Immunotherapy represents a recent breakthrough in cancer treatment fueled by the accelerating mechanistic understanding of how transformed cells subvert our immunosurveillance. Therefore, developing therapeutic strategies that enhance systemic immunosurveillance in a controllable manner has become one of the major interests in this field. Among different agents, small molecule activators (agonists) of the cGAS-STING pathway have recently attracted attention because they are expected to synergize with immunotherapies and enhance anti-cancer immune response by upregulating the interferon response. We previously identified a compound, BDW568, that was shown to activate the interferon pathway in a STING-dependent manner. Surprisingly, follow-up studies demonstrated that cGAS and STING are not direct target(s) of BDW568. Therefore, observed phenotype suggests that BDW568 acts either through (1) binding to an unknown regulator of the cGAS-STING pathway, or (2) by generating an unknown signaling molecule independent of 2?,3?-cGAMP, the endogenous STING agonist. In the proposed study we will identify the cellular target of BDW568 responsible for the previously discovered phenotype using two orthogonal strategies. In the first approach, we will synthesize BDW568-based photoaffinity probes and use them to label any protein that binds to BDW568. Labeled target(s) will be identified by a whole-cell lysate pulldown experiments coupled with mass spectrometry (MS). In the second approach, we will use CRISPR-Cas9 to individually knock out all the genes that are known to interact with STING or associate with interferon pathways. The knockout of the BDW568 target should demonstrate either resistance to the compound or elevation of the basal interferon level without BDW568 through STING. Any target candidate that emerges from either one of these approaches will be rigorously validated according to the standards of the field to establish that observed phenotype is due to on-target engagement. Successful completion of proposed studies will expand the target space within cGAS-STING pathway. These insights will lead to additional opportunities for developing adjuvant immunotherapies as well as expand our understanding of innate immune response in mammalian cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Exploratory Grants (P20)
Project #
5P20GM103638-09
Application #
10242612
Study Section
Special Emphasis Panel (ZGM1)
Program Officer
Davani, Behrous
Project Start
2012-07-15
Project End
2022-06-30
Budget Start
2020-07-01
Budget End
2021-06-30
Support Year
9
Fiscal Year
2020
Total Cost
Indirect Cost
Name
University of Kansas Lawrence
Department
Type
DUNS #
076248616
City
Lawrence
State
KS
Country
United States
Zip Code
66045
Field, Thomas M; Shin, Mimi; Stucky, Chase S et al. (2018) Electrochemical Measurement of Dopamine Release and Uptake in Zebrafish Following Treatment with Carboplatin. Chemphyschem 19:1192-1196
McGill, Jodi L; Kelly, Sean M; Kumar, Pankaj et al. (2018) Efficacy of mucosal polyanhydride nanovaccine against respiratory syncytial virus infection in the neonatal calf. Sci Rep 8:3021
Waters, Renae; Alam, Perwez; Pacelli, Settimio et al. (2018) Stem cell-inspired secretome-rich injectable hydrogel to repair injured cardiac tissue. Acta Biomater 69:95-106
Saylor, Rachel A; Lunte, Susan M (2018) PDMS/glass hybrid device with a reusable carbon electrode for on-line monitoring of catecholamines using microdialysis sampling coupled to microchip electrophoresis with electrochemical detection. Electrophoresis 39:462-469
Zhu, Qingfu; Heon, Mikala; Zhao, Zheng et al. (2018) Microfluidic engineering of exosomes: editing cellular messages for precision therapeutics. Lab Chip 18:1690-1703
Pacelli, Settimio; Basu, Sayantani; Berkland, Cory et al. (2018) Design of a cytocompatible hydrogel coating to modulate properties of ceramic-based scaffolds for bone repair. Cell Mol Bioeng 11:211-217
Wessinger, Carolyn A; Kelly, John K; Jiang, Peng et al. (2018) SNP-skimming: A fast approach to map loci generating quantitative variation in natural populations. Mol Ecol Resour 18:1402-1414
Zhang, Peng; Crow, Jennifer; Lella, Divya et al. (2018) Ultrasensitive quantification of tumor mRNAs in extracellular vesicles with an integrated microfluidic digital analysis chip. Lab Chip 18:3790-3801
Klaus, Jennifer R; Deay, Jacqueline; Neuenswander, Benjamin et al. (2018) Malleilactone Is a Burkholderia pseudomallei Virulence Factor Regulated by Antibiotics and Quorum Sensing. J Bacteriol 200:
Abisado, Rhea G; Benomar, Saida; Klaus, Jennifer R et al. (2018) Bacterial Quorum Sensing and Microbial Community Interactions. MBio 9:

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