Novel nanotechnology-based optical stimulation platform for predictive cardiotoxicity assessment

基于新型纳米技术的光刺激平台，用于预测心脏毒性评估

基本信息

批准号：
9347619
负责人：
ALEX SAVTCHENKO
金额：
$ 32.5万
依托单位：
NANOTOOLS BIOSCIENCE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9347619
关键词：
Academia Achievement Action Potentials Address Adoption Agreement Arrhythmia Benchmarking Biological Biological Assay Calcium Cardiac Cardiac Myocytes Cardiotoxicity Cell Culture Techniques Cell membrane Cells Clinical Communities Custom Data Deposition Detection Development Drug Approval Drug Costs Drug Evaluation Drug Prescriptions Dyes Electricity Ensure Evaluation Event Experimental Models Frequencies Gated Ion Channel Geometry Glycocalyx Goals Heart Human Image Imaging Device In Vitro Industry Industry Standard International Ion Channel Ion Channel Gating Laboratories Light Lighting Membrane Potentials Methods Microscopy Modification Monitor Nanotechnology Optics Performance Pharmaceutical Preparations Phenotype Physiological Preclinical Drug Evaluation Predictive Value Procedures Process Property Protocols documentation Public Health Publishing Quality Control Risk Science Signal Transduction Solvents Source Stem cells Surface Properties Surrogate Markers System Technology Temperature Testing Time Validation base biomaterial compatibility cost culture plates data acquisition design drug candidate drug development drug discovery fluorophore graphene human stem cells in vitro Assay induced pluripotent stem cell instrument instrumentation light intensity novel novel therapeutics photonics pre-clinical preclinical study public health relevance safety study screening tool voltage

项目摘要

ABSTRACT The estimated average industry cost per new prescription drug approval currently now exceeds $2.5 billion. One of the critical factors contributing to the rising costs of drug development is a non-optimal focus of pre- clinical studies for detecting potential cardiac liabilities of novel drug candidates. Recognizing this fact, the biomedical community has recently proposed a new paradigm for cardiac safety studies: the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative. It includes a shift from over-simplified hERG-focused cardiotoxicity screening to evaluation of drug effects on multiple ion channels contributing to action potential in human stem cell-derived cardiomyocytes (CMs). Optical cell-based drug screening assays that monitor the dynamics of intracellular calcium concentrations and the cell membrane potential in stem cell-derived CMs are well-suited for detecting drug candidates with cardiac liabilities. Since drug-induced effects are often dependent on the activation state of cells, it is crucial to be able to stimulate CMs during drug screening. We propose to address this need by offering a revolutionary nanotechnology-based platform for non-invasive optical stimulation of CMs. We designed this optoelectronic platform by taking advantage of the unique properties of graphene, a “wonder material of the 21th century”. These properties ensure excellent biocompatibility of graphene-coated substrates, the ability to efficiently convert light into electricity, and compatibility with optical detection methods. Our preliminary results demonstrate that light can induce fast and reversible changes in contractile and electrical activity of stem cell-derived CMs cultured on graphene-coated substrates. Furthermore, we confirmed that optical stimulation via G-coated substrates can enable all-optical evaluation of use-dependent drug effects on the CM activity. To streamline the customer adoption of our graphene-based optical stimulation platform and incorporation it in into optical drug-screening assays, we are proposing to perform the following activities: a) developing a procedure for graphene deposition into cell culture multi-well plates, following by manufacturing of high-quality graphene-coated plates; b) optimizing the various parameters of voltage and calcium all-optical assays (including a dual-light stimulation protocol) on a customized microscopy system; c) transferring optimized all-optical assays to well-regarded high-through and high-content imaging instruments and performing screening of multiple benchmark compounds on optically stimulated CMs. We expect that optical stimulation of CMs via graphene- based substrates will dramatically increase the predictive value of cardiotoxicity screening assays. The proposed all-optical assays will be instrumental in correctly detecting genuine pro-arrhythmia risks early in the drug discovery process, thus reducing the spending and drug development time.

抽象的目前估计每个新处方药批准的平均行业成本超过 25 亿美元。导致药物开发成本上升的关键因素之一是预审重点不理想。认识到这一事实，检测新候选药物潜在心脏负担的临床研究。生物医学界最近提出了心脏安全研究的新范式：综合研究体外促心律失常测定 (CiPA) 计划包括从过度简化的 hERG 心脏毒性转变。筛选评估药物对多个离子通道的影响，有助于人体干细胞的动作电位细胞源性心肌细胞 (CM) 的光学细胞药物筛选分析，可监测心肌细胞的动态。干细胞衍生的 CM 中的细胞内钙浓度和细胞膜电位非常适合检测具有心脏负担的候选药物，因为药物引起的作用通常取决于细胞的激活状态，在药物筛选过程中能够刺激 CM 至关重要。我们建议通过提供一个革命性的基于纳米技术的非侵入性平台来满足这一需求我们利用独特的优势设计了这个光电平台。石墨烯是“21世纪的神奇材料”，这些特性确保了其卓越的性能。石墨烯涂层基材的生物相容性、有效将光转化为电的能力，以及与光学检测方法的兼容性我们的初步结果表明光可以快速诱导。在石墨烯涂层上培养的干细胞衍生的 CM 的收缩和电活动的可逆变化此外，我们证实通过 G 涂层基底进行光刺激可以实现全光学。评估依赖于使用的药物对 CM 活性的影响。简化客户对我们基于石墨烯的光刺激平台的采用并将其纳入在光学药物筛选分析中，我们建议开展以下活动：a) 开发将石墨烯沉积到细胞培养多孔板中的程序，然后制造高质量的石墨烯 b) 优化电压和钙全光学测定的各种参数（包括双光刺激方案）在定制的显微镜系统上；c）传输优化的全光学测定使用备受推崇的高通量和高内涵成像仪器，并对多个光刺激 CM 的基准化合物我们期望通过石墨烯对 CM 进行光刺激。基底物将显着提高心脏毒性筛选测定的预测价值。全光学检测将有助于在药物早期正确检测真正的促心律失常风险发现过程，从而减少了支出和药物开发时间。