title: Editor's Highlight: Multiparametric Image Analysis of Rat Dorsal Root Ganglion Cultures to Evaluate Peripheral Neuropathy-Inducing Chemotherapeutics.
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.
authors.affiliation: Laboratory of Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, Maryland 21702.
authors.forename: Liang
authors.initials: L
authors.lastname: Guo
authors.affiliation: Laboratory of Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, Maryland 21702.
authors.forename: John
authors.initials: J
authors.lastname: Hamre
authors.affiliation: Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, Maryland 20892.
authors.forename: Sandy
authors.initials: S
authors.lastname: Eldridge
authors.affiliation: Laboratory of Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, Maryland 21702.
authors.forename: Holger P
authors.initials: HP
authors.lastname: Behrsing
authors.affiliation: Laboratory of Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, Maryland 21702.
authors.forename: Facundo M
authors.initials: FM
authors.lastname: Cutuli
authors.affiliation: Laboratory of Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, Maryland 21702.
authors.forename: Jodie
authors.initials: J
authors.lastname: Mussio
authors.affiliation: Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, Maryland 20892.
authors.forename: Myrtle
authors.initials: M
authors.lastname: Davis
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000009:adverse effects*; D015415:Biomarkers / Q000378:metabolism; D066146:Cell Body / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D053144:Cell Nucleus Shape / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods; D019075:Electrophoresis, Capillary; D005455:Fluorescent Antibody Technique; D005727:Ganglia, Spinal / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007091:Image Processing, Computer-Assisted; D007700:Kinetics; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism; D016501:Neurites / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D020258:Neurotoxicity Syndromes / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D053140:Organelle Shape / Q000187:drug effects; D053141:Organelle Size / Q000187:drug effects; D010523:Peripheral Nervous System Diseases / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D051381:Rats
chemical_list: D000970:Antineoplastic Agents; D015415:Biomarkers; D009419:Nerve Tissue Proteins
keywords: automated image analysis; early safety evaluation.; chemotherapy-induced peripheral neuropathy (CIPN); high content analysis (HCA); rat dorsal root ganglion cells (DRGs)