Ion channels are important drug targets-present in every cell, they play key roles in a wide range of physiological processes including the cardiac cycle and neural activity. As a result, unintended drug interactions with ion channels are also of critical importance, requiring the screening of all drug candidates against specific ion channels. Unfortunately, assaying ion channels for pharmaceutical discovery and safety screening is problematic to perform in high throughput because the ion channels must be incorporated into a cell membrane to allow measurement of their ionic transport to determine their functionality. As a result, there are currently no high quality, high throughput assays for ion channel screening. Recent developments of automated patch clamp instrumentation are still over an order of magnitude lower throughput than conventional drug screening for soluble proteins and also require expensive instrumentation, specialized cell lines, and consumables. For existing methods of ion channel screening, there is a large gap in information quality, throughput, and cost. Librede's goal is to develop new technologies that increase the efficiency of early stage pharmaceutical research and development. To this end, Librede is developing an alternative cell-free technology for ion channel screening using artificial cell membranes. Librede's patent pending formulation of cell-free artificial membranes can enable higher throughput and lower consumable costs while requiring less expensive equipment and trained personnel. Librede was founded by UCLA researchers and the inventors of this technology. In Phase I, we measured a 48 membrane array plate simultaneously with a multichannel amplifier and verified that the measurement performance was equal to or exceeded competitive automated patch clamp instruments. In the Phase II work proposed here, we will build on this work by integrating our Phase I instrumentation with fluid handling and motion control hardware to construct an automated workstation for cell-free ion channel screening using Librede's artificial membrane technology. We will consult with a pharmaceutical screening industry automation expert in the design, construction, and operation of our workstation and confirm that it can process and measure Librede's artificial membrane plates similarly to our previous work. To demonstrate its capabilities, we will perform a limited screen of CLIC1, an ion channel implicated in Alzheimer's disease. CLIC1 is particularly difficult to screen with conventional patch clamp, which makes this an ideal proof-of-concept application of our platform. We will work with the Schmidt group at UCLA, who have previously demonstrated the measurement of CLIC1 in artificial membranes. We will run the workstation in full automation mode, measuring ion channel activity as a function of drug concentration for a 72 compound ion channel-targeted drug library. This demonstration is essentially identical to an industry screen and is a major milestone for Librede's development of our artificial membrane technology, which promises an order of magnitude improvement in cost and throughput for ion channel screening.
Librede is developing a novel cell-free ion channel screening platform that has the potential to significantly reduce costs and increase throughput of drug discovery and safety screening for ion channels. We will develop an ion channel measurement workstation that is capable of completely automated processing and measurement of Librede's ion channel plates after the addition of drug compounds modulating the channel activity. We are validating this workstation by screening compounds for activity against the CLIC1 human ion channel which is thought to play an important role in Alzheimer's disease.