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Unbiased Screens for Modifiers of Alpha-Synuclein Toxicity

  • Matthias Höllerhage
  • Marc Bickle
  • Günter U. HöglingerEmail author
Genetics (V Bonifati, Section Editor)
Part of the following topical collections:
  1. Topical Collection on Genetics

Abstract

Purpose of Review

We provide an overview about unbiased screens to identify modifiers of alpha-synuclein (αSyn)-induced toxicity, present the models and the libraries that have been used for screening, and describe how hits from primary screens were selected and validated.

Recent Findings

Screens can be classified as either genetic or chemical compound modifier screens, but a few screens do not fit this classification. Most screens addressing αSyn-induced toxicity, including genome-wide overexpressing and deletion, were performed in yeast. More recently, newer methods such as CRISPR-Cas9 became available and were used for screening purposes. Paradoxically, given that αSyn-induced toxicity plays a role in neurological diseases, there is a shortage of human cell-based models for screening. Moreover, most screens used mutant or fluorescently tagged forms of αSyn and only very few screens investigated wild-type αSyn. Particularly, no genome-wide αSyn toxicity screen in human dopaminergic neurons has been published so far.

Summary

Most unbiased screens for modifiers of αSyn toxicity were performed in yeast, and there is a lack of screens performed in human and particularly dopaminergic cells.

Keywords

Screen Alpha-synuclein Toxicity Parkinson’s disease 

Notes

Funding Information

This work was supported by the BMBF-funded project “HitTau” (01EK1605A to G.U.H.), the Deutsche Forschungsgemeinschaft (DFG, HO2402/18-1, Munich Cluster for Systems Neurology SyNergy) (to G.U.H.), the NOMIS foundation (FTLD project to G.U.H.), and the Parkinson Fonds Deutschland (α-synuclein high-throughput screening to G.U.H. and M.H.).

Compliance with Ethical Standards

Conflict of Interest

Günter Höglinger and Matthias Höllerhage each declare no potential conflict of interest.

Marc Bickle reports a fee for service from DZNE, during the conduct of the study.

Human and Animal Rights and Informed Consent

This article does not contain any studies with human or animal subjects performed by any of the authors.

References

Papers of particular interest, published recently, have been highlighted as: • Of importance •• Of major importance

