Artificial Transcription Factors for Cancer Immunotherapy: This project seeks to develop transcription factor proteins for immunotherapy applications. Our key insight is that ATF proteins can activate intracellular genes of any cell for achieving an appropriate immune function. Our approach uses zinc finger proteins that mediate gene recognition, which we develop through identifying and characterizing successful ATFs with biophysical and biological assays. We are developing two classes of transcription factors: (1) to activate interferon signaling for inducing antitumor inflammation and (2) to activate the expression of low-level endogenous tumor antigens for enhancing immune recognition. We have successfully developed three ATF proteins that are beginning to show antitumor activity in mouse models. We are also launching in-depth mechanistic studies to characterize the biological activity across different species (e.g., human, mouse, etc) and in different cancer cells and other cells.
Synthetic Protein Switches for Expanding the Folding Code: This project seeks to better understand polymorphic protein structures, through the synthesis and characterization of fold-switching proteins with stabilized features. My research group has constructed an automated flow peptide synthesis (AFPS) instrument to gain access to novel proteins with unique structures and properties. We have recently discovered that AFPS enables access to synthetic fold-switching proteins, which pushes the limits of synthesis fidelity and enables the characterization of protein folding at the molecular level. We verify the identity of the products on an in-house LC-MS (Q-TOF) and isolate the purified protein by preparative RP-HPLC. Protein characterization is further achieved through advanced biophysical and biological methods, including circular dichroism (CD), isothermal titration calorimetry (ITC), dot blot and western blot (WB) assays, and enzyme-linked immunosorbent assays (ELISA). We are pursuing NMR spectroscopy studies of these proteins, which is becoming an important for characterizing intricate protein structural details.