OR01
mRNA-based therapies for liver diseases: HNF4A mRNA administration via lipid nanoparticles mitigates liver fibrosis in preclinical models
R Khanal¹ T Yang¹ M Poenisch² Q Hu¹ Z Dai¹ R Li¹ G Song³ Q Yuan¹ Q Yao³ X Shen³ R Taubert¹ B Engel¹ E Jaeckel¹ A Vogel¹ C Falk¹ A Schambach¹ D Gerovska⁴ M J. Araúzo-Bravo⁴ F W.R. Vondran¹ T Cantz¹ N Horscroft² A Balakrishnan¹ F Chevessier² M Ott¹ A Sharma¹
1:Hannover Medical School; 2:CureVac AG; 3:Department of Gastroenterology, Zhongshan Hospital of Fudan University, Shanghai, China; 4:Group of Computational Biology and Systems Biomedicine, Biodonostia Health Research Institute, San Sebastián, Spain
Messenger RNA (mRNA)-based therapeutics are speedily approaching the clinic and hold immense capacity to benefit millions of individuals worldwide. Nevertheless, therapeutic targeting of injuries that demand transient restoration of proteins by mRNA delivery requires more in-depth investigation. Here, we examined therapeutic utility of mRNA delivery in liver fibrosis and cirrhosis, which cause millions of deaths every year. Particularly, we envisioned demonstrating the therapeutic applicability of the human transcription factor, hepatocyte nuclear factor alpha (HNF4A) encoding mRNA, in chronically injured mouse models of liver fibrosis and cirrhosis.
We demonstrated restoration of hepatocyte functions by HNF4A mRNA transfection in vitro, and assessed the attenuation of liver fibrosis and cirrhosis in multiple murine models, by injecting hepatocyte-targeted lipid nanoparticles (LNP) encapsulating HNF4A mRNA. The potential mechanisms of HNF4A mRNA-mediated mitigation of liver fibrosis were unravelled by performing microarray-based gene expression profiling, single cell RNA sequencing, and chromatin immunoprecipitation. Primary liver cells and human liver buds were used for further functional validation.
HNF4A mRNA expression restored metabolic functions of fibrotic primary human hepatocytes in vitro. Repeated in vivo administration of LNP-encapsulated HNF4A mRNA triggered robust suppression of fibrogenesis in four independent murine models of hepatotoxin- and cholestasis-induced liver fibrosis. We identified that paraoxonase 1 is a direct target of HNF4A, which contributes to HNF4A-directed inhibition of liver fibrosis by regulating hepatic stellate cells and liver macrophages.
Our findings provide the first direct pre-clinical evidence that HNF4A mRNA delivery via LNP could be a promising therapeutic option for the treatment of liver fibrosis.