Cell Factory for Small Molecule Inducible Delivery of Low Dose IL-2
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Abstract
Autoimmune and inflammatory diseases are characterized by insufficient regulatory T cells (Treg), which play a central role in maintaining peripheral tolerance, preventing autoimmunity, and limiting chronic inflammation. Treg cells depend on IL-2 for survival and maintenance of their suppressive functions. IL-2, however, not only controls the proliferation, homeostasis, and immunosuppressive function of regulatory T cells, but also promotes the cytotoxic function of pro-inflammatory T cells. Clinical studies demonstrated that low doses of IL-2, delivered through subcutaneous injections, increase the number of Treg in T1D, GVHD, and SLE patients⁵. Even though effective, there are some areas of improvement: medication absorption is slower than vein injection or oral administration, severe pain and redness in site location, tissue damage when injections are given in the same site frequently. We will engineer this genetic control scheme by using a transcription factor, EKRAB, to repress the production of IL-2. This transcription factor can then be sequestered using an FDA approved small molecule drug, Erythromycin, to induce production of IL-2 in a dose-responsive manner. We will integrate the genetic circuitry through piggyBac transposition into ARPE-19 cells and perform selection using Puromycin and Zeocin antibiotics. We will test different ratios between EKRAB plasmid and IL-2 plasmid copy number integrations to find the optimal ratio which reduces leaky expression, and evaluate the systems dose-response behavior to Erythromycin through flow cytometry using fluorophores co-expressed with EKRAB (iRFP) and IL-2 (GFP).
Sponsored by
NSF's Nano Research Experience for Teachers at Rice University- NSF EEC-1406885
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Office of STEM Engagement, Rice University, Houston, TX