Engineered extracellular vesicles for enhanced tissue delivery
Several embodiments relate to engineered extracellular vesicles (EVs) using the membrane cloaking platform technology described herein, the cloaking imparting to the EVs enhanced delivery to tissues of interest, such as damaged or dysfunctional tissue. Several embodiments relate to engineering exosomes derived from cardiosphere-derived cells (CDCs) using the membrane cloaking platform technology described herein to confer enhanced tissue homing specificities, thereby leading to repair and regeneration at sites of injury. Uses of engineered EV compositions to treat diseases are also provided for in several embodiments.
1. An engineered extracellular vesicle, comprising:
a lipid anchor moiety configured to insert at least partially into a lipid bilayer of the extracellular vesicle,
a targeting moiety configured to bind to a target molecule, a spacer, and
a coupling moiety comprising:
a first member of a specific binding pair and a second member of a specific binding pair,
wherein said first member is covalently linked to the targeting moiety,
said second member is covalently linked to the spacer,
said first member and said second member noncovalently bind one another with a dissociation constant less than or equal to 10 −8 mol/L, and
said first and second members of the specific binding pair are a biotin moiety and an avidin moiety.
2. The engineered extracellular vesicle of claim 1 , wherein the lipid anchor moiety comprises an amphipathic lipid having a hydrophobic moiety and a hydrophilic portion.
3. The engineered extracellular vesicle of claim 2 , wherein the amphipathic lipid is selected from the group comprising phospholipids, aminolipids and sphingolipids.
4. The engineered extracellular vesicle of claim 3 , wherein the phospholipid is selected from the group comprising dilauroyl-phosphatidylcholine (DLPC), dimyristoyl-phosphatidyl choline (DMPC), dipalmitoyl-phosphatidyl choline (DPPC), diarachidoyl-phosphatidyl choline (DAPC), distearoyl-phosphatidyl choline (DSPC), dioleoyl-phosphatidyl choline (DOPC), 1,2 Distearoyl-sn-glycero-3-Ethylphosphocholine (Ethyl-DSPC), dipentadecanoyl-phosphatidylcholine (DPDPC), 1-myristoyl-2-palmitoyl-phosphatidylcholine (MPPC), 1-palmitoyl-2-myristoyl-phosphatidylcholine (PMPC), 1-palmitoyl-2-stearoyl-phosphatidylcholine (PSPC), 1-stearoyl-2-palmitoyl-phosphatidylcholine (SPPC), 1-palmitoyl-2-oleylphosphatidyl choline (POC), 1-oleyl-2-palmitoyl-phosphatidylcholine (OPPC), dilauroylphosphatidyl glycerol (DLPG), diarachidoyl phosphatidyl glycerol (DAPG), dimyristoylphosphatidylglycerol (DMPG) and its, dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPC), dioleoyl-phosphatidylglycerol (DOPG), dimyristoyl phosphatidic acid (RMPA), dipalmitoyl phosphatidic acid (DPPA), distearoyl phosphatidic acid (DSPA), diarachidoylphosphatidic acid (DAPA), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoyl phosphatidyl-ethanolamine (DSPE), dioleylphosphatidy lethanolarnine (DOPE), diarachidoy lphosphatidy lethanolarnine (DAPE), dilinoleylphosphatidylethanolamine (DLPE), dimyristoyl phosphatidylserine (DMPS), diarachidoyl phosphatidylserine (DAPS), dipalmitoyl phosphatidylserine (DPPS), distearoylphosphatidylserine (DSPS), dioleoylphosphatidylserine (DOPS), dipalmitoyl sphingomyelin (DPSP), and distearoylsphingomyelin (DSSP), dilauroyl-phosphatidylinositol (DLPI), diarachidoylphosphatidylinositol (DAPI), dimyristoylphosphatidylinositol (DMPI), dipalmitoylphosphatidylinositol (DPPI), distearoylphosphatidylinositol (DSPI), and dioleoyl-phosphatidylinositol (DOM).
5. The engineered extracellular vesicle of claim 4 , wherein the target molecule is a ligand that is over-expressed on target cells, and wherein the targeting moiety is a peptide.
6. The engineered extracellular vesicle of claim 5 , wherein the peptide comprises a monoclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a Fab, a Fab′, a F(ab′)2, an Fv, a single-chain Fv (scFv), minibody, a diabody, and a single-domain antibody, a functional derivative thereof, a variant thereof or a fragment thereof.
7. The engineered extracellular vesicle of claim 5 , wherein the target cells are cells that have incurred damage due to ischemia following acute myocardial infarction.
8. The engineered extracellular vesicle of claim 7 , wherein the peptide is selected from the group consisting of CSTSMLKAC (SEQ ID NO: 1), CKPGTSSYC (SEQ ID NO: 2), and CPDRSVNNC (SEQ H) NO: 3).
9. The engineered extracellular vesicle of claim 1 , wherein the first member of the specific binding pair is a biotin moiety and the second member of the specific binding pair is an avidin moiety.
10. The engineered extracellular vesicle of claim 1 , wherein the spacer is a hydrophilic polymer, wherein the hydrophilic polymer is polyethylene glycol (PEG), polypropylene glycol, methoxypolyethylene glycol (mPEG), polyvinylalcohol, polyvinylpyrrolidone, and copolymers thereof.
11. The engineered extracellular vesicle of claim 1 , wherein the lipid anchor moiety comprises a phosphatidylethanolamine conjugated through covalent, bond to the spacer, wherein the spacer is a hydrophilic polymer.
12. The engineered extracellular vesicle of claim 11 , wherein the lipid anchor moiety comprises DMPE-PEG, DPPE-PEG, DSPE-PEG, or DAPE-PEG.
13. The engineered extracellular vesicle of claim 1 , wherein the extracellular vesicle is derived from the group comprising cardiosphere-derived cells (CDCs), human neural stem cells (hNSCs), bone marrow stem cells, and mononuclear cells (MNCs).
14. The engineered extracellular vesicle of claim 13 , wherein the extracellular vesicle is an exosome derived from CDCs.