Knocking out C9ORF72 Exacerbates Axonal Trafficking Defects Associated with Hexanucleotide Repeat Expansion and Reduces Levels of Heat Shock Proteins

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2020 Feb 22

PMID 32084385

Citations 38

Authors

Masin Abo-Rady

Norman Kalmbach

Arun Pal

Carina Schludi

Antje Janosch

Tanja Richter

Petra Freitag

Marc Bickle

Anne-Karin Kahlert

Susanne Petri

Stefan Stefanov

Hannes Glass

Selma Staege

Walter Just

Rajat Bhatnagar

Dieter Edbauer

Andreas Hermann

Florian Wegner

Jared L Sterneckert

Affiliations

Soon will be listed here.

Abstract

In amyotrophic lateral sclerosis (ALS) motor neurons (MNs) undergo dying-back, where the distal axon degenerates before the soma. The hexanucleotide repeat expansion (HRE) in C9ORF72 is the most common genetic cause of ALS, but the mechanism of pathogenesis is largely unknown with both gain- and loss-of-function mechanisms being proposed. To better understand C9ORF72-ALS pathogenesis, we generated isogenic induced pluripotent stem cells. MNs with HRE in C9ORF72 showed decreased axonal trafficking compared with gene corrected MNs. However, knocking out C9ORF72 did not recapitulate these changes in MNs from healthy controls, suggesting a gain-of-function mechanism. In contrast, knocking out C9ORF72 in MNs with HRE exacerbated axonal trafficking defects and increased apoptosis as well as decreased levels of HSP70 and HSP40, and inhibition of HSPs exacerbated ALS phenotypes in MNs with HRE. Therefore, we propose that the HRE in C9ORF72 induces ALS pathogenesis via a combination of gain- and loss-of-function mechanisms.

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References

Ciura S, Lattante S, Le Ber I, Latouche M, Tostivint H, Brice A . Loss of function of C9orf72 causes motor deficits in a zebrafish model of amyotrophic lateral sclerosis. Ann Neurol. 2013; 74(2):180-7. DOI: 10.1002/ana.23946. View

Hruscha A, Krawitz P, Rechenberg A, Heinrich V, Hecht J, Haass C . Efficient CRISPR/Cas9 genome editing with low off-target effects in zebrafish. Development. 2013; 140(24):4982-7. DOI: 10.1242/dev.099085. View

Li Y, King O, Shorter J, Gitler A . Stress granules as crucibles of ALS pathogenesis. J Cell Biol. 2013; 201(3):361-72. PMC: 3639398. DOI: 10.1083/jcb.201302044. View

Chew J, Gendron T, Prudencio M, Sasaguri H, Zhang Y, Castanedes-Casey M . Neurodegeneration. C9ORF72 repeat expansions in mice cause TDP-43 pathology, neuronal loss, and behavioral deficits. Science. 2015; 348(6239):1151-4. PMC: 4692360. DOI: 10.1126/science.aaa9344. View

Ganassi M, Mateju D, Bigi I, Mediani L, Poser I, Lee H . A Surveillance Function of the HSPB8-BAG3-HSP70 Chaperone Complex Ensures Stress Granule Integrity and Dynamism. Mol Cell. 2016; 63(5):796-810. DOI: 10.1016/j.molcel.2016.07.021. View

Waite A, Baumer D, East S, Neal J, Morris H, Ansorge O . Reduced C9orf72 protein levels in frontal cortex of amyotrophic lateral sclerosis and frontotemporal degeneration brain with the C9ORF72 hexanucleotide repeat expansion. Neurobiol Aging. 2014; 35(7):1779.e5-1779.e13. PMC: 3988882. DOI: 10.1016/j.neurobiolaging.2014.01.016. View

Pal A, Glass H, Naumann M, Kreiter N, Japtok J, Sczech R . High content organelle trafficking enables disease state profiling as powerful tool for disease modelling. Sci Data. 2018; 5:180241. PMC: 6233479. DOI: 10.1038/sdata.2018.241. View

Wainger B, Kiskinis E, Mellin C, Wiskow O, Han S, Sandoe J . Intrinsic membrane hyperexcitability of amyotrophic lateral sclerosis patient-derived motor neurons. Cell Rep. 2014; 7(1):1-11. PMC: 4023477. DOI: 10.1016/j.celrep.2014.03.019. View

Mordes D, Prudencio M, Goodman L, Klim J, Moccia R, Limone F . Dipeptide repeat proteins activate a heat shock response found in C9ORF72-ALS/FTLD patients. Acta Neuropathol Commun. 2018; 6(1):55. PMC: 6031111. DOI: 10.1186/s40478-018-0555-8. View

10.

