Modified Foxp3 mRNA protects against asthma through an IL-10–dependent mechanism

Abstract

Chemically modified mRNA is capable of inducing therapeutic levels of protein expression while circumventing the threat of genomic integration often associated with viral vectors. We utilized this novel therapeutic tool to express the regulatory T cell transcription factor, FOXP3, in a time-and site-specific fashion in murine lung, in order to prevent allergic asthma in vivo. We show that modified Foxp3 mRNA rebalanced pulmonary T helper cell responses and protected from allergen-induced tissue inflammation, airway hyperresponsiveness, and goblet cell metaplasia in 2 asthma models. This protection was conferred following delivery of modified mRNA either before or after the onset of allergen challenge, demonstrating its potential as both a preventive and a therapeutic agent. Mechanistically, FOXP3 induction controlled Th2 and

In vivo genome editing using nuclease-encoding mRNA corrects SP-B deficiency

“…Compliance was determined by using an ex vivo model of the isolated perfused lung as described previously (IPL, Harvard Apparatus) 4,18 . To lower the variability, all mice were treated, and subsequently lungs were isolated within a defined time period.…”

“…These requirements have not yet been achieved simultaneously by any nuclease delivery vector. We and others have used modified mRNA, which is non-integrating and provides a transient pulse of protein expression, as an alternative to traditional viral vectors [1][2][3][4][5] . This approach allowed us to deliver therapeutic proteins in mouse models of Surfactant Protein B (SP-B) deficiency 3 and experimental asthma 4 .…”

“…We and others have used modified mRNA, which is non-integrating and provides a transient pulse of protein expression, as an alternative to traditional viral vectors [1][2][3][4][5] . This approach allowed us to deliver therapeutic proteins in mouse models of Surfactant Protein B (SP-B) deficiency 3 and experimental asthma 4 . Here we apply it to deliver site-specific nucleases, demonstrating the value of nuclease-encoding chemically modified (nec) mRNA as a tool for in vivo genome editing.…”

“…Successful site-specific HDR would allow mice to survive and produce SP-B in the absence of doxycycline. As it is critical to deliver the donor template in excess to ensure it is favored over the homologous chromosome during HDR, we used a vector known to transduce lung cells with high efficiency 4 , AAV6…”

Update in Asthma 2013

“…Jin and colleagues showed that the E3 ubiquitin ligase, Itch, restrains Treg cells from producing Th2 cytokines (49). To define the therapeutic potential of increasing Treg cells in the lung, Mays and colleagues administered to the airways of mice chemically modified FoxP3 mRNA and showed increased Treg cells and reduced allergic airway inflammation, leading to a "rebalancing" of the helper T-cell response (50).…”