Reprinted from the January/February 2014 Claymore.

While there haven’t been any earth-shattering discoveries this year, research into the health problems that affect Deerhounds has made some slow and steady progress on several fronts. There are also a couple of new projects.

Osteosarcoma

Two researchers are actively working on osteosarcoma in Deerhounds. Dr. Carlos Alvarez, of Nationwide Children’s Hospital in Columbus, Ohio, is researching osteosarcoma in Greyhounds and is including in his research Deerhound samples he received from Dr. Marlene Hauck of North Carolina State University. Dr. Guillermo Couto has retired from Ohio State University, although he is still involved in this project.

This summer, a report on the Internet stated that Drs. Couto and Alvarez had “found” the gene that caused osteosarcoma in Greyhounds. I of course asked about this, and it turns out the report was a bit premature. Rather, they had found candidate genome regions, each implicating one or more genes. Dr. Alvarez has been doing genetic mapping to validate their findings and is now working with researchers at the Broad Institute, who have also been working on Greyhound osteosarcoma, to combine data and compare findings. Once the mutation(s) are discovered, Dr. Alvarez has promised to test his Deerhound samples to see if they have the same mutation(s) as Greyhounds.

The other researcher who is looking at osteosarcoma in Deerhounds (as well as other breeds) is Dr. Marlene Hauck at North Carolina State University. She very nicely wrote up a report of her project:

Sequencing of the transcriptiome (all transcribed genes) in the osteosarcoma of a Scottish Deerhound demonstrated an increased level of expression in a pathway named “Hedgehog.” This is a known oncogenic pathway in other tumor types, and there are drugs in various stages of development that will target this pathway. We have evaluated this pathway in cell lines, and there does appear to be inhibition of the growth of early passage osteosarcoma cells from tumors when the Hedgehog pathway is inhibited: we treated cells that grew from a tumor biopsy with Hedgehog inhibitors, and some of the tumors grew more slowly than untreated control tumors.

In evaluating canine osteosarcoma, we looked at messenger RNA (mRNA) levels for multiple genes in this pathway, and we found that one-third of the dogs appear to have increased expression of this pathway. We are currently looking at protein expression of the most commonly expressed gene with immunohistochemistry (using an antibody against the protein in tissue sections) to ensure that the increase in mRNA correlates with protein expression.

Previous write-ups on their research can be found at http://sdcahealth.wordpress.com/research/osteosarcoma-research-2/.

SAMPLES NEEDED: Please note that tumor and/or DNA samples are still needed from dogs affected with osteosarcoma. Sample collection instructions can be found at http://sdcahealth.wordpress.com/samples-needed/osteosarcoma-sample-collection-instructions/.

In the meantime, there are two clinical trials underway that test new treatments for dogs with osteosarcoma: There’s a study at Auburn University that is using a virus to kill cancer cells, and a researcher at the University of Pennsylvania who is exploring the use of a vaccine that causes the dog’s own immune system to kill cancer cells. More information on these studies can be found at http://sdcahealth.wordpress.com/category/clinical-trials/.

So while we wait…..and wait…..and wait for a genetic test, there may be some new treatments that will extend both the quality and quantity of life of our dogs with osteosarcoma. That would be huge.

Two more researchers have been interested in osteosarcoma in Deerhounds in past years:

Van Andel Institute

As I reported last year, researchers at Van Andel decided to focus on osteosarcoma in Great Pyrenees, as that was the breed where they had enough samples to move forward with their research. As of this summer, they had not yet reduced the candidate mutations in the Great Pyrenees to a single mutation.  Until they do, they are unable to perform surveys in other breeds. So their Deerhound research is on hold.

Jeff Phillips

Repeated attempts to contact Jeff have not gotten a response. I am going to keep trying and will post something on our web site when I have an update.

Cardiomyopathy

Last year, we started to work with Dr. Kate Meurs of North Carolina State University to begin a project studying dilated cardiomyopathy (DCM) in Deerhounds. To do this, she needs samples – 20 affected dogs would be a bare minimum. So far, she has collected a total of one sample.

SAMPLES NEEDED: If you have a dog affected with DCM, PLEASE participate in this study. The sooner we get enough samples, the sooner we can start the search for the genetic basis of this disease. To send a sample from an affected dog, go to http://sdcahealth.files.wordpress.com/2013/12/dna-collection-form-for-deerhound-oct-2013.pdf.

