Photo by Katrina Richardson.
Our research is still being impacted by COVID. Many projects continue to be delayed. But we have made a bit of progress on some fronts.
Dr. Carlos Alvarez, of Nationwide Children’s Hospital and The Ohio State University Colleges of Medicine and Veterinary Medicine, is studying the genetics of osteosarcoma in Deerhounds and other breeds. We have supported his work with both a direct grant and donating some of our CHF funds to his Canine Health Foundation project.
Dr. Alvarez’s 2020 experimental lab’s hiatus due to the COVID pandemic appeared to be over in early 2021. However, despite the lab opening again for work, the unanticipated problem was human resources. Although funding to hire three staff members was in place, the lab was only able to fill one of those positions. As a result, Dr. Alvarez’s lab subcontracted two other principal investigators to remotely support its work in statistics and DNA processing (Isain Zapata, Rocky Vista University, and Heather Huson, Cornell). Dr. Alvarez’s lab thus operated remotely for all of 2021. The two missing positions were recently posted again with the hopes of returning to normal operations in 2022.
Dr. Alvarez’s primary avenue of progress on canine osteosarcoma in 2021 was establishing a new collaboration with a complementary group of Italian veterinary oncologists. The collaboration was initiated by Dr. Marcella Massimini, Faculty of Veterinary Medicine, University of Teramo. She is a young investigator who works with well-established scientists who are also involved in the collaboration: Profs. Mariarita Romanucci and Leonardo Della Salda. Together they submitted a small grant application on the development of canine osteosarcoma FGF Receptor inhibition studies.
Fibroblast growth factors (FGF) are a family of cell signaling proteins that are involved in a wide variety of processes, most notably as part of normal development in animal cells. Some are released as a response to injury at injury sites. Fibroblast growth factor receptors (FGFR) are receptors that bind to fibroblast growth factor proteins. The FGF/FGFR signaling pathway is involved in a variety of cancers, including osteosarcoma.
Dr. Alvarez’s group reported in their 2019 paper the discovery of an FGF gene variant associated with osteosarcoma in Greyhounds, Irish Wolfhounds, and Rottweilers. Dr. Alvarez also found the FGF variant in the small initial Deerhound study he did last year. However, the variant is common across distantly related breeds and presumed to be associated with increased osteosarcoma risk in all large to giant breeds. Prospective studies are necessary to measure actual osteosarcoma risk, and therapeutic studies must be done to determine whether the FGF receptors can be drug targets.
Dr. Alvarez’s role in the new collaboration is to design and oversee the genetic studies and to train Italian scientists to perform those techniques. That would include Dr. Massimini visiting Dr. Alvarez’s lab and performing molecular studies there. The long-term goal of that proposal and another planned for 2021 is to validate FGF Receptor inhibition therapy and to determine whether most cases of osteosarcoma are likely to respond or whether genetic testing of likely responders is necessary (for example, in patient dogs with acquired amplification of FGFR1/2 gene loci). This is a promising opportunity because of the biological relevance and because there is great interest in FGF Receptor inhibitors for use as cancer therapies. That includes two FDA-approved compounds and several more in the pharma pipeline.
The collaboration is also going to edit a special issue of the journal Animals that will be published in 2022 on Veterinary Clinical Studies. The special issue title is “Frontiers in Comparative Oncology in Animals.” The collaborative group will also publish an article on canine osteosarcoma in that issue.
- Krook MA, Reeser JW, Ernst G, Barker H, Wilberding M, Li G, Chen HZ, Roychowdhury S: “Fibroblast growth factor receptors in cancer: genetic alterations, diagnostics, therapeutic targets and mechanisms of resistance.” British Journal of Cancer 2021, 124(5):880-892.
Anesthesia Sensitivities/Delayed Bleeding Syndrome
Research by Dr. Michael Court at Washington State University is studying the genetic causes of several health problems in Deerhounds, including Delayed [postoperative] Bleeding Syndrome (hyperfibrinolysis), hyperthermia (anesthesia and/or stress related), and slow drug metabolism (anesthesia and sedative sensitivity). We have supported Dr. Court both with a direct grant as well as with some of our CHF funds to support this CHF project.
