FB 62. It has always perplexed me that scientific discoveries so often come out in twos, from different labs. I suppose people are asking the same questions based on what is known at the time, and therefore are liable to come up with the same ideas and results. And so it would seem with our Boxer stuff. Professor Amos says he has the identified the chromosome on which the JKD gene lies, and almost immediately the Swedish/American labs say the same thing. Could this be true and have they each have found the same chromosome? Either way, it seems some collaboration may result. And then no sooner do we have Dr Hamilton talking about his bio-marker for ARVC when I hear that a British cardiologist, David Connolly, who I also know from former times, is asking for ARVC cases and controls to test out something a bit similar. And just like the JKD groups, these two ARVC groups may join forces. They might even use the same dogs/blood samples. It’s all progress I suppose, but Boxers' problems are getting their share of attention. Wouldn’t you agree?
FB 61. Life has its ups and downs. The latest ‘down’ for me was having to put to sleep my nearly 10 year old girl and favourite, my last show bobtail, Sunny or Steynmere Sunshine Girl. Maybe by having taken up violent tug-of-war games with her much bigger brother she had damaged a disc in her neck or maybe it was a cancer symptoms but she started going through periods of agony in her neck region. There only one thing to do at her age. So she now rests among the apple tress in the garden with so many other Steynmere Boxers. I only showed her as a puppy as I was so wrapped up with cardiomyopathy that I never got her and her brother socialised normally and she did not like the show ring. Nevertheless she qualified for the Pup of the Year competition easily as a baby despite being the youngest. This is the end of an era with a vengeance.
But there have been ‘ups’, with both the ARVC and JKD research. A few days ago I visited paediatric cardiologist Robert Hamilton from Canada who was attending a cardiology conference in Wales and got an update on his bio-marker for ARVC. In a remarkable piece of outside-the-box thinking he had come up with an idea for detecting the presence of the disease before it was overtly recognisable. He had tested it out on number of his ARVC patients and all showed positive. Then he turned to Boxers affected with this same heart disease. All affected Boxers (American) also showed positive while a sample of UK Boxers, deduced by pedigree to be free of the gene responsible, proved negative. More intriguingly, but in a separate study, several dogs from a further UK sample, including some that were seemingly totally normal, although at risk by pedigree of carrying the gene, showed weakly positive suggesting they carried the gene and the test was offering an early diagnosis.
Finding this bio-marker represents a stunning discovery and it has since been probed and checked. The human work, supplemented by the main Boxer result, have been submitted for publication in a top American medical journal. I believe I have got it right when I say that only 40% of papers submitted to this journal get through the first editorial stage and go out to formal review. Dr Hamilton’s paper cleared this first hurdle immediately and I have great expectations for full reviewer acceptance as I think this a major milestone in human, and also Boxer cardiomyopathy research. For both species it opens up the possibility for a pre-clinical test which will serve virtually as a gene test for the disease. Much work still needs to be done to ascertain how early in life, and how effectively it picks up dogs which carry the gene but are apparently normal still needs to be ascertained; further investigation is needed.
For this further Boxer work, a first requirement is funding, and application has already been made to the AKC (about $50,000 is needed) and, given its importance for American breeders, I would hope that the ABC will step in to help. The second very obvious requirement is full breeder support without which nothing can be done. Here about 50 new affected cases will be needed plus a like number of clear-by-pedigree controls, plus as many close family relatives as possible, all with pedigrees (in total confidence). Breeders interested in helping would best contact me (email@example.com) in the first place. But no work or even sample collection will start until after the needed funding is obtained. In conclusion, the opportunity to get a sound working test for ARVC is now possible, but it is entirely up to Boxer breeders to help with the needed investigations. I do hope there will be major breeder support as an ARVC test is desperately needed in NA and, I’m afraid also in the UK.
