1. Editor's Highlight: Multiparametric Image Analysis of Rat Dorsal Root Ganglion Cultures to Evaluate Peripheral Neuropathy-Inducing Chemotherapeutics.
- Author
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Guo L, Hamre J 3rd, Eldridge S, Behrsing HP, Cutuli FM, Mussio J, and Davis M
- Subjects
- Animals, Biomarkers metabolism, Cell Body drug effects, Cell Body metabolism, Cell Body pathology, Cell Nucleus Shape drug effects, Cell Shape drug effects, Cells, Cultured, Drug Evaluation, Preclinical methods, Electrophoresis, Capillary, Fluorescent Antibody Technique, Ganglia, Spinal metabolism, Ganglia, Spinal pathology, Image Processing, Computer-Assisted, Kinetics, Molecular Weight, Nerve Tissue Proteins chemistry, Nerve Tissue Proteins metabolism, Neurites drug effects, Neurites metabolism, Neurites pathology, Neurons metabolism, Neurons pathology, Neurotoxicity Syndromes etiology, Neurotoxicity Syndromes metabolism, Neurotoxicity Syndromes pathology, Organelle Shape drug effects, Organelle Size drug effects, Peripheral Nervous System Diseases etiology, Peripheral Nervous System Diseases metabolism, Peripheral Nervous System Diseases pathology, Rats, Antineoplastic Agents adverse effects, Ganglia, Spinal drug effects, Neurons drug effects
- Abstract
Chemotherapy-induced peripheral neuropathy (CIPN) is a major, dose-limiting adverse effect experienced by cancer patients. Advancements in mechanism-based risk mitigation and effective treatments for CIPN can be aided by suitable in vitro assays. To this end, we developed a multiparametric morphology-centered rat dorsal root ganglion (DRG) assay. Morphologic alterations in subcellular structures of neurons and non-neurons were analyzed with an automated microscopy system. Stains for NeuN (a neuron-specific nuclear protein) and Tuj-1 (β-III tubulin) were used to identify neuronal cell nuclei and neuronal cell bodies/neurites, respectively. Vimentin staining (a component of Schwann cell intermediate filaments) was used to label non-neuronal supporting cells. Nuclei that stained with DAPI, but lacked NeuN represented non-neuronal cells. Images were analyzed following 24 h of continuous exposure to CIPN-inducing agents and 72 h after drug removal to provide a dynamic measure of recovery from initial drug effects. Treatment with bortezomib, cisplatin, eribulin, paclitaxel or vincristine induced a dose-dependent loss of neurite/process areas, mimicking the 'dying back' degeneration of axons, a histopathological hallmark of clinical CIPN in vivo. The IC50 for neurite loss was within 3-fold of the maximal clinical exposure (Cmax) for all five CIPN-inducing drugs, but was >4- or ≥ 28-fold of the Cmax for 2 non-CIPN-inducing agents. Compound-specific effects, eg, neurite fragmentation by cisplatin or bortezomib and enlarged neuronal cell bodies by paclitaxel, were also observed. Collectively, these results support the use of a quantitative, morphologic evaluation and a DRG cell culture model to inform risk and examine mechanisms of CIPN., (Published by Oxford University Press on behalf of the Society of Toxicology 2017. This work is written by US Government employees and is in the public domain in the US.)
- Published
- 2017
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