Penetrating Traumatic Brain Injury Triggers Dysregulation of Cathepsin B Protein Levels Independent of Cysteine Protease Activity in Brain and Cerebral Spinal Fluid

Boutté, Angela M.; Hook, Vivian; Thangavelu, Bharani; Sarkis, George Anis; Abbatiello, Brittany N; Hook, Gary E. R.; Jacobsen, J. Steven; Robertson, Claudia S.; Gilsdorf, Janice; Yang, Zhihui; Wang, Kevin; Shear, Deborah A.

doi:10.1089/neu.2019.6537

Cited by 19 publications

(21 citation statements)

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“…Portions of the dissected right lateral neocortices of 6 h–4w sham and injured animals were homogenized in 50 μM citric acid at a pH 6.0, spun at 12,000 xg at 4° C for 10 min and the supernatant of the whole homogenate was collected. Protein concentrations were measured using a NanoDrop Lite (Thermo Fisher Scientific, Wilmington, DE) and Cathepsin B activity was measured in a 96-well plate, each well containing 2x Assay Buffer (100 mM sodium acetate pH 5.5, 2 mM EDTA, 200 mM sodium chloride, 8 mM DTT), 2 μg of neocortex whole homogenate, and Z-Phe-Arg-7-amino-4-(trifluoromethyl) coumarin (ZFR-AMC), a substrate for cysteine proteases that fluoresces upon cleavage by Cathepsin B ( Hook et al, 2013 ; Boutté et al, 2020 ; Yoon et al, 2021 ). Assay specificity for Cathepsin B was validated using Cathepsin B specific inhibitor, CA-074, and general cystine protease inhibitor, E64c, pre-incubated in brain homogenate ( Supplementary Figure 2 ).…”

Section: Methodsmentioning

confidence: 99%

“…While Cath B usually functions within the acidic environment of the lysosome, it also has endopeptidase activity at the more neutral pH of the cytosol, unlike other lysosomal proteases. Within the context of TBI, Cath B has been found to be upregulated in rodent TBI models of focal lesions, such as the controlled cortical impact (CCI) and penetrating ballistic-like brain injury (PBBI) ( Luo et al, 2010 ; Hook et al, 2013 ; Boutté et al, 2020 ). Additional findings within patient cerebrospinal fluid also indicated an upregulation of Cath B within hours or days post-penetrating TBI compared to non-TBI controls, providing further evidence that Cath B could play a role in TBI pathology ( Boutté et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

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Cathepsin B Relocalization in Late Membrane Disrupted Neurons Following Diffuse Brain Injury in Rats

et al. 2022

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Traumatic brain injury (TBI) has consequences that last for years following injury. While TBI can precipitate a variety of diffuse pathologies, the mechanisms involved in injury-induced neuronal membrane disruption remain elusive. The lysosomal cysteine protease, Cathepsin B (Cath B), and specifically its redistribution into the cytosol has been implicated in cell death. Little is known about Cath B or neuronal membrane disruption chronically following diffuse TBI. Therefore, the current study evaluated Cath B and diffuse neuronal membrane disruption over a more chronic post-injury window (6 h–4 w). We evaluated Cath B in adult male Sprague-Dawley rats following central fluid percussion injury (CFPI). Expression of Cath B, as well as Cath B-associated pro (Bak and AIF) and anti-apoptotic (Bcl-xl) proteins, were assessed using western blot analysis. Cath B activity was also assessed. Localization of Cath B was evaluated in the membrane disrupted and non-disrupted population following CFPI using immunohistochemistry paired with quantitative image analysis and ultrastructural verification. There was no difference in expression or activity of Cath B or any of the associated proteins between sham and CFPI at any time post-injury. Immunohistological studies, however, showed a sub-cellular re-localization of Cath B at 2 w and 4 w post-injury in the membrane disrupted neuronal population as compared to the time-point matched non-disrupted neurons. Both membrane disruption and Cath B relocalization appear linked to neuronal atrophy. These observations are indicative of a late secondary pathology that represents an opportunity for therapeutic treatment of these neurons following diffuse TBI. Summary Statement Lysosomal cathepsin B relocalizes to the cytosol in neurons with disrupted plasmalemmal membranes weeks following diffuse brain injury. Both the membrane disrupted and cathepsin B relocalized neuronal subpopulations displayed smaller soma and nucleus size compared to non-pathological neurons, indicating atrophy.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cathepsin B Relocalization in Late Membrane Disrupted Neurons Following Diffuse Brain Injury in Rats

et al. 2022

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“…This model has been extensively characterized; several studies have found that key features are generally highest at 1, 3, or 7 days post-injury. For instance, blood brain barrier disruption is greatest within 1 day ( Cunningham et al, 2014 ), while indices of inflammation, cell loss, and protein fragmentation are prominent at 3–7 days post-injury ( Boutte et al, 2016 , 2020 ; Cartagena et al, 2016 ; DeDominicis et al, 2018 ). Collectively, these features are involved in severe TBI progression for which miRNA may be involved.…”