  1. 1.
    •• Uversky VN. Looking at the recent advances in understanding α-synuclein and its aggregation through the proteoform prism. F1000Res. 2017;6:525.  https://doi.org/10.12688/f1000research.10536.1 A recent review about the function of alpha-synuclein. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Alafuzoff I, Hartikainen P. Alpha-synucleinopathies. Handb Clin Neurol. 2017;145:339–53.  https://doi.org/10.1016/B978-0-12-802395-2.00024-9.CrossRefPubMedGoogle Scholar
  3. 3.
    Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M. Alpha-synuclein in Lewy bodies. Nature. 1997;388:839–40.  https://doi.org/10.1038/42166.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Spillantini MG, Crowther RA, Jakes R, Cairns NJ, Lantos PL, Goedert M. Filamentous alpha-synuclein inclusions link multiple system atrophy with Parkinson’s disease and dementia with Lewy bodies. Neurosci Lett. 1998;251:205–8.CrossRefGoogle Scholar
  5. 5.
    Tysnes O-B, Storstein A. Epidemiology of Parkinson’s disease. J Neural Transm (Vienna). 2017;124:901–5.  https://doi.org/10.1007/s00702-017-1686-y.CrossRefGoogle Scholar
  6. 6.
    Petrucci S, Ginevrino M, Valente EM. Phenotypic spectrum of alpha-synuclein mutations: new insights from patients and cellular models. Parkinsonism Relat Disord. 2016;22(Suppl 1):S16–20.  https://doi.org/10.1016/j.parkreldis.2015.08.015.CrossRefPubMedGoogle Scholar
  7. 7.
    Chartier-Harlin M-C, Kachergus J, Roumier C, Mouroux V, Douay X, Lincoln S, et al. Alpha-synuclein locus duplication as a cause of familial Parkinson’s disease. Lancet. 2004;364:1167–9.  https://doi.org/10.1016/S0140-6736(04)17103-1.CrossRefPubMedGoogle Scholar
  8. 8.
    Singleton AB, Farrer M, Johnson J, Singleton A, Hague S, Kachergus J, et al. alpha-Synuclein locus triplication causes Parkinson’s disease. Science. 2003;302:841.  https://doi.org/10.1126/science.1090278.CrossRefPubMedGoogle Scholar
  9. 9.
    Nalls MA, Pankratz N, Lill CM, Do CB, Hernandez DG, Saad M, et al. Large-scale meta-analysis of genome-wide association data identifies six new risk loci for Parkinson’s disease. Nat Genet. 2014;46:989–93.  https://doi.org/10.1038/ng.3043.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Oertel WH. Recent advances in treating Parkinson’s disease. F1000Res. 2017;6:260.  https://doi.org/10.12688/f1000research.10100.1.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Dijkstra AA, Voorn P, Berendse HW, Groenewegen HJ, Rozemuller AJM, van de Berg WDJ. Stage-dependent nigral neuronal loss in incidental Lewy body and Parkinson’s disease. Mov Disord. 2014;29:1244–51.  https://doi.org/10.1002/mds.25952.CrossRefPubMedGoogle Scholar
  12. 12.
    Goetz CG, Pal G. Initial management of Parkinson’s disease. BMJ. 2014;349:g6258.  https://doi.org/10.1136/bmj.g6258.CrossRefPubMedGoogle Scholar
  13. 13.
    Reichmann H, Brandt MD, Klingelhoefer L. The nonmotor features of Parkinson’s disease: pathophysiology and management advances. Curr Opin Neurol. 2016;29:467–73.  https://doi.org/10.1097/WCO.0000000000000348.CrossRefPubMedGoogle Scholar
  14. 14.
    Fanciulli A, Wenning GK. Multiple-system atrophy. N Engl J Med. 2015;372:249–63.  https://doi.org/10.1056/NEJMra1311488.CrossRefPubMedGoogle Scholar
  15. 15.
    Velayudhan L, Ffytche D, Ballard C, Aarsland D. New therapeutic strategies for Lewy body dementias. Curr Neurol Neurosci Rep. 2017;17:68.  https://doi.org/10.1007/s11910-017-0778-2.CrossRefPubMedGoogle Scholar
  16. 16.
    Lashuel HA, Overk CR, Oueslati A, Masliah E. The many faces of α-synuclein: from structure and toxicity to therapeutic target. Nat Rev Neurosci. 2013;14:38–48.  https://doi.org/10.1038/nrn3406.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Levin J, Maaß S, Schuberth M, Respondek G, Paul F, Mansmann U, et al. The PROMESA-protocol: progression rate of multiple system atrophy under EGCG supplementation as anti-aggregation-approach. J Neural Transm (Vienna). 2016;123:439–45.  https://doi.org/10.1007/s00702-016-1507-8.CrossRefGoogle Scholar
  18. 18.
    Hamamichi S, Rivas RN, Knight AL, Cao S, Caldwell KA, Caldwell GA. Hypothesis-based RNAi screening identifies neuroprotective genes in a Parkinson’s disease model. Proc Natl Acad Sci U S A. 2008;105:728–33.  https://doi.org/10.1073/pnas.0711018105.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Gonçalves SA, Macedo D, Raquel H, Simões PD, Giorgini F, Ramalho JS, et al. shRNA-based screen identifies endocytic recycling pathway components that act as genetic modifiers of alpha-synuclein aggregation, secretion and toxicity. PLoS Genet. 2016;12:e1005995.  https://doi.org/10.1371/journal.pgen.1005995.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Goffeau A, Barrell BG, Bussey H, Davis RW, Dujon B, Feldmann H, et al. Life with 6000 genes. Science. 1996;274(546):563–7.Google Scholar
  21. 21.
