EXTRACELLULAR VESICLES TARGETING DENDRITIC CELLS AND USES THEREOF
The present disclosure relates to modified extracellular vesicles, e.g., exosomes, comprising a targeting moiety, wherein the targeting moiety can specifically bind to markers expressed on distinct immune cells (e.g., dendritic cells). Also provided herein are methods for using the exosomes to treat and/or prevent a range of medical disorders.
1 . An extracellular vesicle (EV) comprising an exogenous targeting moiety that specifically binds to a marker for a dendritic cell.
2 . (canceled)
3 . The EV of claim 1 , wherein the dendritic cell comprises a plasmacytoid dendritic cell (pDC), a myeloid/conventional dendritic cell 1 (cDC1), a myeloid/conventional dendritic cell 2 (cDC2), or any combination thereof.
4 . (canceled)
5 . The EV of claim 1 , wherein the marker comprises a C-type lectin domain family 9 member A (Clec9a) protein, a dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN), CD207, CD40, Clec6, dendritic cell immunoreceptor (DCIR), DEC-205, lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), MARCO, Clec12a, DC-asialoglycoprotein receptor (DC-ASGPR), DC immunoreceptor 2 (DCIR2), Dectin-1, macrophage mannose receptor (MMR), BDCA-1 (CD303, Clec4c), Dectin-2, Bst-2 (CD317), or any combination thereof.
6 . (canceled)
7 . The EV of claim 5 , wherein the marker comprises a C-type lectin like domain of the Clec9a protein, an extracellular region of the Clec9a protein, or both.
8 . The EV of claim 7 , wherein (i) the C-type lectin like domain of the Clec9a protein comprises the amino acid sequence comprising amino acids 120 to 233 of SEQ ID NO: 1; (ii) the extracellular region of the Clec9a protein comprises the amino acid sequence comprising amino acids 57 to 241 of SEQ ID NO: 1; or (iii) both (i) and (ii).
9 - 10 . (canceled)
11 . The EV of claim 1 , wherein the exogenous targeting moiety comprises a peptide, an antibody or an antigen-binding fragment thereof, a chemical compound, a microprotein, a designed ankyrin repeat protein (darpin), an anticalin, an adnectin, an aptamer, a peptide mimetic molecule, a natural ligand for a receptor, a camelid nanobody, or any combination thereof.
12 - 15 . (canceled)
16 . The EV of claim 1 , wherein the EV further comprises a Scaffold X protein linking the exogenous targeting moiety to the EV.
17 - 18 . (canceled)
19 . The EV of claim 16 , wherein the Scaffold X protein comprises (i) the amino acid sequence set forth in SEQ ID NO: 33; or (ii) an amino acid sequence having at least about 70% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1.
20 . (canceled)
21 . The EV of claim 16 , further comprising a Scaffold Y protein.
22 . The EV of claim 21 , wherein the Scaffold Y protein comprises 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 fragment thereof, and or any combination thereof.
23 - 45 . (canceled)
46 . The EV of claim 1 , further comprising a therapeutic molecule, an immune modulator, an adjuvant, or any combination thereof.
47 - 54 . (canceled)
55 . The EV of claim 46 , wherein the therapeutic molecule, immune modulator, adjuvant, or any combination thereof, is linked to the EV via a Scaffold X protein, Scaffold Y protein, or both.
56 - 57 . (canceled)
58 . The EV of claim 1 , wherein the EV is an exosome.
59 - 64 . (canceled)
65 . A pharmaceutical composition comprising the EV of claim 1 and a pharmaceutically acceptable carrier.
66 . A cell that produces the EV of claim 1 .
67 . A cell comprising one or more vectors, wherein the vectors comprise a nucleic acid sequence encoding the targeting moiety of claim 1 .
68 . A kit comprising the EV of claim 1 and instructions for use.
69 . A method of making EVs comprising culturing the cell of claim 66 under a suitable condition and obtaining the EVs.
70 . A method of treating a disease in a subject in need thereof, comprising administering to the subject the EV of claim 1 .
71 . (canceled)
72 . A method of delivering an EV to a subject, comprising administering to the subject the EV of claim 1 .
73 - 74 . (canceled)