Therapeutic crosslinking of cytokine receptors

Abstract

The present invention relates to a compound comprising (1) a first binding moiety that binds to an Interleukin 4 receptor; and (2) a second binding moiety that binds to a cytokine receptor (e.g., an Interleukin 10 receptor, Interleukin 13 receptor, interleukin 27 receptor, interleukin 33 receptor, or transforming growth factor beta 1/2 receptor). Compounds of the disclosure cluster or crosslink the IL4 receptor and a cytokine receptor, and surprisingly, elicit unique responses in the nervous system, including unique signaling and gene expression profiles. The signaling and gene expression profiles generated IL4-cytokine fusion proteins of the disclosure are distinct from those observed in response to the combination of IL4 and the cytokine, and contribute to the superior therapeutic effects over the combination of the component parts.

1 - 256 . (canceled) 
     
     
         257 . A method of normalizing a response of a sensitized human neuron to a stimulus, the method comprising contacting the sensitized human neuron with a compound that comprises an interleukin 4 polypeptide attached to a cytokine, wherein the compound is present at a concentration of at least 1 pM during the contacting, wherein the response of the sensitized human neuron to the stimulus is normalized relative to before the contacting. 
     
     
         258 . The method of  claim 257 , wherein the compound normalizes the response of the sensitized human neuron to the stimulus to a greater degree than a ten-fold higher concentration of the interleukin 4 polypeptide and the cytokine that are not attached to each other. 
     
     
         259 . The method of  claim 257 , wherein the compound is present at a concentration of at most 100 μM during the contacting. 
     
     
         260 . The method of  claim 257 , wherein the human neuron has been sensitized by a pro-inflammatory mediator. 
     
     
         261 . The method of  claim 260 , wherein the pro-inflammatory mediator comprises a prostaglandin. 
     
     
         262 . The method of  claim 260 , wherein the pro-inflammatory mediator comprises an inflammatory cytokine. 
     
     
         263 . The method of  claim 257 , wherein the stimulus induces neuronal action potential firing. 
     
     
         264 . The method of  claim 257 , wherein the stimulus comprises a pro-inflammatory mediator, a drug, a toxicant, or a chemical stimulus. 
     
     
         265 . The method of  claim 257 , wherein the response to the stimulus is not significantly altered in a non-sensitized human neuron contacted with the compound. 
     
     
         266 . The method of  claim 257 , wherein the response comprises a magnitude of a calcium flux response. 
     
     
         267 . The method of  claim 257 , wherein the response comprises depolarization. 
     
     
         268 . The method of  claim 257 , wherein the response comprises action potential frequency. 
     
     
         269 . The method of  claim 257 , wherein the response comprises a signal transduction response. 
     
     
         270 . The method of  claim 257 , wherein the response is normalized as demonstrated by a lower magnitude of a calcium flux response to a predetermined amount of capsaicin. 
     
     
         271 . The method of  claim 257 , wherein the contacting decreases ectopic neuronal activity relative to before the contacting. 
     
     
         272 . A method of modulating signaling in a human nervous system cell, the method comprising contacting the human nervous system cell with a compound that comprises an interleukin 4 polypeptide attached to a cytokine under conditions that induce heterologous clustering of a receptor for the interleukin 4 polypeptide and a receptor for the cytokine. 
     
     
         273 . The method of  claim 272 , wherein the heterologous clustering of the receptor for the interleukin 4 polypeptide and the receptor for the cytokine is as determined by a protein proximity assay. 
     
     
         274 . The method of  claim 272 , wherein the heterologous clustering of the receptor for the interleukin 4 polypeptide and the receptor for the cytokine is as determined by a proximity ligation assay 
     
     
         275 . The method of  claim 272 , wherein the receptor for the interleukin 4 polypeptide and the receptor for the cytokine are clustered within 51 nm of each other. 
     
     
         276 . The method of  claim 272 , wherein the heterologous clustering of the receptor for the interleukin 4 polypeptide and the receptor for the cytokine is not induced by equivalent concentrations of the interleukin 4 polypeptide and the cytokine that are not attached to each other. 
     
     
         277 . The method of  claim 272 , wherein the heterologous receptor clustering reduces sensitization of the human nervous system cell. 
     
     
         278 . The method of  claim 272 , wherein the heterologous receptor clustering protects the human nervous system cell against neurotoxicity as determined by a neurite outgrowth assay. 
     
     
         279 . The method of  claim 272 , wherein the heterologous receptor clustering protects the human nervous system cell against neurotoxicity as determined by an intraepidermal nerve fiber density assay. 
     
     
         280 . The method of  claim 272 , wherein the heterologous receptor clustering decreases ectopic neuronal activity. 
     
     
         281 . The method of  claim 272 , wherein the cytokine is a regulatory cytokine. 
     
     
         282 . The method of  claim 272 , wherein the heterologous receptor clustering induces an altered kinomic profile compared to a human nervous system cell that is contacted with equivalent amounts of the interleukin 4 polypeptide and the cytokine that are not attached to each other as determined by kinase array profiling. 
     
     
         283 . The method of  claim 282 , wherein altered kinomic profile comprises altered JAK-STAT signaling. 
     
     
         284 . The method of  claim 283 , wherein a level of activity of JAK1, c-Kit, or c-Met is modulated.