    Wagner J, Ryazanov S, Leonov A, Levin J, Shi S, Schmidt F, et al. Anle138b: a novel oligomer modulator for disease-modifying therapy of neurodegenerative diseases such as prion and Parkinson’s disease. Acta Neuropathol. 2013;125:795–813.  https://doi.org/10.1007/s00401-013-1114-9.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Tardiff DF, Lindquist S. Phenotypic screens for compounds that target the cellular pathologies underlying Parkinson’s disease. Drug Discov Today Technol. 2013;10:e121–8.  https://doi.org/10.1016/j.ddtec.2012.02.003.CrossRefPubMedGoogle Scholar
  23. 23.
    Botstein D, Chervitz SA, Cherry JM. Yeast as a model organism. Science. 1997;277:1259–60.CrossRefGoogle Scholar
  24. 24.
    Giaever G, Nislow C. The yeast deletion collection: a decade of functional genomics. Genetics. 2014;197:451–65.  https://doi.org/10.1534/genetics.114.161620.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Fleming MS, Gitler AD. High-throughput yeast plasmid overexpression screen. J Vis Exp. 2011.  https://doi.org/10.3791/2836.
  26. 26.
    Outeiro TF, Lindquist S. Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science. 2003;302:1772–5.  https://doi.org/10.1126/science.1090439.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Willingham S, Outeiro TF, DeVit MJ, Lindquist SL, Muchowski PJ. Yeast genes that enhance the toxicity of a mutant huntingtin fragment or alpha-synuclein. Science. 2003;302:1769–72.  https://doi.org/10.1126/science.1090389.CrossRefPubMedGoogle Scholar
  28. 28.
    Cooper AA, Gitler AD, Cashikar A, Haynes CM, Hill KJ, Bhullar B, et al. Alpha-synuclein blocks ER-Golgi traffic and Rab1 rescues neuron loss in Parkinson’s models. Science. 2006;313:324–8.  https://doi.org/10.1126/science.1129462.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Yeger-Lotem E, Riva L, Su LJ, Gitler AD, Cashikar AG, King OD, et al. Bridging high-throughput genetic and transcriptional data reveals cellular responses to alpha-synuclein toxicity. Nat Genet. 2009;41:316–23.  https://doi.org/10.1038/ng.337.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Gitler AD, Chesi A, Geddie ML, Strathearn KE, Hamamichi S, Hill KJ, et al. Alpha-synuclein is part of a diverse and highly conserved interaction network that includes PARK9 and manganese toxicity. Nat Genet. 2009;41:308–15.  https://doi.org/10.1038/ng.300.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Flower TR, Clark-Dixon C, Metoyer C, Yang H, Shi R, Zhang Z, et al. YGR198w (YPP1) targets A30P alpha-synuclein to the vacuole for degradation. J Cell Biol. 2007;177:1091–104.  https://doi.org/10.1083/jcb.200610071.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Liang J, Clark-Dixon C, Wang S, Flower TR, Williams-Hart T, Zweig R, et al. Novel suppressors of alpha-synuclein toxicity identified using yeast. Hum Mol Genet. 2008;17:3784–95.  https://doi.org/10.1093/hmg/ddn276.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    • Chen Y-C, Farzadfard F, Gharaei N, Chen WCW, Cao J, Lu TK. Randomized CRISPR-Cas transcriptional perturbation screening reveals protective genes against alpha-synuclein toxicity. Mol Cell. 2017;68:247–257.e5.  https://doi.org/10.1016/j.molcel.2017.09.014 A screen identifying genes differentially expressed upon alpha-synuclein overexpression by an innovative CRISPR-Cas approach to pertube transcriptional networks.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Griffioen G, Duhamel H, van Damme N, Pellens K, Zabrocki P, Pannecouque C, et al. A yeast-based model of alpha-synucleinopathy identifies compounds with therapeutic potential. Biochim Biophys Acta. 2006, 1762:312–8.  https://doi.org/10.1016/j.bbadis.2005.11.009.
  35. 35.
    Williams RB, Gutekunst WR, Joyner PM, Duan W, Li Q, Ross CA, et al. Bioactivity profiling with parallel mass spectrometry reveals an assemblage of green tea metabolites affording protection against human huntingtin and alpha-synuclein toxicity. J Agric Food Chem. 2007;55:9450–6.  https://doi.org/10.1021/jf072241x.CrossRefPubMedGoogle Scholar
  36. 36.
    Su LJ, Auluck PK, Outeiro TF, Yeger-Lotem E, Kritzer JA, Tardiff DF, et al. Compounds from an unbiased chemical screen reverse both ER-to-Golgi trafficking defects and mitochondrial dysfunction in Parkinson’s disease models. Dis Model Mech. 2010;3:194–208.  https://doi.org/10.1242/dmm.004267.CrossRefPubMedGoogle Scholar
  37. 37.
    Kim J, Sasaki Y, Yoshida W, Kobayashi N, Veloso AJ, Kerman K, et al. Rapid cytotoxicity screening platform for amyloid inhibitors using a membrane-potential sensitive fluorescent probe. Anal Chem. 2013;85:185–92.  https://doi.org/10.1021/ac302442q.CrossRefPubMedGoogle Scholar
  38. 38.