Cystinuria

Last year, the SDCA gave Dr. Paula Henthorn of the University of Pennsylvania $8000 to do a full-genome analysis of a cystinuric Deerhound. This was done, and the data compiled. Currently, Dr. Henthorn is analyzing the data, looking for the needle in the genetic haystack that will help to explain why cystinuria occurs in Deerhounds. To give you an idea of the scale of this type of search, here is a partial list of only some of the data:

Homozygous unique variants (almost 12,000)

Homozygous coding variants (over 16,000)

Homozygous unique coding variants  (53)

Dr. Henthorn is currently sifting through this data and continues to give us progress reports.

SAMPLES NEEDED: In the meantime, she still needs DNA and urine samples from affected dogs and their relatives. In addition, a few more samples were provided last year from dogs that participated in the neutering study, and so far, it still appears as though neutering prevents stone formation. However, they have not collected information from enough individuals to say this with any certainty, so if you have a dog that has formed stones or tested positive on a nitroprusside test and are going to neuter the dog, please collect the samples requested so your dog can participate in the neutering study. Sample collection information can be found at

http://sdcahealth.wordpress.com/samples-needed/cystinuria-samples/.

If you are planning on attending the 2014 national specialty, we will be collecting blood and urine samples for the cystinuria study there. See below for details.

Bleeding Problems

Fibrinolysis: Dr. Guillermo Couto, who was spearheading the fibrinolysis research at Ohio State University, has retired, and there is no one there who is continuing this research. We are currently weighing our options as to what is the best way to proceed so we can keep moving forward with this project. In the meantime, we can keep acquiring anecdotal information about this problem. If you have a dog that is experiencing post-surgical bleeding, and the administration of aminocaproic acid stops the bleeding, we would be very interested to hear that. Conversely, if you administered aminocaproic acid and your dog did not stop bleeding, we would be interested to hear about that, too. As always, this is for research purposes only and your information will be kept confidential. For more information on using aminocaproic acid for post-operative bleeding, go to http://sdcahealth.wordpress.com/health-issues/bleeding-problems/fibrinolysis/.

Factor VII: Even though we still don’t fully understand how Factor VII deficiency affects Deerhounds, because we have a genetic test for this condition, there has been little motivation to research this problem further, so no progress has been made.

This is a good place for a reminder that genetic tests don’t do anyone any good if they aren’t used: you should know the Factor VII status of your dog, and if you have to operate on a Factor VII affected dog, please make sure your vet is prepared for potential bleeding problems.

The other thing to remember about genetic tests is it is only through using all of the lines we have that we’ll be able to maximize the effectiveness of the tests and improve the health of our breed. Dr. Jerold Bell, the geneticist who spoke at one of our specialties (was it Kentucky?), writes, “A genetic test should not alter WHO gets bred, only WHO the dog gets BRED TO.” Factor VII is an autosomal recessive gene, so the breeding goal should be to never produce an affected individual. Factor VII clear dogs can be bred to any dog. Factor VII carriers and affecteds should only be bred to Factor VII clear dogs, although you should discuss with your vet the potential risks involved with breeding a Factor-VII-affected bitch.

If you are planning on attending the 2014 national specialty, the club will be offering deeply discounted Factor VII testing at the show. See below for details.

NEW! Portosystemic Shunt Research

I am happy to report that we were approached this year by a vet student in the United Kingdom who wants to do an initial study on portosystemic shunts in Deerhounds. In January, we sent an email via the various lists to Deerhound breeders asking them to complete a survey on the incidence of liver shunts in their dogs. This survey has also been sent to Deerhound breeders in the U.K. Here is the letter from the researcher, Lydia Kerridge, that accompanied the survey:

My name is Lydia Kerridge, a final year veterinary student at Cambridge Veterinary School, working under the supervision of Dr Penny Watson. I am undertaking a final year elective project looking at the incidence of portosystemic shunts in Deerhounds.

The aim of this questionnaire is to find out the incidence of portosystemic shunts in Deerhounds to see if it is increased in comparison to other breeds. If this research finds an increased incidence in Deerhounds, then further work can be done to look at why this is happening and possible ways of treating/preventing it in the future.

