Compositions and methods for organ specific delivery of nucleic acids
The present disclosure provides compositions which shown preferential targeting or delivery of a nucleic acid composition to a particular organ. In some embodiments, the composition comprises a steroid or sterol, an ionizable cationic lipid, a phospholipid, a PEG lipid, and a permanently cationic lipid which may be used to deliver a nucleic acid.
1. A composition for modifying a target gene or transcript in an organ or a cell therein, the composition comprising a gene or transcript editing composition assembled with a selective organ targeting (SORT) lipid composition, wherein the SORT lipid composition comprises an ionizable cationic lipid and a SORT lipid separate from the ionizable cationic lipid, wherein the SORT lipid is selected from the group consisting of a cationic SORT lipid, a zwitterionic SORT lipid, and an anionic SORT lipid; and
wherein the gene or transcript editing composition comprises:
(i) a polynucleotide-guided nuclease, or a polynucleotide comprising a sequence that encodes the polynucleotide-guided nuclease; and
(ii) a guide polynucleotide configured to complex with at least a portion of the target gene or transcript in the organ or the cell therein, or a polynucleotide comprising a sequence that encodes the guide polynucleotide; and
wherein the gene or transcript editing composition is non-naturally occurring.
2. The composition of claim 1 , wherein (i) and (ii) are not in one molecule.
3. The composition of claim 1 , wherein (i) is the polynucleotide-guided nuclease.
4. The composition of claim 1 , wherein (i) is a polynucleotide comprising a sequence that encodes the polynucleotide-guided nuclease.
5. The composition of claim 4 , wherein (i) is a messenger ribonucleic acid (mRNA) comprising a sequence that encodes the polynucleotide-guided nuclease.
6. The composition of claim 1 , wherein the polynucleotide-guided nuclease of (i) is a clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease.
7. The composition of claim 1 , wherein the guide polynucleotide of (ii) comprises a CRISPR-RNA (crRNA), a trans-activating CRISPR ribonucleic acid (tracrRNA), or a combination thereof.
8. The composition of claim 2 , wherein said gene or transcript editing composition further comprises: (iii) a donor polynucleotide configured to repair a modified target gene or transcript, or a polynucleotide sequence encoding said donor polynucleotide.
9. The composition of claim 1 , wherein said organ is liver.
10. The composition of claim 1 , wherein said organ is a non-liver organ.
11. The composition of claim 10 , wherein said non-liver organ comprises a lung, spleen, a bone marrow, a lymph node, or any combination thereof.
12. The composition of claim 1 , wherein said cationic SORT lipid is a permanently cationic SORT lipid.
13. The composition of claim 12 , wherein said permanently cationic SORT lipid comprises a quaternary ammonium ion.
14. The composition of claim 12 , wherein said permanently cationic SORT lipid has a structure of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof:
wherein, in Formula (I):
R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;
R 3 , R 3 ′, and R 3 ″ are each independently alkyl (C≤6) or substituted alkyl (C≤6) ; and
X − is a monovalent anion; or
wherein, in Formula (II):
R 4 and R 4 ′ are each independently alkyl (C6-C24) , alkenyl (C6-C24) , or a substituted version of either group;
R 4 ″ is alkyl (C≤24) , alkenyl (C≤24) , or a substituted version of either group;
R 4 ′″ is alkyl (C1-C8) , alkenyl (C2-C8) , or a substituted version of either group; and
X 2 − is a monovalent anion; or
wherein, in Formula (III):
R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;
R 3 , R 3 ′, and R 3 ″ are each independently alkyl (C≤6) or substituted alkyl (C≤6) ;
R 4 is alkyl (C≤6) or substituted alkyl (C≤6) ; and
X − is a monovalent anion.
15. The composition of claim 1 , wherein said cationic SORT lipid is an ionizable cationic SORT lipid.
16. The composition of claim 15 , wherein said ionizable cationic SORT lipid is 1,2-dioleoyl-3-dimethylammonium propane (DODAP).
17. The composition of claim 1 , wherein said SORT lipid composition comprises said cationic SORT lipid at a molar percentage from about 5% to about 65%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.
18. The composition of claim 1 , wherein said zwitterionic SORT lipid is a zwitterionic phospholipid.
19. The composition of claim 1 , wherein said SORT lipid composition comprises said zwitterionic SORT lipid at a molar percentage from about 8% to about 23%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.
20. The composition of claim 1 , wherein said anionic SORT lipid comprises a phosphate group.
21. The composition of claim 1 , wherein said anionic SORT lipid has a structure of Formula (IV):
wherein, in Formula (IV):
R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;
R 3 is hydrogen, alkyl (C≤6) , or substituted alkyl (C≤6) , or —Y 1 —R 4 , wherein:
Y 1 is alkanediyl (C≤6) or substituted alkanediyl (C≤6) ; and
R 4 is acyloxy (C≤8-24) or substituted acyloxy (C≤8-24) .
22. The composition of claim 1 , wherein said SORT lipid composition comprises said anionic SORT lipid at a molar percentage from about 5% to about 50%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.
23. The composition of claim 1 , wherein said ionizable cationic lipid is a dendrimer, a dendron, or a pharmaceutically acceptable salt of either of the foregoing.
24. The composition of claim 1 , wherein said SORT lipid composition comprises said ionizable cationic lipid at a molar percentage from about 5% to about 30%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.
25. The composition of claim 1 , wherein said SORT lipid composition comprises a lipid nanoparticle.
26. The composition of claim 1 , wherein said SORT lipid composition comprises a plurality of lipid nanoparticles.
27. The composition of claim 1 , wherein said SORT lipid composition further comprises a phospholipid, a polymer-conjugated lipid, a steroid or steroid derivative, or any combination thereof.
28. The composition of claim 27 , wherein said polymer-conjugated lipid is a polyethylene glycol (PEG) conjugated lipid.
29. The composition of claim 27 , wherein said one or more hydrophobic moieties each comprises 6 to 24 carbon atoms.
30. The composition of claim 27 , wherein said polymer-conjugated lipid comprises a polymer component having an average molecular weight from about 100 Daltons (Da) to about 15,000 Daltons.