Granted Patent B2

US 12,668,814 · App. 17/614,327 · Granted Jun 30, 2026

Inventors: Pardis Sabeti (Cambridge, MA); Mohammadsharif Tabebordbar (Cambridge, MA); Simon Ye (Cambridge, MA); Kim Lagerborg (Cambridge, MA); Alexandra Stanton (Cambridge, MA); Amy Wagers (Cambridge, MA)

Assignees: The Broad Institute, Inc.; President and Fellows of Harvard College; Massachusetts Institute of Technology

C12N15/86A61P21/00A61K48/00C12N2750/14122C12N2750/14142C12N2750/14143C12N2750/14145C12N2750/14171C12N2830/008

Abstract

Claims (48)

1 . A composition comprising:

a modified AAV capsid polypeptide comprising a targeting moiety capable of conferring a muscle tropism to a capsid or a viral particle

wherein the targeting moiety comprises one or more n-mer motifs inserted after amino acids 453, 587, and 588 of a wild-type AAV-9 capsid polypeptide, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype,

wherein each of the one or more n-mer motifs comprises an amino acid sequence of

X m3 X m2 X m1 RGD X 1 X 2 X 3 X 4 ,

wherein X m3 , X m2 , and X m1 are each independently selected from any one of amino acids A, D, E, G, K, N, R, S, L, and T,

wherein X 1 , X 2 , and X 3 are independently selected from any amino acid, and

wherein X 4 is L.

2 . The composition of claim 1 , wherein X 1 is selected from any one of amino acids F, H, L, Q, and Y.

3 . The composition of claim 1 , wherein X 2 is selected from any one of amino acids A, D, E, G, I, K, N, Q, R, S, and T.

4 . The composition of claim 1 , wherein X 3 is selected from any one of amino acids A, D, E, G, K, L, N, R, S, and T.

5 . The composition of claim 1 , further comprising a cargo, wherein the cargo is coupled to or is otherwise associated with the modified AAV capsid polypeptide.

6 . The composition of claim 5 , wherein the cargo is capable of treating or preventing a muscle disease or disorder, wherein the muscle disease or disorder comprises

a. an autoimmune disease;

b. a cancer;

c. a muscular dystrophy;

d. a neuro-muscular disease;

e. a sugar or glycogen storage disease;

f. an expanded repeat disease;

g. a dominant negative disease;

h. a cardiomyopathy;

i. a viral disease;

j. a progeroid disease; or

k. any combination thereof.

7 . The composition of claim 6 , wherein

(a) the expanded repeat disease comprises Huntington's disease, Myotonic Dystrophy, or Facioscapulohumeral muscular dystrophy (FSHD),

(b) wherein the muscular dystrophy comprises Duchenne muscular dystrophy, Becker Muscular dystrophy, a Limb-Girdle muscular dystrophy, an Emery Dreifuss muscular dystrophy, a myotonic dystrophy, or FSHD, optionally wherein the myotonic dystrophy is Type 1 or Type 2,

(c) wherein the cardiomyopathy comprises dilated cardiomyopathy, hypertrophic cardiomyopathy, DMD-associated cardiomyopathy, or Dannon disease,

(d) wherein the sugar or glycogen storage disease comprises a MPS type III disease or Pompe disease, optionally wherein the MPS type III disease comprises MPS Type IIIA, IIIB, IIIC, or HID,

(e) wherein the neuro-muscular disease comprises Charcot-Marie-Tooth disease or Friedreich's Ataxia, or

(f) any combination of (a)-(e).

8 . The composition of claim 5 , wherein the cargo is a morpholino, a peptide-linked morpholino, an antisense oligonucleotide, a PMO, a therapeutic transgene, a polynucleotide encoding a therapeutic polypeptide or peptide, a PPMO, one or more peptides, one or more polynucleotides encoding a CRISPR-Cas protein, a guide RNA, or both, a ribonucleoprotein, wherein the ribonucleoprotein comprises a CRISPR-Cas system molecule, a therapeutic transgene RNA, or other gene modifying or therapeutic RNA and/or protein, or any combination thereof.

9 . The composition of claim 5 , wherein the cargo is (a) capable of inducing exon skipping in a gene, optionally a dystrophin gene, or (b) a mini- or micro-dystrophin gene, optionally wherein the mini- or micro-dystrophin gene comprises spectrin-like repeats 1, 2, 3, and 24, and optionally an nNOS domain.

10 . The composition of claim 1 , wherein the wild-type AAV capsid polypeptide of a different serotype is selected from the group consisting of a wild-type AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV rh.74, and AAV rh.10 VP1 capsid polypeptide.

11 . The composition of claim 1 , wherein the modified AAV capsid polypeptide comprises one or more mutations at positions 267, 269, 504, 505, or 590, or any combination thereof, of a wild-type AAV9 VP1 capsid polypeptide, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, and wherein the modified AAV capsid polypeptide is capable of conferring on a capsid or a viral particle, a reduced tropism for non-muscle cells, as compared to a corresponding wild-type AAV capsid polypeptide.

12 . The composition of claim 11 , wherein the one or more mutations of the wild-type AAV9 VP1 capsid polypeptide

in position 267, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a G or X to A mutation, wherein X is any amino acid, or

in position 269, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is an S or X to T mutation, wherein X is any amino acid, or

in position 504, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a G or X to A mutation, wherein X is any amino acid, or

in position 505, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a P or X to A mutation, wherein X is any amino acid, or

in position 590, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a Q or X to A mutation, wherein X is any amino acid, or

in position 267, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a G to A mutation, or

in position 269, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is an S to T mutation, or

in position 509, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a Q to A mutation, or

in position 504, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a G to A mutation, or

in position 505, or an equivalent position in a wild-type AAV capsid polypeptide of a different serotype, is a P to A mutation, or

any combination of the mutations in positions 267, 269, 504, 505, or 590, or equivalent positions in a wild-type AAV capsid polypeptide of a different serotype.

13 . The composition of claim 11 , wherein the non-muscle cell comprises a liver cell.

Assignments (8)

Continuity (4)

References Cited (243)