Today was the Northwest Cancer Research annual symposium held in the stunning, Harry Potter-esque, Victoria Gallery on the
University of Liverpool campus.
Lots of excellent, inspiring talks including two of my long-term favourites; actin remodelling and RhoGTPases, and integrin signalling. The notebook is full of the next set of experiments and grant ideas! (Thanks Mark)
There was also a thoroughly impressive poster session, displaying the diverse, excellent cancer research work going on in Liverpool, Bangor and Lancaster.
Included in that poster session, Lee Troughton and Valentina Barrera
presented our new splice regulation story in LAMA3 that is beginning to really take off. Unfortunately, I missed the opportunity to grab a photo with Vale so its just Lee on this edition of the obligatory “they grow up so fast”.
Lee moaned extensively about his quads hurting while squatting down to take this picture so I made him pose for 10 more, saying it was out of focus…
I also captured his response to finding out about the unnecessary extras. Totally worth it
Mini (or squatting) Lee
April time again and the end of another block of MRes Clinical Sciences projects. This time the Hamill lab hosted three students; Kareem Hassanin, Conro Conro Sugden and Tobi Oyewole. Today the boys had their presentations/poster session.
First up; Conro told his story of developing our new minigene construct to investigate LaNt regulation and testing a few mutant versions of it to prove it works. Conor, with Lee Troughton’s help, did some excellent work; generating flow cytometry,
RT-PCR, western blotting and fluorescence microscopy images that demonstrate that our new system is effective for studying intron retention and alternative polyadenylation and showing that the exon 9 splice site has a big role to play in determining splicing efficiency. His data has gone straight into a grant application and should form the starting point for the next stage of the LaNt project.
Next, Tobi under the joint supervision of myself and Colin Willoughby and working alongside Fight For Sight student Thanos, worked on determining the optimal delivery mechanism and conditions for delivering small RNA molecules into corneal epithelial cells. This work is critical first step for the the next stage of Thanos’ PhD studies and, although Tobi was frustrated at times, makes a big difference to the lab. Optimisation is the biggest part of all experiments, without these steps we wouldn’t be able to ask the big questions.
Finally, Kareem working alongside Dr Valentina Barrera and using samples kindly provided by Prof George Bou Gharios, optimised our new rabbit anti-mouse LaNt antibody staining protocol and then did the first staining of embryos and various tissues in the adult. These are really short projects and filled with frustration as just at the very end, Kareem finally had conditions nailed down to get super clean, specific, staining that has opened up a whole range of new options and research questions. One thousand emails bounced back and forward yesterday as he tried to identify all the structures that were and were not stained. Although super cool, really this is just the beginning; knowing where the protein is automatically generates the question of what it is doing in that location as we predict that LaNt function is context specific, so these data too will lay the groundwork for the next grant(s) and next paper.
All in all, a really productive 3 months. Looking forward to the next block…
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!
This week PhD student Lee Troughton and I have been attending the British Society for Investigative Dermatology meeting in exotic Manchester.
Lee presented some of our recent findings in poster form. This is the first time we have rolled out this new story and, so far, the response has been very positive.
Loads of data including contributions from Valentina Barrera and Conro Sugden. If anyone in the IACD was wondering why Lee was frantically running 1000s of qPCRs last week, check out the graphs in the bottom left. Nothing like the impending deadline to give a sense of urgency! However, the findings are really interesting and have opened up some really nice questions.
It’s also cool to be back at the BSID after many years away. I think my first “real” conference presentation was at a BSID in Oxford during my PhD where I presented some of our very preliminary data describing the first identification of the alternative splice isoforms that we now call the LaNts. It’s really cool to be back and to see many of the people I know in the skin field and learn about all the excellent derm research going on in the UK.
Lee looking chuffed at his bold colour choices