It’s Valentina here, the first PhD student to have joined the Hamill’s lab. I am writing my first article on a very special day…on my way to Dundee to defend my thesis! It’s been a really cool adventure, full of incredible discoveries.
When I have started, the only available data on LaNts were related to the skin; after three years and many exciting experiments we now know the effects of LaNt a31 in the cornea…which may seem only a little organ of the body, but how amazing is it to be able to see the world?! We have loads of beautiful videos of live laminin’s deposition from corneal epithelial cells and we have established a LaNt a31 overexpression animal model, the first one ever!
Working in this lab has been fantastic and hopefully today I will be able to transmit my excitement to the examiners 🙂 If my work could open at least one more door to future projects and ideas, then I’ll be doubly proud.
Going on, the Hamill lab, laminins and LaNts are waiting to be completely undercovered 🙂 much more to follow on this exciting topic…
Soon to be (hopefully) Dr. Vale
Just back from another conference and Lee has added his recent poster to our growing collection. A quick rearrange and we have the wall of LaNt research!
Yes, we have gone too far, taking over a bunch of the poster boards on our floor but it is so cool seeing the work from the various members of my group coming together into a really big story. I recommend working from left to right as you look at them to get a picture from the gene to the protein to the whole tissue.
It starts at the transcript level with Lee’s poster showing LaNt regulation of laminin expression. Also including images of changing protein expression in squamous cell carcinoma from Valentina Barrera and Conro’s minigene analysis of critical residues involved in splicing regulation.
Next we move on to some of our cell biology work looking at LaNt a31 in the front of the eye where Vale B demonstrates differential distribution accacross the corneal epithelium and Lee and Valentina Iorio show knockdown and overexpression affecting cell migration/spreading
After that its into the molecular biology, with Vale I using fluorescently tagged LaNts and laminins to study their interplay in corneal epithelial cells in culture and also the effects LaNt overexpression has on junctional complex assembly and matrix organisation.
In Poster 4 Tobi and Vale B’s work brings us back to whole tissues, this time using limbal explants as a 3D model of corneal wound repair and includes our data demonstrating changes in LaNt distribution as wounds heal.
Finally Umar and Vale B show some quite preliminary work of LaNt roles in angiogenesis and the effect of overexpression/knockdown on HUVEC tube formation.
I will now be sending all new students to study all these posters before starting work in the group. Seems like an easy way to get up to speed!
My first post and its big news! The Biotechnology and Bioscience Research Council have decided to fund my application “Characterisation of LaNt regulation of basement membrane organisation in wound repair and angiogenesis.” Exciting times!
Below is the summary from the proposal. Please feel free to comment with ideas, suggestions, feedback, collaboration ideas etc.
Characterisation of LaNt regulation of basement membrane organisation in wound repair and angiogenesis In this work I propose to study the LaNt family of proteins which were recently identified and which I believe are important for processes such as wound repair, blood vessel growth and the spread of tumours. Through these studies, a deeper understanding of these processes will be obtained and this, in turn, may lead to identification of new treatment approaches for conditions such as chronic or slow-healing ulcers and cancer. The different tissues of the body are composed of defined combinations of specialised cell types and a mixture of proteins and sugars outside the cells, termed the extracellular matrix (ECM). Some of the cell types reside in and contribute to the production of this ECM, whereas others cover the outer (epithelial) and inner (endothelial) surfaces of regions of ECM as sheets of cells. Directly beneath these cell sheets, as well as surrounding nerves and muscles, there is an organised region of ECM termed basement membrane (BM). BMs provide the anchorage point for cells and are therefore important for stress resistance and structural integrity. In addition, BMs support the different behavioural requirements of a wide range of cell type at different times, including acting as the road upon which the skin cells migrate to close wounds. A major component of all BMs is the laminin family of proteins. Laminins assemble into cross shaped molecules that associate with one another to form a network. Formation of this network has been shown to involve a small region at the very end of the short arms of the laminin cross, which is termed a LN domain. The importance of this interaction is exemplified by a number of genetic diseases where specific defects in LN domains impact the laminin network and BM organisation resulting in skin blistering, eye defects, kidney failure or muscular dystrophy. However, despite this knowledge, the ways in which laminin networks form, how network organisation changes during different cellular processes and what drives those changes is yet to be fully understood. This project will focus on the LaNts which have been demonstrated to play a role in cell attachment and migration and which my preliminary data indicate is likely to be through regulating BM formation. Like the laminins, the LaNt (Laminin N-terminus) also contain a LN domain, this suggests that they can interact directly with laminins and modify the ways in which laminin networks are organised. Importantly, there are also tissue specific differences within the laminin family and these differences are likely to mean that the impact of the LaNts is cell type specific. This may also mean that LaNts play different roles during blood vessel growth or wound repair than during normal tissue function. In order to characterise the roles of LaNts in BM formation and the impact they have on cell behaviour and tissue function, this project will pursue three aims.
- In Aim 1, I will use proteins in solution to directly assess the ability of LaNts to interact with laminins and other BM proteins and to determine their impact on network formation.
- In Aim 2, I will use skin and corneal epithelial cells and blood vessel endothelial cells to study the impact of changing LaNt protein levels on cell behaviour. Specifically, we will determine if the BM deposited by cells changes in response to increasing or decreasing LaNt levels and we will assess cell movement speeds, how strongly they attach and how rapidly they divide on the different substrates.
- In Aim 3, I will use three dimensional models of skin, eye and of blood vessel growth to study LaNt roles in these more complex tissue models.
Together the data obtained from these studies will dramatically expand what is known about LaNts, about laminin network formation, about BM organisation and ultimately about wound repair, blood vessel growth and tumour progression. In the longer term, this may translate into identification of new strategies for therapy development .