Conference over, time to go home

Fountain in the local park where I went for my runs

So the conference is over and it is time to go home and say goodbye to Holland. I have had a fantastic time in Groningen and the conference certainly lived up to my lofty expectations. I have spent so much time in the last few days hearing about Dupuytren's and Ledderhose from the best experts and doctors in the world that I am on information overload, hopefully that is a good thing and hopefully these doctors will stay in touch and feel they are able to use the patient community to enhance their research. 

My talk was apparently received very well and I have had doctors and other attendees telling me I did a good job and hopefully when it comes on YouTube everyone else will agree. I feel like I did myself and the patients proud although I forgot to mention that 92.5% of Ledderhose patients that responded that were over 60 had Dupuytren's, not overly important but that is a shockingly high percentage. 

The final part of the conference was a discussion on collaboration, people working together to get the best funding that they can for this kind of research. It was great to see people working together like that. There seemed to be some great ideas of what is needed to really progress Dupuytren's research and further the work that is going on. 

I certainly feel that with the minds that are looking into this and the way they all want to work together for the benefit of patients there is hope for patients that we may one day have a cure for this. Sure it may not be in the next 5 or 10 years but I am hoping that if nothing else it would be available before my daughter would develop this condition (hopefully she won't get it). 

Dupuytren's cells used in another scientific paper

Fibroblasts from phenotypically normal palmar fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture: 

BMC Med Genomics. 2012; 5: 15.Published online 2012 May 4. doi:  10.1186/1755-8794-5-15

PMCID: PMC3375203

The introduction to this paper starts with the normal spiel about the things associated with this disease and I didn't really learn anything new there. There was also the usual talk about it coming back and this again is of unknown cause but likely because of genetics or sufficient disease tissue being left behind to cause regrowth. 

In this paper they are looking at cells from Dupuytren's and carpel tunnel patients as well as normal uninvolved tissue to use as a comparison. For those with limited understanding of science or those that fancy a refresh have a quick look at this post  -- Analysis of Duputren's cells -- I will try and explain the science here but reading the previous post will probably help. 

In this paper they are using a technique that is called a micro-array this allows them compare the levels of thousands of different read outs from the different types of cells are compare them. This means that if they have 4 different cells lines (i.e. 4 different groups of cells derived from different patients) they can build up a profile of what the cells from patients with this condition look like and see how they are the same and how they are different from 'normal' cells. This is a big deal for something like Dupuytren's where of course they do not know what the cause is, they have some idea of the pathways involved 

• Can scar formation be used as a useful model for Ledderhose and Dupuytren's?

• IGF-2 and its relationship to Dupuytren's

The advantage of a micro-array is that it allows you to look at allow the changes that are happening in all the pathways in the cells that have come directly from patients. 

The Results:

I will stick to looking at what they found for the Dupuytren's tissue rather than bog down the post with the carpel tunnel stuff, When comparing DC and normal cells they found out that there were 308 transcripts that were different between the two cell types, they also show lots of other data saying that there is some difference between the two different sets of cells. 

They next go on to look at identification of biological pathways that are altered among these different cell types. When looking at the pathways altered in the different cell types for Dupuytren's and normal cells were related to cell death, cell growth and proliferation (speed and control of growth) which of course makes sense when you see less cell death and too much cell growth and division in Dupuytren's cells. The analysis also suggested that there was some involvement of the regulation of things called microRNAs. These results were then validated using another technique which showed that the results were reliable. 

They conclude by saying that the progression and recurrence of DC could be based upon similar theories to those applied to tumour biology (in no way are they suggesting it is cancerous). They say that perhaps there is an inherited mutation but that it takes a second mutation to cause the disease to become active, this kind of fits with the fact that the disease appears to be dominant but with a variable hit rate as the dominant factor is the initial mutation but the variable bit is acquiring the second one. Again this also fits with the disease normally happening later in life as this would give time for the additional mutation to occur.  

They have a lot more information in that paper which is why I have linked to it above. Hopefully research continues into these conditions so that a better and more successful treatment / cure can be found. 

Analysis of Dupuytren's Cells

So I thought it was about time for another science post, don't worry it is only going to be a short one and is again based on a chapter from the book, Dupuytren's Disease and Hyperproliferative Disorders. C. Eaton et al, Chapter 14.

Right so for those who don't know (I have explained it before but hey and see the picture below from the previous post) DNA is a constant in the cell and it is converted to mRNA in a regulated manner which is then converted into proteins which have an effect.

The central dogma 

So one way of analysing cells and differences in them is to look at the levels of mRNA. There is a technique called a Microarray and whilst I will not going into detail here what it basically does is give you a read out of the levels of mRNA for pretty much every gene, so what that means is that you have a read out of how much there is of the things that are responsible for what happens in the cell.

