β-catenin mRNA encapsulated in SM-102 lipid nanoparticles enhances bone formation in a murine tibia fracture repair model

Anna Laura Nelson; Chiara Mancino; Xueqin Gao; Joshua A. Choe; Laura Chubb; Katherine Williams; Molly Czachor; Ralph Marcucio; Francesca Taraballi; John P. Cooke; Johnny Huard; Chelsea Bahney; Nicole Ehrhart

Bioactive Materials (Sep 2024)

β-catenin mRNA encapsulated in SM-102 lipid nanoparticles enhances bone formation in a murine tibia fracture repair model

Anna Laura Nelson,
Chiara Mancino,
Xueqin Gao,
Joshua A. Choe,
Laura Chubb,
Katherine Williams,
Molly Czachor,
Ralph Marcucio,
Francesca Taraballi,
John P. Cooke,
Johnny Huard,
Chelsea Bahney,
Nicole Ehrhart

Affiliations

Anna Laura Nelson: Steadman Philippon Research Institute (SPRI), Center for Regenerative and Personalized Medicine, Vail, CO, USA; Colorado State University, School of Biomedical Engineering, Fort Collins CO, USA
Chiara Mancino: Houston Methodist Research Institute, Center for Musculoskeletal Regeneration, Houston TX, USA
Xueqin Gao: Steadman Philippon Research Institute (SPRI), Center for Regenerative and Personalized Medicine, Vail, CO, USA
Joshua A. Choe: University of Wisconsin-Madison, Department of Orthopedics and Rehabilitation, Department of Biomedical Engineering, Medical Scientist Training Program, Madison, WI, USA
Laura Chubb: Colorado State University, Department of Clinical Sciences, Fort Collins CO, USA
Katherine Williams: Colorado State University, Department of Microbiology, Immunology, and Pathology, Fort Collins, CO, USA
Molly Czachor: Steadman Philippon Research Institute (SPRI), Center for Regenerative and Personalized Medicine, Vail, CO, USA
Ralph Marcucio: University of California, San Francisco (UCSF), Orthopaedic Trauma Institute, San Francisco, CA, USA
Francesca Taraballi: Houston Methodist Research Institute, Center for Musculoskeletal Regeneration, Houston TX, USA
John P. Cooke: Houston Methodist Research Institute, Center for RNA Therapeutics, Department of Cardiovascular Sciences, Houston, TX, USA
Johnny Huard: Steadman Philippon Research Institute (SPRI), Center for Regenerative and Personalized Medicine, Vail, CO, USA; Colorado State University, Department of Clinical Sciences, Fort Collins CO, USA
Chelsea Bahney: Steadman Philippon Research Institute (SPRI), Center for Regenerative and Personalized Medicine, Vail, CO, USA; Colorado State University, Department of Clinical Sciences, Fort Collins CO, USA; University of California, San Francisco (UCSF), Orthopaedic Trauma Institute, San Francisco, CA, USA; Corresponding author. Colorado State University, Department of Clinical Sciences, Fort Collins CO, USA.
Nicole Ehrhart: Colorado State University, School of Biomedical Engineering, Fort Collins CO, USA; Colorado State University, Department of Clinical Sciences, Fort Collins CO, USA; Colorado State University, Department of Microbiology, Immunology, and Pathology, Fort Collins, CO, USA; Corresponding author. Colorado State University, Department of Clinical Sciences, Fort Collins CO, USA.

Journal volume & issue: Vol. 39
pp. 273 – 286

Abstract

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Fractures continue to be a global economic burden as there are currently no osteoanabolic drugs approved to accelerate fracture healing. In this study, we aimed to develop an osteoanabolic therapy which activates the Wnt/β-catenin pathway, a molecular driver of endochondral ossification. We hypothesize that using an mRNA-based therapeutic encoding β-catenin could promote cartilage to bone transformation formation by activating the canonical Wnt signaling pathway in chondrocytes. To optimize a delivery platform built on recent advancements in liposomal technologies, two FDA-approved ionizable phospholipids, DLin-MC3-DMA (MC3) and SM-102, were used to fabricate unique ionizable lipid nanoparticle (LNP) formulations and then tested for transfection efficacy both in vitro and in a murine tibia fracture model. Using firefly luciferase mRNA as a reporter gene to track and quantify transfection, SM-102 LNPs showed enhanced transfection efficacy in vitro and prolonged transfection, minimal fracture interference and no localized inflammatory response in vivo over MC3 LNPs. The generated β-cateninGOF mRNA encapsulated in SM-102 LNPs (SM-102-β-cateninGOF mRNA) showed bioactivity in vitro through upregulation of downstream canonical Wnt genes, axin2 and runx2. When testing SM-102-β-cateninGOF mRNA therapeutic in a murine tibia fracture model, histomorphometric analysis showed increased bone and decreased cartilage composition with the 45 μg concentration at 2 weeks post-fracture. μCT testing confirmed that SM-102-β-cateninGOF mRNA promoted bone formation in vivo, revealing significantly more bone volume over total volume in the 45 μg group. Thus, we generated a novel mRNA-based therapeutic encoding a β-catenin mRNA and optimized an SM-102-based LNP to maximize transfection efficacy with a localized delivery.

Published in Bioactive Materials

ISSN: 2452-199X (Online)
Publisher: KeAi Communications Co., Ltd.
Country of publisher: China
LCC subjects: Technology: Electrical engineering. Electronics. Nuclear engineering: Materials of engineering and construction. Mechanics of materials; Science: Biology (General)
Website: https://www.keaipublishing.com/en/journals/bioactive-materials/

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