan Med Sch, Worcester, MA 01605 USA
[7] Wesleyan Univ, Dept Chem, Neurosci & Behav Program, Middletown, CT 06459 USA
关键词
amyotrophic lateral sclerosis (ALS) (Lou Gehrig disease); antibody engineering; neurite outgrowth; protein misfolding; superoxide dismutase (SOD); AMYOTROPHIC-LATERAL-SCLEROSIS; SUPEROXIDE-DISMUTASE; MOTOR-NEURONS; WILD-TYPE; ADENOASSOCIATED VIRUS; MOUSE MODEL; ALS; MICE; AGGREGATION; ANTIBODIES;
D O I
10.3390/ijms232416013
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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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页数:24
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