So what they have decided to do in this experiment is to get cells from patients with Dupuytren's disease from healthy and Dupuytren's tissue and they look at what differences there are between the 2 different types of cells using a microarray. This is then repeated, in this case on 20 patients and they look at what mRNAs are at different levels in ALL patients and from this they come up with a list of genes (genes are the DNA that codes for the mRNA) that might be linked to Dupuytren's disease.

They then cut this already small list down to ones that are likely to play a role in the development of Dupuytren's and in particular they are looking at the ones that are likely to play a role in contracture. They come up with several different proteins which I won't go into as they don't mean anything to me but they come up with things that are involved with the extracellular matrix (ECM) which plays an important role in connective tissue and I have mentioned it in my past post along with collagen.

They then go on to basically say that there are some mRNAs that are at a lower level in the disease tissue and this might be linked to the development of contracture but do not speculate as to how and how this might be useful. I guess knowing the cause if the first step towards a sure and this information helps in the understanding of the cause.

The British Dupuytren's Society

The British Dupuytren's Society is a small UK charity for people affected by Dupuytren's Contracture, Ledderhose Disease and Peyronie's disease:

Today I am going to do a blog post on the above charity, as you can see from the description I have posted they do work for people with Ledderhose as well as Dupuytren’s and Ledderhose so I urge you to visit their site.

Firstly I would also urge you to follow them on facebook and twitter …

Facebook –  http://www.facebook.com/britdupsoc

Twitter - @Dupuytren

Ok so I am not going to make a huge post as I am hoping to do an interview type thing with them in order to get a better and more in depth idea of what they do and where the money goes. For now though I do have the following from an e-mail that my wife received.

“The British Dupuytren's Society will be giving a talk to the British Chiropody and Podiatry Association later this month, hoping to spark a dialogue on creating a national approved treatment protocol for Ledderhose. This is an example of where we use donations…”

Xiaflex in cells - Trying to help you understand, maybe

So today I am going to try and review this paper In Vitro Study of Novel Collagenase (XIAFLEX®) on Dupuytren's Disease Fibroblasts Displays Unique Drug Related Properties. This paper is open access so you might like to take a look for yourself or have it open at the same time as you read this and it might help. I don't understand all of it so good luck!

I say try because there is a lot of scientific content in this paper, a lot of stuff that I have not done in the lab before and this makes it harder for me to work out exactly what it means, I will however try my best to understand it and where possible try and explain it in as simple terms as possible.

To start with for those who don't know Xiaflex is a treatment for Dupuytren's disease (DD) and potentially Ledderhose disease (LD), it is a collagenase and the main component of the lumps in DD and LD is collagen and so it breaks them down.

Right onto the paper, where possible I shall try and bullet point it and I am going to place a summary at the end where I will put what I consider to be the key points.

Introduction:

• DD is a common and benign disorder. 
• The cause(s) of DD and LD are unknown however DD has been linked to genetics, smoking, diabetes, alcohol, anti-epilepsy medication, occupation and local trauma (so all the usual stuff). 
• The group of people at the most risk are Caucasian men of North European descent that are 40 or over with highest incidence in USA, Scandinavia, Britain and Australia. 
• The disease shows cell proliferation, high levels of collagen and extra-cellular matrix remodelling. 

They also make a key point which is what frustrates many of us with the disease and this is that:

"Treatment of DD is not curative" and this means that rather than trying to rid the body of the disease they are just trying to remove the symptoms.

One current treatment that is showing promise is Xiaflex which is a collagenase that digest the triple helical structure of collagen and therefore removal the lumps and whilst this is significantly less invasive than surgery it does have a much higher level of recurrence.

Aim: Their aim is to investigate the functional effects of Xiaflex in comparison to Collagenase A when used on cells cultured from DD nodules, cord, fat and skin. They look into various different aspects of the cells including looking at the cell growth and at gene expression.

So I am going to try and go through this and explain the key points one figure at a time to make it easier to follow.

Figure 1: Here they just show the process through which they have derived the cells and then the experiments that they are planning to do on them.

Figure 2 (and Figure 3): Here they are measuring the ability of the cells from the DD nodules and cords to grow in the presence of  Xiaflex / Collagenase A and whether they are able to recover from this treatment in 24 hours by removing the drug and continuing to monitor the cells. In Figure 3: They also look at fat and skin cells from DD patients in a similar way.

• They find that without treatment there is no significant different between the nodules and cords compared to the skin and fat although the DD nodules and cords do show a faster growth rate. This makes sense as it is a proliferative disease.
• Treatment with  Xiaflex / Collagenase A decreases all of the things that they are measuring in ALL cell types in a dose dependent manner. 
• 24 hours after removal of the drug the cells recover, this happens even at the highest dose and may help to explain why DD comes back so frequently when treated with  Xiaflex. 

