Phenotypic Screening of Small-Molecule Inhibitors: Implications for Therapeutic Discovery and Drug Target Development in Traumatic Brain Injury

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1462)

Abstract

The inability of central nervous system (CNS) neurons to regenerate damaged axons and dendrites following traumatic brain injury (TBI) creates a substantial obstacle for functional recovery. Apoptotic cell death, deposition of scar tissue, and growth-repressive molecules produced by glia further complicate the problem and make it challenging for re-growing axons to extend across injury sites. To date, there are no approved drugs for the treatment of TBI, accentuating the need for relevant leads. Cell-based and organotypic bioassays can better mimic outcomes within the native CNS microenvironment than target-based screening methods and thus should speed the discovery of therapeutic agents that induce axon or dendrite regeneration. Additionally, when used to screen focused chemical libraries such as small-molecule protein kinase inhibitors, these assays can help elucidate molecular mechanisms involved in neurite outgrowth and regeneration as well as identify novel drug targets. Here, we describe a phenotypic cellular (high content) screening assay that utilizes brain-derived primary neurons for screening small-molecule chemical libraries.

Key words

High-content screening Primary neurons Cell-based assay Axon regeneration CNS injury Kinase inhibitor Drug discovery 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Hassan Al-Ali
    • 1
  • Vance P. Lemmon
    • 1
    • 2
    • 3
  • John L. Bixby
    • 1
    • 2
    • 3
    • 4
  1. 1.Miami Project to Cure ParalysisUniversity of Miami Miller School of MedicineMiamiUSA
  2. 2.Center for Computational ScienceUniversity of Miami Miller School of MedicineMiamiUSA
  3. 3.Departments of Neurological SurgeryUniversity of Miami Miller School of MedicineMiamiUSA
  4. 4.Molecular & Cellular PharmacologyUniversity of Miami Miller School of MedicineMiamiUSA

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