Investigating Calcium-Dependent Depolarisation of Uterine Smooth Muscle and the Role of ANO1

Programme overview:
This MRC-funded doctoral training partnership (DTP) brings together cutting-edge molecular and analytical sciences with innovative computational approaches in data analysis to enable students to address hypothesis-led biomedical research questions. This is a 4-year programme whose first year involves a series of taught modules and two laboratory-based research projects that lead to an MSc in Interdisciplinary Biomedical Research. The first two terms consist of a selection of taught modules that allow students to gain a solid grounding in multidisciplinary science. Students also attend a series of masterclasses led by academic and industry experts in areas of molecular, cellular and tissue dynamics, microbiology and infection, applied biomedical technologies and artificial intelligence and data science. During the third and summer terms students conduct two eleven-week research projects in labs of their choice.

Project:
During pregnancy, the uterus must not contract in a coordinated fashion until the time of delivery. The initiation of labour is a complex process, driven by both systemic and local humoral influences as well as neurological control in response to mechanical forces. However, it remains unclear how these stimuli are converted into the electrical activity needed to bring about contractions. Previous work from the Blanks lab (Atia el al. 2016) has suggested that the Calcium-activated Chloride Channel (CaCC), formed by the protein Anoctamin-1 (ANO1), may play a key role in initiating spontaneous depolarisation of Uterine Smooth Muscle Cells (USMCs). Due to early lethality associated with ANO1 deletion, experimental validation of this hypothesis in an animal model requires an alternative strategy. Our collaborator at the University of Columbia (Dr George Gallos) has created lines of ANO1-Cre mice, allowing for inducible, non-lethal gene knockdown.

In this project, the phenotype of USMCs from these mutant mice will be characterised at the electrophysiological level. CaCC activation by stochastic calcium release (Calcium Sparking) is proposed to induce spontaneous action potentials within the cells, that ultimately spread to adjacent cells, and effect contraction throughout the uterus. By employing a combination of calcium imaging and electrophysiological techniques, and correlating the findings to simulations, a detailed insight into the role of the CaCC will be established.

Computationally modelling the processes observed enables the generation of quantitative hypothesis, and optimisation of experimental design. By determining the role of this channel within USMCs, and integrating this with current understanding of the electrical network of the uterus, a better understanding of gross uterine function during parturition can be developed. This work is potentially highly translatable, and may provide evidence that ANO1 is a viable therapeutic target for tocolytic agents.