EXTRACELLULAR VESICLE-ASO CONSTRUCTS TARGETING STAT6
The present disclosure relates to extracellular vesicles. e.g., exosomes, comprising an antisense oligonucleotide (ASO), wherein the ASO comprises a contiguous nucleotide sequence of 10 to 30 nucleotides in length that is complementary to a nucleic acid sequence within a STAT6 transcript. Also provided herein are methods for producing the exosomes and methods for using the exosomes to treat and/or prevent diseases or disorders.
1 . An extracellular vesicle comprising an antisense oligonucleotide (ASO) which comprises a contiguous nucleotide sequence of 10 to 30 nucleotides in length that is complementary to a nucleic acid sequence within a STAT6 transcript (SEQ ID NO: 1 or SEQ ID NO: 3), wherein the ASO is not TGAGCGAATGGACAGGTCTT (SEQ ID NO: 89).
2 . (canceled)
3 . The extracellular vesicle of claim 1 , which targets a cell selected from the group consisting of a macrophage, a myeloid-derived suppressor cell (MDSC), a monocyte, a basophil, a neutrophil, an eosinophil, and any combination thereof.
4 . The extracellular vesicle of claim 1 , wherein the ASO comprises a contiguous nucleotide sequence of 10 to 30 nucleotides in length that is at least 90% complementary to a nucleic acid sequence within nucleotides 1 to 2056 of a STAT6 transcript corresponding to a nucleotide sequence as set forth in SEQ ID NO: 3 or nucleotides 2059 to 3963 of a STAT6 transcript corresponding to a nucleotide sequence as set forth in SEQ ID NO: 3.
5 . (canceled)
6 . The extracellular vesicle of claim 1 , wherein the ASO is capable of reducing STAT6 mRNA or protein expression in a human cell, wherein the human cell expresses the STAT6 mRNA or protein.
7 - 10 . (canceled)
11 . The extracellular vesicle of claim 1 , wherein the ASO comprises one or more nucleoside analogs.
12 - 19 . (canceled)
20 . The extracellular vesicle of claim 1 , wherein the contiguous nucleotide sequence is complementary to a nucleic acid sequence comprising (i) nucleotides 1-700 of SEQ ID NO: 3; (ii) nucleotides 1000-1500 of SEQ ID NO: 3; (iii) nucleotides 1500-2000 of SEQ ID NO: 3; (iv) nucleotides 2000-2500 of SEQ ID NO: 3; (v) nucleotides 2500-3000 of SEQ ID NO: 3; (vi) nucleotides 3000-3700 of SEQ ID NO: 3, (vii) nucleotides 413-803 of SEQ ID NO: 3; (viii) nucleotides 952-1688 of SEQ ID NO: 3; (ix) nucleotides 1726-2489 of SEQ ID NO: 3; (x) nucleotides 2682-2912 of SEQ ID NO: 3; (xi) nucleotides 2970-3203 of SEQ ID NO: 3; (xii) nucleotides 3331-3561 of SEQ ID NO: 3; (xiii) nucleotides 463-753 of SEQ ID NO: 3; (xiv) nucleotides 1002-1638 of SEQ ID NO: 3; (xv) nucleotides 1776-2439 of SEQ ID NO: 3; (xvi) nucleotides 2682-2862 of SEQ ID NO: 3; (xvii) 3020-3153 of SEQ ID NO: 3; (xviii) nucleotides 3381-3511 of SEQ ID NO: 3; (xix) nucleotides 503-713 of SEQ ID NO: 3; (xx) nucleotides 1042-1598 of SEQ ID NO: 3; (xxi) nucleotides 1816-2399 of SEQ ID NO: 3; (xxii) nucleotides 2722-2822 of SEQ ID NO: 3; (xxiii) nucleotides 3060-3113 of SEQ ID NO: 3; or (xxiv) nucleotides 3421-3471 of SEQ ID NO: 3.
21 . The extracellular vesicle of claim 1 , wherein the contiguous nucleotide sequence is complementary to a nucleic acid sequence within (i) (i) nucleotides 513-703 of SEQ ID NO: 3; (ii) nucleotides 1052-1588 of SEQ ID NO: 3; (iii) nucleotides 1826-2389 of SEQ ID NO: 3; (iv) nucleotides 2732-2812 of SEQ ID NO: 3; (v) nucleotides 3070-3103 of SEQ ID NO: 3; or (vi) nucleotides 3431-3461 of SEQ ID NO: 3.
22 . (canceled)
23 . The extracellular vesicle of claim 1 , wherein the continuous nucleotide sequence is fully complementary to a nucleotide sequence within the STAT6 transcript.
24 . (canceled)
25 . The extracellular vesicle of claim 1 , wherein the ASO has a design selected from the group consisting of the designs in FIG. 1 , wherein the upper-case letter is a sugar modified nucleoside and the lower case letter is DNA.
26 . (canceled)
27 . The extracellular vesicle of claim 4 , wherein the contiguous nucleotide sequence comprises one or more modified internucleoside linkages.
28 - 30 . (canceled)
31 . The extracellular vesicle of claim 1 , which further comprises an anchoring moiety.
32 . (canceled)
33 . The extracellular vesicle of claim 1 , further comprising an exogenous targeting moiety.
34 - 40 . (canceled)
41 . The extracellular vesicle of claim 33 , wherein the EV comprises a scaffold moiety linking the exogenous targeting moiety to the EV.
42 . The extracellular vesicle of claim 41 , wherein the scaffold moiety is a Scaffold X.
43 . The extracellular vesicle of claim 41 , wherein the scaffold moiety is a Scaffold Y.
44 . (canceled)
45 . The extracellular vesicle of claim 42 , wherein the Scaffold X is selected from the group consisting of prostaglandin F2 receptor negative regulator (the PTGFRN protein); basigin (the BSG protein); immunoglobulin superfamily member 2 (the IGSF2 protein); immunoglobulin superfamily member 3 (the IGSF3 protein); immunoglobulin superfamily member 8 (the IGSF8 protein); integrin beta-1 (the ITGB1 protein); integrin alpha-4 (the ITGA4 protein); 4F2 cell-surface antigen heavy chain (the SLC3A2 protein); a class of ATP transporter proteins (the ATP1A1, ATP1A2, ATP1A3, ATPIA4, ATPIB3, ATP2B1, ATP2B2, ATP2B3, ATP2B4 proteins); a functional fragment thereof; and any combination thereof.
46 . The extracellular vesicle of claim 31 , wherein the scaffold moiety is PTGFRN protein or a functional fragment thereof.
47 - 49 . (canceled)
50 . The extracellular vesicle of claim 43 , wherein the Scaffold Y is selected from the group consisting of myristoylated alanine rich Protein Kinase C substrate (the MARCKS protein), myristoylated alanine rich Protein Kinase C substrate like 1 (the MARCKSL1 protein), brain acid soluble protein 1 (the BASP1 protein), a functional fragment thereof, and any combination thereof.
51 - 77 . (canceled)
78 . The extracellular vesicle of claim 31 , wherein the ASO is linked to the scaffold moiety by a linker.
79 . The extracellular vesicle of claim 1 , wherein the ASO is linked to the EV by a linker.
80 - 142 . (canceled)