Multiplexed single-cell contractility assay to develop therapeutics for asthma and hypertension - SUMMARY / ABSTRACT
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.