Developments with JKD research are equally exciting. I wrote earlier about Professor Amos’ new findings, and progress continues He is quite confident that he has found the chromosome on which the gene for JKD lies, this indicated by an association between the disease and two of his micro-satellite markers on the chromosome. Moreover they mark the site to a small region in the chromosome where a number of genes lie, mutations in which could be responsible for the disease. He is now working through each of these genes, sequencing them for mutations. One gene has been eliminated and results on a second should be available by the end of November, and so it will continue until the gene with the causal mutation is found. If a mutation cannot be found then it all gets difficult; the mutation may lie in an inter-genic or regulatory region, which would make it very hard to find, but let’s hope for something simple.
Interestingly, we hear that the Swedish and Norwegian work has now also found what they believe is the location of the gene, and discussions on the possibility of all researchers working together henceforth are now in progress. However, although the Scandinavian interests are currently focussed on the diagnostic aspects of the work, any future collaboration will depend on agreement by everyone on the candidate region. Nobody is offering this information yet, and there are also practical problems to be resolved such as the partial duplication of material caused by the AHT contribution of swabs from UK dogs with the blood samples I sent to Norway earlier. With or without any collaboration this could bias the Scandinavian GWAS results. But it is all very exciting and illustrates the commitment of all researchers and their confidence that JKD has a basically simple inheritance. Professor Amos is still eagerly accepting swabs from affected normal dogs, and may be contacted directly for swab kits (firstname.lastname@example.org).
FB 60. It may seem that nothing is happening with JKD at the moment. As always this is far from the truth. In part this has been due to my change in email address which caused a number of reports on new cases temporarily to go astray. My first news , therefore, is that this has been rectified and all the new pedigrees are now up on the boxerjkd.com website, with few more yet to come in. They include JKD cases from countries other than the UK and the whole makes informative reading. The same pattern of inheritance is becoming evident everywhere.
Disappointingly, critical personal comments on the boxerjkd group written by Tim Hutchings have appeared in the British canine newspaper, Our Dogs. The seriousness of the JKD situation is also dismissed. Responding to this has taken up some time but the whole story to date can be viewed in the boxerjkd.com blog. It all makes for sad reading.
It seems that much of the problem is due a lack of understanding of the genetics of the disease and full appreciation of the pedigree evidence. Therefore boxerjkd.com is now trying to alleviate this by annotating the pedigrees to provide more key information; this is based on the published evidence. However, as not all pedigrees have been reported to us, notably those held by the Breed Council health committee, the annotations are incomplete. The annotated pedigrees can now be viewed on the boxerjkd website.
It should also be noted that the Norway list is incomplete. The Norwegian Boxer Club has decided that it wants to publish its own pedigrees, although with our annotation system. This may be appropriate and would save us much work. We are therefore closing our Norway list and replacing it with a link to the Norwegian one. Little difference will be noted in practice.
Of far greater importance however are the new research findings also reported in the boxerjkd blog. These comprise the incredible news from Professor Amos that he is confident that he has identified the chromosome on which the gene for JKD lies. Finding the chromosomal location of the gene is a major milestone in identifying the gene itself. More detail is given in the blog.
FB 58. There must be few people in Boxers who have not heard of the inherited juvenile kidney disease (JKD) that has risen to the fore in recent years. I have tried to keep everyone aware of the situation and as a professional geneticist and former Boxer breeder I have done my best, along with others (www.boxerjkd.com) to help breeders deal with the problem. Critical to this has been the publication of pedigrees of affected dogs on the boxerjkd website. Together with appropriate breeding guidelines, this is the only available option for breeders, and will continue to be, until the gene responsible is found. But clearly not everyone agrees with this, preferring instead to keep all available information hidden and leaving breeders with only ringside gossip to help them avoid walking into the disease. Typically my efforts have involved describing other people’s experiences which is not pleasant, but when JKD comes close to me, as it now has, the tragedy of it all becomes so much more poignant. So this message gives an account of a personal experience with JKD, but the situation is relevant for all.
