Inhibition of innate immune response

Abstract

The present invention provides methods, kits, and compositions for reducing an innate immune system response in a human or animal cell, tissue or organism. One embodiment comprises: introducing an Agent mRNA comprising in vitro-synthesized mRNA encoding one or more proteins that affect the induction, activity or response of an innate immune response pathway; whereby, the innate immune response in the cell, tissue or organism is reduced compared to the innate immune response in the absence of the Agent mRNA. Other embodiments are methods, compositions and kits for using an Agent mRNA for treating a disease or medical condition in a human or animal that exhibits symptoms of an elevated innate immune system, or for reducing an innate immune response that is induced in a human or animal cell, tissue or organism by a Foreign Substance that is administered to the cell, tissue or organism.

We claim: 
     
         1 . A method for reducing an innate immune response in a human or animal cell, tissue or organism, comprising:
 introducing into the cell, tissue or organism an Agent mRNA comprising in vitro-synthesized mRNA encoding one or more proteins that affect the induction, activity and/or response of an innate immune response pathway;   whereby the innate immune response in the cell, tissue or organism is reduced compared to the innate immune response that results or would have resulted in the absence of introducing said Agent mRNA.   
     
     
         2 . The method of  claim 1 , wherein said Agent mRNA encodes one or more proteins that inhibits the activity of an innate immune effector protein in a signaling pathway mediated by a TLR selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 and TLR10, or a biologically active fragment, analog or variant of any of said effector proteins. 
     
     
         3 . The method of  claim 1 , wherein said Agent mRNA encodes one or more proteins that is a regulator or inhibitor of type I-interferon signaling, induction, or response selected from the group consisting of:
 a) a biologically inactive fragment, mutant, analog or variant or a dominant negative functional inhibitor of TP53, TLR3, TLR4, TLR7, TLR8, RARRES3, IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNK, IFNB1, IL6, TICAM1, TICAM2, MAVS, STAT1, STAT2, EIF2AK2, IRF3, TBK1, CDKN1A, CDKN2A, RNASEL, IFNAR1, IFNAR2, OAS1, OAS2, OAS3, OASL, RB1, ISG15, MX1, IRF9, ISG20, IFIT1, IFIT2, IFIT3, IFIT5, PKR, RIG-1, MDA5, NF-κB, TRIF, Tyk2 and IRF7; and   b) Vaccinia virus B18R protein, Vaccinia virus E3L protein, Vaccinia virus K3L protein, Influenza A virus NS1 protein, human papilloma virus 18 protein E6, human interferon alpha/beta binding proteins, a soluble form of a human interferon alpha receptors, or a biologically active fragment, analog or variant of any of said proteins.   
     
     
         4 . The method of  claim 1 , wherein the cell is a human or animal cell t selected from the group consisting of: a fibroblast cell, a fetal fibroblast, a neonatal fibroblasts, adult fibroblasts, an hematopoietic cell, a B cell, a T cell, a dendritic cell, a macrophage cell, a Langerhans cell, a Kuppfer cell, an artificial APC, a monocyte, mononuclear cells, a keratinocyte cell, a primary keratinocyte, a keratinocyte derived from hair, an adipose cell, an epithelial cell, an epidermal cell, a chondrocyte, a cumulus cell, a neural cell, a glial cell, an astrocyte, a cardiac cell, an esophageal cell, a muscle cell, a melanocyte, and an osteocyte. 
     
     
         5 . The method of  claim 1 , wherein said introducing is into said organism, wherein said organism is a human, has a disease or medical condition comprising an elevated type I IFN-mediated innate immune response. 
     
     
         6 . The method of  claim 5 , wherein said human has psoriasis or systemic lupus erythematosus (SLE). 
     
     
         7 . The method of  claim 1 , wherein said innate immune response that is reduced is caused by introduction of a Foreign Substance that is capable of causing an innate immune response in said cell, tissue or organism by affecting the induction, activity and/or response of an innate immune response pathway in said cell, tissue or organism;
 wherein said Foreign Substance is selected from the group consisting of:   a) Exogenous RNA;   b) Exogenous siRNA or Exogenous miRNA;   c) dsRNA that is transfected into said cell, tissue or organism; and   d) a lipopolysaccharide (LPS) that is introduced to said cell, tissue or organism.   
     
