Murine liver repair via transient activation of regenerative pathways in hepatocytes using lipid nanoparticle-complexed nucleoside-modified mRNA

Fatima Rizvi,Elissa Everton,Hua Liu,Mitchell Beattie,Drew Weissman,Norbert Pardi,Anna R Smith,Valerie Gouon-Evans,Elizabeth Osota,Ying Tam

doi:10.1038/s41467-021-20903-3

Abstract

Induction of intrinsic liver regeneration is an unmet need that can be achieved by temporally activating key hepatocyte regenerative pathways. Here, we establish an efficient, safe, non-integrative method to transiently express hepatocyte-growth-factor (HGF) and epidermal-growth-factor (EGF) in hepatocytes via nucleoside-modified, lipid-nanoparticle-encapsulated mRNA (mRNA-LNP) delivery in mice. We confirm specific hepatotropism of mRNA-LNP via intravenous injection of firefly luciferase encoding mRNA-LNP, with protein expression lasting about 3 days. In the liver, virtually all hepatocytes are transfected along with a subpopulation of endothelial and Kupffer cells. In homeostasis, HGF mRNA-LNP efficiently induce hepatocyte proliferation. In a chronic liver injury mouse model recapitulating non-alcoholic fatty liver disease, injections of both HGF and EGF mRNA-LNP sharply reverse steatosis and accelerate restoration of liver function. Likewise, HGF and EGF mRNA-LNP accelerate liver regeneration after acetaminophen-induced acute liver injury with rapid return to baseline ALT levels. This study introduces mRNA-LNP as a potentially translatable safe therapeutic intervention to harness liver regeneration via controlled expression of endogenous mitogens in vivo.

Highlights

Induction of intrinsic liver regeneration is an unmet need that can be achieved by temporally activating key hepatocyte regenerative pathways
To test whether mRNA-LNP are effective tools for activating hepatocyte mitogen pathways in vivo, here we comprehensively identify the liver as the major target organ of mRNA-LNPproduced proteins following a single injection of firefly luciferase (Luc)-encoding mRNA-LNP, and show that robust protein expression lasts for about 3 days. mRNA-LNP transfect virtually all hepatocytes in the liver along with a subpopulation of endothelial cells and Kupffer cells
As a proof-of-principle of therapeutic intervention, we demonstrate that HGF mRNA-LNP efficiently induce proliferation of hepatocytes in homeostasis, and that the combination of HGF and EGF mRNA-LNP accelerates the restoration of liver function in the chronic Choline-Deficient Ethionine-supplemented (CDE) diet-induced non-alcoholic fatty liver disease (NAFLD) mouse model as well as in the acute acetaminophen-induced liver injury mouse model

Summary

Introduction

Induction of intrinsic liver regeneration is an unmet need that can be achieved by temporally activating key hepatocyte regenerative pathways. We demonstrate the application of transient, yet robust, protein production via mRNA-LNP to promptly activate regenerative pathways in hepatocytes to accelerate intrinsic liver regeneration. To test whether mRNA-LNP are effective tools for activating hepatocyte mitogen pathways in vivo, here we comprehensively identify the liver as the major target organ of mRNA-LNPproduced proteins following a single injection of firefly luciferase (Luc)-encoding mRNA-LNP, and show that robust protein expression lasts for about 3 days. As a proof-of-principle of therapeutic intervention, we demonstrate that HGF mRNA-LNP efficiently induce proliferation of hepatocytes in homeostasis, and that the combination of HGF and EGF mRNA-LNP accelerates the restoration of liver function in the chronic Choline-Deficient Ethionine-supplemented (CDE) diet-induced non-alcoholic fatty liver disease (NAFLD) mouse model as well as in the acute acetaminophen-induced liver injury mouse model

Methods

Results

Conclusion