From the outside in: Modular bespoke artificial cells for drug discovery

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I propose to create a paradigm shift in the way that drugs are tested early in the discovery process by building artificial cells as bio-analytical tools to quantify transport into and out of cells. My research proposes a way to build bespoke artificial cells from the bottom up that contain the major membrane-based cellular components and organelles to study molecular transport. By allowing both passive and active molecular transport to be quantified at the same time, this has the potential to be the first technique that can be used to answer this fundamental question in drug discovery. Over the last five years my research group have developed a unique new method to quantify how molecules enter cells by building custom-made human-mimetic artificial cell membranes on a microfluidic platform. We developed microfluidic technologies that allow the formation of cell-sized artificial cell membranes based on droplet interface bilayers (DIBs) made from the main phospholipids found in human intestinal cells. We used the platform to create a new type of pharmacokinetic compartment model and show that DIBs have a threefold improvement in the prediction of molecular absorption than the commercial in vitro technique. Since little is known about what different types of lipids can be used to build DIBs, we then investigated the effect of temperature on the formation of these lipid-based artificial cell membranes to enable us to create more biomimetic DIBs from a wide range of phospholipids. Then we built asymmetric DIBs to mimic a structural feature of the molecules that compose the cell membrane. We quantified how the transport of molecules is affected by the changes the molecular asymmetry of membranes, potentially highlighting a new mechanism for chemoresistance. Our work suggests that our bespoke artificial membranes will allow us to discover new aspects of cell biology and how they affect molecular transport. In the next five years, we will add the remaining membrane-based components to our artificial cells. We will use these systems to perform fundamental studies into the effect that different membrane components have on molecular passive membrane permeability. These include lipid domain formation, and the effect of sex on permeability. Then, we will insert human membrane transport proteins to be able to quantify active transport. Thirdly, since each compartment in a cell is delineated by a membrane, we will mimic these inside our droplet interface bilayer (DIB)-based artificial cells using liposomes. Finally, we will develop detection methods that allow us to quantify the kinetics of drug transport accurately in each location of the compartmentalised artificial cells. This research will significantly contribute to the field by providing a new technology to predict whether molecular transport occurs via passive or active mechanisms, which is a fundamental scientific question in drug discovery. This will allow Canada to develop drugs faster and cheaper.