Prediction of computational pathway circuits
Described herein are methods of making recombinant cells using an in silico-generated pathway map of an endogenous pathway in a cell of interest to determine, in silico, a predicted effect of incorporating a recombinant nucleic acid into the endogenous pathway; and based on the predicted effect, incorporating the recombinant nucleic acid into the endogenous pathway in the cell of interest such that activation of the endogenous pathway regulates expression of the recombinant nucleic acid.
1. A method for making a recombinant cell, the method comprising:
(i) generating, in silico via PARADIGM, a pathway map of an endogenous IL-8 signaling pathway and a HIF-1a signaling pathway in a cell of interest;
(ii) determining, in silico, an expected effect of incorporating a recombinant nucleic acid on pathways of interest within the cell, wherein the recombinant nucleic acid comprises a promotor that is activated upon sensing IL-8 and HIF-1a signaling, and wherein the promotor controls expression of a sequence encoding a therapeutic molecule; and
(iii) transfecting the cell of interest with the recombinant nucleic acid, thereby making a recombinant cell that expresses the therapeutic molecule only upon IL-8 and HIF-1a signaling.
2. The method of claim 1 , wherein the recombinant cell is a recombinant therapeutic cell.
3. The method of claim 1 , wherein the therapeutic molecule is selected from the group consisting of a pro-apoptotic protein, a therapeutic antibody, a chimeric antigen receptor, an antisense RNA, an immune stimulating cytokine, a chemokine, a cytotoxic protein, an immunostimulating protein, a suicide gene, and a sodium iodide symporter (NIS).
4. The method of claim 1 , wherein the recombinant cell is a recombinant sensor cell and the recombinant nucleic acid encodes a sensor molecule that senses a molecule of interest.
5. The method of claim 4 , wherein the sensor molecule senses cell density, pH, hypoxia, radio signal, MRI, heat, presence of a molecule of interest, or concentration of a molecule of interest.
6. The method of claim 4 , wherein the molecule of interest is a cytokine, a chemokine, a metabolite, an exosome, an enzyme, a sugar, an intracellular component, a soluble checkpoint inhibitor, a signaling factor, a virus, a yeast cell, or a bacterial cell.
7. The method of claim 1 , wherein the recombinant nucleic acid is incorporated into the cell of interest if the expected effect would not impact one or more additional endogenous pathways in the cell.
8. The method of claim 1 , wherein the recombinant nucleic acid is incorporated into the cell of interest if the expected effect would have a large impact on one or more additional endogenous pathways in the cell.