2. Academic Validation
  3. Exceptional Uptake, Limited Protein Expression: Liver Macrophages Lost in Translation of Synthetic mRNA

Exceptional Uptake, Limited Protein Expression: Liver Macrophages Lost in Translation of Synthetic mRNA

  • Adv Sci (Weinh). 2025 Mar;12(9):e2409729. doi: 10.1002/advs.202409729.
Cheng Lin 1 2,  Adrian Kuzmanović 1,  Nan Wang 1 3,  Liangliang Liao 1 4,  Sabrina Ernst 5,  Christian Penners 1,  Alexander Jans 1,  Thomas Hammoor 6 7,  Petra Bumnuri Stach 1,  Mona Peltzer 1,  Ines Volkert 1,  Elisabeth Zechendorf 8,  Reham Hassan 9 10,  Maiju Myllys 9,  Christian Liedtke 1,  Andreas Herrmann 6 7,  Gurudas Chakraborty 6,  Christian Trautwein 9,  Jan Hengstler 9,  Gerhard Müller-Newen 11,  Junqing Wang 3,  Ahmed Ghallab 9 10,  Matthias Bartneck 1 6 7
Affiliations

Affiliations

  • 1 Department of Internal Medicine III, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074, Aachen, Germany.
  • 2 Department of Rheumatology and Shanghai Institute of Rheumatology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200240, China.
  • 3 Department of General Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200240, China.
  • 4 Japan Union Hospital of Jilin University, 130033, Changchun, China.
  • 5 Confocal Microscopy Facility, Interdisciplinary Center for Clinical Research IZKF, University Hospital RWTH Aachen, 52074, Aachen, Germany.
  • 6 DWI - Leibniz Institute for Interactive Materials, Forckenbeckstraße 50, 52074, Aachen, Germany.
  • 7 Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 2, 52074, Aachen, Germany.
  • 8 Department of Intensive and Intermediate Care, University Hospital RWTH Aachen, 52074, Aachen, Germany.
  • 9 Leibniz Research Centre for Working Environment and Human Factors, 44139, Dortmund, Germany.
  • 10 Department of Forensic and Veterinary Toxicology, Faculty of Veterinary Medicine, South Valley University, 83523, Qena, Egypt.
  • 11 Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
PMID: 39792811 DOI: 10.1002/advs.202409729
Abstract

Most gene therapies exert their actions via manipulation of hepatocytes (parenchymal cells) and the reasons behind the suboptimal performance of synthetic mRNA in non-parenchymal cells (NPC) such as Kupffer cells (KC), and liver macrophages, remain unclear. Here, the spatio-temporal distribution of mRNA encoding enhanced green fluorescent protein (Egfp), siRNA, or both co-encapsulated into lipid nanoparticles (LNP) in the liver in vivo using real-time intravital imaging is investigated. Although both KC and hepatocytes demonstrate comparable high and rapid uptake of mRNA-LNP and siRNA-LNP in vivo, the translation of Egfp mRNA occurs exclusively in hepatocytes during intravital imaging. Despite attempts such as inhibiting intracellular ribonuclease, substituting uridine Bases in mRNA with pseudouridine, and using a different ionizable lipid in the LNP mixture, no substantial increase in Egfp translation by NPC is possible. The investigation reveals that hepatocytes, which are distinct from Other liver cells due to their polyploidy, exhibit significantly elevated levels of total RNA and protein, along with a higher proportion of ribosomal protein per individual cell. Consequently, fundamental cellular differences account for the low mRNA translation observed in NPC. The findings therefore suggest that cellular biology imposes a natural limitation on synthetic mRNA translation that is strongly influenced by cellular ploidy.

Keywords

hepatocytes; lipid nanoparticles; non‐parenchymal cells; ploidy; synthetic mRNA.

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