Asthmatic bronchoconstriction and hypertensive vasoconstriction are extremely common disease states in which excessive contractile cellular forces directly contribute to the pathophysiology. Existing treatments for these diseases, which affect 25 million and 75 million Americans, respectively, have severe side-effects, become desensitized over prolonged use, or lack efficacy altogether. In particular, LABAs used in asthma management carry a ?black-box? warning, and 15-20% of hypertensive patients require >3 drugs to control blood pressure. Despite understanding the role of cellular force in these scenarios, drug developers have lacked the drug discovery tools that directly target this critically important phenotype. Instead, many new drug development efforts continue to focus on known pathways. Clearly, there is a significant clinical unmet need in treating resistant asthma and hypertension, and there are large associated (>$20B) markets worldwide. Specifically, there is need to develop new classes of drugs with molecular mechanisms of action that are orthogonal to existing therapies that promote smooth muscle cell relaxation causing bronchodilation or vasodilation. Forcyte Biotechnologies is an early-stage bio-pharmaceutical company incubating at UCLA that is leveraging a microtechnology known as FLECS ? a high-throughput screening (HTS) platform that measures contractility of single-cells in a 384-wellplate format ? to identify and bring to market new compound classes that act on force-generating pathways within cells. This is the first and only reported assay that obtains functional force generation data for single cells, at HTS scales. Our initial programs will focus on treatment resistant asthma and hypertension, but can extend to other diseases associated with abnormal cellular force. In this proposal, we seek to implement novel multiplexing strategies to extend FLECS?s screening bandwidth from 1 to 8 cell types simultaneously. Specifically, we will exploit the single cell nature of our platform, and combine cell patterning with cell-barcoding to enable discrimination of individual cell types from a large mixed population in each well on our well- plates. We will assess the robustness and separation of the multiplexed signals and develop protocols to maximize signal-to-noise and reduce both optical and biological cross-talk. Completion of our proposed aims will enrich the data generated in screens and substantially reduce the cost per data point. These enhancements will lay the foundation for successful phase II screens of a proprietary 200,000-compound library to identify potential new therapeutics for asthma and hypertension.
In the proposed work, we will continue development of the first high-throughput single-cell contractility screening platform, based on fluorescently-labeled elastomeric contractible surfaces (?FLECS?), as a means for bolstering the pharmaceutical pipeline for asthma and hypertension therapeutics. FLECS currently integrates with 384-wellplate formats to facilitate high-throughput screening of compounds that act on cellular contractility. This proposal aims to implement novel spatial and temporal multiplexing strategies to extend FLECS?s screening bandwidth to 8 cell types simultaneously to i) enrich data generation ii) substantially reduce cost per data point and 3) reduce time-to-market for potential therapeutics. By the conclusion of this Phase I, the platform will be ready to screen a proprietary 200,000-compound library for compounds targeting airway and vascular smooth muscle cell contractility in a follow-on Phase II SBIR.
| Noordraven, Ernst L; Wierdsma, André I; Blanken, Peter et al. (2018) Medical and social costs after using financial incentives to improve medication adherence: results of a 1 year randomised controlled trial. BMC Res Notes 11:655 |