     
         8 . The method of  claim 7 , wherein said Exogenous RNA comprises mRNA, and wherein the expression of said Agent mRNA:
 a) increases the translation of said Exogenous RNA in said cells; and/or   b) decreases the cell toxicity to said cells; and/or   c) increases the survival of the cells.   
     
     
         9 . The method of any  claim 7 , wherein said cell, tissue or organism comprises an antigen-presenting cell selected from the group consisting of: a dendritic cell, a macrophage, a Langerhans cell, a Kuppfer cell, and an artificial APC, from a human or animal patient, and wherein said Foreign Substance that is transfected into said cell is Exogenous mRNA comprising or consisting of one or multiple mRNAs derived from a cancer cell from a human or animal patient by in vitro transcription (IVT) of cDNA generated from substantially all of the mRNA isolated from one or more cancer cells. 
     
     
         10 . The method of  claim 7 , wherein said Foreign Substance comprises Exogenous mRNA encoding a non-defective form of a protein that is defective or lacking in a cell of a human or animal patient, and said method further comprises transfecting said Exogenous mRNA into a cell of said cell, tissue or organism. 
     
     
         11 . The method of  claim 7 , wherein said Foreign Substance is Exogenous mRNA encoding one or more proteins selected from the group consisting of: OCT3/4, SOX2, KLF4, c-MYC, c-MYC(T58A), L-MYC, NANOG, LIN28, SV40 Large-T antigen, hTERT, E-Cadherin, MYOD1, SHH, GLI1, RARγ, LRH1, GLIS1, NURR1, MASH1, LMX1A, BRN2, MYT1L, GATA4, MEF2C, TBX5, HAND2, FOXA1, FOXA2, FOXA3, HNF1α, HNF4α, PAX3 and PAX7. 
     
     
         12 . The method of  claim 1 , wherein, prior to introducing to the cell, tissue or organism said Agent mRNA, the method comprises the step of contacting the cell, tissue, or organism with an effective amount of a protein that is capable of reducing an innate immune response due to said Agent mRNA. 
     
     
         13 . A system comprising:
 a) an Agent mRNA that reduces the innate immune response in a cell that is induced a Foreign Substance, and   b) a Foreign Substance that induces said innate immune response, particularly.   
     
     
         14 . The system of  claim 13 , wherein said Agent mRNA encodes one or more proteins that inhibits the activity of an innate immune effector protein in a signaling pathway mediated by a TLR selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 and TLR10, or a biologically active fragment, analog or variant of any of said proteins. 
     
     
         15 . The system of  claim 13 , wherein said Agent mRNA encodes one or more proteins is selected from the group consisting of:
 a) a biologically inactive fragment, mutant, analog or variant or a dominant negative functional inhibitor of TP53, TLR3, TLR4, TLR7, TLR8, RARRES3, IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNK, IFNB1, IL6, TICAM1, TICAM2, MAVS, STAT1, STAT2, EIF2AK2, IRF3, TBK1, CDKN1A, CDKN2A, RNASEL, IFNAR1, IFNAR2, OAS1, OAS2, OAS3, OASL, RB1, ISG15, MX1, IRF9, ISG20, IFIT1, IFIT2, IFIT3, IFIT5, PKR, RIG-1, MDA5, NF-κB, TRIF, Tyk2, IRF7, or a biologically active fragment, analog or variant of any of said proteins; and   b) Vaccinia virus B18R protein, Vaccinia virus E3L protein, Vaccinia virus K3L protein, Influenza A virus NS1 protein, human papilloma virus 18 protein E6, human interferon alpha/beta binding proteins, a soluble form of a human interferon alpha receptors (e.g., INFAR1, INFAR2), or a biologically active fragment, analog or variant of any of said proteins.   
     
     
         16 . The system of  claim 13 , further comprising a human or animal cell selected from the group consisting of: a fibroblast cell, a fetal fibroblast, a neonatal fibroblast, adult fibroblast, an hematopoietic cell, a B cell, a T cell, a dendritic cell, a macrophage cell, a Langerhans cell, a Kuppfer cell, an artificial APC, a monocyte, a mononuclear cell, a keratinocyte cell, a primary keratinocyte, a keratinocyte derived from hair, an adipose cell, an epithelial cell, an epidermal cell, a chondrocyte, a cumulus cell, a neural cell, a glial cell, an astrocyte, a cardiac cell, an esophageal cell, a muscle cell, a melanocyte, and an osteocyte. 
     
