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

作　　者：Anna Laura Nelson Chiara Mancino Xueqin Gao Joshua AChoe Laura Chubb Katherine Williams Molly Czachor Ralph Marcucio Francesca Taraballi John P.Cooke Johnny Huard Chelsea Bahney Nicole Ehrhart 

机构地区：[1]Steadman Philippon Research Institute(SPRI),Center for Regenerative and Personalized Medicine,Vail,CO,USA [2]Houston Methodist Research Institute,Center for Musculoskeletal Regeneration,Houston TX,USA [3]University of Wisconsin-Madison,Department of Orthopedics and Rehabilitation,Department of Biomedical Engineering,Medical Scientist Training Program,Madison,WI,USA [4]Colorado State University,School of Biomedical Engineering,Fort Collins CO,USA [5]Colorado State University,Department of Clinical Sciences,Fort Collins CO,USA [6]Colorado State University,Department of Microbiology,Immunology,and Pathology,Fort Collins,CO,USA [7]Houston Methodist Research Institute,Center for RNA Therapeutics,Department of Cardiovascular Sciences,Houston,TX,USA [8]University of Californi1,San Francisco(UCSF),Orthopaedic Trauma Institute,San Francisco,CA,USA

出　　处：《Bioactive Materials》2024年第9期273-286,共14页生物活性材料（英文）

基　　金：supported by National Institute of Arthritis and Musculoskeletal and Skin Diseases(NIAMS)of the National Institutes of Health(NIH)under award number R01 AR077761;support from the Musculoskeletal Regeneration Partnership Fund by Mary Sue and Michael Shannon and by Project Number 20-166 from the Orthoregeneration Network for Kick-Starter Grant;supported by the National Institute on Aging of the National Institutes of Health under Award Number F30AG077748 and the University of Wisconsin-Madison Medical Scientist Training Program:T32GM140935.All content is solely the responsibility of the authors and does not represent the official views of National Institutes of Health,Orthoregeneration Network or Shannon Foundation.

摘　　要：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 mRNAbased therapeutic encodingβ-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 generatedβ-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 45μg 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 45μg group.Thus,we generated a novel mRNA-based therapeutic encoding aβ-catenin mRNA and optimized an SM-102-based LNP to maximize transfection efficacy with a localized delivery.

关 键 词：Gene therapy Fracture healing Lipid nanoparticles MRNA Canonical Wnt 

分 类 号：R68[医药卫生—骨科学] TB383[医药卫生—外科学]