ErbB2 regulates autophagic flux to modulate the proteostasis of APP-CTFs in Alzheimer’s disease

Significance We demonstrate that ErbB2 can regulate autophagic flux through its direct interaction with Beclin-1, which effectively blocks autophagy initiation. Although the expression of ErbB2 becomes dormant during adulthood, it becomes reactivated during the pathogenesis of Alzheimer’s disease (AD), blocking the autophagy-mediated clearance of amyloid precursor protein (APP) C-terminal fragments (CTFs) [99-residue CTF (C99)]. Consequently, the accumulated APP-C99 can be further processed by γ-secretase, resulting in augmented production of amyloid-β and the APP intracellular domain. The chemical inhibition of ErbB2 by CL-387,785 effectively rescues the cognitive impairment of APP/presenilin-1 (PS1) transgenic AD mice. The present study thus defines a molecular basis by which the aberrant expression of ErbB2 could instigate the pathogenesis of AD. These findings provide the proof-of-principle evidence for rational design of ErbB2-targeted therapeutics for AD. Proteolytic processing of amyloid precursor protein (APP) C-terminal fragments (CTFs) by γ-secretase underlies the pathogenesis of Alzheimer’s disease (AD). An RNA interference screen using APP-CTF [99-residue CTF (C99)]- and Notch-specific γ-secretase interaction assays identified a unique ErbB2-centered signaling network that was predicted to preferentially govern the proteostasis of APP-C99. Consistently, significantly elevated levels of ErbB2 were confirmed in the hippocampus of human AD brains. We then found that ErbB2 effectively suppressed autophagic flux by physically dissociating Beclin-1 from the Vps34–Vps15 complex independent of its kinase activity. Down-regulation of ErbB2 by CL-387,785 decreased the levels of C99 and secreted amyloid-β in cellular, zebrafish, and mouse models of AD, through the activation of autophagy. Oral administration of an ErbB2-targeted CL-387,785 for 3 wk significantly improves the cognitive functions of APP/presenilin-1 (PS1) transgenic mice. This work unveils a noncanonical function of ErbB2 in modulating autophagy and establishes ErbB2 as a therapeutic target for AD.

[1]  W. Whetsell,et al.  Immunoreactive Epidermal Growth Factor Receptors in Neuritic Plaques from Patients with Alzheimer's Disease , 1988, Journal of neuropathology and experimental neurology.

[2]  J. Hardy,et al.  Amyloid deposition as the central event in the aetiology of Alzheimer's disease. , 1991, Trends in pharmacological sciences.

[3]  Y. Suh,et al.  Neurotoxicity of a Carboxyl‐Terminal Fragment of the Alzheimer's Amyloid Precursor Protein , 1996, Journal of neurochemistry.

[4]  Raphael Kopan,et al.  Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain , 1998, Nature.

[5]  S. Angers,et al.  Detection of beta 2-adrenergic receptor dimerization in living cells using bioluminescence resonance energy transfer (BRET). , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[6]  T. Russo,et al.  Signal Transduction through Tyrosine-phosphorylated C-terminal Fragments of Amyloid Precursor Protein via an Enhanced Interaction with Shc/Grb2 Adaptor Proteins in Reactive Astrocytes of Alzheimer's Disease Brain* , 2002, The Journal of Biological Chemistry.

[7]  M. Sliwkowski,et al.  Identification of a Region within the ErbB2/HER2 Intracellular Domain That Is Necessary for Ligand-independent Association* , 2002, The Journal of Biological Chemistry.

[8]  T. Issad,et al.  The use of resonance energy transfer in high-throughput screening: BRET versus FRET. , 2002, Trends in pharmacological sciences.

[9]  Michael S. Wolfe,et al.  γ-Secretase is a membrane protein complex comprised of presenilin, nicastrin, aph-1, and pen-2 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. Carroll,et al.  Neuregulin–1 and ErbB4 Immunoreactivity Is Associated with Neuritic Plaques in Alzheimer Disease Brain and in a Transgenic Model of Alzheimer Disease , 2003, Journal of neuropathology and experimental neurology.

[11]  B. Karolewski,et al.  Comparison of transfection conditions for a lentivirus vector produced in large volumes. , 2003, Human gene therapy.

[12]  M. Wolfe,et al.  Tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma stimulate gamma-secretase-mediated cleavage of amyloid precursor protein through a JNK-dependent MAPK pathway. , 2004, The Journal of biological chemistry.

[13]  M. Duchen,et al.  Toxicity of amyloid beta peptide: tales of calcium, mitochondria, and oxidative stress. , 2004, Neurochemical research.

[14]  M. Wolfe,et al.  Tumor Necrosis Factor-α, Interleukin-1β, and Interferon-γ Stimulate γ-Secretase-mediated Cleavage of Amyloid Precursor Protein through a JNK-dependent MAPK Pathway* , 2004, Journal of Biological Chemistry.

[15]  W. Talbot,et al.  erbb3 and erbb2 Are Essential for Schwann Cell Migration and Myelination in Zebrafish , 2005, Current Biology.