Figure 4: I can't really follow this figure very well so rather than get it wrong and make mistakes I am going to leave this figure out but their key point from this seems to be that Xiaflex / Collagenase A induce membrane leakage and reduce cellular viability and metabolic activity in DD cells.

Figure 5: Here they are looking at the toxicity of the 2 different drugs and want to see which is more toxic to cells and if there is any specificity towards DD nodules and cords. 

• Xiaflex / Collagenase A both cause cells to die. 
• Collagenase A causes both necrosis (premature cell death) and apoptosis (programmed cell death) 
• Xiaflex only causes necrosis.

As far as I can tell from reading this (and correct me if I am wrong) they do not comment on whether this is a big deal or anything? But I think they say in the discussion that it may mean that there are likely to be viable DD cells and therefore there is an increased chance of disease recurrence. 

Figure 6: Here they are looking in the different types of cells at the levels of mRNA for key components in the production of collagen to see if this is altered by the presence of Xiaflex / Collagenase A. For those who don't know about the central dogma of life I have tried to outline it in the diagram below. But the key thing you need to know to understand these results are that mRNA is produced from DNA and that the levels of mRNA vary so that it can be used as an indicator of protein levels in the cell as protein is produced from mRNA. (Below image is VERY simplified)

The central dogma: DNA is where the information is stored and this is processed into RNA and then proteins. 

• They are looking in DD disease and see that  Xiaflex treatment causes a decrease in levels of collagen I and collagen III as well as fibronectin (forms part of ECM and interacts with collagen), alpha SMA and TGF-B1. If you remember from my previous post a decrease in TGFB1 would be useful. Explained in the image below. 

Central blue boxes show state in DD cells, right hand side is reported influence of Xiapex in DD cells

• They also say that levels of collagen are at their highest in the DD nodule which is exactly what you would expect. 

Figures 7 & 8: In figures 7 and 8 they are looking at the protein levels of Collagen I and III in the different cells types to see a) is there any difference in their profiles between the different sources (skin, fat, nodule and cord) and b) does Xiaflex / Collagenase A treatment cause any changes to these levels. 

• They show that the levels of Collagen I and III in the nodules are very high with Collagen III in particular very, very high (basically two times more than seen in anywhere else). 
• When looking at the effects of Xiaflex and Collagenase A they see that: 
• Collagenase A has an influence on Collagen I by causing a decrease on all samples at the highest dose only. 
•  Collagenase A only decreases Collagen III at the highest 2 doses in the nodules, cords and fat.
• Xiaflex causes a large decrease on all samples at all doses for both Collagen I & III with the largest decrease observed in the Nodules and cords.  

Figure 9: Here they are trying to show that the decreased levels of Collagen and other ECM factors was not just at the mRNA level but that they were also decreased at the protein level when DD tissue was treated with Xiaflex. 

• Xiaflex does down regulate the protein levels of fibronectin, Collagen and CTGF (and more) at  the protein level. 

Figure 10: I am just going to say that they see that Xiaflex and Collagenase causes a decrease in cell cycle markers. This makes sense as when the drug is added you see a slowing on cell growth and this means that the cell cycle is slower and therefore you should have a decrease in cell cycle related proteins (though in this case they are look at mRNA).

Right so that is all that I am going to cover for this paper, as I said it is a long post and it is very science based so here is the summary for all those who just want the key points or who are just too lazy to read it all (which is fair enough). 

Summary: 

• DD nodules and cords do show a faster growth rate.
• Xiaflex and collagenase A both decrease the growth rate in DD cells. 
• Xiaflex treatment causes a decrease in levels of collagen I and collagen III as well as fibronectin  and TGF-B1 (see the diagram as to why this might be good). 
• Levels of Collagen I and III in the DD nodules.

Reference:
Syed F, Thomas AN, Singh S, Kolluru V, Emeigh Hart SG, et al. (2012) In Vitro Study of Novel Collagenase (XIAFLEX®) on Dupuytren's Disease Fibroblasts Displays Unique Drug Related Properties. PLoS ONE 7(2): e31430. doi:10.1371/journal.pone.0031430

IGF-2 and its relationship to Dupuytren's Contracture

Today was a hard day, I was not on my feet a great amount but apparently just the thought of going back to play badminton (which I am awesome at by the way) is enough to trigger my foot into going nooooooo. From about 2:30pm today until the end of work I was in pain when sitting that just increased when I was walking and went into overdrive when I was standing. I am not sure what to make of it, on the one hand I really want and need to lose weight but I am not in a position to cycle and I am very self concious about swimming and always get ear infections (though would start using ear plugs if I started again). Does anybody in a similar position have any suggestions? I used to like running but clearly that is so far out of the question it is not even funny. I would also like to know if anyone out there has any questions about the disease or anything that I do? 

