A robust platform for high-throughput screening of therapeutic strategies for acute and chronic spinal cord injury

Vaibhav Patil; Enda O'Connell; Leo R Quinlan; Howard Fearnhead; Siobhan McMahon; Abhay Pandit

doi:10.1016/j.isci.2021.102182

A robust platform for high-throughput screening of therapeutic strategies for acute and chronic spinal cord injury

iScience. 2021 Feb 12;24(3):102182. doi: 10.1016/j.isci.2021.102182. eCollection 2021 Mar 19.

Authors

Vaibhav Patil¹, Enda O'Connell², Leo R Quinlan^{1

3}, Howard Fearnhead⁴, Siobhan McMahon^{1

5}, Abhay Pandit¹

Affiliations

¹ CÚRAM, SFI Research Centre for Medical Devices, National University of Ireland, Galway, Ireland.
² Genomics and Screening Core Facility, National University of Ireland, Galway, Ireland.
³ Physiology, National University of Ireland, Galway, Ireland.
⁴ Pharmacology and Therapeutics, National University of Ireland, Galway, Ireland.
⁵ Anatomy, National University of Ireland, Galway, Ireland.

Abstract

Astrocytes and microglia are critical regulators of inflammatory cascade after spinal cord injury (SCI). Existing glial in vitro studies do not replicate inflammatory phases associated with SCI. Here, we report an in vitro model of mixed glial culture where inflammation is induced by the administration of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and interleukin-6) to promote pathologically relevant "acute" and "chronic" inflammatory phases. We observed SCI relevant differential modulation of inflammatory pathways, cytokines, chemokines, and growth factors over 21 days. Mitochondrial dysfunction was associated with a cytokine combination treatment. Highly expressed cytokine induced neutrophil chemoattractant (CINC-3) chemokine was used as a biomarker to establish an enzyme-linked immunosorbent assay-based high-throughput screening (HTS) platform. We screened a 786-compound drug library to demonstrate the efficacy of the HTS platform. The developed model is robust and will facilitate in vitro screening of anti-reactive glial therapeutics for the treatment of SCI.

Keywords: Cellular Neuroscience; Immunology; Molecular Neuroscience; Proteomics.