[16]  H. Kornblum,et al.  Developmental profile of ErbB receptors in murine central nervous system: Implications for functional interactions , 2005, Journal of neuroscience research.

[17]  P. Emson,et al.  Chronic stress accelerates learning and memory impairments and increases amyloid deposition in APP V717I -CT100 transgenic mice, an Alzheimer’s disease model , 2006 .

[18]  J. Rothman,et al.  Autophagy-mediated clearance of huntingtin aggregates triggered by the insulin-signaling pathway , 2006, The Journal of cell biology.

[19]  A. Citri,et al.  EGF–ERBB signalling: towards the systems level , 2006, Nature Reviews Molecular Cell Biology.

[20]  C. Vorhees,et al.  Morris water maze: procedures for assessing spatial and related forms of learning and memory , 2006, Nature Protocols.

[21]  P. Greengard,et al.  Regulation of Alzheimer's disease amyloid-beta formation by casein kinase I. , 2007, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Wolfe,et al.  APP at a glance , 2007, Journal of Cell Science.

[23]  Yvonne S. Eisele,et al.  Gleevec increases levels of the amyloid precursor protein intracellular domain and of the amyloid-beta degrading enzyme neprilysin. , 2007, Molecular biology of the cell.

[24]  D. Curtis,et al.  Presenilin diversifies its portfolio. , 2007, Trends in genetics : TIG.

[25]  R. Tibshirani,et al.  Classification and prediction of clinical Alzheimer's diagnosis based on plasma signaling proteins , 2007, Nature Medicine.

[26]  P. Greengard,et al.  Regulation of Alzheimer's disease amyloid-β formation by casein kinase I , 2007, Proceedings of the National Academy of Sciences.

[27]  Ming‐Kuan Hu,et al.  Sodium selenite inhibits γ-secretase activity through activation of ERK , 2008, Neuroscience Letters.

[28]  Pablo Moscato,et al.  Identification of a 5-Protein Biomarker Molecular Signature for Predicting Alzheimer's Disease , 2008, PloS one.

[29]  SUBSTRATE-TARGETING GAMMA-SECRETASE MODULATORS , 2008 .

[30]  M. Wolfe,et al.  Substrate-targeting γ-secretase modulators , 2008, Nature.

[31]  Brian Spencer,et al.  The autophagy-related protein beclin 1 shows reduced expression in early Alzheimer disease and regulates amyloid beta accumulation in mice. , 2008, The Journal of clinical investigation.

[32]  S. Ojeda,et al.  Differential erbB signaling in astrocytes from the cerebral cortex and the hypothalamus of the human brain , 2009, Glia.

[33]  Yong Shen,et al.  Alzheimer's disease-like pathological features in transgenic mice expressing the APP intracellular domain , 2009, Proceedings of the National Academy of Sciences.

[34]  P. Wong,et al.  The β-Secretase Enzyme BACE in Health and Alzheimer's Disease: Regulation, Cell Biology, Function, and Therapeutic Potential , 2009, The Journal of Neuroscience.

[35]  B. De Strooper,et al.  The Orphan G Protein–Coupled Receptor 3 Modulates Amyloid-Beta Peptide Generation in Neurons , 2009, Science.

[36]  E. Martegani,et al.  Activation of Amyloid Precursor Protein Processing by Growth Factors is Dependent on Ras GTPase Activity , 2011, Neurochemical Research.

[37]  P. Greengard,et al.  Gamma-secretase activating protein, a therapeutic target for Alzheimer's disease , 2010, Nature.

[38]  Xulun Zhang,et al.  Modulation of γ-Secretase Reduces β-Amyloid Deposition in a Transgenic Mouse Model of Alzheimer's Disease , 2010, Neuron.

[39]  W. Xia.  Exploring Alzheimer's disease in zebrafish. , 2010, Journal of Alzheimer's disease : JAD.

[40]  D. Westaway,et al.  Lysosomal Proteolysis and Autophagy Require Presenilin 1 and Are Disrupted by Alzheimer-Related PS1 Mutations , 2010, Cell.

[41]  M. Citron,et al.  Alzheimer's disease: strategies for disease modification , 2010, Nature Reviews Drug Discovery.

[42]  Stephen T. C. Wong,et al.  Phenotypic analysis of images of zebrafish treated with Alzheimer's γ-secretase inhibitors , 2010, BMC biotechnology.

[43]  E. Lundberg,et al.  Towards a knowledge-based Human Protein Atlas , 2010, Nature Biotechnology.

[44]  P. Greengard,et al.  A small‐molecule enhancer of autophagy decreases levels of Aβ and APP‐CTF via Atg5‐dependent autophagy pathway , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[45]  John Kuriyan,et al.  Regulation of the catalytic activity of the EGF receptor. , 2011, Current opinion in structural biology.

[46]  B. Strooper,et al.  The amyloid cascade hypothesis for Alzheimer's disease: an appraisal for the development of therapeutics , 2011, Nature Reviews Drug Discovery.

