Subcutaneous delivery of messenger RNA
The present invention provides, among other things, methods of formulating nucleic acid-containing nanoparticles with an enzyme to afford efficient delivery of payload to a cell or tissue of interest via subcutaneous administration. In some embodiments, the present invention provides a process in which mRNA-loaded lipid nanoparticles are co-mixed with various amounts of hyaluronidase and administered via subcutaneous administration. The resulting payload can be efficiently delivered to the liver and other organs or tissues of a treated subject.
1. A method of delivering an mRNA to a cell in vivo comprising administering via subcutaneous or intramuscular injection to a subject in need of treatment
a) an mRNA encoding a protein, at a concentration of greater than 1 mg/kg, wherein the mRNA is encapsulated in a lipid-based nanoparticle (LNP) and
b) a hyaluronidase enzyme, at a concentration of at least 200 U.
2. The method of claim 1 , wherein the subcutaneous or intramuscular injection results in delivery of the mRNA in the liver.
3. The method of claim 1 , wherein the subcutaneous or intramuscular injection results in expression of the protein in the liver of the subject.
4. The method of claim 1 , wherein the subcutaneous or intramuscular injection results in expression of the protein in the serum of the subject.
5. The method of claim 1 , wherein the lipid-based nanoparticle is a liposome.
6. The method of claim 5 , wherein the liposome comprises a PEGylated lipid.
7. The method of claim 6 , wherein the PEGylated lipid constitutes at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10% of the total lipids in the liposome.
8. The method of claim 1 , wherein the hyaluronidase enzyme is at a concentration of at least 220 U.
9. The method of claim 1 , wherein the hyaluronidase enzyme is at a concentration of at least 260 U.
10. A method of treating a disease, disorder or condition comprising delivering messenger RNA (mRNA) to a subject in need of treatment according to a method of claim 1 , wherein the mRNA encodes a protein deficient in the subject.
11. A composition for treating ornithine transcarbamylase (OTC deficiency), comprising
a) an mRNA encoding an ornithine transcarbamylase (OTC) protein, at a concentration of greater than 1 mg/kg, wherein the mRNA is encapsulated in a lipid-based nanoparticle (LNP) and
b) a hyaluronidase enzyme, at a concentration of at least 200 U.
12. The composition of claim 11 , wherein the mRNA and/or the hyaluronidase enzyme is encapsulated within a nanoparticle.
13. The composition of claim 11 , wherein the mRNA and the hyaluronidase enzyme are encapsulated within the same nanoparticle.
14. The composition of claim 11 , wherein the mRNA and the hyaluronidase enzyme are encapsulated within separate nanoparticles.
15. The composition of claim 11 , wherein the separate nanoparticles encapsulating the mRNA and the hyaluronidase enzyme comprise the same formulation.
16. The composition of claim 11 , wherein the separate nanoparticles encapsulating the mRNA and the hyaluronidase enzyme comprise different formulations.
17. The composition of claim 11 , wherein the mRNA is encapsulated within the nanoparticle and the hyaluronidase enzyme is not encapsulated.