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1. A light inducible gene activation system toward controllable cell‐based therapeutics
Author(s) -
Huang Ziliang,
Wu Yiqian,
Pan Yijia,
Allen Molly,
Chang YaJu,
Chien Shu,
Wang Yingxiao
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.804.62
Subject(s) - hek 293 cells , jurkat cells , cytotoxicity , microbiology and biotechnology , cancer immunotherapy , optogenetics , cancer cell , cell culture , cancer research , cell , chemistry , immunotherapy , biology , t cell , cancer , immune system , immunology , biochemistry , in vitro , genetics , neuroscience
T cells engineered to express chimeric antigen receptors (CARs) on cell surface can recognize and engage with target cancer cells with redirected specificity. This CAR T cell‐mediated immunotherapy is under intensive investigation as a revolutionary therapy for cancer. However, adverse effects have been reported, including on‐target off‐tumor cytotoxicity, cytokine release syndrome and tumor lysis syndrome. One of the solutions is to engineer on/off‐switchable cells to achieve high‐precision control over their activation. Optogenetics utilizing genetically encoded dimerizers has been applied to remotely activate gene expression. Here, we developed a new blue light‐mediated l ight‐ i nducible n uclear t ranslocation a nd d imerization (LINTAD) system for gene regulation to control cell activation, by integrating a LOV2‐based light‐inducible nuclear localization signal with CRY2‐CIB1 dimerization. We demonstrated light‐controllable gene expression and functional modulation in HEK 293T and Jurkat T cell lines. We further applied the LINTAD system in primary human CAR T cells and showed that the light‐stimulated cells possessed a significantly stronger cytotoxicity against target cancer cells than the non‐stimulated ones. We also extended the LINTAD system to activate the expression of the TNF‐related apoptosis‐inducing ligand (TRAIL) in HEK 293T cells, and observed a gain of cytotoxicity upon light stimulation. Therefore, our newly developed LINTAD system can serve as an efficient and general tool to remotely and non‐invasively control gene activation in live cells for therapeutic applications. Support or Funding Information This work is supported by grants from NIH HL121365, GM125379 (S.C., Y. Wang), CA204704, CA209629, NSF CBET1360341, DMS1361421 (Y. Wang), and UC San Diego. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .