Development of a systems‐based in situ multiplex biomarker screening approach for the assessment of immunopathology and neural tissue plasticity in male rats after traumatic brain injury

Abstract

Traumatic brain injuries (TBI) pose a massive burden of disease and continue to be a leading cause of morbidity and mortality throughout the world. A major obstacle in developing effective treatments is the lack of comprehensive understanding of the underlying mechanisms that mediate tissue damage and recovery after TBI. As such, our work aims to highlight the development of a novel experimental platform capable of fully characterizing the underlying pathobiology that unfolds post-TBI. This platform encompasses an empirically optimized multiplex immunohistochemistry staining and imaging system customized to screen for a myriad of biomarkers required to comprehensively evaluate the extent of neuroinflammation, neural tissue damage and repair in response to TBI. Herein, we demonstrate that our multiplex biomarker screening platform is capable of evaluating changes in both the topographical location and functional states of resident and infiltrating cell types that play a role in neuropathology after controlled cortical impact injury to the brain in male Sprague-Dawley rats. Our results demonstrate that our multiplex biomarker screening platform lays the groundwork for the comprehensive characterization of changes that occur within the brain after TBI. Such work may ultimately lead to the understanding of the governing pathobiology of TBI, thereby fostering the development of novel therapeutic interventions tailored to produce optimal tissue protection, repair and/or regeneration with minimal side effects and may ultimately find utility in wide variety of other neurological injuries, diseases and disorders that share components of TBI pathobiology.

Whole-brain tissue mapping toolkit using large-scale highly multiplexed immunofluorescence imaging and deep neural networks

“…3a). Then, signal corrections are performed, providing seamless 10-100 plex mosaics of whole-brain slices that provide sufficient spatial extent and spatial resolution to reveal cellular structures 12,13 . Importantly, they are suitable for automated scoring, for identifying a rich diversity of cell types (Fig.…”

Severe Spinal Cord Injury in Rats Induces Chronic Changes in the Spinal Cord and Cerebral Cortex Metabolism, Adjusted by Thiamine That Improves Locomotor Performance

“…This work has shown that in both the spinal cord and brain, a key mitochondrial multienzyme complex, OGDHC, is downregulated as a chronic consequence of severe SCI, with the downregulation linked to decreased activity of the immediate metabolic partner of OGDHC—GDH. The extension of the spinal trauma consequences to the distant area of CNS should be taken into account in the studies which develop the damage markers of neurotrauma, using non-damaged area of the same organism as a matched control ( Bogoslovsky et al, 2018 ).…”

A novel in situ multiplex immunofluorescence panel for the assessment of tumor immunopathology and response to virotherapy in pediatric glioblastoma reveals a role for checkpoint protein inhibition

“…22 More advanced computational solutions are also being developed that will employ machine learning algorithms customized to our imaging paradigm, ultimately enabling easily scalable image data analysis to support systems biology and translational research into devising optimal treatment strategies to reduce the burden of neurological disease/ disorders and/or traumatic insults. 23 Computational advances, in combination with future mechanical automation of the sample preparation, can improve the scalability of this methodology so that it may be routinely performed on patients monitored for treatment response after virotherapy or other immunotherapies. 24 Increased application of our novel in situ multiplex immunofluorescence will also promote an improved understanding of the range of immune and tumor cell response to virotherapy, and with clinical correlation will provide biological insights in patient stratification and monitoring response to treatment.…”