SBIR Phase I: Optoelectronic Microplates: Disruptive Optical Stimulation Technology for Drug Discovery Screening Assays

这个小企业创新研究（SBIR）项目的更广泛的影响/商业潜力是开发专门的细胞培养板，可以在动态离子通道药物筛选试验中提供细胞的动态光学刺激。提出的细胞刺激技术有望提高药物发现的效率，并导致更多具有广泛作用机制的有前途的候选药物。由于药物的作用往往取决于药物靶点的功能状态，体外高通量测定必须能够在药物筛选过程中重建不同的功能状态。目前，筛选分析要么从一种功能状态获取数据，要么从异质状态的总体平均值获取数据，这并不代表体内环境。所提出的基于微板的细胞刺激技术将有助于药物发现公司更快地引入更有效的药物，同时降低开发成本，因为所提出的技术可以通过不同的功能状态动态“循环”离子通道药物靶点，同时在动力学分析中评估药物效果。这些微创全光学检测将增加信息含量和增强预测价值。该SBIR一期项目建议开发基于纳米技术的光电微孔板，该微孔板可以作为光控致动器，实现全光药物筛选试验中细胞的动态光学刺激。目前的细胞刺激技术要么需要对细胞进行基因修饰（这会影响药物筛选结果），要么需要电场刺激（这需要专门的仪器，可能会损害细胞）。所提出的技术是一种基于非侵入性光学刺激板的平台，可以在基因和结构完整的细胞上工作，并将与现有的筛选仪器兼容。材料科学领域的研究计划包括石墨烯材料沉积方案的开发以及石墨烯涂层微孔板在无细胞和基于细胞模式下的综合表征。生物研究计划包括开发全光学筛选试验，其中包括石墨烯涂层微孔板，以实现光学刺激。具体来说，目标是专注于l型电压门控钙离子通道的钙成像分析，该通道将通过使用光电板的光照射“循环”通过不同的功能状态。验证过程将包括评估已知状态依赖性钙通道阻滞剂的药理作用。预计光电微孔板将与现有的细胞培养方案和成像仪器兼容。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is to develop specialized cell culture plates that can provide dynamic optical stimulation of cells during kinetic ion channel drug screening assays. The proposed cell stimulation technology is expected to improve the efficiency of drug discovery, and lead to more promising drug candidates with a wide range of mechanisms of actions. Since effects of drugs often depend on functional states of drug targets, in vitro high-throughput assays must be able to re-create different functional states during drug screening. Currently, screening assays acquire the data either from one functional state or an ensemble average of heterogeneous states, which is not representative of in vivo settings. The proposed microplate-based cell stimulation technology will help drug discovery companies to introduce more efficient drugs faster while reducing the development costs, because the proposed technology can dynamically "cycle" ion channel drug targets through different functional states while evaluating drug effects in kinetic assays. These minimally-invasive all-optical assays will have increased information content and enhanced predictive values. This SBIR Phase I project proposes to develop nanotechnology-based optoelectronic microplates that can serve as a light-controlled actuator enabling dynamic optical stimulation of cells for all-optical drug screening assays. Current cell stimulation technologies require either genetic modifications of cells (which can affect the drug screening results) or electric-field stimulation (which requires specialized instrumentation and can damage cells). The proposed technology is a non-invasive optical stimulation plate-based platform that can work on genetically and structurally intact cells, and will be compatible with existing screening instruments. The research plan in the material science area covers the development of protocols for deposition of graphene materials and comprehensive characterization of graphene-coated microplates in cell-free and cell-based modes. The biological research plan includes the development of all-optical screening assays that incorporate graphene-coated microplates for enabling optical stimulation. Specifically, the goal is to focus on calcium imaging assays for L-type voltage-gated calcium ion channels that will be "cycled" through different functional states by light illumination using optoelectronic plates. The validation process will include the evaluation of pharmacological effects of known state-dependent calcium channel blockers. It is anticipated that optoelectronic microplates will be compatible with existing cell culture protocols and imaging instrumentation.