Cargando…

Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury

INTRODUCTION: A variety of methods have been used to study inflammatory changes in the acutely injured spinal cord. Recently novel multiplex assays have been used in an attempt to overcome limitations in numbers of available targets studied in a single experiment. Other technical challenges in devel...

Descripción completa

Detalles Bibliográficos
Autores principales: Light, Matthew, Minor, Kenneth H, DeWitt, Peter, Jasper, Kyle H, Davies, Stephen JA
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3439361/
https://www.ncbi.nlm.nih.gov/pubmed/22687332
http://dx.doi.org/10.1186/1742-2094-9-122
_version_ 1782242990082752512
author Light, Matthew
Minor, Kenneth H
DeWitt, Peter
Jasper, Kyle H
Davies, Stephen JA
author_facet Light, Matthew
Minor, Kenneth H
DeWitt, Peter
Jasper, Kyle H
Davies, Stephen JA
author_sort Light, Matthew
collection PubMed
description INTRODUCTION: A variety of methods have been used to study inflammatory changes in the acutely injured spinal cord. Recently novel multiplex assays have been used in an attempt to overcome limitations in numbers of available targets studied in a single experiment. Other technical challenges in developing pre-clinical rodent models to investigate biomarkers in cerebrospinal fluid (CSF) include relatively small volumes of sample and low concentrations of target proteins. The primary objective of this study was to characterize the inflammatory profile present in CSF at a subacute time point in a clinically relevant rodent model of traumatic spinal cord injury (SCI). Our other aim was to test a microarray proteomics platform specifically for this application. METHODS: A 34 cytokine sandwich ELISA microarray was used to study inflammatory changes in CSF samples taken 12 days post-cervical SCI in adult rats. The difference between the median foreground signal and the median background signal was measured. Bonferroni and Benjamini-Hochburg multiple testing corrections were applied to limit the False Discovery Rate (FDR), and a linear mixed model was used to account for repeated measures in the array. RESULTS: We report a novel subacute SCI biomarker, elevated levels of matrix metalloproteinase-8 protein in CSF, and discuss application of statistical models designed for multiplex testing. CONCLUSIONS: Major advantages of this assay over conventional methods include high-throughput format, good sensitivity, and reduced sample consumption. This method can be useful for creating comprehensive inflammatory profiles, and biomarkers can be used in the clinic to assess injury severity and to objectively grade response to therapy.
format Online
Article
Text
id pubmed-3439361
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-34393612012-09-12 Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury Light, Matthew Minor, Kenneth H DeWitt, Peter Jasper, Kyle H Davies, Stephen JA J Neuroinflammation Research INTRODUCTION: A variety of methods have been used to study inflammatory changes in the acutely injured spinal cord. Recently novel multiplex assays have been used in an attempt to overcome limitations in numbers of available targets studied in a single experiment. Other technical challenges in developing pre-clinical rodent models to investigate biomarkers in cerebrospinal fluid (CSF) include relatively small volumes of sample and low concentrations of target proteins. The primary objective of this study was to characterize the inflammatory profile present in CSF at a subacute time point in a clinically relevant rodent model of traumatic spinal cord injury (SCI). Our other aim was to test a microarray proteomics platform specifically for this application. METHODS: A 34 cytokine sandwich ELISA microarray was used to study inflammatory changes in CSF samples taken 12 days post-cervical SCI in adult rats. The difference between the median foreground signal and the median background signal was measured. Bonferroni and Benjamini-Hochburg multiple testing corrections were applied to limit the False Discovery Rate (FDR), and a linear mixed model was used to account for repeated measures in the array. RESULTS: We report a novel subacute SCI biomarker, elevated levels of matrix metalloproteinase-8 protein in CSF, and discuss application of statistical models designed for multiplex testing. CONCLUSIONS: Major advantages of this assay over conventional methods include high-throughput format, good sensitivity, and reduced sample consumption. This method can be useful for creating comprehensive inflammatory profiles, and biomarkers can be used in the clinic to assess injury severity and to objectively grade response to therapy. BioMed Central 2012-06-11 /pmc/articles/PMC3439361/ /pubmed/22687332 http://dx.doi.org/10.1186/1742-2094-9-122 Text en Copyright ©2012 Light et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License(http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Light, Matthew
Minor, Kenneth H
DeWitt, Peter
Jasper, Kyle H
Davies, Stephen JA
Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury
title Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury
title_full Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury
title_fullStr Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury
title_full_unstemmed Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury
title_short Multiplex array proteomics detects increased MMP-8 in CSF after spinal cord injury
title_sort multiplex array proteomics detects increased mmp-8 in csf after spinal cord injury
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3439361/
https://www.ncbi.nlm.nih.gov/pubmed/22687332
http://dx.doi.org/10.1186/1742-2094-9-122
work_keys_str_mv AT lightmatthew multiplexarrayproteomicsdetectsincreasedmmp8incsfafterspinalcordinjury
AT minorkennethh multiplexarrayproteomicsdetectsincreasedmmp8incsfafterspinalcordinjury
AT dewittpeter multiplexarrayproteomicsdetectsincreasedmmp8incsfafterspinalcordinjury
AT jasperkyleh multiplexarrayproteomicsdetectsincreasedmmp8incsfafterspinalcordinjury
AT daviesstephenja multiplexarrayproteomicsdetectsincreasedmmp8incsfafterspinalcordinjury