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Molecular Neurodegeneration
Published in: Molecular Neurodegeneration 1/2016

Open Access 01-12-2016 | Methodology

ZNStress: a high-throughput drug screening protocol for identification of compounds modulating neuronal stress in the transgenic mutant sod1G93R zebrafish model of amyotrophic lateral sclerosis

Authors: Alexander McGown, Dame Pamela J. Shaw, Tennore Ramesh

Published in: Molecular Neurodegeneration | Issue 1/2016

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Abstract

Background

Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative disease with death on average within 2–3 years of symptom onset. Mutations in superoxide dismutase 1 (SOD1) have been identified to cause ALS. Riluzole, the only neuroprotective drug for ALS provides life extension of only 3 months on average. Thishighlights the need for compound screening in disease models to identify new neuroprotective therapies for this disease. Zebrafish is an emerging model system that is well suited for the study of diseasepathophysiology and also for high throughput (HT) drug screening. The mutant sod1 zebrafish model of ALS mimics the hallmark features of ALS. Using a fluorescence based readout of neuronal stress, we developed a high throughput (HT) screen to identify neuroprotective compounds.

Results

Here we show that the zebrafish screen is a robust system that can be used to rapidly screen thousands ofcompounds and also demonstrate that riluzole is capable of reducing neuronal stress in this model system. The screen shows optimal quality control, maintaining a high sensitivity and specificity withoutcompromising throughput. Most importantly, we demonstrate that many compounds previously failed in human clinical trials, showed no stress reducing activity in the zebrafish assay.

