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3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes
OBJECTIVES/SPECIFIC AIMS: We previously demonstrated that products released by cultured B cells from patients with Multiple Sclerosis (MS) are cytotoxic to neurons and oligodendrocytes, while minimal toxicity was observed in response to B cell secretory products from age- and sex-matched normal cont...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cambridge University Press
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6799518/ http://dx.doi.org/10.1017/cts.2019.265 |
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author | Zuroff, Leah Touil, Hanane Romer, Micah Nedelkoska, Liljana Benjamins, Joyce A. Lisak, Robert P. Grinspan, Judith B. Bar-Or, Amit |
author_facet | Zuroff, Leah Touil, Hanane Romer, Micah Nedelkoska, Liljana Benjamins, Joyce A. Lisak, Robert P. Grinspan, Judith B. Bar-Or, Amit |
author_sort | Zuroff, Leah |
collection | PubMed |
description | OBJECTIVES/SPECIFIC AIMS: We previously demonstrated that products released by cultured B cells from patients with Multiple Sclerosis (MS) are cytotoxic to neurons and oligodendrocytes, while minimal toxicity was observed in response to B cell secretory products from age- and sex-matched normal controls. The goal of this proposal is to identify the range of brain cells susceptible to MS B cell-mediated cytotoxicity, to define the cytotoxic factor(s) released by MS B cells, and to determine whether particular subset(s) of MS B cells harbor the greatest pathogenic potential. METHODS/STUDY POPULATION: The toxicity of B cell products will be demonstrated by incubating primary rat cultures of neurons, oligodendrocytes, and oligodendrocyte progenitor cells (OPCs) with B cell supernatants. B cells will be isolated from the peripheral circulation of untreated relapse-remitting MS (RRMS) patients and age- and sex-matched normal controls. The identification of specific toxic factor(s) in MS B cell supernatants will be achieved through a combination of exosome-depletion/enrichment of conditioned media, proteomics, next generation sequencing, and lipidomics. Determining pathogenic B cell subsets will be achieved by cell sorting into memory and naïve B cell subsets prior to collection of supernatants. RESULTS/ANTICIPATED RESULTS: We hypothesize that the toxicity of MS B cell products is mediated, at least in part, by extracellular vesicles, such as exosomes. We expect depletion of these exosomes from the B cell conditioned media or inhibition of their biogenesis will mitigate the observed toxicity. Furthermore, differences in B cell-derived exosomal content, such as proteins, (mi)RNAs, or lipids, likely explain the differences in observed toxicity. Lastly, we hypothesize that memory B cells, which are enriched in the CNS of MS patients and demonstrate a more pro-inflammatory profile than naïve B cells, are responsible for the toxicity observed in supernatants of total B cells. DISCUSSION/SIGNIFICANCE OF IMPACT: MS is the most prevalent chronic inflammatory disease of the CNS, affecting more than 2 million people worldwide. Although over a dozen disease-modifying therapies are approved for the treatment of RRMS, none are meaningfully effective at limiting disease progression. This proposal will provide new insight into immune-CNS interactions in progressive MS and provide much-needed novel targets for therapeutic intervention, either via blocking identified toxic molecule(s) or by selectively depleting pathogenic B cell subsets. |
format | Online Article Text |
id | pubmed-6799518 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Cambridge University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-67995182019-10-28 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes Zuroff, Leah Touil, Hanane Romer, Micah Nedelkoska, Liljana Benjamins, Joyce A. Lisak, Robert P. Grinspan, Judith B. Bar-Or, Amit J Clin Transl Sci Mechanistic Basic to Clinical OBJECTIVES/SPECIFIC AIMS: We previously demonstrated that products released by cultured B cells from patients with Multiple Sclerosis (MS) are cytotoxic to neurons and oligodendrocytes, while minimal toxicity was observed in response to B cell secretory products from age- and sex-matched normal controls. The goal of this proposal is to identify the range of brain cells susceptible to MS B cell-mediated cytotoxicity, to define the cytotoxic factor(s) released by MS B cells, and to determine whether particular subset(s) of MS B cells harbor the greatest pathogenic potential. METHODS/STUDY POPULATION: The toxicity of B cell products will be demonstrated by incubating primary rat cultures of neurons, oligodendrocytes, and oligodendrocyte progenitor cells (OPCs) with B cell supernatants. B cells will be isolated from the peripheral circulation of untreated relapse-remitting MS (RRMS) patients and age- and sex-matched normal controls. The identification of specific toxic factor(s) in MS B cell supernatants will be achieved through a combination of exosome-depletion/enrichment of conditioned media, proteomics, next generation sequencing, and lipidomics. Determining pathogenic B cell subsets will be achieved by cell sorting into memory and naïve B cell subsets prior to collection of supernatants. RESULTS/ANTICIPATED RESULTS: We hypothesize that the toxicity of MS B cell products is mediated, at least in part, by extracellular vesicles, such as exosomes. We expect depletion of these exosomes from the B cell conditioned media or inhibition of their biogenesis will mitigate the observed toxicity. Furthermore, differences in B cell-derived exosomal content, such as proteins, (mi)RNAs, or lipids, likely explain the differences in observed toxicity. Lastly, we hypothesize that memory B cells, which are enriched in the CNS of MS patients and demonstrate a more pro-inflammatory profile than naïve B cells, are responsible for the toxicity observed in supernatants of total B cells. DISCUSSION/SIGNIFICANCE OF IMPACT: MS is the most prevalent chronic inflammatory disease of the CNS, affecting more than 2 million people worldwide. Although over a dozen disease-modifying therapies are approved for the treatment of RRMS, none are meaningfully effective at limiting disease progression. This proposal will provide new insight into immune-CNS interactions in progressive MS and provide much-needed novel targets for therapeutic intervention, either via blocking identified toxic molecule(s) or by selectively depleting pathogenic B cell subsets. Cambridge University Press 2019-03-27 /pmc/articles/PMC6799518/ http://dx.doi.org/10.1017/cts.2019.265 Text en © The Association for Clinical and Translational Science 2019 http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article, distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives licence (http://creativecommons.org/licenses/by-ncnd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is unaltered and is properly cited. The written permission of Cambridge University Press must be obtained for commercial re-use or in order to create a derivative work. |
spellingShingle | Mechanistic Basic to Clinical Zuroff, Leah Touil, Hanane Romer, Micah Nedelkoska, Liljana Benjamins, Joyce A. Lisak, Robert P. Grinspan, Judith B. Bar-Or, Amit 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes |
title | 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes |
title_full | 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes |
title_fullStr | 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes |
title_full_unstemmed | 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes |
title_short | 3285 Toxicity of Released B Cell Products in Multiple Sclerosis: Effects on Neurons and Oligodendrocytes |
title_sort | 3285 toxicity of released b cell products in multiple sclerosis: effects on neurons and oligodendrocytes |
topic | Mechanistic Basic to Clinical |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6799518/ http://dx.doi.org/10.1017/cts.2019.265 |
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