Cross-Platform Synaptic Network Analysis of Human Entorhinal Cortex Identifies TWF2 as a Modulator of Dendritic Spine Length

• Type of Material: Article
• Authors: Courtney K Walker, University of Alabama Birmingham; Kelsey M Greathouse, University of Alabama Birmingham; Jennifer J Tuscher, University of Alabama Birmingham; Eric Dammer, Emory University; Audrey J Weber, University of Alabama Birmingham; Evan Liu, University of Alabama Birmingham; Kendall A Curtis, University of Alabama Birmingham; Benjamin D Boros, University of Alabama Birmingham; Cameron D Freeman, University of Alabama Birmingham; Jung Vin Seo, University of Alabama Birmingham; Raksha Ramdas, University of Alabama Birmingham; Cheyenne Hurst, Emory University; Duc M Duong, Emory University; Marla Gearing, Emory University; Charles F Murchison, University of Alabama Birmingham; Jeremy J Day, University of Alabama Birmingham; Nicholas Seyfried, Emory University; Jeremy H Herskowitz, University of Alabama Birmingham
• Language: English
• Date: 2023-05-17
• Publisher: SOC NEUROSCIENCE
• Publication Version: Final Published Version
• Copyright Statement: © 2023 Walker et al.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1523/JNEUROSCI.2102-22.2023
• Title of Journal or Parent Work: JOURNAL OF NEUROSCIENCE
• Volume: 43
• Issue: 20
• Start Page: 3764
• End Page: 3785
• Grant/Funding Information: This work was supported by the National Institutes of Health National Institute of Neurological Disorders and Stroke (NINDS) Grant T32 NS061788 (o C.K.W.), National Institute on Aging(NIA) Grant F31 AG067635 (to C.K.W.), NINDS Grant T32 NS095775 (to A.J.W.), NIA Grant AG061800 (to J.H.H. and N.T.S.), NIA Grant AG054719 (to J.H.H.), National Institute on Drug Abuse Grant DA039650 (to J.J.D.), NIA Grant AG063755, NIA Grant AG068024, and Grant U01AG061357 (to N.T.S.).
• Keywords: Neurosciences; TWINFILIN ISOFORMS; DYSFUNCTION; Alzheimer ' s disease; ACTIN; TAU PATHOLOGY; dendritic spines; COEXPRESSION; synapse; proteomics; ALZHEIMERS-DISEASE; systems biology; Neurosciences & Neurology; FILAMENT BARBED ENDS; entorhinal cortex; NEUROPATHOLOGIC ASSESSMENT; LOCALIZATION; Science & Technology; AMYLOID-BETA; Life Sciences & Biomedicine
• Research Categories: Chemistry, Biochemistry; Health Sciences, Medicine and Surgery

Abstract

Proteomic studies using postmortem human brain tissue samples have yielded robust assessments of the aging and neurodegenerative disease(s) proteomes. While these analyses provide lists of molecular alterations in human conditions, like Alzheimer’s disease (AD), identifying individual proteins that affect biological processes remains a challenge. To complicate matters, protein targets may be highly understudied and have limited information on their function. To address these hurdles, we sought to establish a blueprint to aid selection and functional validation of targets from proteomic datasets. A cross-platform pipeline was engineered to focus on synaptic processes in the entorhinal cortex (EC) of human patients, including controls, preclinical AD, and AD cases. Label-free quantification mass spectrometry (MS) data (n=2260 proteins) was generated on synaptosome fractionated tissue from Brodmann area 28 (BA28; n=58 samples). In parallel, dendritic spine density and morphology was measured in the same individuals. Weighted gene co-expression network analysis was used to construct a network of protein co-expression modules that were correlated with dendritic spine metrics. Module-trait correlations were used to guide unbiased selection of Twinfilin-2 (TWF2), which was the top hub protein of a module that positively correlated with thin spine length. Using CRISPR-dCas9 activation strategies, we demonstrated that boosting endogenous TWF2 protein levels in primary hippocampal neurons increased thin spine length, thus providing experimental validation for the human network analysis. Collectively, this study describes alterations in dendritic spine density and morphology as well as synaptic proteins and phosphorylated tau from the entorhinal cortex of preclinical and advanced stage AD patients.