Conclusion

We conclude that HT drug screening using a mutant sod1 zebrafish is a reliable model system which supplemented with secondary assays would be useful in identifying drugs with potential for neuroprotective efficacy in ALS.
Appendix
Available only for authorised users
Literature
1.
Turner BJ, Talbot K. Transgenics, toxicity and therapeutics in rodent models of mutant SOD1-mediated familial ALS. Prog Neurobiol. 2008;85(1):94–134.CrossRefPubMed
2.
Da Costa MM, Allen CE, Higginbottom A, Ramesh T, Shaw PJ, McDermott CJ. A new zebrafish model produced by TILLING of SOD1-related amyotrophic lateral sclerosis replicates key features of the disease and represents a tool for in vivo therapeutic screening. Dis Model Mech. 2014;7(1):73–81.CrossRefPubMed
3.
McGown A, McDearmid JR, Panagiotaki N, Tong H, Al Mashhadi S, Redhead N, et al. Early interneuron dysfunction in ALS: insights from a mutant sod1 zebrafish model. Ann Neurol. 2013;73(2):246–58.CrossRefPubMed
4.
Sakowski SA, Lunn JS, Busta AS, Oh SS, Zamora-Berridi G, Palmer M, et al. Neuromuscular effects of G93A-SOD1 expression in zebrafish. Mol Neurodegener. 2012;7:44.CrossRefPubMedPubMedCentral
5.
Ramesh T, Lyon AN, Pineda RH, Wang C, Janssen PM, Canan BD, et al. A genetic model of amyotrophic lateral sclerosis in zebrafish displays phenotypic hallmarks of motoneuron disease. Dis Model Mech. 2010;3(9-10):652–62.CrossRefPubMedPubMedCentral
6.
Miller RG, Mitchell JD, Moore DH. Riluzole for amyotrophic lateral sclerosis (ALS)/motor neuron disease (MND). Cochrane Database Syst Rev. 2012;3:CD001447.PubMed
7.
Lee JA, Berg EL. Neoclassic drug discovery: the case for lead generation using phenotypic and functional approaches. J Biomol Screen. 2013;18(10):1143–55.CrossRefPubMed
8.
Lee JA, Uhlik MT, Moxham CM, Tomandl D, Sall DJ. Modern phenotypic drug discovery is a viable, neoclassic pharma strategy. J Med Chem. 2012;55(10):4527–38.CrossRefPubMed
9.
Swinney DC, Anthony J. How were new medicines discovered? Nat Rev Drug Discov. 2011;10(7):507–19.CrossRefPubMed
10.
11.
Zhang M, Luo G, Zhou Y, Wang S, Zhong Z. Phenotypic screens targeting neurodegenerative diseases. J Biomol Screen. 2014;19(1):1–16.CrossRefPubMed
12.
Patten SA, Armstrong GA, Lissouba A, Kabashi E, Parker JA, Drapeau P. Fishing for causes and cures of motor neuron disorders. Dis Model Mech. 2014;7(7):799–809.CrossRefPubMedPubMedCentral
13.
Babin PJ, Goizet C, Raldua D. Zebrafish models of human motor neuron diseases: advantages and limitations. Prog Neurobiol. 2014;118:36–58.CrossRefPubMed
14.
15.
16.
Baxendale S, Holdsworth CJ, Meza Santoscoy PL, Harrison MR, Fox J, Parkin CA, et al. Identification of compounds with anti-convulsant properties in a zebrafish model of epileptic seizures. Dis Model Mech. 2012;5(6):773–84.CrossRefPubMedPubMedCentral
17.
Ramesh TM, Shaw DP, McDearmid J. A zebrafish model exemplifies the long preclinical period of motor neuron disease. J Neurol Neurosurg Psychiatry. 2014;85(11):1288–9.
18.
Eisen A, Kiernan M, Mitsumoto H, Swash M. Amyotrophic lateral sclerosis: a long preclinical period? J Neurol Neurosurg Psychiatry. 2014;85(11):1232–8.
19.
Eisen A. Response to a letter by Dr T Ramesh. J Neurol Neurosurg Psychiatry. 2014;85(11):1289.
20.
Turner MR, Kiernan MC. Does interneuronal dysfunction contribute to neurodegeneration in amyotrophic lateral sclerosis? Amyotroph Lateral Scler. 2012;13(3):245–50.CrossRefPubMed
21.
Carew JD, Nair G, Andersen PM, Wuu J, Gronka S, Hu X, et al. Presymptomatic spinal cord neurometabolic findings in SOD1-positive people at risk for familial ALS. Neurology. 2011;77(14):1370–5.CrossRefPubMedPubMedCentral
22.
Vucic S, Nicholson GA, Kiernan MC. Cortical hyperexcitability may precede the onset of familial amyotrophic lateral sclerosis. Brain. 2008;131(Pt 6):1540–50.CrossRefPubMed
23.
24.
25.
26.
Liu J, Shinobu LA, Ward CM, Young D, Cleveland DW. Elevation of the Hsp70 chaperone does not effect toxicity in mouse models of familial amyotrophic lateral sclerosis. J Neurochem. 2005;93(4):875–82.CrossRefPubMed
27.
28.
Zhang XD, Ferrer M, Espeseth AS, Marine SD, Stec EM, Crackower MA, et al. The use of strictly standardized mean difference for hit selection in primary RNA interference high-throughput screening experiments. J Biomol Screen. 2007;12(4):497–509.CrossRefPubMed
29.
Zhang XD. Illustration of SSMD, z score, SSMD*, z* score, and t statistic for hit selection in RNAi high-throughput screens. J Biomol Screen. 2011;16(7):775–85.CrossRefPubMed
30.
Benatar M. Lost in translation: treatment trials in the SOD1 mouse and in human ALS. Neurobiol Dis. 2007;26(1):1–13.CrossRefPubMed
31.
Gill A, Kidd J, Vieira F, Thompson K, Perrin S. No benefit from chronic lithium dosing in a sibling-matched, gender balanced, investigator-blinded trial using a standard mouse model of familial ALS. PLoS One. 2009;4(8):e6489.CrossRefPubMedPubMedCentral
32.
Ludolph AC, Bendotti C, Blaugrund E, Hengerer B, Loffler JP, Martin J, et al. Guidelines for the preclinical in vivo evaluation of pharmacological active drugs for ALS/MND: report on the 142nd ENMC international workshop. Amyotroph Lateral Scler. 2007;8(4):217–23.CrossRefPubMed
33.
Scott S, Kranz JE, Cole J, Lincecum JM, Thompson K, Kelly N, et al. Design, power, and interpretation of studies in the standard murine model of ALS. Amyotroph Lateral Scler. 2008;9(1):4–15.CrossRefPubMed
34.
Vieira FG, LaDow E, Moreno A, Kidd JD, Levine B, Thompson K, et al. Dexpramipexole Is Ineffective in Two Models of ALS Related Neurodegeneration. PLoS One. 2014;9(12):e91608.CrossRefPubMedPubMedCentral
35.
Kalmar B, Novoselov S, Gray A, Cheetham ME, Margulis B, Greensmith L. Late stage treatment with arimoclomol delays disease progression and prevents protein aggregation in the SOD1 mouse model of ALS. J Neurochem. 2008;107(2):339–50.CrossRefPubMed
36.
Kieran D, Kalmar B, Dick JR, Riddoch-Contreras J, Burnstock G, Greensmith L. Treatment with arimoclomol, a coinducer of heat shock proteins, delays disease progression in ALS mice. Nat Med. 2004;10(4):402–5.CrossRefPubMed
37.
38.
Zou ZY, Liu CY, Che CH, Huang HP. Toward precision medicine in amyotrophic lateral sclerosis. Ann Transl Med. 2016;4(2):27.PubMedPubMedCentral
39.
Ludolph AC, Bendotti C, Blaugrund E, Chio A, Greensmith L, Loeffler JP, et al. Guidelines for preclinical animal research in ALS/MND: A consensus meeting. Amyotroph Lateral Scler. 2010;11(1-2):38–45.CrossRefPubMed
Metadata
Title
ZNStress: a high-throughput drug screening protocol for identification of compounds modulating neuronal stress in the transgenic mutant sod1G93R zebrafish model of amyotrophic lateral sclerosis
Authors
Alexander McGown
Dame Pamela J. Shaw
Tennore Ramesh
Publication date
01-12-2016
Publisher
BioMed Central
Published in
Molecular Neurodegeneration / Issue 1/2016
Electronic ISSN: 1750-1326
DOI
https://doi.org/10.1186/s13024-016-0122-3

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