Matrix stiffness regulates lipid nanoparticle-mRNA delivery in cell-laden hydrogels

Avathamsa Athirasala; Siddharth Patel; Paula P Menezes; Jeonghwan Kim; Anthony Tahayeri; Gaurav Sahay; Luiz E Bertassoni

doi:10.1016/j.nano.2022.102550

Matrix stiffness regulates lipid nanoparticle-mRNA delivery in cell-laden hydrogels

Nanomedicine. 2022 Jun:42:102550. doi: 10.1016/j.nano.2022.102550. Epub 2022 Mar 12.

Authors

Avathamsa Athirasala¹, Siddharth Patel², Paula P Menezes³, Jeonghwan Kim², Anthony Tahayeri⁴, Gaurav Sahay⁵, Luiz E Bertassoni⁶

Affiliations

¹ Department of Biomedical Engineering, Collaborative Life Sciences Building, Oregon Health and Science University, Portland, OR, USA.
² Department of Pharmaceutical Sciences, College of Pharmacy, Collaborative Life Science Building, Oregon State University, Portland, OR, USA.
³ Division of Biomaterials and Biomechanics, Department of Restorative Dentistry, School of Dentistry, Oregon Health and Science University, Portland, OR, USA; Department of Pharmacy, Federal University of Sergipe, Aracaju, Sergipe, Brazil.
⁴ Division of Biomaterials and Biomechanics, Department of Restorative Dentistry, School of Dentistry, Oregon Health and Science University, Portland, OR, USA.
⁵ Department of Biomedical Engineering, Collaborative Life Sciences Building, Oregon Health and Science University, Portland, OR, USA; Department of Pharmaceutical Sciences, College of Pharmacy, Collaborative Life Science Building, Oregon State University, Portland, OR, USA; Department of Ophthalmology, Casey Eye Institute, Oregon Health & Science University, Portland, OR, USA.
⁶ Department of Biomedical Engineering, Collaborative Life Sciences Building, Oregon Health and Science University, Portland, OR, USA; Division of Biomaterials and Biomechanics, Department of Restorative Dentistry, School of Dentistry, Oregon Health and Science University, Portland, OR, USA; Center for Regenerative Medicine, Oregon Health and Science University, Portland, OR, USA; Cancer Early Detection Advanced Research (CEDAR) Center, Knight Cancer Institute, Oregon Health and Science University, Portland, OR, USA. Electronic address: bertasso@ohsu.edu.

Abstract

mRNA therapeutics have increased in popularity, largely due to the transient and fast nature of protein expression and the low risk of off-target effects. This has increased drastically with the remarkable success of mRNA-based vaccines for COVID-19. Despite advances in lipid nanoparticle (LNP)-based delivery, the mechanisms that regulate efficient endocytic trafficking and translation of mRNA remain poorly understood. Although it is widely acknowledged that the extracellular matrix (ECM) regulates uptake and expression of exogenous nano-complexed genetic material, its specific effects on mRNA delivery and expression have not yet been examined. Here, we demonstrate a critical role for matrix stiffness in modulating both mRNA transfection and expression and uncover distinct mechano-regulatory mechanisms for endocytosis of mRNA through RhoA mediated mTOR signaling and cytoskeletal dynamics. Our findings have implications for effective delivery of therapeutic mRNA to targeted tissues that may be differentially affected by tissue and matrix stiffness.

Keywords: 3D hydrogels; LNPs; Mechanotransduction; RhoA; mRNA delivery; mTOR.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

COVID-19 Vaccines
COVID-19* / therapy
Humans
Hydrogels
Lipids / genetics
Liposomes
Nanoparticles*
RNA, Messenger / genetics

Substances

COVID-19 Vaccines
Hydrogels
Lipid Nanoparticles
Lipids
Liposomes
RNA, Messenger

Abstract

Publication types

MeSH terms

Substances

Grants and funding