Laser Diode for Ion Channel Stimulation

用于离子通道刺激的激光二极管

• 批准号: 7275476
• 负责人: Mikhail I. Nemenov
• 金额: $ 60.96万
• 依托单位: LASMED, LLC
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-15 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7275476
• 关键词: Action Potentials; Analgesics; Asses; Binding; C Fiber; Calcium; Calibration; Capsaicin; Cells; Certification; Class; Computer software; Cultured Cells; Data; Development; Devices; Diagnosis; Electronics; Evaluation; Eye; Family; Fiber; Funding; Gated Ion Channel; Goals; Heating; Human; Image; In Vitro; Invasive; Ion Channel; Lasers; Magnetism; Marketing; Measurement; Measures; Mediating; Microscope; Modeling; Molecular; Monitor; Nerve; Nerve Fibers; Nerve Tissue; Neurons; Nociceptors; Pain; Patients; Phase; Physiologic pulse; Process; Protein Family; Proteins; Protocols documentation; Pulse taking; Rate; Rattus; Ruthenium Red; Shapes; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Standards of Weights and Measures; TRPV1 gene; Temperature; Testing; Tissues; United States Food and Drug Administration; base; capsazepine; concept; desensitization; design; drug discovery; in vivo; patch clamp; prototype; receptor; research study; tool

DESCRIPTION (provided by applicant): The goal of this Phase II SBIR project is to develop and validate a laser diode stimulator to assess heat-sensitive Transient Receptor Potential (TRP) ion channels for in vitro and in vivo studies, using patch clamp recording and calcium imaging and compound action potential recording, respectively. This laser will be very useful for new drug discovery and evaluation, as there is currently no commercially available tool of similar capabilities. Evaluation of TRP family proteins can be useful in describing the potential utility of putative analgesic drugs. The ability to selectively (by temperature thresholds), reliably and reproducibly activate TRPs may also provide unique utility in understanding the mechanisms of antagonist action (binding) and provide additional information about mechanisms of interaction of different TRP proteins in cells and nerve tissue. There is also a potential utility of being able to differentially evaluate pain mediated by TRPV1 or TRPV2 proteins in the diagnosis and monitoring of progression of pain patients. Thus, it is our overall goal to establish stimulation protocols for use with a laser stimulator for in vitro and in vivo activation of heat gated ion channels. Laser stimulation is used for selective activation of C and A delta fibers in humans and rats (in our parallel projects), allowing reproducible protocols to test putative analgesic, from cell culture to humans. Thus, we hypothesize that the laser ion channel stimulator will prove safe and reliable action and will meet the FDA requirements for a laser device. To do this, we will develop an additional electronic circuit that allows the blockade of laser emission through the microscope, protecting eyes from any damage. We will also submit the laser and its components for certification of electrical and magnetic interference standards, and make any device changes that are necessary to meet the requirements. In addition, we will develop a protocol of direct temperature measurement of laser-induced temperature and modify the device to allow a tunable rate of heating. We will also insure the compatibility of the laser with calcium imaging and will continue to test the laser with TRPV1 DRG and HEK293 cells in order to study sensitization/desensitization processes. To do this we will apply brief and prolonged laser pulses and capsazepine. We will test the concept of TRP-associated activation of C and A delta fibers by direct laser activation of nerve fibers and application of the TRPV1 and TRPV2 antagonists capsazepine and ruthenium red. In vitro and in vivo protocols of non-invasive repeatable and reproducible activation of heat gated ion channels (TRP) can be useful tool for discovery and developing of analgesics. The ability to reliably, activate TRPV1/TRPV2 proteins in cell cultures and in tissue (in vivo) may also provide more accurate prediction of the efficacy of pain treatments based on TRP's antagonists. The brief heating allow investigating of molecular bases of activation of heat gated ion channels. There is currently no commercially available tool of similar capabilities on the market.

描述（由申请人提供）：这个二期SBIR项目的目标是开发和验证激光二极管刺激器，以评估热敏瞬时受体电位（TRP）离子通道，用于体外和体内研究，分别使用膜片钳记录和钙成像和复合动作电位记录。这种激光器将对新药发现和评估非常有用，因为目前还没有类似能力的商用工具。TRP家族蛋白的评估可用于描述推定的镇痛药物的潜在效用。选择性地（通过温度阈值）、可靠地和可重复地激活TRP的能力也可能为理解拮抗剂作用（结合）的机制提供独特的实用工具，并提供关于细胞和神经组织中不同TRP蛋白相互作用机制的额外信息。在疼痛患者的诊断和监测中，TRPV1或TRPV2蛋白介导的疼痛也有潜在的效用。因此，我们的总体目标是建立与激光刺激器一起用于体外和体内热门控离子通道激活的刺激方案。激光刺激用于人类和大鼠的C和A δ纤维的选择性激活（在我们的平行项目中），允许可重复的方案来测试从细胞培养到人类的推定镇痛药。因此，我们假设激光离子通道刺激器将证明安全可靠的行动，并将满足FDA对激光设备的要求。为了做到这一点，我们将开发一个额外的电子电路，可以阻止激光通过显微镜发射，保护眼睛免受任何伤害。我们还将提交激光器及其组件进行电磁干扰标准认证，并进行必要的设备更改以满足要求。此外，我们将开发一种激光诱导温度的直接温度测量方案，并修改设备以允许可调的加热速率。我们还将确保激光与钙成像的兼容性，并将继续在TRPV1 DRG和HEK293细胞上测试激光，以研究增敏/脱敏过程。为了做到这一点，我们将使用短暂和长时间的激光脉冲和辣椒素。我们将通过直接激光激活神经纤维和应用TRPV1和TRPV2拮抗剂辣椒平和钌红来测试trp相关的C和A δ纤维激活的概念。热门控离子通道（TRP）的非侵入性、可重复和可复制激活的体外和体内研究方案可作为发现和开发镇痛药的有用工具。在细胞培养物和组织（体内）中可靠地激活TRPV1/TRPV2蛋白的能力也可以更准确地预测基于TRP拮抗剂的疼痛治疗效果。短暂的加热可以研究热门控离子通道激活的分子基础。目前市场上还没有类似功能的商业工具。