Tag: cell signalling

How saying ‘yes’ to a quick side project lead to one of my main research interests!

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It was the final year of my PhD, and I was presenting a poster at a conference, alongside my supervisor Dr Kathy Gately. We were showing off our new panel of PI3K inhibitor resistant lung cancer cell lines, which we had developed and begun to characterize. We were excited to tease out which signalling pathways might be playing a role in resistance to these drugs.

Along came Dr Michael O’Neill, the co-founder of Inflection Bioscience, who had recently licenced a drug that targeted the PIM kinases. At the time, I had never heard of PIM. He saw our poster, and suggested we should test their drug in our cell lines. It seemed straight forward enough.

After a couple of quick ‘look see’ experiments, we ended up submitting a grant.

Then another.

Then some student projects.

Some posters….

That escalated quickly meme | Memes catholiques, Humour, Débile

Before we knew it, this ‘quick win’ was becoming a driving interest for Kathy, and she was gathering researchers along the way (notably Dr Gillian Moore). I had left Kathy’s lab at this stage, but as a wider team we were beginning to build up a picture of how best we could potentially develop these drugs in the lung cancer space.

PIM research didn’t stop for Kathy, and it didn’t stop for me either.

When interviewing for a postdoc position in University College London with Dr Hayley Whitaker, I was asked ‘if you had access to human prostate cancer specimens, what would you do with them?’ On a whim, and with interview pressure weighing down on me, I responded ‘well there’s this really exciting drug target called PIM in lung cancer, I think it looks like it might be promising in prostate cancer too, so I’d probably run some experiments on that’.

I arrived home to Dublin that night, exhausted after a long day of travel & interviewing, and found out immediately that I’d been invited to a second round interview. This was great – but it would be in London again, in just a few days! I purchased a second pair of flights, cried over my bank balance for a moment, and then hunkered down in our basement office for the weekend, trying to pull together a presentation that had been assigned for the second round. The challenge that had been set was of course ‘if you had access to human prostate cancer specimens, what would you do with them?’ How could I present on anything other than PIM after suggesting it in my previous interview?!

I rushed a project pitch, which by chance turned out quite promising. There were a good few papers looking at PIM in prostate cancer, but not many looking at drug treatments, and none looking at the same co-targets that we were working on in lung cancer. I checked with Kathy if it was ok with her for me to present this, while rushing out of the building to get to the airport – but our conversation got slightly side-tracked when she told me she was expecting a baby! Safe to say PIM got a bit overlooked that lovely day.

The presentation went well, I got the job, and to my delight I was offered the chance to actually work on the project that I had pitched in the interview. What a wonderful opportunity for a postdoc to be given that level of freedom!

In order to differentiate my new prostate cancer project from the work Kathy was leading on, I set out to investigate a wider panel of drugs, including the PIM inhibitors but also quite a few others. The aim was to test promising late stage pre-clinical drugs in human prostate cancer tissue, using ex vivo culture and new omics technologies. I gathered some preliminary data and submitted it as a fellowship proposal, trying to position myself as someone who worked on drug development in general. Thankfully, I was successful.

It wasn’t mean to be a ‘PIM project’. But as luck would have it, PIM wasn’t going away.

One by one, the other drugs dropped off for one reason or another. Some couldn’t be investigated in an ex vivo model because they needed to be metabolised within the body, some needed to build up for a few weeks before an effect would be seen, some failed during concurrent animal testing, and some just showed disappointingly little activity in my model. By the time the work was close to publication, we were down to just 4 different treatments, and they were a very similar panel to what Kathy was leading on in lung cancer. I hope she forgives me!

Now, years later, we’ve just had our first original article come out on PIM in prostate cancer1. This is our first ‘flag in the sand’ where we put forward the idea of co-targeting PIM with the PI3K pathway. There are bigger and more detailed works to come from this in the future. If you’d like to read about the paper itself, I wrote a tweetorial that you can read unfurled here: https://threadreaderapp.com/thread/1300721602854871040

This paper came off the back of a couple of reviews on PIM as a drug target3,4, and there is of course more on the way.

Now, plans are brewing for wider PIM collaborations, and who knows, maybe PIM will stick around in my world even longer.

Did I ever set out to become a PIM researcher? No, not particularly.

But I suppose the lessons learned here are to say yes to opportunities, and to follow the data – if something isn’t your ‘plan A’ but it might make a difference to cancer patients in the future, then why wouldn’t you follow it?

Extra credit to my friend AJ (@AyoksAJ) for his very inspiring ‘Say Yes’ presentation to our postdoc networking group a few years ago, which still sticks around in my mind, and lead me to say YES to an opportunity that came my way this morning – let’s see where this one goes!

Thank you to Kathy, and to all the PIM friends I’ve made over the years.

1 https://www.nature.com/articles/s41598-020-71263-9

2 https://www.sciencedirect.com/science/article/pii/S0163725819302062

3 https://www.nature.com/articles/s41392-020-0109-y

What role does NFkB play in chemoresistance?

