Deciphering glycomics and neuroproteomic alterations in experimental traumatic brain injury: Comparative analysis of aspirin and clopidogrel treatment

Abstract

As populations age, the number of patients sustaining traumatic brain injury (TBI) and concomitantly receiving preinjury antiplatelet therapy such as aspirin (ASA) and clopidogrel (CLOP) is rising. These drugs have been linked with unfavorable clinical outcomes following TBI, where the exact mechanism(s) involved are still unknown. In this novel work, we aimed to identify and compare the altered proteome profile imposed by ASA and CLOP when administered alone or in combination, prior to experimental TBI. Furthermore, we assessed differential glycosylation PTM patterns following experimental controlled cortical impact model of TBI, ASA, CLOP, and ASA + CLOP. Ipsilateral cortical brain tissues were harvested 48 h postinjury and were analyzed using an advanced neuroproteomics LC-MS/MS platform to assess proteomic and glycoproteins alterations. Of interest, differential proteins pertaining to each group (22 in TBI, 41 in TBI + ASA, 44 in TBI + CLOP, and 34 in TBI + ASA + CLOP) were revealed. Advanced bioinformatics/systems biology and clustering analyses were performed to evaluate biological networks and protein interaction maps illustrating molecular pathways involved in the experimental conditions. Results have indicated that proteins involved in neuroprotective cellular pathways were upregulated in the ASA and CLOP groups when given separately. However, ASA + CLOP administration revealed enrichment in biological pathways relevant to inflammation and proinjury mechanisms. Moreover, results showed differential upregulation of glycoproteins levels in the sialylated N-glycans PTMs that can be implicated in pathological changes. Omics data obtained have provided molecular insights of the underlying mechanisms that can be translated into clinical bedside settings.

LC–MS/MS glycomics of idiopathic rapid eye movement sleep behavior disorder

“…Student's t ‐tests were applied to absolute intensities acquired on C18‐LC–MS and PGC‐LC–MS to compare glycans and N‐glycan isomers between control and idiopathic RBD cohorts. Two‐tailed t ‐tests were performed, similar to what we have used in our previous studies using Markerview ™ software (AB Sciex, Framingham, MA) .…”

Neuroproteomics and Systems Biology Approach to Identify Temporal Biomarker Changes Post Experimental Traumatic Brain Injury in Rats

“…The injured rats were sacrificed by decapitation. From each group, we used five rats for our proteomics analysis, which has been validated to be statistically significant (26, 35, 51–53). For our control animals, sham-injured rats were subjected to identical surgical procedures but did not receive an impact injury.…”

“…The potential of neuroproteomics platforms have been explored using acute paradigms of TBI (26, 27, 29–35), spinal cord injury (36–44), and cerebral ischemia or stroke (34, 45–50). Our group has previously reported TBI effects upon the global proteome where we combined cyanine labeling with SDS PAGE–capillary LC–MS/MS to study hippocampal tissue (30).…”

Characterization of isomeric glycan structures by LC‐MS/MS

“…The same method was also used to profile glycans from mouse brain tissue . Glycomics changes associated with trumatic brain injury were also assessed using the above LC‐MS/MS analysis of permethylated glycans . Glycans derived from glioma stem cell xenografts , neuroblastoma and breast cancer cell lines, IgG , prostate‐specific antigen , and human milk, bovine milk, and goat milk were also analyzed using RPLC‐MS/MS of reduced and permethylated glycans.…”