Platform for generating safe cell therapeutics

Abstract

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.

What is claimed is: 
     
         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 .