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This was one of the very first research questions I ever set out to answer, way back as a final year undergraduate in Trinity College Dublin, in 2010/2011.

Since then, over many years, a few papers, and with an ever expanding global team, it has remained at least a side project (and in some cases a driving interest) for some of my oldest friends in research!

For me, it began with my final year thesis project, which involved comparing a panel chemosensitive and chemoresistant lung cancer cell lines, developed by Dr Martin Barr as a tool to investigate response to chemotherapy.

We wanted to understand how lung tumours develop resistance to chemotherapy over time, so that we could find better ways to treat them!

We screened the cell lines using qPCR arrays, and identified a few potential genes of interest including NFKBIA, which is involved in regulating NFkB.

NFkB is a well studied transcription factor that seems to play a flexible and complex role in many biological processes.

This initial finding caught the eye of my project supervisor Dr Kathy Gately, who immediately began to wonder if NFkB could be a potential therapeutic target in chemoresistant lung cancer. The project ended up winning the Margaret Ciotti medal as the highest marked thesis in the School, and with the excitement of a new finding (and this early recognition boosting my confidence), it was an easy decision to stick around in Dr Gately’s lab to begin my PhD.

While the PhD project itself was focused more upstream on PI3K/AKT/mTOR, we did further develop our NFkB finding, and it ended up becoming both my first research paper and the first chapter of my PhD thesis. In this work, we used a drug called DHMEQ, which is an inhibitor of NFkB translocation developed by a Japanese collaborator, and we found that it was more effective in treating chemoresistant lung cells than chemosensitive ones.

I have fond memories of this work, as I got to try out a range of new molecular techniques for the first time, including the obligatory ones like cell culture, drug treatments, qPCR and Western blotting, but also things like high content immunofluorescence imaging, proliferation and apoptosis assays, and I even got to run some Sanger sequencing on a machine named ‘Spongebob’!

https://www.tandfonline.com/doi/pdf/10.4161/cbt.29841

It was clear from the data that this work opened up a whole range of possibilities, and it was time to expand. My colleagues Peter Godwin and Dr Anne-Marie Baird each took aspects of this forward, with Peter publishing a great review on the topic that remains the most highly cited paper on my Google Scholar profile (thanks Peter!) and Anne-Marie being awarded a fellowship from the International Association for the Study of Lung Cancer to investigate a new take on the work down in Brisbane, Australia.

https://www.frontiersin.org/articles/10.3389/fonc.2013.00120/full

Anne-Marie’s work (alongside Dr Sarah-Louise Ryan) brought a fresh and exciting angle, interrogating the role of inflammatory pathways in NFkB mediated cisplatin resistance, as well as confirming that the NFkB translocation inhibitor wasn’t just more effective in chemoresistant cells, but in fact could be used to resensitise those cells to the effects of the chemotherapy itself.

https://www.sciencedirect.com/science/article/pii/S016950021930529X

So where are we now?

Well, we’re certainly not the only ones investigating NFkB in chemoresistance…

With groups all over the world also teasing out the role that NFkB is playing in this setting, things are becoming increasingly clear, and also increasingly complex.

It turns out NFkB is quite a promiscuous player in cancer development and aggresivity!

With seemingly endless feedback loops, regulation and cross-talk with other pathways, it seems to have the capacity to drive wide ranging and even opposing phenotypes. Equally, it often appears to be a mere passenger, caught up in attempts at cell survival during times of stress.

With these flexible abilities, can we truly say that NFkB itself is a viable target for drug development?

Sadly, probably not.

But we do hope that our growing understanding of the role that it plays in chemoresistance will help us to identify better targets that work alongside it, and ultimately better ways to treat these tumours.

As ever, we keep searching.

Big thanks to Kathy, Anne-Marie, Martin, Peter and Sarah-Louise, as well as all the other researchers involved in these projects!

Are the PIM kinases promising targets in prostate cancer?

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We recently published a review on this topic, and we’ll soon have our first data out investigating this in the lab too!

There are a few drugs that target PIM kinases, that have been investigated pre-clinically, or in early stage clinical trials. PIM kinases do seem like promising targets in prostate cancer – they are proto-oncogenes, they seem to be expressed more often in prostate cancer tissue than benign tissue, and perhaps also associate with particularly aggressive disease. But sadly the drugs available to date haven’t skyrocketed through trials to approval and use in the clinic. This is partly due to the drugs themselves (maybe the next generation of drugs will be better, we hope) but we also suspect it might be a better strategy to co-target PIM with other interconnected pathways, using combinations of drugs, or even individual multi-targeted drugs.

In our review, we summarized some of the drugs available to carry out research on, that directly target PIM, as well as ones that we think could be used in combination with PIM inhibition:

https://www.nature.com/articles/s41392-020-0109-y

I was also involved in another review which took an in depth look at the role of PIM in resistance to other treatments and in immune evasion, across various cancer types.

https://www.sciencedirect.com/science/article/pii/S0163725819302062

Overall, yes we think PIM is a promising target in prostate cancer as well as other cancers, and we suspect it will be best targeted as part of a combination therapy approach, and perhaps in the resistance setting.