Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons
AuthorsKumar, Meenakshi Sundaram
Fowler-Magaw, Megan E
Gadd, Del Hayden
Brown, Robert H
O'Neil, Alison L
Bosco, Daryl A
Academic ProgramBiochemistry and Molecular Biotechnology; Neuroscience
UMass Chan AffiliationsBiochemistry and Molecular Biotechnology
Horae Gene Therapy Center
Morningside Graduate School of Biomedical Sciences
Document TypeJournal Article
Keywordsamyotrophic lateral sclerosis (ALS) (Lou Gehrig disease)
superoxide dismutase (SOD)
MetadataShow full item record
AbstractALS-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.
SourceKumar MS, Fowler-Magaw ME, Kulick D, Boopathy S, Gadd DH, Rotunno M, Douthwright C, Golebiowski D, Yusuf I, Xu Z, Brown RH Jr, Sena-Esteves M, O'Neil AL, Bosco DA. Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons. Int J Mol Sci. 2022 Dec 16;23(24):16013. doi: 10.3390/ijms232416013. PMID: 36555655; PMCID: PMC9784173.
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/51735
Funding and AcknowledgementsThis project was also funded by the Angel Fund for ALS Research for R.H.B.J.; the Pilot Project Program grant UL1TR001453 from the UMass Center for Clinical and Translational Science (UMCCTS) for D.A.B., M.E.-S., and R.H.B.J.; and NIH/NINDS R01 NS067206 for D.A.B.
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Except where otherwise noted, this item's license is described as Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).