Self-Regulating Device for Modulating Inflammation

Abstract

A bioreactor is provided which contains cells capable of producing cytokine inhibitors in response to cytokines, in a manner regulated by the local or systemic milieu of an individual patient and predicted by mechanistic computational simulations. The bioreactor transfers the cytokine inhibitors to a patient in need of control of the inflammation process as part of a disease or condition in the patient, such as sepsis, trauma, traumatic brain injury, or wound healing. Related methods also are provided.

1 . A bioreactor comprising a compartment comprising cells comprising a chimeric gene comprising a response element operably linked to a sequence encoding a cytokine or an inhibitor of a cytokine, in which the response element causes expression of the cytokine or inhibitor of the cytokine when the cells are contacted with the cytokine, the bioreactor comprising a selectively permeable membrane in contact with the cells. 
     
     
         2 . The bioreactor of  claim 1 , in which the gene expresses a cytokine inhibitor of one of TNF, IL-1, TGF-β1 and IL-6. 
     
     
         3 . The bioreactor of  claim 1 , in which the gene expresses an inhibitor of a cytokine selected from the group consisting of soluble TNF receptor, IL-1 receptor agonist, TGF-β1 LAP (latency-associated peptide) and an IL-6Ralpha/gp130 fusion protein. 
     
     
         4 . The bioreactor of  claim 1 , in which the selectively permeable membrane is a selectively-permeable hollow fiber. 
     
     
         5 . The bioreactor of  claim 1 , wherein the compartment comprising the cells comprises a vessel having a selectively permeable wall. 
     
     
         6 . The bioreactor of  claim 1 , wherein the cells are xenogeneic, syngeneic, allogeneic, or autologous cells to a patient. 
     
     
         7 . The bioreactor of  claim 1 , wherein the cells are transfected or transduced hepatocytes or a hepatocyte cell line. 
     
     
         8 . The bioreactor of  claim 7  in which the cells are a hepatocyte cell line. 
     
     
         9 . The bioreactor of  claim 8 , in which the hepatocyte cell line is HepG2. 
     
     
         10 . The bioreactor of  claim 1 , further comprising a cell comprising a sequence encoding a fluorescent protein that either is:
 a. operably linked to the response element and the sequence encoding the cytokine or inhibitor of the cytokine is attached to and in frame with the sequence encoding the fluorescent protein and a self-cleaving polypeptide sequence between the sequence encoding the cytokine or inhibitor of the cytokine and the sequence encoding the fluorescent protein; or   b. under control of a second response element that causes expression of the fluorescent protein when the cells are contacted with the cytokine.   
     
     
         11 . The bioreactor of  claim 1 , in which one or more of the cytokines or inhibitors of cytokines encoded by the one or more non-native inducible genes comprises a fluorescent tag. 
     
     
         12 . The bioreactor of  claim 1 , in which the compartment comprising the cells comprises at least one wall that is the selectively permeable membrane. 
     
     
         13 . The bioreactor of  claim 12  in which the compartment comprises a plurality of selectively permeable hollow fibers passing through the compartment through which one or both of a gas and a fluid comprising nutrients for the cells can be passed. 
     
     
         14 . The bioreactor of  claim 1 , in which the compartment comprising the cells comprises a plurality of selectively permeable hollow fibers passing through the compartment in which the plurality of hollow fibers are fluidly connected to a plasma or blood circulation system in which blood or plasma from the patient can be circulated through the hollow fibers and into a patient. 
     
     
         15 . A method of modulating wound healing, sepsis, trauma, or traumatic brain injury (TBI), comprising, contacting a bodily fluid of a patient with the selectively permeable membrane of the bioreactor of  claim 1  such that a cytokine in the bodily fluid can pass through the selectively permeable membrane and a cytokine or cytokine inhibitor produced by the cells can pass into the bodily fluid, and returning the bodily fluid to the patient. 
     
     
         16 . The method of  claim 15 , in which the cells are selected by use of a computer model of an inflammatory response characteristic of a disease or condition in the patient. 
     
     
         17 . The method of  claim 15 , comprising modeling inflammation associated with sepsis and determining one or more cytokines to inhibit or produce to control inflammation in the patient associated with sepsis. 
     
     
         18 . The method of  claim 17 , comprising determining levels of one or more cytokines in the patient and modeling inflammation using the one or more levels of cytokines in the patient and determining a cytokine level to be controlled in the patient to determine a chimeric gene construct to place in the bioreactor based on an outcome of the modeling. 
     
     
         19 . The method of  claim 18 , comprising modeling inflammation associated with TBI and determining one or more cytokines to inhibit or produce to control inflammation in the patient associated with TBI. 
     
     
         20 . The method of  claim 19 , comprising determining levels of one or more cytokines in the patient and modeling inflammation using the one or more levels of cytokines in the patient and determining a cytokine level to be controlled in the patient to determine a chimeric gene construct to place in the bioreactor based on an outcome of the modeling. 
     
     
         21 . The method of  claim 15 , in which the patient is a TBI patient. 
     
     
         22 . The method of  claim 21 , in which one or both of an inhibitor of TNF and an inhibitor of IL-6 are produced by the cells. 
     
     
         23 . The method of  claim 15 , in which the gene expresses an inhibitor of a cytokine selected from the group consisting of soluble TNF receptor, IL-1 receptor agonist, and TGF-β1 LAP (latency-associated peptide). 
     
     
         24 . The method of  claim 15 , in which the cells comprise one or more genes that express an inhibitor of one or both of TNF and IL-1α or IL-1β. 
     
     
         25 . The method of  claim 15 , in which the cells comprise one or more genes that express one or both of soluble TNF receptor and IL-1 receptor agonist. 
     
     
         26 . The method of  claim 15 , in which the compartment comprising the cells and comprises a plurality of selectively permeable hollow fibers passing through the compartment in which the plurality of hollow fibers are fluidly connected to a plasma or blood circulation system in which blood or plasma from the patient is circulated through the hollow fibers and into the patient. 
     
     
         27 . The method of  claim 15 , in which the compartment comprising the cells has at least one wall that is the selectively permeable membrane, in which the selectively permeable membrane is placed in contact with a wound on the patient or a bodily fluid in situ in the patient. 
     
     
         28 . The method of  claim 24 , the compartment comprising a plurality of selectively permeable hollow fibers passing through the compartment through which one or both of a gas and a fluid comprising nutrients for the cells is passed.