The Irish wolfhound breed has already been shown to have increased incidence of congenital portosystemic shunts, and some work has been done to ascertain why this is the case and how it can be reduced.

The questionnaires are anonymous unless breeders wish to provide their contact details at the end. All information will be held in the strictest confidence and be used only by myself for statistical purposes.

If you haven’t already completed the survey, the link is https://www.surveymonkey.com/s/FSVBJCM.

Although we in the Deerhound community feel as though we have an increased incidence of liver shunts, we don’t know that for certain, so this survey, provided it is completed by enough breeders, will go a long way toward pinpointing the actual incidence of this problem in Deerhounds. It’s an important first step before any more research can be done on this problem in our breed. Every breeder should complete the survey for as many litters as possible. Even if you’ve never had a liver shunt puppy, it is still important to complete the survey; researchers can’t figure out the incidence of a problem unless the survey is completed both by those who have had affected puppies and those who have not. You can complete the survey anytime between now and April 1. For more information, please contact Lydia at lck30 “at” cam.ac.uk.

SAMPLES NEEDED: If you have a puppy that is affected with a shunt, please collect some blood and store it in the CHIC DNA bank, so when we get to actually looking at DNA from affected individuals we have some samples available from affected dogs. For information on our CHIC DNA bank, go to http://sdcahealth.wordpress.com/samples-needed/chic-dna-bank-samples/.

Also, don’t forget that the club has an arrangement with Texas A&M University, which offers bile-acid testing at a reasonable cost. For more information, go to

http://sdcahealth.wordpress.com/health-issues/portosystemic-liver-shunt/liver-shunt-testing/.

NEW! Bloat/Torsion Research

In 2013, on the recommendation on the H&G Committee, the SDCA Board voted to support the Canine Health Foundation’s (CHF) Bloat Initiative with $2500 of our CHF Fund monies. The money in this fund must be spent on CHF projects, and this seemed to be a great match for our breed. In October, the first two projects were announced:

“Abnormalities in the Stomach’s Ability to Contract Predisposes Large-Breed Dogs to Bloat”

Principle Investigator: Laura L. Nelson, DVM; Michigan State University

Grant Amount: $233,774.00

Abstract: Gastric dilatation-volvulus (GDV or “bloat”) is a devastating disease common in large and giant-breed dogs. Occurring most frequently in older dogs with a close relative who has also suffered the condition, the stomach becomes both displaced and distended with air. Without emergency medical stabilization and surgical intervention, affected dogs quickly experience shock, damage to the stomach wall, and death. Most of the research relating to GDV has described risk factors for the disease, determinants of outcome with treatment, and the effectiveness of preventive surgery (gastropexy). However, the underlying cause of GDV remains unknown.  Abnormalities in the ability of the stomach to contract have been documented in dogs after naturally-occurring GDV. An analogous stomach condition in cattle, left-sided displacement of the abomasum (LDA) has been shown to, in some instances, be associated with abnormalities in the motilin gene. Motilin is an important driver of stomach contraction. This suggests that LDA and potentially GDV may be primarily caused by a stomach that does not properly contract, and that this condition may be inherited. The goals of Dr. Nelson’s study are to determine the relationship of abnormal stomach contraction with GDV and to define the biochemical and genetic alterations that may be associated with these stomach abnormalities. In the long term, they hope to develop a test to identify dogs at high risk for GDV that would allow selective breeding to eliminate the condition and to determine which dogs will benefit most from prophylactic gastropexy or other preventive therapies.

“Evaluating the Complex Genetic Basis of Bloat”

Principle Investigator: Claire Rebecca Sharp, BVMS; Tufts University

Grant Amount: $251,097.00

Abstract: Gastric dilatation and volvulus (GDV), or bloat, is a common condition in large and giant breed dogs with an unacceptably high morbidity and mortality rate. Due to the importance of GDV in many dog breeds, several previous studies have investigated potential risk factors for the development of GDV. It is known that there is no single cause for GDV, rather its occurrence is multifactorial, with both genetic and environmental factors contributing. Dr. Sharp proposes to further investigate how these risk factors cause GDV through the application of genomic and molecular methods. She will do this by analyzing samples from purebred dogs with GDV and comparing them to dogs of similar age and breed that have not developed GDV. She will perform a genome wide association study (GWAS) to identify differences in the genetic makeup of dogs with GDV, and see which genes are turned on and off in GDV (epigenomics). She will also determine if dogs with GDV have different types or amounts of proteins, hormones and other molecules in their blood and tissues (transcriptomics, proteomics and metabolomics). She and collaborators hypothesize that only when we put all of this information together (genomic, epigenomic, transcriptomic, proteomic and metabolomic) will we truly understand what causes GDV. The ultimate aim of understanding what causes GDV is to allow us to best intervene to prevent the disease from occurring.

