Dual mRNA therapy restores metabolic function in long-term studies in mice with propionic acidemia

Lei Jiang,Steven Fortucci,Jinsong Yang,Cosmin Mihai,Kerry E Benenato,Ji-Sun Park,Shi Liang,Rodrigo Laureano,Jenny Zhuo,Xinwen Zhu ,Christopher Tunkey,Kara M Hoar ,Ling Yin,Vladimir Presnyak ,Eric Jacquinet,Andrea Frassetto,Christine Lukacs ,Xinhua Yan,Paolo G.v Martini ,Lin T Guey

doi:10.1038/s41467-020-19156-3

Lei Jiang, Steven Fortucci + Show 18 more

Open Access

https://doi.org/10.1038/s41467-020-19156-3

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Journal: Nature Communications	Publication Date: Oct 21, 2020
Citations: 83	License type: open-access

Affiliation: Moderna Therapeutics (United States)

Abstract

Propionic acidemia/aciduria (PA) is an ultra-rare, life-threatening, inherited metabolic disorder caused by deficiency of the mitochondrial enzyme, propionyl-CoA carboxylase (PCC) composed of six alpha (PCCA) and six beta (PCCB) subunits. We herein report an enzyme replacement approach to treat PA using a combination of two messenger RNAs (mRNAs) (dual mRNAs) encoding both human PCCA (hPCCA) and PCCB (hPCCB) encapsulated in biodegradable lipid nanoparticles (LNPs) to produce functional PCC enzyme in liver. In patient fibroblasts, dual mRNAs encoded proteins localize in mitochondria and produce higher PCC enzyme activity vs. single (PCCA or PCCB) mRNA alone. In a hypomorphic murine model of PA, dual mRNAs normalize ammonia similarly to carglumic acid, a drug approved in Europe for the treatment of hyperammonemia due to PA. Dual mRNAs additionally restore functional PCC enzyme in liver and thus reduce primary disease-associated toxins in a dose-dependent manner in long-term 3- and 6-month repeat-dose studies in PA mice. Dual mRNAs are well-tolerated in these studies with no adverse findings. These studies demonstrate the potential of mRNA technology to chronically administer multiple mRNAs to produce large complex enzymes, with applicability to other genetic disorders.

Highlights

Propionic acidemia/aciduria (PA) is an ultra-rare, life-threatening, inherited metabolic disorder caused by deficiency of the mitochondrial enzyme, propionyl-CoA carboxylase (PCC) composed of six alpha (PCCA) and six beta (PCCB) subunits
Transfection of dual messenger RNAs (mRNAs) in patient fibroblasts obtained from both PA subtypes resulted in dose-dependent increases in PCC enzyme activity and protein subunits (Fig. 1a–c and Supplementary Fig. 1a–c)
PCC enzyme activity was 5–24× greater in fibroblasts transfected with dual mRNAs vs. human PCCA (hPCCA) or hPCCB mRNA alone (Fig. 1a), suggesting that both mRNAencoded PCC protein subunits contribute to increasing PCC activity

Summary

Introduction

Propionic acidemia/aciduria (PA) is an ultra-rare, life-threatening, inherited metabolic disorder caused by deficiency of the mitochondrial enzyme, propionyl-CoA carboxylase (PCC) composed of six alpha (PCCA) and six beta (PCCB) subunits. We report an enzyme replacement approach to treat PA using a combination of two messenger RNAs (mRNAs) (dual mRNAs) encoding both human PCCA (hPCCA) and PCCB (hPCCB) encapsulated in biodegradable lipid nanoparticles (LNPs) to produce functional PCC enzyme in liver. Dual mRNAs are well-tolerated in these studies with no adverse findings These studies demonstrate the potential of mRNA technology to chronically administer multiple mRNAs to produce large complex enzymes, with applicability to other genetic disorders. Preclinical experiments that include 3- and 6-month studies in a relevant murine disease model demonstrate the long-term therapeutic potential of a combination mRNA therapy approach for a rare metabolic disorder

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