Microorganisms programmed to produce immune modulators and anti-cancer therapeutics in tumor cells

Abstract

Genetically programmed microorganisms, such as bacteria or virus, pharmaceutical compositions thereof, and methods of modulating and treating cancers are disclosed.

1 - 136 . (canceled) 
     
     
         137 . A genetically engineered bacterium comprising at least one non-native gene encoding an enzyme capable of producing a stimulator of interferon gene (STING) agonist, wherein the STING agonist is c-di-AMP,
 wherein the non-native gene is a dacA (diadenylate cyclase) gene and encodes a polypeptide comprising a sequence that has at least 90% identity to SEQ ID NO: 1209,   wherein the dacA gene is operatively linked to an inducible promoter, and   wherein the bacterium is an auxotroph in dapA (4-hydroxy-tetrahydrodipicolinate synthase) and thyA (thymidylate synthase).   
     
     
         138 . The genetically engineered bacterium of  claim 137 , wherein the polypeptide comprises a sequence that has at least 95% identity to SEQ ID NO: 1209. 
     
     
         139 . The genetically engineered bacterium of  claim 138 , wherein the polypeptide comprises a sequence that has at least 97% identity to SEQ ID NO: 1209. 
     
     
         140 . The genetically engineered bacterium of  claim 139 , wherein the polypeptide comprises a sequence that has at least 99% identity to SEQ ID NO: 1209. 
     
     
         141 . The genetically engineered bacterium of  claim 137 , wherein the dacA gene is integrated into a chromosome of the bacterium or is present on a plasmid in the bacterium. 
     
     
         142 . The genetically engineered bacterium of  claim 137 , wherein the inducible promoter is induced by low-oxygen or anaerobic conditions, or wherein the inducible promoter is induced by a hypoxic environment of a tumor. 
     
     
         143 . The genetically engineered bacterium of  claim 137 , wherein the bacterium is non-pathogenic. 
     
     
         144 . The genetically engineered bacterium of  claim 143 , wherein the bacterium is  Escherichia coli  Nissle. 
     
     
         145 . A pharmaceutical composition comprising the genetically engineered bacterium of  claim 137  and a pharmaceutically acceptable carrier. 
     
     
         146 . A method of treating cancer in a subject, the method comprising administering the pharmaceutical composition of  claim 145  to the subject, thereby treating cancer in the subject. 
     
     
         147 . The method of  claim 146 , wherein the administering is via intratumoral injection. 
     
     
         148 . A genetically engineered bacterium comprising at least one non-native gene encoding an enzyme capable of producing a stimulator of interferon gene (STING) agonist, wherein the STING agonist is c-di-AMP,
 wherein the non-native gene is a dacA (diadenylate cyclase) gene that comprises a sequence having at least 90% identity to SEQ ID NO: 1210,   wherein the dacA gene is operatively linked to an inducible promoter, and   wherein the bacterium is an auxotroph in dapA (4-hydroxy-tetrahydrodipicolinate synthase) and thyA (thymidylate synthase).   
     
     
         149 . The genetically engineered bacterium of  claim 148 , wherein the dacA gene is integrated into a chromosome of the bacterium or is present on a plasmid in the bacterium. 
     
     
         150 . The genetically engineered bacterium of  claim 148 , wherein the inducible promoter is induced by low-oxygen or anaerobic conditions, or wherein the inducible promoter is induced by a hypoxic environment of a tumor. 
     
     
         151 . The genetically engineered bacterium of  claim 148 , wherein the bacterium is non-pathogenic. 
     
     
         152 . The genetically engineered bacterium of  claim 151 , wherein the bacterium is  Escherichia coli  Nissle. 
     
     
         153 . A pharmaceutical composition comprising the genetically engineered bacterium of  claim 148  and a pharmaceutically acceptable carrier. 
     
     
         154 . A method of treating cancer in a subject, the method comprising administering the pharmaceutical composition of  claim 153  to the subject, thereby treating cancer in the subject. 
     
     
         155 . The method of  claim 154 , wherein the administering is via intratumoral injection.