[47]  M. Lotze,et al.  The Beclin 1 network regulates autophagy and apoptosis , 2011, Cell Death and Differentiation.

[48]  Hsinyu Lee,et al.  Autophagy: A double-edged sword in Alzheimer’s disease , 2012, Journal of Biosciences.

[49]  H. Park,et al.  High Cleavage Efficiency of a 2A Peptide Derived from Porcine Teschovirus-1 in Human Cell Lines, Zebrafish and Mice , 2011, PloS one.

[50]  M. Goedert,et al.  Stimulation of autophagy reduces neurodegeneration in a mouse model of human tauopathy , 2012, Brain : a journal of neurology.

[51]  Xinsheng Yao,et al.  Epidermal growth factor receptor is a preferred target for treating Amyloid-β–induced memory loss , 2012, Proceedings of the National Academy of Sciences.

[52]  Robert Clarke,et al.  Guidelines for the use and interpretation of assays for monitoring autophagy , 2012 .

[53]  N. Grishin,et al.  EGFR-Mediated Beclin 1 Phosphorylation in Autophagy Suppression, Tumor Progression, and Tumor Chemoresistance , 2013, Cell.

[54]  Simon C Watkins,et al.  Interaction between Her2 and Beclin-1 Proteins Underlies a New Mechanism of Reciprocal Regulation* , 2013, The Journal of Biological Chemistry.

[55]  M. Vaccaro,et al.  The VMP1-Beclin 1 interaction regulates autophagy induction , 2013, Scientific Reports.

[56]  Liang-Jun Yan,et al.  Rapamycin, Autophagy, and Alzheimer's Disease. , 2013, Journal of biochemical and pharmacological research.

[57]  M. Gallas,et al.  Truncated p110 ERBB2 induces mammary epithelial cell migration, invasion and orthotopic xenograft formation, and is associated with loss of phosphorylated STAT5 , 2012, Oncogene.

[58]  G. Her,et al.  Presenilin-1 Regulates the Expression of p62 to Govern p62-dependent Tau Degradation , 2013, Molecular Neurobiology.

[59]  K. Brunden,et al.  Lewy Body-like α-Synuclein Aggregates Resist Degradation and Impair Macroautophagy*♦ , 2013, The Journal of Biological Chemistry.

[60]  R. Nixon,et al.  The role of autophagy in neurodegenerative disease , 2013, Nature Medicine.

[61]  Maria Xilouri,et al.  Boosting chaperone-mediated autophagy in vivo mitigates α-synuclein-induced neurodegeneration. , 2013, Brain : a journal of neurology.

[62]  S. Pedersen,et al.  Constitutively Active ErbB2 Regulates Cisplatin-Induced Cell Death in Breast Cancer Cells via Pro- and Antiapoptotic Mechanisms , 2014, Molecular Cancer Research.

[63]  L. Yang,et al.  Identification of prolidase as a high affinity ligand of the ErbB2 receptor and its regulation of ErbB2 signaling and cell growth , 2014, Cell Death and Disease.

[64]  W. Annaert,et al.  Early etiology of Alzheimer’s disease: tipping the balance toward autophagy or endosomal dysfunction? , 2015, Acta Neuropathologica.

[65]  E. Siemers,et al.  Phase 3 trials of solanezumab for mild-to-moderate Alzheimer's disease. , 2014, The New England journal of medicine.

[66]  Nick C Fox,et al.  Two phase 3 trials of bapineuzumab in mild-to-moderate Alzheimer's disease. , 2014, The New England journal of medicine.

[67]  P. Aisen,et al.  Advances in Alzheimer’s Disease Drug Development , 2015, BMC Medicine.

[68]  Davide Heller,et al.  STRING v10: protein–protein interaction networks, integrated over the tree of life , 2014, Nucleic Acids Res..

[69]  Xiaojun Tan,et al.  A Kinase-Independent Role for EGF Receptor in Autophagy Initiation , 2015, Cell.

[70]  E. Holzbaur,et al.  Autophagosome dynamics in neurodegeneration at a glance , 2015, Journal of Cell Science.

[71]  Johannes E. Schindelin,et al.  The ImageJ ecosystem: An open platform for biomedical image analysis , 2015, Molecular reproduction and development.

[72]  G. von Heijne,et al.  Tissue-based map of the human proteome , 2015, Science.

[73]  C. Kenific,et al.  Cellular and metabolic functions for autophagy in cancer cells. , 2015, Trends in cell biology.

[74]  Angeleen Fleming,et al.  Compromised autophagy and neurodegenerative diseases , 2015, Nature Reviews Neuroscience.

[75]  Sangeeta Khare,et al.  Guidelines for the use and interpretation of assays formonitoring autophagy (3rd edition) , 2016 .

[76]  K. Rhodes,et al.  The antibody aducanumab reduces Aβ plaques in Alzheimer’s disease , 2016, Nature.

[77]  J. Hardy,et al.  The amyloid hypothesis of Alzheimer's disease at 25 years , 2016, EMBO molecular medicine.