Extracellular vesicles from non-pathogenic amoebae useful as vehicle for transferring a molecule of interest to an eukaryotic cell
View Patent ↗The invention relates to membrane vesicles from non-pathogenic amoeba, in particular from the amoeba Dictyostelium discoideum , containing a molecule of therapeutic interest, to a method of preparing such vesicles and to the use of said vesicles as a vehicle for transferring the molecule therapeutic interest to an eukaryotic mammalian cell.
1. A method for preparing a non-pathogenic amoeba vesicle for delivering a molecule of interest to an eukaryotic target cell, which method comprises the steps consisting of:
a) culturing a non-pathogenic amoeba cell in a culture medium comprising said molecule of interest, under conditions sufficient to allow the non-pathogenic amoeba cell to release vesicles; and
b) recovering a vesicle released by said non-pathogenic amoeba cell, which vesicle contains said molecule of interest,
wherein said molecule of interest is selected from the group consisting of a therapeutic molecule, an imaging agent and a diagnostic agent, with the proviso that said vesicle is not a Hoechst 33342-containing Dictyostelium discoideum vesicle.
2. The method according to claim 1 , wherein said nonpathogenic amoeba is Dictyostelium discoideum.
3. The method of claim 1 , wherein said molecule of interest is a therapeutic molecule.
4. The method of claim 1 ,
wherein said therapeutic molecule is selected from the group consisting of an antibiotic, an antimicrobial agent, an antimycobacterial, antifungal, or antiviral agent, an agent affecting the immune response, a blood calcium regulator, an agent useful in glucose regulation, an anticoagulant, an antithrombotic, an antihyperlipidemic agent, a cardiac drug, a thyromimetic or antithyroid drug, an adrenergic, an antihypertensive agents, a cholinergic, an anticholinergic, an antispasmodic, an antiulcer agent, a skeletal and smooth muscle relaxant, a prostaglandin, a general inhibitor of the allergic response, an antihistamine, a local anesthetic, an analgesic, a narcotic antagonist, an antitussive, a sedative-hypnotic agent, an anticonvulsant, an antipsychotic, an anti-anxiety agent, an antidepressant agent, an anorexigenic, a non-steroidal anti-inflammatory agent, a steroidal anti-inflammatory agent, an antioxidant, a vasoactive agent, a bone-active agent, an anti-arthritic agent and an anti-neoplastic agent.
5. The method of claim 1 , wherein said therapeutic molecule is an anti-neoplastic agent.
6. The method of claim 1 , wherein said therapeutic molecule is an antibiotic.
7. The method of claim 1 , wherein said molecule of interest is an imaging agent.
8. The method of claim 1 , wherein said molecule of interest is a diagnostic agent.
9. The method of claim 1 , wherein said molecule of interest is selected from the group consisting of a small chemical molecule, a small organic or inorganic compound and a nucleic acid.
10. The method of claim 1 , wherein said target cell is a human cell.
11. The method of claim 1 , wherein said molecule of interest is not Hoechst 33342.
12. The method of claim 2 , wherein said molecule of interest is a therapeutic molecule.
13. The method of claim 2 ,
wherein said therapeutic molecule is selected from the group consisting of an antibiotic, an antimicrobial agent, an antimycobacterial, antifungal, or antiviral agent, an agent affecting the immune response, a blood calcium regulator, an agent useful in glucose regulation, an anticoagulant, an antithrombotic, an antihyperlipidemic agent, a cardiac drug, a thyromimetic or antithyroid drug, an adrenergic, an antihypertensive agents, a cholinergic, an anticholinergic, an antispasmodic, an antiulcer agent, a skeletal and smooth muscle relaxant, a prostaglandin, a general inhibitor of the allergic response, an antihistamine, a local anesthetic, an analgesic, a narcotic antagonist, an antitussive, a sedative-hypnotic agent, an anticonvulsant, an antipsychotic, an anti-anxiety agent, an antidepressant agent, an anorexigenic, a nonsteroidal anti-inflammatory agent, a steroidal anti-inflammatory agent, an antioxidant, a vasoactive agent, a bone-active agent, an anti-arthritic agent and an anti-neoplastic agent.
14. The method of claim 2 , wherein said therapeutic molecule is an anti-neoplastic agent.
15. The method of claim 2 , wherein said therapeutic molecule is an antibiotic.
16. The method of claim 2 , wherein said molecule of interest is an imaging agent.
17. The method of claim 2 , wherein said molecule of interest is a diagnostic agent.
18. The method of claim 2 , wherein said molecule of interest is selected from the group consisting of a small chemical molecule, a small organic or inorganic compound and a nucleic acid.
19. The method of claim 2 , wherein said target cell is a human cell.
20. An isolated non-pathogenic amoeba vesicle for delivering a molecule of interest to an eukaryotic target cell, wherein said vesicle contains a molecule of interest selected from the group consisting of a therapeutic molecule, an imaging agent and a diagnostic agent, with the proviso that said vesicle is not a Hoechst 33342-containing Dictyostelium discoideum vesicle.
21. The vesicle according to claim 20 , wherein said non-pathogenic amoeba is Dictyostelium discoideum.
22. The isolated non-pathogenic amoeba vesicle of claim 20 , wherein said molecule of interest is a therapeutic molecule.
23. The isolated non-pathogenic amoeba vesicle of claim 20 , wherein said molecule of interest is an imaging agent.
24. The isolated non-pathogenic amoeba vesicle of claim 20 , wherein said molecule of interest is a diagnostic agent.
25. The isolated non-pathogenic amoeba vesicle of claim 20 , wherein said molecule of interest is selected from the group consisting of a small chemical molecule, a small organic or inorganic compound, and a nucleic acid.
26. The isolated non-pathogenic amoeba vesicle of claim 20 , wherein said target cell is a human cell.
27. The isolated non-pathogenic amoeba vesicle of claim 20 , wherein said molecule of interest is not Hoechst 33342.
28. The isolated non-pathogenic amoeba vesicle of claim 21 , wherein said molecule of interest is a therapeutic molecule.
29. The isolated non-pathogenic amoeba vesicle of claim 21 , wherein said molecule of interest is an imaging agent.
30. The isolated non-pathogenic amoeba vesicle of claim 21 , wherein said molecule of interest is a diagnostic agent.
31. The isolated non-pathogenic amoeba vesicle of claim 21 , wherein said molecule of interest is selected from the group consisting of a small chemical molecule, a small organic or inorganic compound, and a nucleic acid.
32. The isolated non-pathogenic amoeba vesicle of claim 21 , wherein said target cell is a human cell.