    •• Höllerhage M, Moebius C, Melms J, Chiu W-H, Goebel JN, Chakroun T, et al. Protective efficacy of phosphodiesterase-1 inhibition against alpha-synuclein toxicity revealed by compound screening in LUHMES cells. Sci Rep. 2017;7:11469.  https://doi.org/10.1038/s41598-017-11664-5 The first and only unbiased screen of small compounds as modifiers of alpha-synuclein induced toxicity in human dopaminergic cells. CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Kritzer JA, Hamamichi S, McCaffery JM, Santagata S, Naumann TA, Caldwell KA, et al. Rapid selection of cyclic peptides that reduce alpha-synuclein toxicity in yeast and animal models. Nat Chem Biol. 2009;5:655–63.  https://doi.org/10.1038/nchembio.193.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Cheruvara H, Allen-Baume VL, Kad NM, Mason JM. Intracellular screening of a peptide library to derive a potent peptide inhibitor of α-synuclein aggregation. J Biol Chem. 2015;290:7426–35.  https://doi.org/10.1074/jbc.M114.620484.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    • Mittal S, Bjørnevik K, Im DS, Flierl A, Dong X, Locascio JJ, et al. β2-Adrenoreceptor is a regulator of the α-synuclein gene driving risk of Parkinson’s disease. Science. 2017;357:891–8.  https://doi.org/10.1126/science.aaf3934 A screen identifying β-receptor blockers as enhancers of alpha-synuclein expression, important due to its possible clinical implications. CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Cao S, Gelwix CC, Caldwell KA, Caldwell GA. Torsin-mediated protection from cellular stress in the dopaminergic neurons of Caenorhabditis elegans. J Neurosci. 2005;25:3801–12.  https://doi.org/10.1523/JNEUROSCI.5157-04.2005.CrossRefPubMedGoogle Scholar
  43. 43.
    Lan A, Smoly IY, Rapaport G, Lindquist S, Fraenkel E, Yeger-Lotem E. ResponseNet: revealing signaling and regulatory networks linking genetic and transcriptomic screening data. Nucleic Acids Res. 2011;39:W424–9.  https://doi.org/10.1093/nar/gkr359.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Basha O, Tirman S, Eluk A, Yeger-Lotem E. ResponseNet2.0: revealing signaling and regulatory pathways connecting your proteins and genes—now with human data. Nucleic Acids Res. 2013;41:W198–203.  https://doi.org/10.1093/nar/gkt532.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Vekrellis K, Xilouri M, Emmanouilidou E, Stefanis L. Inducible over-expression of wild type alpha-synuclein in human neuronal cells leads to caspase-dependent non-apoptotic death. J Neurochem. 2009;109:1348–62.  https://doi.org/10.1111/j.1471-4159.2009.06054.x.CrossRefPubMedGoogle Scholar
  46. 46.
    Lotharius J, Barg S, Wiekop P, Lundberg C, Raymon HK, Brundin P. Effect of mutant alpha-synuclein on dopamine homeostasis in a new human mesencephalic cell line. J Biol Chem. 2002;277:38884–94.  https://doi.org/10.1074/jbc.M205518200.CrossRefPubMedGoogle Scholar
  47. 47.
    Lotharius J, Falsig J, van Beek J, Payne S, Dringen R, Brundin P, et al. Progressive degeneration of human mesencephalic neuron-derived cells triggered by dopamine-dependent oxidative stress is dependent on the mixed-lineage kinase pathway. J Neurosci. 2005;25:6329–42.  https://doi.org/10.1523/JNEUROSCI.1746-05.2005.CrossRefPubMedGoogle Scholar
  48. 48.
    Höllerhage M, Goebel JN, de Andrade A, Hildebrandt T, Dolga A, Culmsee C, et al. Trifluoperazine rescues human dopaminergic cells from wild-type α-synuclein-induced toxicity. Neurobiol Aging. 2014;35:1700–11.  https://doi.org/10.1016/j.neurobiolaging.2014.01.027.CrossRefPubMedGoogle Scholar
  49. 49.
    Jung HJ, Kwon HJ. Target deconvolution of bioactive small molecules: the heart of chemical biology and drug discovery. Arch Pharm Res. 2015;38:1627–41.  https://doi.org/10.1007/s12272-015-0618-3.CrossRefPubMedGoogle Scholar
  50. 50.
    Lee H, Lee JW. Target identification for biologically active small molecules using chemical biology approaches. Arch Pharm Res. 2016;39:1193–201.  https://doi.org/10.1007/s12272-016-0791-z.CrossRefPubMedGoogle Scholar
  51. 51.
    Eckermann K, Kügler S, Bähr M. Dimerization propensities of synucleins are not predictive for synuclein aggregation. Biochim Biophys Acta. 1852;2015:1658–64.  https://doi.org/10.1016/j.bbadis.2015.05.002.CrossRefGoogle Scholar
  52. 52.
    Leestemaker Y, de Jong A, Witting KF, Penning R, Schuurman K, Rodenko B, et al. Proteasome activation by small molecules. Cell Chem Biol. 2017;24:725–736.e7.  https://doi.org/10.1016/j.chembiol.2017.05.010.CrossRefPubMedGoogle Scholar
  53. 53.
    Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, et al. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science. 2014;343:84–7.  https://doi.org/10.1126/science.1247005.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Matthias Höllerhage
    • 1
    • 2
  • Marc Bickle
    • 3
  • Günter U. Höglinger
    • 1
    • 2
    • 4
    Email author
  1. 1.Department of Translational NeurodegenerationGerman Center for Neurodegenerative Diseases (DZNE)MunichGermany
  2. 2.Department of NeurologyTechnical University of Munich (TUM)MunichGermany
  3. 3.HT-Technology Development StudioMax Planck Institute of Molecular Cell Biology and GeneticsDresdenGermany
  4. 4.Munich Cluster for Systems Neurology (SyNergy)Ludwig Maximilians University (LMU)MunichGermany

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