TNFα affects CREB-mediated neuroprotective signaling pathways of synaptic plasticity in neurons as revealed by proteomics and phospho-proteomics

// Pia Jensen 1 , Christa L. Myhre 2 , Pernille S. Lassen 1 , Athanasios Metaxas 2 , Asif M. Khan 2, 5 , Kate L. Lambertsen 2, 3, 4 , Alicia A. Babcock 2 , Bente Finsen 2, 4, * , Martin R. Larsen 1, * and Stefan J. Kempf 1 1 Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark 2 Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark 3 Department of Neurology, Odense University Hospital, Odense, Denmark 4 BRIDGE, Brain Research–Inter-Disciplinary Guided Excellence, Department of Clinical Research, University of Southern Denmark, Odense, Denmark 5 Current address: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark * These authors are equally contributed to these work Correspondence to: Stefan J. Kempf, email: stefank@bmb.sdu.dk Keywords: mTOR, neuroinflammation, alzheimer’s disease, post translational modification, LPS Received: March 21, 2017      Accepted: July 11, 2017      Published: July 21, 2017 ABSTRACT Neuroinflammation is a hallmark of Alzheimer’s disease and TNFα as the main inducer of neuroinflammation has neurodegenerative but also pro-regenerative properties, however, the dose-dependent molecular changes on signaling pathway level are not fully understood. We performed quantitative proteomics and phospho-proteomics to target this point. In HT22 cells, we found that TNFα reduced mitochondrial signaling and inhibited mTOR protein translation signaling but also led to induction of neuroprotective MAPK-CREB signaling. Stimulation of human neurons with TNFα revealed similar cellular mechanisms. Moreover, a number of synaptic plasticity-associated genes were altered in their expression profile including CREB . SiRNA-mediated knockdown of CREB in human neurons prior to TNFα stimulation led to a reduced number of protein/phospho-protein hits compared to siRNA-mediated knockdown of CREB or TNFα stimulation alone and countermeasured the reduced CREB signaling. In vivo data of TNFα knockout mice showed that learning ability did not depend on TNFα per se but that TNFα was essential for preserving the learning ability after episodes of lipopolysaccharide-induced neuroinflammation. This may be based on modulation of CREB/CREB signaling as revealed by the in vitro / in vivo data. Our data show that several molecular targets and signaling pathways induced by TNFα in neurons resemble those seen in Alzheimer’s disease pathology.