Last year, as things appeared to be quietening down a bit with JKD, I had the wild thought of venturing back into the show ring after a Boxer cardiomyopathy- and JKD-forced long absence. The idea was triggered by an impressive photo of a young bobtail puppy. I enquired about her to learn of a further attraction; she was the granddaughter of a champion dog of mine I collected semen from in the 1970s. Expressing an interest I was offered her on condition I would show and have a litter from her. However, to be fair, she was not as good as her photograph, but a deal is a deal and I did my best on showing, not realising the difficulties of her exuberant wild temperament and my loss of fitness after 10 and more years sitting in front of a computer dealing with Boxer health problems. In the meantime the breeder had some very bad luck and decided to give up Boxers, which I really hoped would mean I would not have to breed her. She did have obvious JKD risks in her pedigree but fairly far back as is the case for so many Boxers these days.
But the next thing that happened was that I was contacted by an owner of a young bitch that had died of JKD earlier this year and who, on replacing the youngster with a pup from a known person in the breed, learned about my battles with JKD. But she also thought that I might have the sister. It turned out that I did. It was my young bitch. So, she could now be deduced not only to being at high risk of being a carrier, but also at risk of developing the disease herself at some point. This obviously put paid to any ideas on breeding from her, for which I was actually quite thankful, but of course left me with concerns for her future health. However what really had me fuming was that although the owner of the affected sister had reported the case to the health committee, the situation had not been reported to me. Had my interest in showing and maybe breeding not eased off I could have gone ahead in ignorance of the risks, and bred this high risk bitch, so to propagate the disease gene.
This is hardly a novel situation. Let me be clear. This note is not about me or my dog; it is about the whole breed situation. The central point is that the Breed Council health committee, supported by Council itself has a policy of NOT publishing information that could help everyone avoid JKD. How many other people, and dogs, must unknowingly be in this position? It is preposterous. I have been dismayed about this for a long time, but as it could have dragged me into the mess, I become totally disgusted. I find it quite criminal and have felt so strongly about the health committee avoiding basic action to limit propagation of the disease that I have written explaining the situation to the Kennel Club. What they can do, I don’t know, but I can say that I got a reply within the hour and am sure enquiries will be made. As it is I feel that my efforts and those of boxerjkd to help breeders avoid JKD are in effect just a waste of time. We have been handicapped by the health committee throughout, but the current withholding of pedigree information is far more serious. It should be up to breeders to decide what to do.
FB 55. We have a result for the prospective new ARVC test. It looks to be working. The new group of American dogs affected by ARVC all gave a positive response to the test as also did one British case that I was fortunate enough to find. More importantly for me personally was the uniformly negative response from the UK group deduced by pedigree to be free of the gene. So all in all we have a clear indication that the test serves to detect the disease. But the hope and expectation was that it would serve as a pre-clinical test. In other words would it pick up evidence of the disease before clinical signs appear and hopefully even before Holter can detect increased numbers of VPCs in runs etc to signify developing ARVC? I had included several dogs in the above study that could be deduced by pedigree to be at risk of having the gene. In other words, they had known transmitting dogs quite close up in their pedigrees and therefore might actually carry the gene although outwardly perfectly normal and free of the disease. What did they show? Amazingly, two of them gave what one might call ambiguous results. They showed positive, but only in a weak way. The simplest explanation is that we have the answer we really seek. The test is picking up normal dogs with hidden low expressing ARVC which is exactly what we hoped the test would do. It would look to be more sensitive than Holter.
It has to be proven that this is the correct interpretation but here I am on home territory as I know exactly what to do to establish the genetic facts. However, I don’t think I could convince owners and breeders to pay for more blood tests and no doubt also for Holter information. So looking forward, what can be done? And with ARVC all but extinct in the UK, what can UK breeders offer other than controls.
I conclude that any further investigations really will have to be done in NA. But, even though the Canadian researchers seem to expect me to resolve the genetic issues, I am unhappy about intruding into other country’s affairs and it is quite probable that NA breeders would not want me interfering in their business either. Would they indeed help me by reporting cases and the breeding. Would owners of related dogs submit with them for analyses? And then there is still the funding issue.
So here we are. I believe we have an incredibly valuable test within reach, one that is probably more sensitive than Holter and almost as good as a gene test, but we need the critical supporting evidence to establish this, and it will need the cooperation of numbers of NA breeders who would be willing to submit their show dogs for study. Will any step forward to help the verification of the test? Let me know (email@example.com). Pedigrees and full identification of dogs would be essential, but of course everything would be in the strictest confidence with no identities ever being disclosed. And then on top of this, there is the funding issue. Here we have a hold up, although funding is being applied for. Does anyone have any ideas? Otherwise we are pretty well stuck for the moment. So near and yet so far.