Parone P, Da Cruz S, Han J, McAlonis-Downes M, Vetto A, Lee S . Enhancing mitochondrial calcium buffering capacity reduces aggregation of misfolded SOD1 and motor neuron cell death without extending survival in mouse models of inherited amyotrophic lateral sclerosis. J Neurosci. 2013; 33(11):4657-71. PMC: 3711648. DOI: 10.1523/JNEUROSCI.1119-12.2013. View

11.

Puls I, Jonnakuty C, LaMonte B, Holzbaur E, Tokito M, Mann E . Mutant dynactin in motor neuron disease. Nat Genet. 2003; 33(4):455-6. DOI: 10.1038/ng1123. View

12.

Zhang Z, Almeida S, Lu Y, Nishimura A, Peng L, Sun D . Downregulation of microRNA-9 in iPSC-derived neurons of FTD/ALS patients with TDP-43 mutations. PLoS One. 2013; 8(10):e76055. PMC: 3797144. DOI: 10.1371/journal.pone.0076055. View

13.

Niblock M, Smith B, Lee Y, Sardone V, Topp S, Troakes C . Retention of hexanucleotide repeat-containing intron in C9orf72 mRNA: implications for the pathogenesis of ALS/FTD. Acta Neuropathol Commun. 2016; 4:18. PMC: 4766718. DOI: 10.1186/s40478-016-0289-4. View

14.

Prudencio M, Belzil V, Batra R, Ross C, Gendron T, Pregent L . Distinct brain transcriptome profiles in C9orf72-associated and sporadic ALS. Nat Neurosci. 2015; 18(8):1175-82. PMC: 4830686. DOI: 10.1038/nn.4065. View

15.

Naumann M, Pal A, Goswami A, Lojewski X, Japtok J, Vehlow A . Impaired DNA damage response signaling by FUS-NLS mutations leads to neurodegeneration and FUS aggregate formation. Nat Commun. 2018; 9(1):335. PMC: 5780468. DOI: 10.1038/s41467-017-02299-1. View

16.

Devlin A, Burr K, Borooah S, Foster J, Cleary E, Geti I . Human iPSC-derived motoneurons harbouring TARDBP or C9ORF72 ALS mutations are dysfunctional despite maintaining viability. Nat Commun. 2015; 6:5999. PMC: 4338554. DOI: 10.1038/ncomms6999. View

17.

Simon-Sanchez J, Dopper E, Cohn-Hokke P, Hukema R, Nicolaou N, Seelaar H . The clinical and pathological phenotype of C9ORF72 hexanucleotide repeat expansions. Brain. 2012; 135(Pt 3):723-35. DOI: 10.1093/brain/awr353. View

18.

Chitiprolu M, Jagow C, Tremblay V, Bondy-Chorney E, Paris G, Savard A . A complex of C9ORF72 and p62 uses arginine methylation to eliminate stress granules by autophagy. Nat Commun. 2018; 9(1):2794. PMC: 6052026. DOI: 10.1038/s41467-018-05273-7. View

19.

Sareen D, ORourke J, Meera P, Muhammad A, Grant S, Simpkinson M . Targeting RNA foci in iPSC-derived motor neurons from ALS patients with a C9ORF72 repeat expansion. Sci Transl Med. 2013; 5(208):208ra149. PMC: 4090945. DOI: 10.1126/scitranslmed.3007529. View

20.

Frick P, Sellier C, Mackenzie I, Cheng C, Tahraoui-Bories J, Martinat C . Novel antibodies reveal presynaptic localization of C9orf72 protein and reduced protein levels in C9orf72 mutation carriers. Acta Neuropathol Commun. 2018; 6(1):72. PMC: 6091050. DOI: 10.1186/s40478-018-0579-0. View