Efficient protein expression in vivo using modified RNA (MOD-RNA)
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, cardiomyoctes, and tumors.
1. A method for increasing capillary density in cardiac tissue in a subject by expressing a VEGF-A protein in a cardiac tissue in vivo, the method comprising contacting the cardiac tissue in vivo with a composition comprising a synthetic, modified RNA molecule encoding a VEGF-A polypeptide,
wherein the synthetic, modified RNA molecule comprises one or more modifications selected from the group consisting of 5-methylcytidine (5mC), N6-methyladenosine (m6A), 3,2′-O-dimethyluridine (m4U), 2-thiouridine (s2U), 2′ fluorouridine, pseudouridine, 2′-O-methyluridine (Um), 2′deoxy uridine (2′ dU), 4-thiouridine (s4U), 5-methyluridine (m5U), 2′-O-methyladenosine (m6A), N6,2′-O-dimethyladenosine (m6Am), N6,N6,2′-O-trimethyladenosine (m62Am), 2′-O-methylcytidine (Cm), 7-methylguanosine (m7G), 2′-O-methylguanosine (Gm), N2,7-dimethylguanosine (m2,7G), N2, N2, 7-trimethylguanosine (m2,2,7G), and inosine (I),
such that introducing said synthetic, modified RNA molecule to a cell in the cardiac tissue in vivo results in increased capillary density in the cardiac tissue, and also results in a reduced innate immune response relative to a cell in the cardiac tissue in vivo contacted with a synthetic RNA molecule encoding the polypeptide not comprising said one or more modifications.
2. The method of claim 1 , wherein the cardiac tissue is in a subject having a disease or disorder.
3. The method of claim 2 , wherein the disease or disorder is selected from the group consisting of: acute coronary syndrome, congestive heart failure, cardiomyopathy, myocardial infarction, tissue ischemia, cardiac ischemia, vascular disease, acquired heart disease, atherosclerosis, dysfunctional conduction systems, pulmonary heart hypertension, structural heart disease, or disease or disorder characterized by insufficient cardiac function, coronary artery disease, myocardial ischemia, atherosclerosis, idiopathic cardiomyopathy, cardiac arrhythmias, muscular dystrophy, muscle mass abnormality, muscle degeneration, infective myocarditis, drug- or toxin-induced muscle abnormalities, hypersensitivity myocarditis, an autoimmune endocarditis and congenital heart disease or a symptom of hypertension; blood flow disorders; symptomatic arrhythmia; pulmonary hypertension; dysfunction in conduction system; dysfunction in coronary arteries; dysfunction in coronary arterial tree and coronary artery colaterization.
4. The method of claim 1 , wherein the cardiac tissue has a reduced capillary density and is in a subject with, or at risk of having heart failure.
5. The method of claim 1 , wherein the VEGF-A protein is a VEGF-A 165 protein.