Mapping tissue immunity in the human urinary bladder across lifespan

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The urinary bladder is a hollow spherical organ situated in the lower abdomen. It forms part of the urinary tract, which is responsible for expelling waste and surplus water from the body. Urine, containing waste and excess fluid, is produced by the kidneys, and transported to the bladder where it is stored prior to voiding. This means that the bladder is in relatively close proximity to the external environment, and may be attacked by bugs (microbes) coming in from body surfaces, including microbes originating in the gut. We know that all organs, including the bladder, contain some resident defence (immune) cells that help to fight off microbes. Despite this, infection of the bladder is common, affecting 1 in 2 women in their lifetime and 1 in 20 men. Susceptibility to bladder infection increases with age, as do other bladder diseases, like inflammation (that can give symptoms such as pain or incontinence) and cancer. Despite the fact that immune cells play an important role in all of these conditions, we currently don't understand the details of how bladder defence is set-up, how this goes wrong or changes with age, and how this differs between men and women. Our research project aims to address this knowledge gap. Our overall aim is to produce a map or atlas of all the immune cells in the human bladder across lifespan, in men and women.We will take two experimental approaches: The first is to take a piece of human bladder, and to mash it up so that we can analyse individual cells, including immune cells. Different cells have different functions and identification marks because of differences in their genetic material (called genes), and differences in how their genetic code is translated into the working parts of cells, called proteins. Every cell contains thousands of proteins, but these are hard to measure at scale in a single cell. However, we can measure the molecule that acts as an intermediate between genes and proteins - this is called messenger RNA or mRNA. In the past 10 years, technology has advanced such that we can measure thousands of mRNA molecules in a single cell, this is what we plan to do with the mashed bladder samples. We already have ethical permissions in place to take human bladder samples across lifespan, including from embryos (from the Human Development Biology Resource at Newcastle University), and adults (from organ donors in Cambridge that have consented for their tissues to be used for research).Our second experimental approach is to take a piece of bladder and keep it intact so that the relationship of the cells in space is maintained. We will then make slices of the piece of bladder so that they are thin enough to be visualised under a microscope. We will use the mRNA information from our first experiment to identify protein markers that can be visualised by the microscope using probes that are fluorescently labelled and we can also map mRNA molecules in space. This will allow us to investigate the position of cells relative to each other and to determine whether the position of cells differs between men and women, and in the adult and developing bladder.Making a complete map of cells in the bladder across lifespan will provide a critical reference atlas that will help researchers in the future to understand which cells become abnormal in different bladder diseases and how it might be possible to prevent unhelpful age-related changes.