Research | Nanoengineering ion channels & Optogenetic immunomodulation 

Our goal is to achieve noninvasive control of cellular signaling through engineering ion channels or other signalling molecules with multidisciplinary approaches including chemical biology, protein engineering and nanotechnology. Specially, we will explore the use of light, magnet and radiowave to remotely control ion channel activities and their downstream effectors.

Developing novel optical tools to interrogate cellular signaling 

The neuroscience field has witnessed the successful use of channelrhodopsin (ChR)-derived optogenetic tools to perturb and observe the activity of excitable neurons at precise locations and times. Nevertheless, ChR lacks ion selectivity and often causes intracellular pH alteration. There is an urgent need to expand the repertoire of light-switchable or optogenetic tools with tailored ion selectivity that can be applied to study cellular signaling in non-excitable cells. Efforts will be directed to further screen calcium-selective ChR variants by taking site-directed mutagenesis and protein engineering approaches. More importantly, built upon our prior findings, we will attempt to engineer the highly calcium-selective ORAI1-STIM1 pathway for non-invasive manipulation of calcium signaling by light. Efforts will also be directed to engineer T cells with novel functionality to kill tumor cells. 


Opto-CRAC | Optical control of calcium signaling and immune response

Converting SOC to LOC: light-operated calcium entry

OptoCRAC | Illuminating calcium signaling with LOV domain

We are installing light sensitivity into signaling nodes to control cellular event by harnessing the power of light. When paired with NIR-stimulable nanoparticles, our optogenetic tools can be further extended to in vivo applications, such as remote control of innate and adaptive immune responses. 

The application of current channelrhodopsin-based optogenetic tools is limited by the lack of strict ion selectivity and the inability to extend the spectra sensitivity into the near-infrared (NIR) tissue transmissible range. We aim to develop an NIR-stimulable optogenetic platform (termed “Opto-CRAC”) that selectively and remotely controls Ca2+ oscillations and Ca2+-responsive gene expression to regulate the function of non-excitable cells, including T lymphocytes, macrophages and dendritic cells. When coupled to upconversion nanoparticles, the optogenetic operation window will be shifted from the visible range to NIR wavelengths to enable wireless photoactivation of Ca2+-dependent signaling and optogenetic modulation of immunoinflammatory responses. 


In a mouse model of melanoma by using ovalbumin as surrogate tumor antigen, Opto-CRAC has been shown to act as a genetically-encoded “photoactivatable adjuvant” to improve antigen-specific immune responses to specifically destruct tumor cells. Our study represents a solid step forward towards the goal of achieving remote control of Ca2+-modulated activities with tailored function.


Similar strategies will be extended to photo-engineer the innate immunity and aid temporal and spatial control of therapeutic T cells in immunotherapies.


Reference:

He L*, Zhang Y*, Ma G*, Tan P*, Li Z, Zang S, Wu X, Jing J, Fang S, Zhou L, Wang Y, Huang Y, Hogan PG, Han G, Zhou Y. Near-infrared photoactivatable control of ca2+ signaling and optogenetic immunomodulation. eLife. 2015 (accepted)


LiMETER |  A novel optical tool to photo-label and manipulate membrane contact sites

Reference:

Jing J*, He L*, Sun A*, Quintana A*, Ding Y*, Ma G, Tan P, Liang X, Zheng X, Chen L, Shi X, Zhang SL, Zhong L, Huang Y, Dong MQ, Walker CL, Hogan PG, Wang Y and Zhou Y. Proteomic mapping of ER-PM junctions identifies STIMATE as a regulator of Ca2+ influxNature Cell Biology. 2015,17:1339-4. 


More optical tools to come |  LiSPIDER-LiNano-LiPoptosis-LiMAG-LiMETERv2-OptoImmune...