Engineered muscle targeting compositions

Patent metadata

Field	Value
Publication number	US-2025043309-A1
Application number	US-202418915615-A
Country	US
Kind code	A1
Filing date	Oct 15, 2024
Priority date	Jul 22, 2020
Publication date	Feb 6, 2025
Grant date	—

Abstract

Described herein are muscle-specific targeting moieties and compositions including the muscle specific targeting motifs. Also described herein are uses of the muscle-specific targeting motifs and compositions including the muscle specific targeting moieties. In some embodiments, the muscle-specific targeting moieties and compositions including the muscle specific targeting moieties can be used to direct delivery of a cargo to a muscle cell.

First claim

What is claimed is:

1 . A composition comprising a targeting moiety effective to target a muscle cell, wherein the targeting moiety comprises an n-mer motif comprising X m RGDX 1 X 2 X 3 X 4 wherein X is selected from any amino acid, and wherein m is 0-4, and wherein X 4 is L or T.

2 . The composition of claim 1 , wherein X 4 is L and wherein
(a) X 1 is Q, H, or Y; (b) X 2 is T, G, A, or S; (c) X 3 is T, S, N, R, A; (d) (a) and (b); (e) (a) and (c); (f) (b) and (c); or (g) (a), (b), and (c).

3 . The composition of claim 1 , wherein X 4 is L and wherein X 1 is Q, wherein X 2 is T or G, and wherein X 3 is T or R; or
wherein X 1 is Q, wherein X 2 is T, and wherein X 3 is T; or wherein X 1 is Q, wherein X 2 is G, and wherein X 3 is R.

4 . The composition of claim 1 , wherein X 4 is L and wherein X 1 is H, wherein X 2 is G, A, or S, and wherein X 3 is T, S, or A; or
wherein X 1 is H, wherein X 2 is G, and wherein X 3 is T or A; or wherein X 1 is H, wherein X 2 is G, and wherein X 3 is T; or wherein X 1 is H, wherein X 2 is G, and wherein X 3 is A.

5 . The composition of claim 1 , wherein X 4 is L and wherein X 1 is H, wherein X 2 is A, and wherein X 3 is S; or
wherein X 1 is H, wherein X 2 is S, and wherein X 3 is S; or wherein X 1 is Y, wherein X 2 is A, and wherein X 3 is N

6 . The composition of claim 1 , wherein X 4 is L and wherein the first three amino acids of the n-mer motif are GPG, TSV, AMS, GAV, TPS, DSR, NAS, or SPV.

7 . The composition of claim 1 , wherein the n-mer motif comprises ENRRGDFNNT (SEQ ID NO: 32).

8 . The composition of claim 1 , wherein the targeting moiety is an engineered adeno associated virus (AAV) capsid polypeptide.

9 . The composition of claim 8 , wherein the n-mer motif is inserted into a surface variable region of the engineered AAV capsid polypeptide.

10 . The composition of claim 9 , wherein the surface variable region is selected from VR-IV, VR-V, or VR-VIII.

11 . The composition of claim 8 , wherein the n-mer motif is inserted between any two amino acids between amino acids 262-269, 327-332, 382-386, 452-460, 488-505, 527-539, 545-558, 581-593, 704-714, or any combination thereof, in an AAV9 capsid polypeptide or in an analogous position in an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh.74, or AAV rh.10 capsid polypeptide; or is inserted between amino acids 588 and 589, in an AAV9 capsid polypeptide or in an analogous position in an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh.74, or AAV rh.10 capsid polypeptide.

12 . The composition of claim 1 , further comprising a cargo.

13 . The composition of claim 12 , wherein the cargo is
(a) capable of treating or preventing a muscle disease or disorder; (b) 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 or polypeptides; 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; or (c) both (a) and (b).

14 . The composition of claim 13 , wherein the muscle disease or disorder is an auto immune disease; a cancer; a muscular dystrophy; a neuro-muscular disease; a sugar or glycogen storage disease; an expanded repeat disease; a dominant negative disease; a cardiomyopathy; a viral disease; a progeroid disease; or any combination thereof.

15 . An engineered particle comprising composition of claim 1 .

16 . The engineered particle of claim 15 , wherein the wherein the targeting moiety is an engineered adeno associated virus (AAV) capsid polypeptide.

17 . The engineered particle of claim 16 , wherein the n-mer motif is inserted into a surface variable region of the engineered AAV capsid polypeptide.

18 . The engineered particle of claim 17 , wherein the surface variable region is selected from VR-IV, VR-V, or VR-VIII.

19 . The engineered particle of claim 16 , wherein the n-mer motif is inserted between any two amino acids between amino acids 262-269, 327-332, 382-386, 452-460, 488-505, 527-539, 545-558, 581-593, 704-714, or any combination thereof, in an AAV9 capsid polypeptide or in an analogous position in an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh.74, or AAV rh.10 capsid polypeptide; or is inserted between amino acids 588 and 589, in an AAV9 capsid polypeptide or in an analogous position in an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh.74, or AAV rh.10 capsid polypeptide.

20 . The engineered particle of claim 15 , wherein the engineered particle is an adeno associated virus (AAV) capsid.

21 . The engineered particle of claim 15 , further comprising a cargo.

22 . The engineered particle of claim 21 , wherein the cargo is
(a) capable of treating or preventing a muscle disease or disorder; (b) 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 or polypeptides; 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; or (c) both (a) and (b).

23 . The engineered particle of claim 22 , wherein the muscle disease or disorder is an auto immune disease; a cancer; a muscular dystrophy; a neuro-muscular disease; a sugar or glycogen storage disease; an expanded repeat disease; a dominant negative disease; a cardiomyopathy; a viral disease; a progeroid disease; or any combination thereof.

24 . A vector system comprising:
a polynucleotide encoding the composition of claim 1 .

25 . The vector system of claim 24 , further comprising a cargo polynucleotide.

26 . The vector system of claim 25 , wherein the cargo polynucleotide
(a) is capable of treating or preventing a muscle disease or disorder; (b) is or encodes 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 or polypeptides; 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; or (c) both (a) and (b).

27 . A method of treating or preventing a muscle disease or disorder in a subject in need thereof, comprising:
administering a composition of claim 1 or an engineered particle comprising the composition to a subject in need thereof.

28 . The method of claim 27 , wherein the composition or the engineered particle comprises a cargo.

29 . The method of claim 28 , wherein the cargo is
(a) capable of treating or preventing the muscle disease or disorder; (b) a morpholino; a peptide-linked morpholino; an antisense oligonucleotide; a P

Assignees

• Broad Inst Inc
• Harvard College
• Massachusetts Inst Technology

Inventors

• Sabeti Pardis
• Tabebordbar Mohammadsharif
• Ye Simon
• Lagerborg Kim
• Stanton Alexandra
• Wagers Amy

Classifications

• C12N2830/008

  cell type or tissue specific enhancer/promoter combination · CPC title

• C12N2750/14171

  Demonstrated in vivo effect · CPC title

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title

• C12N2750/14142

  virus or viral particle as vehicle, e.g. encapsulating small organic molecule · CPC title

• C12N2750/14122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title