Following FB 54, I am happy to announce that despite a blip, the blood samples for the ARVC trial arrived safely and in apparently good shape in Toronto. The blip was missing a transport connection in Canada which delayed their arrival by a day. It is just as well we used a large box with 30 Kg of dry ice. Work starts on them almost immediately, and I’ll report the results if they arrive before I go on holiday next week. One trouble for me is that I don’t know what to expect; I don’t know whether the bio-marker marks the start of ARVC or only a secondary but early consequence of the disease. The test did not work perfectly in humans, but the dog disease and controls are much more clearly defined, so I have wild hopes for a 100% distinction between the two groups, although I am absolutely sure nobody else has such expectations. We’ll see.
FB 54. As I said in my last note, collecting blood samples for the ARVC trial has not been easy but, finally, samples from some 25 dogs were submitted to me for shipment to Canada. But what a palaver! This time the research group wanted me to take every possible precaution for safe transport. The list included creating a partial vacuum in every tube using a syringe and needle, sealing with parafilm and then wrapping each tube in its own plastic bag so that any leakages could be rescued. This morning with all finally achieved, the 60 plus tubes were loaded into a very large polystyrene traveling box filled with dry ice at -80 degrees centigrade and delivered to my old lab for collection by Fedex. What a relief to be rid of them, but now I wait in trepidation for the results. Will the test work and, more importantly for me, will my selection of dogs be correct? But I think this is the last time for me. I have had enough and it seems pretty clear that UK breeders have had enough too, as maybe also their vets. Regarding the latter, most have been very good and have collected the blood without charge. True, some have charged a standard surgery fee, but I have heard that one vet that wanted £100 for collecting from a single dog; and this exercise is for research, not for the owner. I was irate enough about this to write to the Chairman of the Veterinary Ethics committee. It is not right that owners have to pay to help research and, to be fair, neither is it right that vets should sacrifice time with their paying patients to facilitate this work. Something more formal needs to be organised. However, maybe on behalf of the various research groups involved I could thank all the breeders, owners and vets around the world who have done all that they can to help this and the other ARVC and JKD projects. Now let’s hope that the researchers can succeed in their quests for finding the Boxer ARVC and JKD genes and get this pre-clinical test for the ARVC successfully trialed.
FB 53. I write today, not yet to impart any new findings but just to keep you aware what is happening and also to make a couple of difficult points
Collecting UK controls for the ARVC preclinical diagnosis trial has not been easy. Either there is no real interest among UK breeders because ARVC is no longer a problem for the show section of the breed, or people have become fed up with me asking for one thing after another, whether blood, swabs, pedigrees or whatever. I thought at one point that I would have to write to Dr Hamilton in Canada and indicate that I could not raise the blood samples that were essential for his trial. But then a couple of noted breeders stepped in to boost the numbers that are being provided by old time friends who are still eager to help. I think we will make it now. When I have all the samples promised I’ll be shipping them off to Canada. I hope to hear the results within weeks or days.
On JKD, Professor Amos is working all hours, and I get frequent updates on how many samples he has extracted and entered on his database, and how many probes he now has working. His enthusiasm helps balance the dismal bits that one meets a bit too often at the non-scientific level.
One such dispiriting bit has been learning that one or two breeders abroad are threatening legal action if owners of JKD cases allow the pedigrees of their sick puppies to be published. Let me make the vital point that publication is to help breeders, all breeders, giving everyone a better chance of avoiding production of more affected pups. It is not an attack on anybody. How dismal can all this get?
At another somewhat dismaying level, it seems that some people think the various research groups are in competition and that one should support only one and not another. So, here, can I make the point that while the research groups are each trying to achieve the same results – to find the JKD gene – but they are doing so by different means, and these should, if successful, complement and validate each other. And yet other groups may come forward with new ideas, and these too should be supported. Handicapping one or other group by withholding research material will not help overall, and JKD presents challenge enough in its own right. So, please send blood samples, swabs or whatever is wanted to every research group seeking such material.
