In utero delivery of mRNA to the heart, diaphragm and muscle with lipid nanoparticles

Kewa Gao; Jie Li; Hengyue Song; Hesong Han; Yongheng Wang; Boyan Yin; Diana L Farmer; Niren Murthy; Aijun Wang

doi:10.1016/j.bioactmat.2023.02.011

In utero delivery of mRNA to the heart, diaphragm and muscle with lipid nanoparticles

Bioact Mater. 2023 Feb 17:25:387-398. doi: 10.1016/j.bioactmat.2023.02.011. eCollection 2023 Jul.

Authors

Kewa Gao^{1

2}, Jie Li³, Hengyue Song^{1

2

4}, Hesong Han³, Yongheng Wang^{1

2

5}, Boyan Yin^{1

2}, Diana L Farmer^{1

2}, Niren Murthy³, Aijun Wang^{1

2

5}

Affiliations

¹ Department of Surgery, School of Medicine, University of California, Davis, Sacramento, CA, 95817, United States.
² Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, CA, 95817, United States.
³ Department of Bioengineering, University of California, Berkeley, CA, 94704, United States.
⁴ Department of Burns and Plastic Surgery, The Third Xiangya Hospital of Central South University, Hunan, 410013, China.
⁵ Department of Biomedical Engineering, University of California, Davis, CA, 95616, United States.

Abstract

Nanoparticle-based drug delivery systems have the potential to revolutionize medicine, but their low vascular permeability and rapid clearance by phagocytic cells have limited their medical impact. Nanoparticles delivered at the in utero stage can overcome these key limitations due to the high rate of angiogenesis and cell division in fetal tissue and the under-developed immune system. However, very little is known about nanoparticle drug delivery at the fetal stage of development. In this report, using Ai9 CRE reporter mice, we demonstrate that lipid nanoparticle (LNP) mRNA complexes can deliver mRNA in utero, and can access and transfect major organs, such as the heart, the liver, kidneys, lungs and the gastrointestinal tract with remarkable efficiency and low toxicity. In addition, at 4 weeks after birth, we demonstrate that 50.99 ± 5.05%, 36.62 ± 3.42% and 23.7 ± 3.21% of myofiber in the diaphragm, heart and skeletal muscle, respectively, were transfected. Finally, we show here that Cas9 mRNA and sgRNA complexed to LNPs were able to edit the fetal organs in utero. These experiments demonstrate the possibility of non-viral delivery of mRNA to organs outside of the liver in utero, which provides a promising strategy for treating a wide variety of devastating diseases before birth.

Keywords: CRISPR/Cas9; Gene editing; In utero; Nanoparticles; mRNA delivery.

Abstract

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