Methods for systemically delivering polypeptides and microorganisms therefor

Abstract

Methods and microorganisms for systemically introducing a polypeptide in the bloodstream of a subject. The methods of the invention include administering into the gastrointestinal tract of a subject a bacterium configured to express and produce and release the polypeptide. The bacterium is administered in an amount effective to introduce the polypeptide in the bloodstream of the subject, preferably in a detectable amount. The microorganisms of the invention include lactic acid bacteria, such as Lactobacillus reuteri, that comprise a recombinant gene configured to express a polypeptide to be systemically introduced.

1 - 20 . (canceled) 
     
     
         21 . A method of systemically introducing a polypeptide into a bloodstream of a subject, the method comprising administering into the gastrointestinal tract of the subject a bacterium that produces and releases the polypeptide, wherein the bacterium comprises a recombinant gene configured to express the polypeptide, wherein the bacterium is administered in an amount effective to introduce the polypeptide in the bloodstream of the subject in a detectable amount without the bacterium being introduced in the bloodstream of the subject in a detectable amount. 
     
     
         22 . The method of  claim 21 , wherein the bacterium is administered in an amount effective to introduce the polypeptide in the bloodstream in an amount effective to induce at least one direct systemic effect in the subject. 
     
     
         23 . The method of  claim 21 , wherein the bacterium is administered in an amount effective to introduce the polypeptide in the bloodstream in an amount effective to induce at least one direct effect in a non-gastrointestinal tissue in the subject. 
     
     
         24 . The method of  claim 21 , wherein the bacterium is administered in an amount effective to introduce the polypeptide in the bloodstream in an amount effective to induce at least one direct effect in a tissue selected from the group consisting of liver, muscles, lungs, kidneys, pancreas, and adipose tissue in the subject. 
     
     
         25 . The method of  claim 21 , wherein the subject suffers from a condition treatable with systemic introduction of the polypeptide and wherein the polypeptide is introduced in the bloodstream of the subject in an amount effective to treat the condition. 
     
     
         26 . The method of  claim 21 , wherein the subject suffers from a condition treatable with systemic introduction of the polypeptide but not treatable with local introduction of the polypeptide to the gastrointestinal tract without systemic introduction of the polypeptide, and wherein the polypeptide is introduced in the bloodstream of the subject in an amount effective to treat the condition. 
     
     
         27 . The method of  claim 21 , wherein the recombinant gene is codon optimized. 
     
     
         28 . The method of  claim 21 , wherein the polypeptide is selected from the group consisting of a cytokine, a hormone, an antibody, an antimicrobial peptide, and an antigenic peptide. 
     
     
         29 . The method of  claim 21 , wherein the polypeptide is selected from the group consisting of interleukin-22 (IL-22), interleukin-35 (IL-35), insulin, leptin, cathelicidin related antimicrobial peptide, a peptide inhibitor of proprotein convertase subtilisin/kexin type 9 (PCSK9), and an endolysin. 
     
     
         30 . The method of  claim 29 , wherein the subject suffers from at least one condition selected from the group consisting of insulin resistance, hyperglycemia, lipid dysregulation, hyperlipidemia, and obesity, and wherein the polypeptide is introduced in the bloodstream of the subject in an amount effective to treat the at least one condition. 
     
     
         31 . The method of  claim 21 , wherein the polypeptide is interleukin-22 (IL-22). 
     
     
         32 . The method of  claim 21 , wherein the bacterium expresses a mucus-binding protein. 
     
     
         33 . The method of  claim 21 , wherein the bacterium expresses CmbA. 
     
     
         34 . The method of  claim 21 , wherein the bacterium has a mutation rate less than about 100×10 −10  mutations per cell per generation. 
     
     
         35 . The method of  claim 21 , wherein the bacterium comprises a member of lactic acid bacteria. 
     
     
         36 . The method of  claim 21 , wherein the bacterium comprises a member of lactic acid bacteria other than a member of the  Lactococcus  genus. 
     
     
         37 . The method of  claim 21 , wherein the bacterium comprises a member of  Lactobacillus.    
     
     
         38 . The method of  claim 21 , wherein the bacterium expresses a mucus-binding protein and has a mutation rate less than about 100×10 −10  mutations per cell per generation. 
     
     
         39 . The method of  claim 38 , wherein the mucus-binding protein is CmbA. 
     
     
         40 . The method of  claim 39 , wherein the recombinant gene is codon optimized.