I think I read somewhere that only a third of efforts to find genes in dogs are successful. There are many technical reasons for this, not enough markers, not in the right place, not enough variability the exact functions of genes not known so one does not know what one is looking for. Add in all the complications with dog breeding, with upset owner and hostile breeders, plus the challenges of others affected, and the whole political scene, and the difficulties beyond the basic genetics are huge. So come on, folks, can we not work together to reduce the difficulties that we actually can control.
FB.52 Professor Amos is working furiously on his huge number of JKD samples and has already found something of very great interest for Boxer genetics, although not yet necessarily connected with JKD, but hugely intriguing all the same. However, this note is not about JKD, but rather about ARVC.
My Canadian cardiologist colleague, Dr Robert Hamilton who was a co-author in the ARVC paper has made a finding that I consider a breakthrough for Boxer breeders. It is what one might call a biochemical test for developing ARVC with the potential of recognising the disease BEFORE clinical symptoms appear. Affected dogs show a single band on an identifying gel while normal dogs, free of the disease, show no sign of this band. This is not a gene test but should serve breeders well as a simple diagnostic test for the disease. The effectiveness in dogs of different ages remains to be checked.
This question is being addressed right now in a trial but requires breeder help. Further ARVC cases are needed. This note is therefore a call for blood samples from more affected dogs. These are to verify that all accurately diagnosed animals have the identifying band. North American Boxers are the main source of such dogs as the disease remains a major problem in this part of the world. I believe that Kate Meurs is going to contribute to this part of the study. However, blood samples from numbers of normal dogs must also be checked to verify that none have the identifying ARVC band. Here lies a major problem in NA as Holter analyses only identifies a proportion of dogs that are at risk of developing ARVC at some time in their lives. Any collection of Holter-normal dogs from NA is therefore liable to be contaminated. This is where UK breeders can help. ARVC is now rare to non-existent in the show section of the breed in the UK, so that such dogs should NOT have the tell-tale ARVC band on gels.
This note is therefore a first appeal to all Boxer breeders for blood samples to contribute to a large scale trial being conducted by Dr Hamilton. Blood samples are wanted from ARVC affected dogs which will be mostly of NA origin and the hope is that UK Boxer breeders will provide samples from their normal clear stock (and any rare ARVC cases that may be found). Whole blood in EDTA, as collected in previous UK Boxer studies, will serve. So, in NA, anyone with fully-diagnosed ARVC cases and, in the UK, anyone who is having their dog’s blood tested for any purpose should request their vet to collect blood samples for this diagnostic trial too. Breeders with ARVC cases in NA should contact Virginia Zurflieh(firstname.lastname@example.org) for details on what to do. UK samples should be sent to me (contact: email@example.com) to freeze and hold for shipment to Dr Hamilton in Canada. Pedigrees (or registered names in the UK) will be needed. The total findings will be published in a paper for a medical/veterinary diagnostic journal; it is already in the process of being written. I should stress that the paper will NOT include any owner or dog identification. All information will be held in total confidence.
I hope there will be major NA and UK breeder support for this fantastic opportunity to find a way out of the general ARVC problem, as well as deal with any remaining threads in the UK.
FB 51. I am afraid that I don’t even try to keep up with all discussions in Facebook groups these days and consequently I am told that I have missed some upsetting correspondence concerning recent publications of pedigrees on BoxerJKD. The key to the problem is that owners and breeders of sick, dying, or dead dogs, diagnosed as having chronic kidney failure, have reported this information to BoxerJKD which has subsequently published the pedigrees as a matter of standard procedure, with owners’ permission. The result of this is that some owners and breeders have met considerable harassment and attempts to diminish the significance of their sad experiences.
