Utility of Survival Motor Neuron ELISA for Spinal Muscular Atrophy Clinical and Preclinical Analyses
Open Access
- 31 August 2011
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 6 (8), e24269
- https://doi.org/10.1371/journal.pone.0024269
Abstract
Genetic defects leading to the reduction of the survival motor neuron protein (SMN) are a causal factor for Spinal Muscular Atrophy (SMA). While there are a number of therapies under evaluation as potential treatments for SMA, there is a critical lack of a biomarker method for assessing efficacy of therapeutic interventions, particularly those targeting upregulation of SMN protein levels. Towards this end we have engaged in developing an immunoassay capable of accurately measuring SMN protein levels in blood, specifically in peripheral blood mononuclear cells (PBMCs), as a tool for validating SMN protein as a biomarker in SMA. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed and validated for measuring SMN protein in human PBMCs and other cell lysates. Protocols for detection and extraction of SMN from transgenic SMA mouse tissues were also developed. The assay sensitivity for human SMN is 50 pg/mL. Initial analysis reveals that PBMCs yield enough SMN to analyze from blood volumes of less than 1 mL, and SMA Type I patients' PBMCs show ∼90% reduction of SMN protein compared to normal adults. The ELISA can reliably quantify SMN protein in human and mouse PBMCs and muscle, as well as brain, and spinal cord from a mouse model of severe SMA. This SMN ELISA assay enables the reliable, quantitative and rapid measurement of SMN in healthy human and SMA patient PBMCs, muscle and fibroblasts. SMN was also detected in several tissues in a mouse model of SMA, as well as in wildtype mouse tissues. This SMN ELISA has general translational applicability to both preclinical and clinical research efforts.This publication has 52 references indexed in Scilit:
- CNS-targeted gene therapy improves survival and motor function in a mouse model of spinal muscular atrophyJCI Insight, 2010
- A degron created by SMN2 exon 7 skipping is a principal contributor to spinal muscular atrophy severityGenes & Development, 2010
- RETRACTED ARTICLE: Rescue of the spinal muscular atrophy phenotype in a mouse model by early postnatal delivery of SMNNature Biotechnology, 2010
- Spinal Muscular Atrophy: New and Emerging Insights from Model MiceCurrent Neurology and Neuroscience Reports, 2010
- Effects of 2,4-diaminoquinazoline derivatives on SMN expression and phenotype in a mouse model for spinal muscular atrophyHuman Molecular Genetics, 2009
- Tetracyclines That Promote SMN2 Exon 7 Splicing as Therapeutics for Spinal Muscular AtrophyScience Translational Medicine, 2009
- Vascular Perfusion Abnormalities in Infants with Spinal Muscular AtrophyThe Journal of Pediatrics, 2009
- Differences in SMN1 allele frequencies among ethnic groups within North AmericaJournal of Medical Genetics, 2009
- DcpS as a Therapeutic Target for Spinal Muscular AtrophyACS Chemical Biology, 2008
- Identification and characterization of the porcine (Sus scrofa) survival motor neuron (SMN1) gene: An animal model for therapeutic studiesDevelopmental Dynamics, 2008