CNS delivery of MRNA and uses thereof
The present invention provides, among other things, methods and compositions for effective delivery of messenger RNA (mRNA) to the central nervous system (CNS). In particular, the present invention provides methods and compositions for administering intrathecally to a subject in need of delivery a composition comprising an mRNA encoding a protein, encapsulated within a liposome, such that the administering of the composition results in the intracellular delivery of mRNA in neurons in the brain and/or spinal cord. The present invention is particularly useful for the treatment of CNS diseases, disorders or conditions, such as spinal muscular atrophy.
1. A method of delivery of messenger RNA (mRNA) to the central nervous system (CNS), comprising
administering intrathecally to a subject in need of delivery a composition comprising an mRNA encoding a protein, encapsulated within a liposome such that the administering of the composition results in the intracellular delivery of mRNA in neurons in the brain and/or spinal cord;
wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more polyethylene glycol (PEG)-modified lipids,
wherein the liposome further comprises sphingomyelin,
wherein the mRNA is codon-optimized and comprises the full length sequence of SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 10 or SEQ ID NO. 11 , and
wherein administering of the composition results in expression of the protein or peptide encoded by the mRNA that is detectable in brain and/or spinal tissue at least 24 hours after administration.
2. The method of claim 1 , wherein the one or more cationic lipids are selected from the group consisting of C12-200, MC3, DLinDMA, DLinkC2DMA, cKK-E12, ICE (Imidazol-based), HGT5000, HGT5001, DODAC, DDAB, DMRIE, DOSPA, DOGS, DODAP, DODMA and DMDMA, DODAC, DLenDMA, DMRIE, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DLinDAP, DLincarbDAP, DLinCDAP, KLin-K-DMA, DLin-K-XTC2- DMA, HGT4003, and combination thereof.
3. The method of claim 2 , wherein the cationic lipid is cKK-E12:
4. The method of claim 1 , wherein the one or more PEG-modified lipids constitute about 1-10% by molar ratio of the total lipid composition.
5. The method of claim 1 , wherein the liposome has a size ranging from about 40-100 nm.
6. The method of claim 1 , wherein the mRNA has a length of or greater than about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, or 5 kb.
7. The method of claim 1 , wherein the protein encoded by the mRNA normally functions in the neurons in the brain and/or spinal cord.
8. The method of claim 1 , wherein the protein encoded by the mRNA is the Survival of Motor Neuron (SMN) protein.
9. The method of claim 1 , wherein the protein encoded by the mRNA is an enzyme.
10. The method of claim 1 , wherein the intracellular delivery of mRNA results in intracellular expression of the protein encoded by the mRNA within the cytosol of the neurons.
11. The method of claim 1 , wherein the intracellular delivery of mRNA results in expression of the protein encoded by the mRNA and secretion extracellularly from the neurons after expression.
12. The method of claim 1 , wherein the mRNA comprises one or more modified nucleotides.
13. The method of claim 1 , wherein the mRNA is unmodified.
14. The method of claim 1 , wherein the mRNA is delivered at an amount ranging from about 0.01 mg/kg to about 10 mg/kg body weight.