I will post any updates or clinical trial announcements on our web page. In the meantime, if you have the misfortune to have a dog that bloats and/or torses, please put a DNA (blood) sample in our CHIC DNA bank so it’s available for the researchers when they need it.

Degenerative Myelopathy

Degenerative Myelopathy (DM) continues to be rare in our breed, but the researchers at the University of Missouri, who are studying this disease in many breeds, would still like to hear from owners whose Deerhounds have been diagnosed with DM. Here is a report from Liz Hansen, who is working on this project:

For degenerative myelopathy (DM), we have DNA tested 39 Scottish Deerhounds for the mutation that we know is commonly responsible for development of DM in dogs, and they have all tested NORMAL – they do not have the mutation. This includes randomly chosen samples from individuals DNA banked, samples collected at dog shows, and samples submitted from three dogs suspected to have DM due to clinical signs that appear to be DM-like, but none of these have the common mutation. We have identified a second mutation in the same gene, also causing DM, but only in a small percentage of Bernese Mountain Dogs (BMDs). The original mutation is much more common in BMDs, present in about 35% of the dogs tested from that breed. The second mutation is present in about 5% of BMDs. We have tested over 1000 dogs from other breeds and have not seen the second mutation present anywhere other than in BMDs. In the equivalent gene in humans, there are over 100 known mutations, all linked to ALS (Lou Gehrig’s Disease), plus there are about a dozen other genes with mutations also responsible for some forms of ALS. In the dog, the one original mutation is clearly responsible for the vast majority of DM in many breeds and in mixed-breed dogs, but as the discovery in BMDs points out, there can be other rare forms of DM in dogs. It is possible that there is a different mutation at work in Deerhounds that nobody has discovered yet. We are interested in identifying additional mutations causing these rare and as yet undefined forms of DM.

We first need to establish that DM is really the cause for clinical signs being seen in Deerhounds, and to do that, post-mortem microscopic examination of a section of the spinal cord is still the “gold standard.” It would be very useful to have tissues sent from a Deerhound suspected to have DM at the time it is euthanized. We have a protocol on the website, (www.CanineGeneticDiseases.net under DEGENERATIVE MYELOPATHY and in the RESEARCH section), or an owner or veterinarian can email me (HansenL@missouri.edu) to request the tissue protocol. Once we have a well-defined, confirmed case, then we can search for the mutation that caused the problem. Other spine and spinal cord problems can mimic the symptoms of DM, so it is important to confirm the disease, and then launch a search.

For more information about this study, go to http://sdcahealth.wordpress.com/research/degenerative-myelopathy-research/.

CHIC DNA Bank

One reason why researchers are willing to work with us is because we have been diligent about making DNA samples available from affected dogs. This continues to be the single most important thing we can do, so it is crucial that we continue to bank samples. Most of our researchers have access to our CHIC DNA bank. In addition, for the health problems for which we don’t have researchers accepting samples yet, such as bloat/torsion, liver shunt, chronic/recurrent pneumonia, and other problems that may have a genetic component, it is crucial that we bank samples from affected individuals now. For one, it would be a great selling point to potential researchers if we already had samples from affected individuals. Secondly, once we have interested researchers, we wouldn’t have to wait for years while we slowly build up samples. So please, bank all of your dogs’ DNA in our DNA bank, but this is especially important if you have an dog affected by a disease seen in Deerhounds. Directions for sample collection and shipment can be found here: http://www.caninehealthinfo.org/dnabank.html. If you are planning on attending the 2014 national specialty in June, we will be offering free DNA bank sample collection and shipment there. See below for details.

Also, be sure to provide health updates on your CHIC samples regularly. In fact, you should have recently been requested to update your CHIC health records. Remember, the samples have zero value unless the health records are current, so please update your records now.

