TY  - JOUR
AU  - Khalin, Igor
AU  - Adarsh, Nagappanpillai
AU  - Schifferer, Martina
AU  - Wehn, Antonia
AU  - Boide-Trujillo, Valeria J
AU  - Mamrak, Uta
AU  - Shrouder, Joshua
AU  - Misgeld, Thomas
AU  - Filser, Severin
AU  - Klymchenko, Andrey S
AU  - Plesnila, Nikolaus
TI  - Nanocarrier Drug Release and Blood-Brain Barrier Penetration at Post-Stroke Microthrombi Monitored by Real-Time Förster Resonance Energy Transfer.
JO  - ACS nano
VL  - 19
IS  - 15
SN  - 1936-0851
CY  - Washington, DC
PB  - Soc.
M1  - DZNE-2025-00554
SP  - 14780 - 14794
PY  - 2025
AB  - Nanotechnology holds great promise for improving the delivery of therapeutics to the brain. However, current approaches often operate at the organ or tissue level and are limited by the lack of tools to dynamically monitor cargo delivery in vivo. We have developed highly fluorescent lipid nanodroplets (LNDs) that enable tracking of nanocarrier behavior at the subcellular level while also carrying a Förster resonance energy transfer (FRET)-based drug delivery detection system (FedEcs) capable of monitoring cargo release in vivo. Using two-photon microscopy, we demonstrate that circulating LNDs in naïve mouse brain vasculature exhibit 3D real-time FRET changes, showing size-dependent stability over 2 h in blood circulation. Further, in the Nanostroke model, dynamic intravital two-photon imaging revealed that LNDs accumulated within cerebral postischemic microthrombi, where they released their cargo significantly faster than in normal blood circulation. Furthermore, the blood-brain barrier (BBB) became permeable at the microclot sites thereby allowing accumulated FedEcs-LNDs to cross the BBB and deliver their cargo to the brain parenchyma. This microthrombi-associated translocation was confirmed at the ultrastructural level via volume-correlative light-electron microscopy. Consequently, FedEcs represents an advanced tool to quantitatively study the biodistribution and cargo release of nanocarriers at high resolution in real-time. By enabling us to resolve passive targeting mechanisms poststroke, specifically, accumulation, degradation, and extravasation via poststroke microthrombi, this system could significantly enhance the translational validation of nanocarriers for future treatments of brain diseases.
KW  - Blood-Brain Barrier: metabolism
KW  - Blood-Brain Barrier: drug effects
KW  - Animals
KW  - Fluorescence Resonance Energy Transfer
KW  - Mice
KW  - Drug Carriers: chemistry
KW  - Stroke: drug therapy
KW  - Stroke: metabolism
KW  - Drug Liberation
KW  - Nanoparticles: chemistry
KW  - Lipids: chemistry
KW  - Thrombosis: drug therapy
KW  - Thrombosis: metabolism
KW  - Mice, Inbred C57BL
KW  - blood-brain barrier (Other)
KW  - correlative light-electron microscopy (Other)
KW  - microthrombosis (Other)
KW  - nanocarriers (Other)
KW  - stroke (Other)
KW  - Drug Carriers (NLM Chemicals)
KW  - Lipids (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:40180319
DO  - DOI:10.1021/acsnano.4c17011
UR  - https://pub.dzne.de/record/278048
ER  -