Section: Introductionmentioning

confidence: 99%

Penetrating Ballistic-Like Brain Injury Leads to MicroRNA Dysregulation, BACE1 Upregulation, and Amyloid Precursor Protein Loss in Lesioned Rat Brain Tissues

et al. 2020

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Severe traumatic brain injury (TBI) is a risk factor for neurodegenerative diseases. Yet, the molecular events involving dysregulated miRNAs that may be associated with protein degradation in the brain remains elusive. Quantitation of more than 800 miRNAs was conducted using rat ipsilateral coronal brain tissues collected 1, 3, or 7 days after penetrating ballistic-like brain injury (PBBI). As a control for each time-point, Sham-operated animals received craniotomy alone. Microarray and systems biology analysis indicated that the amplitude and complexity of miRNAs affected were greatest 7 day after PBBI. Arrays and Q-PCR inferred that dysregulation of miR-135a, miR-328, miR-29c, and miR-21 were associated with altered levels of beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), PSEN1, PSEN2, and amyloid precursor protein (APP) genes. These events were followed by increased levels of mature BACE1 protein and concomitant loss of full length APP within 3-7 days, then elevation of amyloid beta (Aβ)-40 7 days after PBBI. This study indicates that miRNA arrays, coupled with systems biology, may be used to guide study design prior validation of miRNA dysregulation. Associative analysis of miRNAs, mRNAs, and proteins within a proposed pathway are poised for further validation as biomarkers and therapeutic targets relevant to TBI-induced APP loss and subsequent Aβ peptide generation during neurodegeneration.

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“…The ability to predict mortality and injury severity may facilitate the development of clinical guidelines for managing pTBI. 15 Several biomarkers are reported to be altered after PBBI, including GFAP (glial fibrillary acidic protein; astrogliosis/astroglial injury), [16][17][18] brain-derived neurotrophic factor (BDNF; neurogenesis/neuroprotection), 19 ubiquitin C-terminal hydrolase-L1 (UCH-L1; neuronal cell body injury), 20 cathepsin B (CatB; apoptosis/ cell necrosis), 21 and microRNA (miRNA; astrogliosis/ inflammation/neurodegeneration). 22,23 Considering that pTBI and polytrauma often occur concurrently in the setting of emergency medicine, we hypothesized that the use of brain-specific and global injury biomarkers can reliably identify HX+HS and frontal PBBI with and without systemic insults (HX+HS) to diagnose and differentiate combined injury from isolated pTBI.…”

Section: Introductionmentioning

confidence: 99%

“…Several biomarkers are reported to be altered after PBBI, including GFAP (glial fibrillary acidic protein; astrogliosis/astroglial injury), 16–18 brain-derived neurotrophic factor (BDNF; neurogenesis/neuroprotection), 19 ubiquitin C-terminal hydrolase-L1 (UCH-L1; neuronal cell body injury), 20 cathepsin B (CatB; apoptosis/cell necrosis), 21 and microRNA (miRNA; astrogliosis/inflammation/neurodegeneration). 22 , 23 …”

Section: Introductionmentioning

confidence: 99%

Blood-Based Brain and Global Biomarker Changes after Combined Hypoxemia and Hemorrhagic Shock in a Rat Model of Penetrating Ballistic-Like Brain Injury

Pierre

Yang

et al. 2021

Neurotrauma Reports

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Penetrating traumatic brain injury (pTBI) often occurs with systemic insults such as hemorrhagic shock (HS) and hypoxemic (HX). This study examines rat models of penetrating ballistic-like brain injury (PBBI) and HX+HS to assess whether the blood levels of brain and systemic response biomarkers phosphorylated neurofilament-heavy protein (pNF-H), neurofilament-light protein (NF-L), aII-spectrin, heat shock protein (HSP70), and high mobility group box 1 protein (HMGB1) can distinguish pTBI from systemic insults and guide in pTBI diagnosis, prognosis, and monitoring. Thirty rats were randomly assigned to sham, PBBI, HS+HX, and PBBI+HS+HX groups. PBBI and sham groups underwent craniotomy with and without probe insertion and balloon expansion, respectively. HX and HS was then simulated by blood withdrawal and fraction of inspired oxygen (FIO 2 ) reduction. Biomarker serum concentrations were determined at one (D1) and two (D2) days post-injury with enzyme-linked immunosorbent assay (ELISA) methods. Axonal injury-linked biomarkers pNF-H and NF-L serum levels in PBBI groups were higher than those in sham and HX+HS groups at D1 and D2 post-injury. The same was true for PBBI+HX+HS compared with sham (D2 only for pNF-H) and HX+HS groups. However, pNF-H and NF-L levels in PBBI+HX+HS groups were not different than their PBBI counterparts. At D1, aII-spectrin levels in the HX+HS and PBBI+HS+HX groups were higher than the sham groups. aII-spectrin levels in the HX+HS group were higher than the PBBI group. This suggests HX+HS as the common insult driving aII-spectrin elevations. In conclusion, pNF-H and NF-L may serve as specific serum biomarkers of pTBI in the presence or absence of systemic insults. aII-spectrin may be a sensitive acute biomarker in detecting systemic insults occurring alone or with pTBI.

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Penetrating Traumatic Brain Injury Triggers Dysregulation of Cathepsin B Protein Levels Independent of Cysteine Protease Activity in Brain and Cerebral Spinal Fluid

Cited by 19 publications

References 59 publications

Cathepsin B Relocalization in Late Membrane Disrupted Neurons Following Diffuse Brain Injury in Rats

Cathepsin B Relocalization in Late Membrane Disrupted Neurons Following Diffuse Brain Injury in Rats

Penetrating Ballistic-Like Brain Injury Leads to MicroRNA Dysregulation, BACE1 Upregulation, and Amyloid Precursor Protein Loss in Lesioned Rat Brain Tissues

Blood-Based Brain and Global Biomarker Changes after Combined Hypoxemia and Hemorrhagic Shock in a Rat Model of Penetrating Ballistic-Like Brain Injury

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