TY - JOUR
T1 - Uncovering temporospatial sensitive TBI targeting strategies via in vivo phage display
AU - Martinez, Briana I.
AU - Mousa, Gergey Alzaem
AU - Fleck, Kiera
AU - MacCulloch, Tara
AU - Diehnelt, Chris W.
AU - Stephanopoulos, Nicholas
AU - Stabenfeldt, Sarah E.
N1 - Funding Information:
We acknowledge the following facilities and/or personnel for support: ASU Genomics Facility and S. Yang for NGS support, ASU Mass Spectrometry Core and T. Karr for MS services, A. Witten for figure illustrations, and N. Borgogni for IHC assistance. This work was supported by NIH NICHD 1DP2HD084067 (S.E.S.), NIH NICHD 1DP2HD084067-01S1 (S.E.S.), NIH NINDS 1R21NS107985 (S.E.S.), ARCS Scholar Award (B.I.M.), and ASU Biomimicry Seed funding (C.W.D., N.S., and S.E.S.).
Publisher Copyright:
Copyright © 2022 The Authors, some rights reserved.
PY - 2022/7
Y1 - 2022/7
N2 - The heterogeneous pathophysiology of traumatic brain injury (TBI) is a barrier to advancing diagnostics and therapeutics, including targeted drug delivery. We used a unique discovery pipeline to identify novel targeting motifs that recognize specific temporal phases of TBI pathology. This pipeline combined in vivo biopanning with domain antibody (dAb) phage display, next-generation sequencing analysis, and peptide synthesis. We identified targeting motifs based on the complementarity-determining region 3 structure of dAbs for acute (1 day post-injury) and subacute (7 days post-injury) post-injury time points in a preclinical TBI model (controlled cortical impact). Bioreactivity and temporal sensitivity of the targeting motifs were validated via immunohistochemistry. Immunoprecipitation–mass spectrometry indicated that the acute TBI targeting motif recognized targets associated with metabolic and mitochondrial dysfunction, whereas the subacute TBI motif was largely associated with neurodegenerative processes. This pipeline successfully discovered temporally specific TBI targeting motif/epitope pairs that will serve as the foundation for the next-generation targeted TBI therapeutics and diagnostics.
AB - The heterogeneous pathophysiology of traumatic brain injury (TBI) is a barrier to advancing diagnostics and therapeutics, including targeted drug delivery. We used a unique discovery pipeline to identify novel targeting motifs that recognize specific temporal phases of TBI pathology. This pipeline combined in vivo biopanning with domain antibody (dAb) phage display, next-generation sequencing analysis, and peptide synthesis. We identified targeting motifs based on the complementarity-determining region 3 structure of dAbs for acute (1 day post-injury) and subacute (7 days post-injury) post-injury time points in a preclinical TBI model (controlled cortical impact). Bioreactivity and temporal sensitivity of the targeting motifs were validated via immunohistochemistry. Immunoprecipitation–mass spectrometry indicated that the acute TBI targeting motif recognized targets associated with metabolic and mitochondrial dysfunction, whereas the subacute TBI motif was largely associated with neurodegenerative processes. This pipeline successfully discovered temporally specific TBI targeting motif/epitope pairs that will serve as the foundation for the next-generation targeted TBI therapeutics and diagnostics.
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U2 - 10.1126/sciadv.abo5047
DO - 10.1126/sciadv.abo5047
M3 - Article
C2 - 35867794
AN - SCOPUS:85135203272
SN - 2375-2548
VL - 8
JO - Science Advances
JF - Science Advances
IS - 29
M1 - eabo5047
ER -