Therapeutic Cell Systems and Methods for Treating Cancer and Infectious Diseases

Abstract

The present disclosure relates to erythroid cells that have been engineered to include, e.g., at the surface of the cell, one or more exogenous stimulatory polypeptides, wherein the exogenous stimulatory polypeptides presented are sufficient to stimulate an immune killer cell. The engineered enucleated cells of the present disclosure are useful in methods of activating NK cells and/or CD8+ T-cells in a subject in need thereof, such as subjects having cancer or an infectious disease, and in particular cancers or infectious diseases characterized by downregulation of MHC Class I presentation.

1 .- 156 . (canceled) 
     
     
         157 . A method of treating a solid tumor in a subject, the method comprising administering to the subject an engineered enucleated erythroid cell, comprising:
 (i) a first exogenous stimulatory polypeptide, wherein the first exogenous stimulatory polypeptide is a fusion polypeptide comprising an IL-15/IL15RA fusion comprising an IL-15 polypeptide, or a fragment thereof, linked to the extracellular portion of an IL-15RA polypeptide, or a fragment thereof, by a linker; and   (ii) a second exogenous stimulatory polypeptide, wherein the second exogenous stimulatory polypeptide comprises 4-1BBL, or a functional fragment thereof;   wherein one or both of the first exogenous stimulatory polypeptide and the second exogenous stimulatory polypeptide comprise a heterologous transmembrane domain from glycophorin A.   
     
     
         158 . The method of  claim 157 , wherein the solid tumor is an advanced and/or metastatic solid tumor. 
     
     
         159 . The method of  claim 157 , wherein the fusion polypeptide comprises an IL-15 receptor alpha sushi domain. 
     
     
         160 . The method of  claim 159 , wherein the fusion polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 2. 
     
     
         161 . The method of  claim 159 , wherein the fusion polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 31. 
     
     
         162 . The method of  claim 159 , wherein the fusion polypeptide comprises SEQ ID NO: 2 or SEQ ID NO: 31. 
     
     
         163 . The method of  claim 157 , wherein the second exogenous stimulatory polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 41. 
     
     
         164 . The method of  claim 157 , wherein the second exogenous stimulatory polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 43. 
     
     
         165 . The method of  claim 157 , wherein the second exogenous stimulatory polypeptide comprises SEQ ID NO: 41 or SEQ ID NO: 43. 
     
     
         166 . The method of  claim 157 , wherein the solid tumor is colorectal cancer, anal cancer, melanoma, mesothelioma, or sarcoma. 
     
     
         167 . The method of  claim 166 , wherein the sarcoma is a soft tissue sarcoma. 
     
     
         168 . A method of treating acute myeloid leukemia (AML) in a subject, the method comprising administering to the subject an engineered enucleated erythroid cell, comprising:
 (i) a first exogenous stimulatory polypeptide, wherein the first exogenous stimulatory polypeptide is a fusion polypeptide comprising an IL-15/IL15RA fusion comprising an IL-15 polypeptide, or a fragment thereof, linked to the extracellular portion of an IL-15RA polypeptide, or a fragment thereof, by a linker; and   (ii) a second exogenous stimulatory polypeptide, wherein the second exogenous stimulatory polypeptide comprises 4-1BBL, or a functional fragment thereof;   wherein one or both of the first exogenous stimulatory polypeptide and the second exogenous stimulatory polypeptide comprise a heterologous transmembrane domain from glycophorin A.   
     
     
         169 . The method of  claim 168 , wherein the AML is a relapsed/refractory AML. 
     
     
         170 . The method of  claim 168 , wherein the fusion polypeptide comprises an IL-15 receptor alpha sushi domain. 
     
     
         171 . The method of  claim 170 , wherein the fusion polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 2. 
     
     
         172 . The method of  claim 170 , wherein the fusion polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 31. 
     
     
         173 . The method of  claim 170 , wherein the fusion polypeptide comprises SEQ ID NO: 2 or SEQ ID NO: 31. 
     
     
         174 . The method of  claim 168 , wherein the second exogenous stimulatory polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 41. 
     
     
         175 . The method of  claim 168 , wherein the second exogenous stimulatory polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 43. 
     
     
         176 . The method of  claim 168 , wherein the second exogenous stimulatory polypeptide comprises SEQ ID NO: 41 or SEQ ID NO: 43.