Targeted delivery of mitochondria to the liver in rats

J Gastroenterol Hepatol. 2020 Dec;35(12):2241-2247. doi: 10.1111/jgh.15091. Epub 2020 Jun 24.

Abstract

Background and aim: Mitochondrial damage is commonly involved in liver injury. We have previously shown that normal mitochondria can be coated with a carrier protein to form complexes that are specifically taken up by liver cells in culture. The aim of the current study was to determine whether mitochondrial complexes could be specifically delivered to the livers of living rats by intravenous injection.

Methods: Mitochondria were harvested from fresh mouse liver, mixed with an asialoglycoprotein-based carrier, asialoorosomucoid-polylysine (AsOR-PL), and purified to form complexes. To facilitate the release of internalized mitochondria from endosomes, an endosomolytic peptide, listeriolysin O (LLO), was coupled to AsOR to form AsOR-LLO. Mitochondria alone, mitochondrial complexes with AsOR-PL, and mitochondrial complexes plus AsOR-LLO conjugate all containing the same number of mitochondria were injected intravenously. Animals were killed, and organs were removed and analyzed by quantitative polymerase chain reaction of mouse mitochondrial DNA, electron microscopy (EM), and in situ polymerase chain reaction and hybridization followed by immunohistochemical analyses.

Results: Calculations revealed that approximately 27% of the total injected mitochondria was detected in the liver, while less than 2% was found in spleen, and < 1% in lungs. Immunohistochemistry showed that mouse mitochondrial DNA staining was minimal with mitochondrial complexes alone, strong periportal with mitochondrial complexes co-injected with AsOR-LLO, and absent with mitochondria alone.

Conclusions: Targetable mitochondrial complexes can be delivered to rat liver, and the efficiency of that process is greatly enhanced by co-injection of a targetable endosomal release agent, AsOR-LLO.

Keywords: asialoglycoprotein; complex; listeriolysin O; liver targeted; mitochondria; receptor-mediated endocytosis; transplantation.

MeSH terms

  • Animals
  • Asialoglycoproteins / administration & dosage*
  • Bacterial Toxins / administration & dosage*
  • Carrier Proteins
  • Cell Transplantation / methods*
  • Endosomes
  • Female
  • Heat-Shock Proteins / administration & dosage*
  • Hemolysin Proteins / administration & dosage*
  • Hepatocytes / cytology
  • Injections, Intravenous
  • Liver*
  • Mice, Inbred Strains
  • Mitochondria, Liver / transplantation*
  • Orosomucoid / administration & dosage
  • Orosomucoid / analogs & derivatives*
  • Polylysine / administration & dosage*
  • Rats, Sprague-Dawley

Substances

  • Asialoglycoproteins
  • Bacterial Toxins
  • Carrier Proteins
  • Heat-Shock Proteins
  • Hemolysin Proteins
  • Orosomucoid
  • asialoorosomucoid
  • Polylysine
  • hlyA protein, Listeria monocytogenes