     
         17 . The system of  claim 1 , wherein said Foreign Substance is Exogenous mRNA encoding one or more proteins selected from the group consisting of: OCT3/4, SOX2, KLF4, c-MYC, c-MYC(T58A), L-MYC, NANOG, LIN28, SV40 Large-T antigen, hTERT, E-Cadherin, MYOD1, SHH, GLI1, RARγ, LRH1, GLIS1, NURR1, MASH1, LMX1A, BRN2, MYT1L, GATA4, MEF2C, TBX5, HAND2, FOXA1, FOXA2, FOXA3, HNF1α, HNF4α, PAX3 and PAX7. 
     
     
         18 . The system of  claim 13  further comprising an effective amount of a protein that is capable of reducing an innate immune response due to said Agent mRNA. 
     
     
         19 . A composition comprising:
 a) an Agent mRNA that reduces the innate immune response in a cell that is induced by a Foreign Substance, and   b) a Foreign Substance RNA that induces said innate immune response when transfected into said cell.   
     
     
         20 . The composition of  claim 19 , wherein said Agent mRNA encodes one or more proteins that inhibits the activity of an innate immune effector protein in a signaling pathway mediated by a TLR selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 and TLR10, or a biologically active fragment, analog or variant of any of said proteins. 
     
     
         21 . The composition of  claim 19 , wherein said Agent mRNA encodes one or more proteins that is a regulator or inhibitor of type I-interferon signaling, induction, or response selected from the group consisting of:
 a) a biologically inactive fragment, mutant, analog or variant or a dominant negative functional inhibitor of TP53, TLR3, TLR4, TLR7, TLR8, RARRES3, IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNK, IFNB1, IL6, TICAM1, TICAM2, MAVS, STAT1, STAT2, EIF2AK2, IRF3, TBK1, CDKN1A, CDKN2A, RNASEL, IFNAR1, IFNAR2, OAS1, OAS2, OAS3, OASL, RB1, ISG15, MX1, IRF9, ISG20, IFIT1, IFIT2, IFIT3, IFIT5, PKR, RIG-1, MDA5, NF-κB, TRIF, Tyk2, IRF7; and   b) Vaccinia virus B18R protein, Vaccinia virus E3L protein, Vaccinia virus K3L protein, Influenza A virus NS1 protein, human papilloma virus 18 protein E6, human interferon alpha/beta binding proteins, soluble forms of the human interferon alpha receptors, INFAR1, INFAR2, or a biologically active fragment, analog or variant of any of said proteins.   
     
     
         22 . The composition of  claim 19 , wherein said Foreign Substance is Exogenous mRNA encoding one or more proteins selected from the group consisting of: OCT3/4, SOX2, KLF4, c-MYC, c-MYC(T58A), L-MYC, NANOG, LIN28, SV40 Large-T antigen, hTERT, E-Cadherin, MYOD1, SHH, GLI1, RARγ, LRH1, GLIS1, NURR1, MASH1, LMX1A, BRN2, MYT1L, GATA4, MEF2C, TBX5, HAND2, FOXA1, FOXA2, FOXA3, HNF1α, HNF4α, PAX3 and PAX7. 
     
     
         23 . The composition of  claim 19 , further comprising a human or animal cell selected from the group consisting of: a fibroblast cell, a fetal fibroblast, a neonatal fibroblasts, adult fibroblast, a hematopoietic cell, a B cell, a T cell, a dendritic cell, a macrophage cell, a Langerhans cell, a Kuppfer cell, an artificial APC, a monocyte, mononuclear cells, a keratinocyte cell, a primary keratinocyte, a keratinocyte derived from hair, an adipose cell, an epithelial cell, an epidermal cell, a chondrocyte, a cumulus cell, a neural cell, a glial cell, an astrocyte, a cardiac cell, an esophageal cell, a muscle cell, a melanocyte, and an osteocyte.