TY - JOUR
T1 - Metabolomic and Signaling Programs Induced by Immobilized versus Soluble IFN γin Neural Stem Cells
AU - Baumann, Hannah J.
AU - Betonio, Patricia
AU - Abeywickrama, Chathura S.
AU - Shriver, Leah P.
AU - Leipzig, Nic D.
N1 - Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/9/16
Y1 - 2020/9/16
N2 - Neural stem cells (NSCs) provide a strategy to replace damaged neurons following traumatic central nervous system injuries. A major hurdle to translation of this therapy is that direct application of NSCs to CNS injury does not support sufficient neurogenesis due to lack of proper cues. To provide prolonged spatial cues to NSCs IFN-γwas immobilized to biomimetic hydrogel substrate to supply physical and biochemical signals to instruct the encapsulated NSCs to be neurogenic. However, the immobilization of factors, including IFN-γ, versus soluble delivery of the same factor, has been incompletely characterized especially with respect to activation of signaling and metabolism in cells over longer time points. In this study, protein and metabolite changes in NSCs induced by immobilized versus soluble IFN-γat 7 days were evaluated. Soluble IFN-γ, refreshed daily over 7 days, elicited stronger responses in NSCs compared to immobilized IFN-γ, indicating that immobilization may not sustain signaling or has altered ligand/receptor interaction and integrity. However, both IFN-γdelivery types supported increased βIII tubulin expression in parallel with canonical and noncanonical receptor-signaling compared to no IFN-γ. Global metabolomics and pathway analysis revealed that soluble and immobilized IFN-γaltered metabolic pathway activities including energy, lipid, and amino acid synthesis, with soluble IFN-γhaving the greatest metabolic impact overall. Finally, soluble and immobilized IFN-γsupport mitochondrial voltage-dependent anion channel (VDAC) expression that correlates to differentiated NSCs. This work utilizes new methods to evaluate cell responses to protein delivery and provides insight into mode of action that can be harnessed to improve regenerative medicine-based strategies.
AB - Neural stem cells (NSCs) provide a strategy to replace damaged neurons following traumatic central nervous system injuries. A major hurdle to translation of this therapy is that direct application of NSCs to CNS injury does not support sufficient neurogenesis due to lack of proper cues. To provide prolonged spatial cues to NSCs IFN-γwas immobilized to biomimetic hydrogel substrate to supply physical and biochemical signals to instruct the encapsulated NSCs to be neurogenic. However, the immobilization of factors, including IFN-γ, versus soluble delivery of the same factor, has been incompletely characterized especially with respect to activation of signaling and metabolism in cells over longer time points. In this study, protein and metabolite changes in NSCs induced by immobilized versus soluble IFN-γat 7 days were evaluated. Soluble IFN-γ, refreshed daily over 7 days, elicited stronger responses in NSCs compared to immobilized IFN-γ, indicating that immobilization may not sustain signaling or has altered ligand/receptor interaction and integrity. However, both IFN-γdelivery types supported increased βIII tubulin expression in parallel with canonical and noncanonical receptor-signaling compared to no IFN-γ. Global metabolomics and pathway analysis revealed that soluble and immobilized IFN-γaltered metabolic pathway activities including energy, lipid, and amino acid synthesis, with soluble IFN-γhaving the greatest metabolic impact overall. Finally, soluble and immobilized IFN-γsupport mitochondrial voltage-dependent anion channel (VDAC) expression that correlates to differentiated NSCs. This work utilizes new methods to evaluate cell responses to protein delivery and provides insight into mode of action that can be harnessed to improve regenerative medicine-based strategies.
UR - https://www.scopus.com/pages/publications/85090481637
U2 - 10.1021/acs.bioconjchem.0c00338
DO - 10.1021/acs.bioconjchem.0c00338
M3 - Article
C2 - 32820900
AN - SCOPUS:85090481637
SN - 1043-1802
VL - 31
SP - 2125
EP - 2135
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 9
ER -