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A cardiac microphysiological system for screening lipid nanoparticle/mRNA complexes predicts in vivo efficacy for heart transfection

Neiman, G.; Costa, M. W.; Han, H.; Zhao, S.; Ng, T.; Siemons, B.; Nishino, T.; Huang, Y.; Lal, S.; Wu, K.; Judge, L.; Conklin, B.; Srivastava, D.; Murthy, N.; Healy, K. E.

2025-03-12 bioengineering
10.1101/2025.03.07.642075 bioRxiv
Show abstract

Lipid nanoparticle (LNP)/mRNA complexes have tremendous potential for treating cardiac diseases but lack the transfection efficiency required for successful heart therapeutics. A challenge preventing the development of cardiac LNPs is the lack of in vitro screening platforms that maximize transfection and predict efficacy. Here, we demonstrate that a phenotypic cardiac microphysiological system (MPS), constructed from a human induced pluripotent stem cell cardiomyocytes (hiPSC-CM) Cre-reporter line, can identify LNP/mRNA complexes that diffuse efficiently within 3D cardiac micromuscles and transfect cardiomyocytes with high efficiency. Successful LNP formulations contained an acid-degradable PEG (ADP)-lipid, where ADP-LNPs containing 10% PEG had enhanced diffusion and gene editing efficiency in the cardiac MPS. The in vivo delivery of LNP/mRNA complexes, including luciferase and CRE mRNA, into Ai6 mice confirmed the cardiac MPS screening outcomes. It revealed that ADP-LNPs achieved notably superior transfection in the heart with reduced off-target liver uptake compared to a standard LNP formulation. The cardiac MPS showed strong LNP transfection in vitro and pinpointed a promising formulation for in vivo mRNA delivery to the heart.

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