Efficient protein expression in vivo using modified RNA (mod-RNA)
Inventors
Chien, Kenneth R. • Ptaszek, Leon M. • Lui, Kathy Oi-Lan • Zangi, Lior • Ebina, Wataru • Rossi, Derrick J.
Assignees
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Publication Number
US-12357675-B2
Publication Date
2025-07-15
Expiration Date
Abstract
Aspects of the invention described herein relate to synthetic, modified RNAs and their use in vivo to modulate gene expression. Aspects of the invention further relate to the use of these synthetic, modified RNAs in myocytes, cardiomyocytes, and tumors.
Core Innovation
The invention concerns synthetic, modified RNA that is in vitro transcribable and translatable in vivo, and that significantly reduces innate/interferon immune activation relative to unmodified RNA while maintaining translatability in vivo. The disclosed approach uses nucleoside base modifications and other RNA features to reduce innate immune signaling, including reduced response metrics associated with innate immune pathways such as IFNα and IFNβ and reduced signaling associated with RIG-1.
The described modified RNA is used to express a VEGF-A protein in cardiac tissue in vivo, including VEGF-A165 in a subject with myocardial infarction. The cardiac tissue is contacted in vivo by myocardial injection with a composition comprising a synthetic, modified RNA molecule encoding VEGF-A165, and introducing the modified RNA into cardiac tissue results in increasing cardiovascular fate cells and reducing fibrosis in the cardiac tissue.
The synthetic, modified RNA encoding VEGF-A165 promotes differentiation of WT-1+ epicardial progenitor cells into endothelial cells, vascular smooth muscle cells, or cardiomyocytes. The disclosure also links reduced innate immune response with improved regenerative and tissue outcomes in vivo, and describes additional contexts including local VEGF-A modified RNA strategies for neovascularization and chemotherapy enhancement in pancreatic tumor contexts.
Claims Coverage
The independent claim is clm-00001. It covers a myocardial-injection method in a myocardial-infarction subject using a synthetic, modified RNA molecule encoding VEGF-A165, with specified nucleoside base modifications, and it requires reduced innate immune response relative to unmodified synthetic RNA, increasing cardiovascular fate cells, reducing fibrosis, and differentiation of WT-1+ epicardial progenitor cells into endothelial cells, vascular smooth muscle cells, or cardiomyocytes.
Myocardial injection of VEGF-A165 MOD-RNA in myocardial-infarction subject
A method for increasing cardiovascular fate cells in cardiac tissue in a subject with myocardial infarction by expressing a VEGF-A protein in cardiac tissue in vivo, comprising contacting the cardiac tissue in vivo by myocardial injection with a composition comprising a synthetic, modified RNA molecule encoding a VEGF-A165 polypeptide.
Nucleoside base modifications to reduce innate immune response
The synthetic, modified RNA molecule comprises at least one nucleoside base modification selected from pseudouracil, (thio)pseudouracil variants, dithio/pseudouracil variants, alkyl-substituted pseudouracils, aminoalkylaminocarbonylethylenyl pseudouracil variants, and related substituted pseudouracil modifications, such that introducing the modified RNA molecule to a cell in the cardiac tissue in vivo results in a reduced innate immune response relative to a synthetic RNA molecule encoding the polypeptide not comprising the modification.
Increased cardiovascular fate, reduced fibrosis, and WT-1+ epicardial progenitor differentiation
Introducing the synthetic, modified RNA molecule encoding a VEGF-A165 polypeptide results in increasing cardiovascular fate cells and reduces fibrosis in the cardiac tissue, and the modified RNA molecule promotes WT-1+ epicardial progenitor cells to differentiate into endothelial cells, vascular smooth muscle cells, or cardiomyocytes.
Claim coverage centers on an in vivo myocardial-injection method using a synthetic, modified RNA encoding VEGF-A165, where nucleoside base modifications are used to achieve reduced innate immune response and improved cardiac outcomes, including increased cardiovascular fate cells, reduced fibrosis, and differentiation of WT-1+ epicardial progenitor cells into endothelial cells, vascular smooth muscle cells, or cardiomyocytes.
Stated Advantages
Reduces fibrosis in the cardiac tissue.
Reduces innate immune response relative to synthetic RNA without the specified nucleoside base modifications.
Increases cardiovascular fate cells in cardiac tissue.
Promotes WT-1+ epicardial progenitor cells to differentiate into endothelial cells, vascular smooth muscle cells, or cardiomyocytes.
Documented Applications
Treating cardiovascular disease associated with myocardial infarction by increasing cardiovascular fate cells and reducing fibrosis in cardiac tissue via in vivo expression of VEGF-A165 from synthetic, modified RNA after myocardial injection.
Increasing neovascularization in tumors by enhancing angiogenesis using VEGF-based MOD-RNA approaches.
Treating tumors by inhibiting Hedgehog signaling/processing of desmoplastic tissue to improve chemotherapeutic efficacy.
Treating muscular dystrophy and other loss-of-function diseases.
Conducting research applications involving rapid, efficient in vivo protein expression using synthetic, modified RNA with reduced innate immune response.
Using VEGF-A modified RNA for neovascularization and chemotherapy enhancement in pancreatic tumor contexts.
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