Macrophages and Exosomes Employ Brain Inflammation for CNS Delivery of Therapeutics A. Kabanov
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1 Macrophages and Exosomes Employ Brain Inflammation for CNS Delivery of Therapeutics A. Kabanov
2 Targeting Brain Inflammation in Disease Biochemical studies of brains from individuals with many neurologic disorders provide clear evidence for an activation of inflammatory pathways in the brain. Activated microglial cells secrete a wide range of inflammatory factors, including reactive oxygen species, cytokines (IL-1β, IL-6, TNF-α, and INF-γ), chemokines (MIP1α), MIP1β, CXCL8), growth factors, and complement components (C1q, C3, C4, and C9). In addition, selectins, integrins and intercellular cell adhesion molecules (ICAM) are upregulated in the inflamed brain endothelium. Microglia activation in MS brain The inflammatory process in the brain provides an opportunity for targeting of therapeutics to the brain.
3 Use of macrophages (M ) as Trojan horses for delivery of therapeutic nanozymes in the areas of inflammation in the brain Loading ex vivo nanozyme Введение нагруженных макрофагов в кровоток кровь Brain Migration of loaded macrophages to the brain and nanozyme release in the site of inflammation Repackaging on nanozymes in exosomes macrophage Proof of concept using delivery of a catalase nanozyme in a mouse models of Parkinson s disease Increased blood circulation time compared to free nanozyme Delivery of nanozymes in cells to the brain in mouse model of PD Nanozyme in M Nanozyme in M control 7 h 24 h neuron Neuronal survival in the PD mouse model healthy Decreased of brain inflammation astrocytes (GFAP) saline carrier Nanozyme in M Nanozyme Nanozyme in M A. Brynskikh et al. Nanomedicine (Lond.) 2010, 53:379 Y. Zhao et al. Nanomedicine (Lond.) 2011, 6:25 M. Haney et al. Nanomedicine (Lond.) 2012, 7:815
4 Adoptive Transfer of Genetically Modified M to Target Inflammation in the Brain Control media Media from Exosomes GDNF, 100 ng/ml GDNFmacrophages from GDNFmacrophages Haney et al. PLoS ONE (4): e61852 Zhao et al. PLoS ONE (9): e Neuroprotective effects of GDNF-transfected macrophages in Parkin-Q3111X(A) mouse model Wild type mice Parkin-Q311X(A) mice GDNF-mac PBS non-transfected M Time remaining, sec treatment Wire hanging test Age, months Time remaining, sec treatment 0 Rotarod test Age, months Time remaining, sec Escaping activity test treatment Wild type/pbs PD mice/pbs PD mice/gdnf-mac PD mice/empty mac PD mice/exogdnf Age, months
5 Exosome as protein carriers to brain in inflammation Therapeutic proteins Hydrophobic drugs DNA/RNA/siRNA Purification of exosomes from cells conditioned media Incubation with drug naïve exosomes Cell + Extrusion Exosome-based drug formulations Naive Incubation at RT Saponin treatment Freeze/thaw Adhesive proteins Hydrophilic drug Freeze-thaw cycles Drug Sonication Extrusion Strong sonication Mild sonication From pre-loaded cells Increased brain accumulation of exosomes and neurotrophin (BDNF) under brain inflammation Delta Brain/Serum Ratio (ml/g) 0.06 Healthy Mice Brain inflammation 0.04 Ki = ml/(g min) Vi = ml, r = Ki = ml/(g min) Vi = ml, r = Exposure time (min) %ID/g BDNF Healthy mice BDNF ExoBDNF Brain inflammation ExoBDNF Yuan, et al. Biomaterials 142 (2017) 1e12
6 Use of macrophage (M ) exosomes for delivery of proteins to the sites of inflammation in the brain Exosomes Anti-inflammatory and neuroprotective effects of macrophage (М2) exosomes loaded with catalase and administered intranasal in a mouse model of Parkinson s disease (6-OHDA in SNpc) Decrease of brain inflammation microglia anti-cd11b Exosomes are isolated from macrophages (М2) and loaded with catalase substantia nigra pars compacta (SNpc) Particle size before and after loading Journal of Controlled Release 2015, 207:18 30
7 Value proposition A) To deliver therapeutic proteins to the brain by using engineered exosomes to target to target sites of inflammation associated with the disease Potential upside in the works : to deliver DNA, sirna, and mrna using exosomes B) To deliver proteins, nucleic acids and nanoparticles to the brain and develop respective therapeutic using genetically engineered macrophages after adoptive transfer to target to target sites of inflammation associated with the disease
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