Uncovering CNS access of lipidated exendin-4 analogues by quantitative whole-brain 3D light sheet imaging
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Uncovering CNS access of lipidated exendin-4 analogues by quantitative whole-brain 3D light sheet imaging. / Skovbjerg, Grethe; Roostalu, Urmas; Salinas, Casper G.; Skytte, Jacob L.; Perens, Johanna; Clemmensen, Christoffer; Elster, Lisbeth; Frich, Camilla K.; Hansen, Henrik H.; Hecksher-Sørensen, Jacob.
In: Neuropharmacology, Vol. 238, 109637, 2023.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Uncovering CNS access of lipidated exendin-4 analogues by quantitative whole-brain 3D light sheet imaging
AU - Skovbjerg, Grethe
AU - Roostalu, Urmas
AU - Salinas, Casper G.
AU - Skytte, Jacob L.
AU - Perens, Johanna
AU - Clemmensen, Christoffer
AU - Elster, Lisbeth
AU - Frich, Camilla K.
AU - Hansen, Henrik H.
AU - Hecksher-Sørensen, Jacob
N1 - Publisher Copyright: © 2023 Elsevier Ltd
PY - 2023
Y1 - 2023
N2 - Peptide-based drug development for CNS disorders is challenged by poor blood-brain barrier (BBB) penetrability of peptides. While acylation protractions (lipidation) have been successfully applied to increase circulating half-life of therapeutic peptides, little is known about the CNS accessibility of lipidated peptide drugs. Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful method to visualize whole-brain 3D distribution of fluorescently labelled therapeutic peptides at single-cell resolution. Here, we applied LSFM to map CNS distribution of the clinically relevant GLP-1 receptor agonist (GLP-1RA) exendin-4 (Ex4) and lipidated analogues following peripheral administration. Mice received an intravenous dose (100 nmol/kg) of IR800 fluorophore-labelled Ex4 (Ex4), Ex4 acylated with a C16-monoacid (Ex4_C16MA) or C18-diacid (Ex4_C18DA). Other mice were administered C16MA-acylated exendin 9-39 (Ex9-39_C16MA), a selective GLP-1R antagonist, serving as negative control for GLP-1R mediated agonist internalization. Two hours post-dosing, brain distribution of Ex4 and analogues was predominantly restricted to the circumventricular organs, notably area postrema and nucleus of the solitary tract. However, Ex4_C16MA and Ex9-39_C16MA also distributed to the paraventricular hypothalamic nucleus and medial habenula. Notably, Ex4_C18DA was detected in deeper-lying brain structures such as dorsomedial/ventromedial hypothalamic nuclei and the dentate gyrus. Similar CNS distribution maps of Ex4_C16MA and Ex9-39_C16MA suggest that brain access of lipidated Ex4 analogues is independent on GLP-1 receptor internalization. The cerebrovasculature was devoid of specific labelling, hence not supporting a direct role of GLP-1 RAs in BBB function. In conclusion, peptide lipidation increases CNS accessibility of Ex4. Our fully automated LSFM pipeline is suitable for mapping whole-brain distribution of fluorescently labelled drugs.
AB - Peptide-based drug development for CNS disorders is challenged by poor blood-brain barrier (BBB) penetrability of peptides. While acylation protractions (lipidation) have been successfully applied to increase circulating half-life of therapeutic peptides, little is known about the CNS accessibility of lipidated peptide drugs. Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful method to visualize whole-brain 3D distribution of fluorescently labelled therapeutic peptides at single-cell resolution. Here, we applied LSFM to map CNS distribution of the clinically relevant GLP-1 receptor agonist (GLP-1RA) exendin-4 (Ex4) and lipidated analogues following peripheral administration. Mice received an intravenous dose (100 nmol/kg) of IR800 fluorophore-labelled Ex4 (Ex4), Ex4 acylated with a C16-monoacid (Ex4_C16MA) or C18-diacid (Ex4_C18DA). Other mice were administered C16MA-acylated exendin 9-39 (Ex9-39_C16MA), a selective GLP-1R antagonist, serving as negative control for GLP-1R mediated agonist internalization. Two hours post-dosing, brain distribution of Ex4 and analogues was predominantly restricted to the circumventricular organs, notably area postrema and nucleus of the solitary tract. However, Ex4_C16MA and Ex9-39_C16MA also distributed to the paraventricular hypothalamic nucleus and medial habenula. Notably, Ex4_C18DA was detected in deeper-lying brain structures such as dorsomedial/ventromedial hypothalamic nuclei and the dentate gyrus. Similar CNS distribution maps of Ex4_C16MA and Ex9-39_C16MA suggest that brain access of lipidated Ex4 analogues is independent on GLP-1 receptor internalization. The cerebrovasculature was devoid of specific labelling, hence not supporting a direct role of GLP-1 RAs in BBB function. In conclusion, peptide lipidation increases CNS accessibility of Ex4. Our fully automated LSFM pipeline is suitable for mapping whole-brain distribution of fluorescently labelled drugs.
KW - 3D imaging
KW - Biodistribution
KW - Blood-brain barrier
KW - GLP-1 analogue
KW - Light-sheet fluorescence microscopy
KW - Peptide lipidation
U2 - 10.1016/j.neuropharm.2023.109637
DO - 10.1016/j.neuropharm.2023.109637
M3 - Journal article
C2 - 37391028
AN - SCOPUS:85165978049
VL - 238
JO - Neuropharmacology
JF - Neuropharmacology
SN - 0028-3908
M1 - 109637
ER -
ID: 361845362