Methods and compositions for use of recombinant bacterial effector proteins as anti-inflammatory agents

Abstract

Provided herein are methods and compositions comprising a set of paired peptides comprising a first bacterial effector polypeptide or fragment thereof linked to a second bacterial effector polypeptide or fragment thereof. The paired peptides can be linked to a protein transduction domain. The compositions can be formulated as pharmaceuticals. The compositions are useful for the treatment of inflammatory disorders.

1 . A composition comprising a set of paired peptides, wherein the set of paired peptides is linked to a protein transduction domain, and wherein the set of paired peptides comprises a first bacterial effector polypeptide or fragment thereof linked to a second bacterial effector polypeptide or fragment thereof. 
     
     
         2 . The composition of  claim 1 , wherein the first bacterial effector polypeptide or fragment thereof and second bacterial effector polypeptide or fragment thereof are different. 
     
     
         3 . The composition of  claim 1 , wherein a first bacterial effector polypeptide or fragment thereof and the second bacterial effector polypeptide or fragment thereof are immunomodulatory. 
     
     
         4 . The composition of  claim 1 , wherein the first bacterial effector polypeptide or fragment thereof and the second bacterial effector polypeptide or fragment thereof recognize a different molecular target or modulate a different inflammatory pathway. 
     
     
         5 . The composition of  claim 4 , wherein the inflammatory pathway is the NFkB pathway, the JNK pathway, the p38 pathway or the STING pathway. 
     
     
         6 . The composition of  claim 1 , wherein the protein transduction domain and the set of paired peptides comprise a fusion protein. 
     
     
         7 . The fusion protein of  claim 6 , further comprising one or more linkers. 
     
     
         8 . The fusion protein of  claim 7 , wherein the linker is positioned between the first bacterial effector polypeptide or fragment thereof and the second bacterial effector polypeptide or fragment thereof. 
     
     
         9 . The composition of  claim 1 , wherein the protein transduction domain is a YopM protein transduction domain, an SspH1 protein transduction domain, or an lpaH protein transduction domain. 
     
     
         10 . The composition of  claim 9 , wherein the protein transduction domain is a YopM protein transduction domain. 
     
     
         11 . The composition of  claim 10 , wherein the protein transduction domain comprises SECS ID NO. 5. 
     
     
         12 . The composition of  claim 1 , wherein the protein transduction domain is selected from the group consisting of Poly-Arg, Tat and VP22, df Tat, a cyclic CPPs, IMT-P8, seven arginine (R7) and Streptolysin 0 (SLO)-mediated systems, elastin like polypeptide, CPP-adaptor system, 1, 2-Benzisothiazolin-3-one (BIT) and Tat, activatable cell-penetrating peptides, LDP12, BR2, POD, native protein independent of R11-CPP, Poly-arginine/Tat and Tat-PTO, Pep-1, CADY-2, R8, azo-R8, Penetratin, HR9 and IR9 peptides, or pVEC. 
     
     
         13 . The composition of  claim 1 , wherein the first bacterial effector polypeptide or fragment thereof is a polypeptide selected from the group consisting of NleE, NleC, NleD, NleB, NleH, YopM, YopE, YopH, YopJ, YopP, SspH1, OspG, OspF, lpaH9.8, lpaH1.4, lpaH2.5, lpaH4.5, lpaH7.8 and SlrP, and the second bacterial effector polypeptide or fragment thereof is a polypeptide selected from the group consisting of NleE, NleC, NleD, NleB, NleH, YopM, YopE, YopH, YopJ, YopP, SspH1, OspG, OspF, lpaH9.8, lpaH1.4, lpaH2.5, lpaH4.5, lpaH7.8 and SlrP. 
     
     
         14 . The composition of  claim 13 , wherein the first bacterial effector polypeptide or fragment thereof is a polypeptide having 90% sequence identity to an amino acid sequence set forth in the group consisting of SEQ ID NOs 3, 89, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, and 79 and the second bacterial effector polypeptide or fragment thereof is a polypeptide having 90% sequence identity to an amino acid sequence set forth in the group consisting of SEQ ID NOs.3, 89, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, and 79. 
     
     
         15 . The composition of  claim 1 , wherein
 (a) the first bacterial effector polypeptide or fragment thereof is a YopM polypeptide or a fragment thereof or an NLeE polypeptide or a fragment thereof; or   (b) the second bacterial effector polypeptide or fragment thereof is a YopM polypeptide or a fragment thereof or an NLeE polypeptide or a fragment thereof.   
     
     
         16 . (canceled) 
     
     
         17 . The composition of  claim 1 , wherein the first bacterial effector polypeptide or fragment thereof is a YopM polypeptide or a fragment thereof and the second bacterial effector polypeptide or fragment thereof is an NLeE polypeptide or a fragment thereof. 
     
     
         18 . The fusion protein of  claim 6 , wherein the amino acid sequence is at least 85% identical to the sequence set forth in SEQ ID NOs. 10, 13, 16, 19, 22, or 24. 
     
     
         19 - 35 . (canceled) 
     
     
         36 . A fusion protein comprising a set of paired peptides, wherein the set of paired peptides comprises a first bacterial effector polypeptide or fragment thereof linked to a second bacterial effector polypeptide or fragment thereof. 
     
     
         37 - 56 . (canceled) 
     
     
         57 . A composition comprising a protein transduction domain polypeptide linked to two or more bacterial effector polypeptides or fragments thereof. 
     
     
         58 . (canceled)