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Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons
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 t...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784173/ https://www.ncbi.nlm.nih.gov/pubmed/36555655 http://dx.doi.org/10.3390/ijms232416013 |
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author | Kumar, Meenakshi Sundaram Fowler-Magaw, Megan E. Kulick, Daniel Boopathy, Sivakumar Gadd, Del Hayden Rotunno, Melissa Douthwright, Catherine Golebiowski, Diane Yusuf, Issa Xu, Zuoshang Brown, Robert H. Sena-Esteves, Miguel O’Neil, Alison L. Bosco, Daryl A. |
author_facet | Kumar, Meenakshi Sundaram Fowler-Magaw, Megan E. Kulick, Daniel Boopathy, Sivakumar Gadd, Del Hayden Rotunno, Melissa Douthwright, Catherine Golebiowski, Diane Yusuf, Issa Xu, Zuoshang Brown, Robert H. Sena-Esteves, Miguel O’Neil, Alison L. Bosco, Daryl A. |
author_sort | Kumar, Meenakshi Sundaram |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9784173 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-97841732022-12-24 Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons Kumar, Meenakshi Sundaram Fowler-Magaw, Megan E. Kulick, Daniel Boopathy, Sivakumar Gadd, Del Hayden Rotunno, Melissa Douthwright, Catherine Golebiowski, Diane Yusuf, Issa Xu, Zuoshang Brown, Robert H. Sena-Esteves, Miguel O’Neil, Alison L. Bosco, Daryl A. Int J Mol Sci Article 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. MDPI 2022-12-16 /pmc/articles/PMC9784173/ /pubmed/36555655 http://dx.doi.org/10.3390/ijms232416013 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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/). |
spellingShingle | Article Kumar, Meenakshi Sundaram Fowler-Magaw, Megan E. Kulick, Daniel Boopathy, Sivakumar Gadd, Del Hayden Rotunno, Melissa Douthwright, Catherine Golebiowski, Diane Yusuf, Issa Xu, Zuoshang Brown, Robert H. Sena-Esteves, Miguel O’Neil, Alison L. Bosco, Daryl A. Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons |
title | Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons |
title_full | Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons |
title_fullStr | Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons |
title_full_unstemmed | Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons |
title_short | Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons |
title_sort | anti-sod1 nanobodies that stabilize misfolded sod1 proteins also promote neurite outgrowth in mutant sod1 human neurons |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784173/ https://www.ncbi.nlm.nih.gov/pubmed/36555655 http://dx.doi.org/10.3390/ijms232416013 |
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