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Amyloid Precursor Protein Dimerisation Reduces Neurite Outgrowth

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Abstract

The amyloid precursor protein (APP) undergoes extensive metabolism, and its transport and proteolytic processing can be modulated by its ability to form a homodimer. We have investigated the functional consequences of stabilised APP dimer expression in cells by studying the engineered dimerisation of the APPL17C (residue 17 in Aβ sequence) construct, which is associated with a 30% increase in APP dimer expression, on APP’s neurite outgrowth promoting activity. Overexpression of APPL17C in SH-SY5Y cells decreased neurite outgrowth upon retinoic acid differentiation as compared to overexpressing APPWT cells. The APPL17C phenotype was rescued by replacing the APPL17C media with conditioned media from APPWT cells, indicating that the APPL17C mutant is impairing the secretion of a neuritogenic promoting factor. APPL17C had altered transport and was localised in the endoplasmic reticulum. Defining the molecular basis of the APPL17C phenotype showed that RhoA GTPase activity, a negative regulator of neurite outgrowth, was increased in APPL17C cells. RhoA activity was decreased after APPWT conditioned media rescue. Moreover, treatment with the RhoA inhibitor, Y27632, restored a wild-type morphology to the APPL17C cells. Small RNAseq analysis of APPL17C and APPWT cells identified several differentially expressed miRNAs relating to neurite outgrowth. Of these, miR-34a showed the greatest decrease in expression. Lentiviral-mediated overexpression of miR-34a rescued neurite outgrowth in APPL17C cells to APPWT levels and changed RhoA activation. This study has identified a novel link between APP dimerisation and its neuritogenic activity which is mediated by miR-34a expression.

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Funding

This work was supported from funding from the Australia National Health and Medical Research Council (R.C., A.F.H.).

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Authors and Affiliations

  1. Department of Pathology, The University of Melbourne, Melbourne, VIC, 3010, Australia

    Luan Luu, Giuseppe D. Ciccotosto, Laila C. Roisman & Roberto Cappai

  2. Department of Pharmacology & Therapeutics, The University of Melbourne, Melbourne, VIC, 3010, Australia

    Giuseppe D. Ciccotosto & Roberto Cappai

  3. Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Melbourne, VIC, 3010, Australia

    Giuseppe D. Ciccotosto & Laura J. Vella

  4. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC, 3086, Australia

    Lesley Cheng & Andrew F. Hill

  5. Department of Pharmacology & Therapeutics, McGill University, Montreal, QC, H3G 1Y6, Canada

    Gerhard Multhaup & Lisa-Marie Munter

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  1. Luan Luu
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Luu, L., Ciccotosto, G.D., Vella, L.J. et al. Amyloid Precursor Protein Dimerisation Reduces Neurite Outgrowth. Mol Neurobiol 56, 13–28 (2019). https://doi.org/10.1007/s12035-018-1070-4

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