PLATFORM FOR GENERATING SAFE CELL THERAPEUTICS
Provided herein are cytoplasts, compositions comprising cytoplasts, methods of using cytoplasts, and methods of treating a subject, such as providing benefits to a healthy or unhealthy subject, or treating or diagnosing a disease or condition in a subject. In some embodiments, methods of treating a subject include: administering to the subject a therapeutically effective amount of a composition comprising a cytoplast. Also, provided herein are compositions (e.g., pharmaceutical compositions) that include a cytoplast. Also, provided herein are kits comprising instructions for using the compositions or methods.
1 . A method comprising:
administering to a subject a therapeutically effective amount of a composition comprising a first cytoplast expressing or containing at least one therapeutic agent.
2 . The method of claim 1 , wherein the therapeutic agent is selected from the group consisting of: a therapeutic DNA molecule, a therapeutic RNA molecule, a therapeutic protein, a therapeutic peptide, a small molecule therapeutic, a therapeutic gene-editing factor, a small molecule therapeutic, a nanoparticle, bacteria, bacterial spores, bacteriophages, bacterial components, viruses, exosomes, lipids, and ions.
3 . The method of any one of claims 1 or 2 , wherein the cell is or is derived from a cell harvested from the subject, or derived from a cell line, an immortalized cell, or a cancer cell.
4 . The method of any one of claims 1 to 3 , wherein the first cytoplast is fused to a second cytoplast.
5 . The method of any one of claims 1 to 4 , further comprising administering to the subject one or more additional therapies.
6 . The method of any one of claims 1 to 5 , wherein the first cytoplast expresses an immune system-evading moiety.
7 . The method of claim 6 , wherein the immune-system evading moiety is CD47.
8 . The method of any one of claims 1 to 7 , wherein the first cytoplast or cell from which the first cytoplast is obtained has been engineered to express the therapeutic agent.
9 . The method of any one of claims 1 to 8 , wherein the first cytoplast or cell from which the first cytoplast is obtained has not been engineered to express the therapeutic agent.
10 . A cytoplast comprising at least one therapeutic agent.
11 . The cytoplast of claim 10 , wherein the therapeutic agent is selected from the group consisting of: a therapeutic DNA molecule, a therapeutic RNA molecule, a therapeutic protein, a therapeutic peptide, a small molecule therapeutic, a therapeutic gene-editing factor, a small molecule therapeutic, a nanoparticle, bacteria, bacterial spores, bacteriophages, bacterial components, viruses, exosomes, lipids, and ions.
12 . The cytoplast of any one of claims 10 or 11 , wherein the cytoplast further comprises an immune system-evading moiety.
13 . A method of making a cytoplast, the method comprising:
introducing into a cell a therapeutic agent; and
enucleating the cell.
14 . The method of claim 13 , wherein the introducing step precedes the enucleating step.
15 . The method of claim 14 , wherein the introducing step results in a permanent expression of the therapeutic agent.
16 . The method of claim 13 , wherein the enucleation step precedes the introducing step.
17 . The method of any one of claims 13 , 14 , or 16 , wherein the introducing step results in a transient expression of the therapeutic agent.
18 . The method of any one of claims 13 to 17 , wherein introducing comprises transfecting.
19 . The method of any one of claims 13 to 17 , wherein introducing comprises electroporating, microinjecting, cell squeezing, sonoporating, impalecting, or hydrodynamic delivery.
20 . A method of treating a subject comprising:
administering to the subject a therapeutically effective amount of a cytoplast of any one of claims 10 to 12 .