Right so with that over onto the science for today (well the light evening reading for me) I will try and keep it as simple as possible and where I can I will try and include some more background information but the main basis for this post is to talk about IGF-2 and its relationship to Dupuytren's contracture. Remember I am not an expert and I am not a doctor but I am a research scientist so I would like to think that I have a better chance of understanding these things and maybe explaining them to others than if people just tried to read them for themselves. 

For those who don't know about Dupuytren's contracture a quick google search will show you that it is a disease that is thought to share many things with Ledderhose but it occurs in the hand rather than the foot, in fact it is thought that they are basically the same condition in different parts of the body and many people who get one disease will get the other (though Dupuytren's in the hand is much more common). 

A lot of the information that I am getting for this is coming from Dupuytrens Disease Related Hyperproliferative Disorders chapter 20. 

So IGF2 is an insulin like growth factor, all this means is that is a protein that is a bit like insulin and it circulates in your blood stream. The IGFs are known to play a role in many different life processes including ageing and growth and are far to complicated to get into here and now (but feel free to ask me questions as I actually know a bit about that). Anyway it has been shown that IGF2 in particular has a role in collagen production. Collagen has been shown to be one of the main components of the lumps that form in both Dupuytren's disease (DD) and in Ledderhose disease (LD) so of course that there maybe a link between IGF2 and DD and LD is worth looking into. 

Things are not as simple as more IGF2 means more collagen. IGF2 is regulated in the bloodstream by IGF binding proteins or IGFBPs which typically stop the IGFs from binding to their receptors. IGFBP6 in particular is thought to be involved in IGF2 regulation as it binds to IGF2 10 to 100 times more than it binds to IGF1. Anyway what people have shown is that IGFBP6 which remember stops the IGF from performing its function is actually at lower levels in DD tissue. So think about that, IGBP6 which lowers levels of IGF2 is decreased, that means that there are higher active levels of IGF2 and now remember IGF2 causes collagen production and what are the lumps in DD and LD made of, Collagen. 

What some researchers have actually done is grow cells and look at them contracting, they have then looked at cells from DD patients and as a control they have used cells derived from non-diseased portions of the hand. What they find is that if you add IGF2 then you increase the contraction and if you add IGFBP6 then you decrease the contraction. They have also looked at something that is upstream of IGFBP6, so by this I mean that they look at something called TGF-B1 and this is known to decrease the levels of IGFBP6 and so increase IGF2 and so increase levels of collagen and in this case find that there are increased levels of TGF-B1 in DD cells. 

So what is the relevance of all of this? Well what do you think might happen to a DD or LD patient when you add IGFBP6 to the lump? Well of course you should decrease the levels of IGF2 and therefore decrease collagen production and therefore ease symptoms of the disease. Sounds good to me and I hope that the picture below helps to explain this to people that have not followed the above which is quite complicated. 

I got the book, hahahahaha

The book:

Dupuytren's Disease and Related Hyperproliferative Disorders: Principles, Research, and Clinical Perspectives

Today I was given a rather awesome late Birthday present in the form of the above book which I posted about a few weeks ago and was saying how great it would be to have it and now I do. Being a scientist I love to do my research and I really like to know how things work and be up to date with the current understanding. So of course when I found out that I had Ledderhose I wanted to know everything about it and of course there is not that much to find and then I saw this book and whilst it is mainly focused on Duputren's disease a lot of the background science such as the cells that are the main part of the lumps and the cause of both is unknown and in all likelihood the same.

So I have only had this book since late this morning and I can already seen a lot of potential to increase my understanding of the disease and I can see the potential for many useful blog posts on the science of the thing (which I will try and make friendly for the non-science community) and also on lots of other different things that I just didn't have the details on before. As regulars will know along with my usual ranting about my foot I like to post more useful things such as the types of treatments and I think I am going to follow this up by posting about risks and prevalence and symptoms etc as I also think that these will be useful and perhaps I should really have started with those to keep things in chronological order. Still I think that the book is going to be very useful for all of these things and I can't wait to get home and knowing me read it from start to finish making notes and learning as much as I can to share with those that want to know.

Having had a bit more of the book I can see that there is going to be so much that I can post now in top of my usual ramblings (see a bit further down for todays).

So I would like to say a huge thanks to JOA for getting this book for me, it is a great and generous birthday present and I couldn't have picked a better man for the job of best man.

So today was pretty good until I got on the train home and then it was like my foot knew that there was nowhere to sit and I was in a lot of pain for the 8 minute journey, I feel that without some king of proof that I have this thing there is no way I can ask people to move out of their seats on a crowded train but I don't know if anything like that exists, guess it gives me something to do this evening. Well first I am having dinner and a foot spa and then I am going to finish my Surgery post for later but then maybe I will have a look and see if there is anything I can get...