Now all of us can understand the horror of learning that a dog that is one’s pride and joy has transmitted the gene for something nasty like JKD, especially as in a recent case the dog was one of the top dogs in the breed. The natural response is to take a defensive position and question the evidence. But the evidence here is strong. The critical factor is that a Juvenile Kidney Disease (JKD) is present in Boxer s and it is inherited; it is found in family groups, and this is the key element in its diagnosis. In this particular instance, the sire has produced four veterinary diagnosed kidney cases and is the grandsire of another published case. In addition it has a close relative who has produced several JKD cases, so all the evidence needed is there. There is not a doubt in the world that the kidney condition here is the inherited one.
But what else has been said against this conclusion?
1. It is stated, correctly, that a case was found NOT to have Renal Dysplasia (RD) on PM, this defined by the presence of premature cells in the glomerulus of the damaged kidney. The Swedish Veterinary School uses for this criterion for its research into Boxer kidney disease. This is fully justifiable for their purpose, but only a minority (1 in 8 cases) of Boxer kidney deaths are diagnosed as having RD (premature cells detected). The majority don’t, but still have a chronic kidney failure which is inherited (JKD), and this is the important point. One could say that RD is JKD with the premature cells additionally being found, and that RD and JKD are basically one and the same disease, as actually indicated by the finding that kidney cases with these cells evident (RD) and others without (JKD) can be found within the same families and even within the same litters. The absence of a specific RD diagnosis is therefore of little relevance for our purposes. The important point is that these cases have an inherited chronic kidney disease AND belong to a family within which the disease has repeatedly occurred. Evidence of the inheritance is THE vital diagnostic feature above all else.
2. It has been reported that that both parents of one of the JKD cases in question had typed negative with the Canadian Dogenes Renal Dysplasia test, implying they did not carry the responsible mutation. But the Dogenes test has been discredited even by the journal that published the original findings (Expression of concern: Novel allelic variants in the canine cyclooxgenase-2 (Cox-2) promoter are associated with renal dysplasia in dogs. PLOS ONE Editors..PLoS One. 2012;7(11):e49703. doi: 10.1371/journal.pone.0049703. The test would seem to be invalid, as this published case actually indicates. The gene test does screen for a particular mutation (in the Cox-2 gene) which seemingly is present in many Boxers, but this does NOT appear to be the gene responsible for the known inherited Boxer JKD.
3. One bitch of this breeding had a uterine tract infection, and it has therefore been suggested that lack of treatment of UTIs can cause the chronic renal failure. But UTIs are not uncommon in JKD cases. They are thought to be a CONSEQUENCE of the kidney malfunction and in fact have often led to the detection of JKD, but they are NOT the cause.
4. It has also been suggested that antibiotic treatment of young puppies can cause renal disease in adulthood. This concept is without any scientific evidence whatsoever, and it could never be imagined that vets would give treatments to pups that they knew would cause a renal failure later. And were there any risk at all, it would be true for all other breeds, whereas the incidence of chronic kidney disease in Boxers, according to the published insurance statistics, is 45 times higher than that observed in other breeds.
5. It is stressed that there are various environmental causes of kidney failure, such as drinking antifreeze, but one cannot imagine rare accidental events like this notably occurring in family groups of Boxers, in different ownerships, and living in various parts of the world, as seen in this family group.
The above are my comments as a geneticist but I hope to get a renal expert to add a specialist veterinary viewpoint. This will be published on www.BoxerJKD.com. I expect no important disagreement.
I would stress that the reporting of JKD cases is not an attack on any person, dog, or kennel. It is only a presentation of pedigrees from established cases. These, when put together, clearly demonstrate the inheritance of the disease and this will help everyone avoid producing further affected Boxers. The battle is with the disease and to win we all have to work together.
At this time when the magnitude of the problem and distribution of the gene responsible in the breed is barely understood the only breeding advice that can be recommended is that dogs and bitches that have produced JKD should not be bred from further. After all, the only way of being certain of perpetuating the problem is through continuing to breed from such animals.
Finally, I would urge those that have concerns about any of this to write to me (firstname.lastname@example.org) or send a message through www.boxerjkd.com and every effort will be made to resolve questions directly or with the help of specialists in the rapidly developing field of canine renal disease. This should not be a battle between people but rather a battle between all of us in Boxers and this lethal inherited disease.
Communication posted through Facebook