Exosome-Based Delivery of Super-Repressor IκBα Alleviates Alcohol-Associated Liver Injury in Mice

Abstract

Activation of Kupffer cells (KCs) by gut-derived lipopolysaccharide (LPS) instigates nuclear factor-κB (NF-κB)-mediated inflammatory responses in alcohol-associated liver diseases (ALD). Here, we utilized a novel optogenetically engineered exosome technology called ‘exosomes for protein loading via optically reversible protein–protein interactions (EXPLOR)’ to efficiently deliver the super-repressor IκB-loaded exosomes (Exo-srIκB) to the liver and examined its therapeutic potential in acute-on-chronic alcohol-associated liver injury. We detected enhanced uptake of DiI-labeled Exo-srIκB by LPS-treated inflammatory KCs, which suppressed LPS-induced inflammatory gene expression levels. In animal experiments, a single intravenous injection of Exo-srIκB prior to alcohol binge drinking significantly attenuated alcohol-associated hepatic steatosis and infiltration of neutrophils and macrophages but not a liver injury. Notably, three consecutive days of Exo-srIκB injection remarkably reduced alcohol-associated liver injury, steatosis, apoptosis of hepatocytes, fibrosis-related gene expression levels in hepatic stellate cells, infiltration of neutrophils and macrophages, and inflammatory gene expression levels in hepatocytes and KCs. In particular, the above effects occurred with inhibition of nuclear translocation of NF-κB in liver tissues, and these beneficial effects of Exo-srIκB on ALD were shown regardless of doses. Our results suggest an exosome-based modulation of NF-κB activity in KCs by Exo-srIκB as a novel and efficient therapeutic approach in ALD.

Intracellular Protein Delivery: Approaches, Challenges, and Clinical Applications

“…[ 226 ] treated post-ischemic mice with srIκB-loaded exosomes to suppress the NF-κB signaling cascade activated during kidney ischemia–reperfusion injury. Recently, these srIκB-loaded exosomes were also employed to suppress inflammation in models of alcohol-associated liver disease [ 227 ] and neuropathic pain [ 228 ].…”

NF-κB in biology and targeted therapy: new insights and translational implications

“…Emerging preclinical evidence has addressed the wide use of IκBα super-repressor delivered through exosome systems in various diseases, including alcohol-associated liver injury, sepsis-associated organ damage, kidney ischemia-reperfusion injury, and amyotrophic lateral sclerosis (ALS). 566 – 570 By utilizing an engineered exosome technology called “exosomes for protein loading via optically reversible protein–protein interactions (EXPLOR)”, Yim et al, engineered exosomes to load super-repressor IκB (Exo-srIκB) for the efficient intracellular transfer of protein-based therapeutics. 571 Although the application of IκBα super-repressor is currently limited to laboratory research settings, it demonstrates high potential as a promising intervention in clinical practice in the future.…”

Role of Exosomal Modulation of Macrophages in Liver Fibrosis

“…For example, Exo-srIB inhibits NF-κB signaling in KCs to reduce LF through exosomes. 105 They successfully delivered exosomes loaded with the super suppressor IB (Exo-srIB) to the liver using a novel optogenetically engineered exosome technology called “exosomes loaded with proteins via optically reversible protein-protein interactions”. Exo-srIB markedly reduced the levels of fibrosis-associated gene expression in HSCs following three-day injections.…”

“…116 Generally speaking, in the progression of LF, macrophages are the core hub of the regulation network (Table 2). 98,[100][101][102][103][104][105][106]109,110,112,117…”