Compositions and methods related to megakaryocyte-derived extracellular vesicles
Disclosed herein are compositions and methods related to megakaryocyte-derived extracellular vesicles derived from human pluripotent stem cells, where the megakaryocyte-derived extracellular vesicles may be utilized for drug delivery and treating various diseases.
1 . A composition comprising:
a plurality of megakaryocyte-generated extracellular vesicles (MkEVs) comprising a lipid bilayer membrane surrounding a lumen, wherein:
a. the MkEV lumen comprises one or more nucleic acid molecules from the megakaryocytes selected from mRNA, tRNA, rRNA, siRNA, microRNA, regulating RNA, and non-coding and coding RNA; and,
b. the lipid bilayer membrane comprises one or more proteins associated with or embedded within, wherein the one or more proteins are selected from CD21 and GPVI; and,
C. the MkEVs are isolated from megakaryocytes derived from a human pluripotent stem cell (HPSC).
2 . The composition of claim 1 , wherein the lipid bilayer membrane further comprises one or more of CD9, CD63, CD47, CD43, CD54, and CD18.
3 . The composition of claim 1 , wherein the lipid bilayer membrane further comprises one or more of CD41, CD61, CD11b, LAMP-1 (CD107a), CD51, CD31, CD147, phosphatidylserine, CLEC-2, CD62, CD42b, and CD32a.
4 . The composition of claim 1 , wherein the MkEVs are of a diameter in the range between 100 nm to 300 nm.
5 . The composition of claim 1 , wherein the MkEVs are essentially free of:
a. megakaryocytes, and/or
b. platelets
C. organelles, and/or
d. mitochondria or nuclei.
6 . The composition of claim 1 , wherein the MkEVs are suitable for homing to a hematopoietic stem cell, a bone marrow, a lymphatic cell, or a regulatory T cell in vivo and/or in vitro.
7 . The composition of claim 1 , wherein the MkEVs are suitable for loading with cargo into the lumen and/or loading with cargo associated with the surface of the MkEVs.
8 . The composition of claim 7 , wherein the cargo is selected from one or more of a RNA, DNA, protein, carbohydrate, lipid, biomolecule, and small molecule.
9 . The composition of claim 8 , wherein the cargo is one or more therapeutic agents.
10 . The composition of claim 9 , wherein the therapeutic agent is selected from one or more of an antibody or an antibody fragment, recombinant protein, fusion protein, gene-editing protein, cytokine, antigen, and peptide.
11 . The composition of claim 9 , wherein the therapeutic agent is a nucleic acid therapeutic agent selected from one or more non-autologous and/or recombinant nucleic acid constructs selected from mRNA, tRNA, rRNA, siRNA, microRNA, regulating RNA, non-coding and coding RNA, linear DNA, DNA fragments, and DNA plasmids.
12 . The composition of claim 11 , wherein the nucleic acid therapeutic agent encodes a functional protein or a gene-editing protein.