Phenotypic Screening of Small-Molecule Inhibitors: Implications for Therapeutic Discovery and Drug Target Development in Traumatic Brain Injury.
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.
Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, 331365, USA.
Hassan
H
Al-Ali
Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, 331365, USA.|Center for Computational Science, University of Miami Miller School of Medicine, Miami, FL, 331365, USA.|Departments of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, 331365, USA.
Vance P
VP
Lemmon
Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, 331365, USA. jbixby@med.miami.edu.|Center for Computational Science, University of Miami Miller School of Medicine, Miami, FL, 331365, USA. jbixby@med.miami.edu.|Departments of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, 331365, USA. jbixby@med.miami.edu.|Molecular & Cellular Pharmacology, University of Miami Miller School of Medicine, 1400 NW 10th Ave., DT 1205, Miami, FL, 331365, USA. jbixby@med.miami.edu.
John L
JL
Bixby
D000818:Animals; D001369:Axons / Q000187:drug effects / Q000378:metabolism; D015415:Biomarkers; D000070642:Brain Injuries, Traumatic / Q000175:diagnosis / Q000188:drug therapy / Q000209:etiology* / Q000378:metabolism*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology; D057166:High-Throughput Screening Assays; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010641:Phenotype; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D054852:Small Molecule Libraries
D015415:Biomarkers; D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D010770:Phosphotransferases
Axon regeneration; CNS injury; Cell-based assay; Drug discovery; High-content screening; Kinase inhibitor; Primary neurons