Dasa Cizkova1,2,3, Jusal Quanico1, Melodie-Anne Kanoub1,4, Fahed Zahri1,4, Franck Rodet1, Adriana-Natalia Murgoci1,2,3, Veronika Cubinkova3, Isabelle Fournier1 and Michel Salzet1*
1Department of Proteomic Laboratory, Inflammatory Response and Mass Spectrometry, University of Lille 1, France
2Department of Anatomy, Histology and Physiology, University of Veterinary Medicine and Pharmacy, Slovakia
3Department of Neuroimmunology, Institute of Neuroimmunology, Slovak Academy of Sciences, Slovakia
4Department of Neurosurgery, University Regional Hospital of Lille, France
Spinal Cord Injury (SCI) represents currently incurable disorder of the CNS often accompanied by permanent health consequences-disability. In order to mimic a SCI, a balloon-compressive technique was performed at thoracic Th8-9 spinal level in adult rat. Shot-gun proteomic was used to identify proteins in each spinal cord segment-derived conditioned medium along the rostralcaudal axis after SCI with time course. In addition, 3D MALDI Imaging, tissue micro-proteomics were undertaken and combined with confocal imaging, in-vitro and in-vivo functional testing. Our data clearly described the spatial and temporal events which developed in acute phase after SCI. While we found some similarities in the segments above and below the lesion, the caudal segment has been identified as the therapeutic target. We then assessed in a rat SCI model the in vivo impact of a sustained RhoA inhibitor administered in situ via functionalized- alginate scaffold. In order to decipher the underlying molecular mechanisms involved in such a process, an in vitro neuroproteomic-systems biology platform was developed. Here, the pan-proteomic profile of the Dorsal Root Ganglia (DRG) cell line ND7/23 DRG was assessed in a large array of culture conditions using RhoA i and conditioned media obtained from SCI ex-vivo derived spinal cord slices. A fine mapping of the spatio-temporal molecular events of the RhoA i treatment in SCI was performed. The data obtained allow a better understanding of regeneration induced above and below the lesion site. Results notably showed a time- dependent alteration of the transcription factors profile along with the synthesis of growth cone-related factors (receptors, ligands, and signaling pathways) in RhoA i treated DRG cells. Furthermore, we demonstrate the inflammatory response process involvement via immunoglobulin’s by binding to their specific receptors on the DRG cells upon neurite outgrowth initiation and thus modulating the neurite outgrowth process. We then validate our results by an in vivo proteomic studies along the spinal cord segments. Taken together, we expand knowledge on SCI proteins involved in inflammation, neurotransmission, immune response, re growth and repair, and other local responses to injury.
Cizkova D, Quanico J, Kanoub M-A, Zahiri F, Rodet F, Murgoci A-N, et al. Shedding New Light on Spinal Cord Injury via a Spatio-Temporal Proteomic and Physiological Approaches. Ann Trauma Acute Care. 2018; 2(1): 1007.