Web Site

You might have noticed that there have been many references to our web site in this article. That’s because one thing that did get done this year was the complete redesign and reorganization of the Health page on the SDCA web site:

http://www.deerhound.org/health.shtml

You can also access it by going to the SDCA web page (www.deerhound.org) and clicking on the “Health Issues” page. These link you to the WordPress site whose address is given above.

On this web site is one-stop shopping for information about Deerhound health issues. You can find research updates and sample collection instructions for all of the research projects and our DNA bank. There’s a blog that posts interesting studies, clinical trials, and other health tidbits. There are some fun photos on the site, too, that will change periodically. If you want to receive the blog posts automatically, just click the “Follow” button in the lower right corner of the site. You can also bookmark the site, but make sure the full address—http://www.deerhound.org/health.shtml—is listed in the address bar when you bookmark it.

2014 National Specialty

If you are planning on attending the 2014 national specialty, it’s a great opportunity to catch up on health testing and DNA collection on your dogs at the best price, and we ship all samples for you. Here’s what we’ll be offering:

CHIC DNA Bank: we will be drawing blood samples for submission to our CHIC DNA Bank. Cost: FREE! The SDCA and OFA underwrite the processing cost.

Cystinuria Study: We will be drawing blood for DNA and accepting urine samples for NP testing to support our cystinuria research. This year, we would like samples from both dogs and bitches age 2 and over. Testing is FREE (Penn and the SDCA underwrite the processing costs) as long as a blood sample is also provided or Penn already has a blood sample from your dog. Even if your dog has had an NP test before, multiple tests are crucial for the research. Urine collection equipment will be available.

Factor VII: Once again, we will be offering Factor VII blood draws, and the SDCA will subsidize $45 of the $75 fee for Factor VII testing at the University of Pennsylvania. An added bonus is Factor VII samples can then be used by Penn for their research.

Cardiac Testing: We have made arrangements with a cardiologist to do auscultations and cardiac ultrasounds (echocardiograms) at the show. Right now, it looks like the cardiologist will be at the show on Wednesday and Thursday, and we’ll be charging approximately $65 for an exam/auscultation and $175 for an auscultation/echo.

Feel free to bring blood and urine to the show from dogs that you are leaving at home.

For more information on the specialty health testing, feel free to contact me or Laura Studer at lstuder “at” allcreatures24hr.com.

Remember, even if you can’t attend a specialty, the club will help underwrite some of the cost for research samples. For more information on individual sample collections, go to http://sdcahealth.wordpress.com/2013/10/03/sample-collection-expenses-reimbursement/.

Finally, DONATE!

One very exciting thing that has happened is the price of a Whole Genome Sequence (WGS), which examines every gene on every chromosome, has dropped significantly. We are already using this technology in our cystinuria study, and other breeds have had some pretty dramatic discoveries using it. It makes it possible for a breed club to make real progress in research, because a WGS is now within most clubs’ budgets. The other beauty of it is, if you choose your dogs carefully, it is possible to provide data to multiple studies.

While we wait for the initial results to come from our cystinuria study, we are looking into what the possibilities are to use WGS to help us move forward with some of our other health issues. We think there is real potential and will keep everyone posted on progress made in this area or if there’s a WGS we should do to keep us moving forward. In the meantime, we are going to need more money to do WGS: each costs about $5000-7500, and most studies need more than one dog sequenced. So please donate as generously as you can to the Bunny Austin Fund. We are finally making some real, honest-to-goodness progress, but we need to keep going. Although doing WGS—or any research, for that matter—is no guarantee that a huge discovery will be made, we know that if we don’t do anything, nothing definitely will be accomplished.

It’s easy to donate to the Bunny Austin Fund: you can donate when you renew your membership or subscription or send a check to Treasurer Ann Hammond anytime: her address is 1422 Stratton Avenue, Nashville, TN 37206. To use your credit card, go to http://sdcahealth.wordpress.com/donate/ and click on the “Donate” button to pay via PayPal.

Thanks to everyone who made generous donations of samples and funds that support our research program; we would make zero progress without them. If you have any questions about our research projects or sample collection, please don’t hesitate to ask. And let’s all hope that 2014 is the year that brings us one or more earth-shattering discoveries, or at least great progress.

Miranda Levin, Research Liaison (ONB), Health & Genetics Committee