Here is where things stand:
Delayed Bleeding Syndrome (Hyperfibrinolysis): As reported last year, Dr. Court has discovered a genetic mutation in Deerhounds that is associated with delayed postoperative bleeding, based on a retrospective case-control study. The results of this study were reported last June at the SDCA National Specialty in Richland, Washington (watch it here: https://www.youtube.com/watch?v=IuvBC1713Cg), and were submitted for peer review and publication. Since then he has been conducting additional studies to confirm these findings in Deerhounds (including whole genome sequencing), to determine whether the same mutation causes delayed bleeding syndrome in Greyhounds, and to develop a commercial DNA test. When available, this test would identify dogs that should be treated with antifibrinolytic drugs (aminocaproic acid or tranexamic acid) to prevent delayed postoperative bleeding if those dogs need surgery.
Slow Anesthetic Drug Metabolism: Dr. Court recently completed his CHF-funded project investigating the genetic causes of anesthetic drug sensitivity in sighthounds. Previous published work had identified a mutation in a drug metabolizing enzyme gene (CYP2B11-H3) that causes reduced anesthesia drug metabolism in lab-based experiments. This mutation occurs in Greyhounds and Deerhounds. The results from the current study showed that Greyhounds with the mutation metabolize certain drugs more slowly than dogs without the mutation. However, the difference was relatively small and did not completely explain the observed high variability in drug metabolism between dogs. Studies are underway to discover other genetic mutations that may explain the remaining variability.
Hyperthermia: Dr. Court continues to actively recruit cases for the hyperthermia study. Although preliminary studies have identified a mutation in the same gene that causes malignant hyperthermia in people, the numbers of cases in Deerhounds are still too small to make any strong conclusions. If you have a dog that experienced hyperthermia (developed a high temperature) during or after surgery, or other stressful event, or if you have DNA stored on a dog that experienced hyperthermia and have not already been in touch with Dr. Court, please contact him.
Samples Needed: If you have a dog that had an adverse event from a drug used for anesthesia, sedation, or pain relief, experienced a hyperthermic reaction, or experienced delayed postoperative bleeding, please participate in this study. Email Dr. Court for a cheek swab kit.
Five years ago we funded a project for Dr. Paula Henthorn, of the University of Pennsylvania, to continue her work on cystinuria in Deerhounds. That project has been completed (see the final report in the January/February 2019 Claymore), but her work continues on this condition in our breed (and many others!).
As previously reported, Dr. Henthorn and the genetics group at Penn have analyzed whole genome sequences and DNA variations identified by SNP (single nucleotide polymorphism) chips for Deerhound samples and have ruled out changes in the protein-coding portions of the two genes known to be responsible for cystinuria in dogs and people. They have also made progress in examining DNA variations, within and surrounding the two cystinuria-associated genes, that may affect the expression of the genes. In other words, they are looking at the parts of the genome that determine where a gene is expressed (what organ and type of cell), when the gene is expressed (anywhere from what time of day to what time of life, or in response to a different gene or environmental condition) and how much of the protein is made. So far, no specific DNA changes that are associated with cystinuria in Deerhounds have been discovered.
Advances in discovering the regions responsible for gene expression in the human genome continue to appear and can be analyzed in conjunction with available dog genome sequences, obviously including the genome sequences from normal and cystinuric Deerhounds, to aid this search.
In the meantime, Penn continues to receive DNA samples from dogs of different breeds—currently more than 30 breeds and counting—that appear to have Type-3 androgen-dependent cystinuria (based on the observation that all the stone-forming samples are from intact male dogs, although for eight of these breeds, there is only one dog in the study.)
The improvements made in the nitroprusside (NP) test, run by the PennGen Metabolic Laboratory (Drs. Werner and Casal), have now been in place for over two years and have been instrumental, not only for detecting dogs with increased urine cystine levels, but also in following the effects of neutering cystinuric male dogs and in determining if changing from a prescription diet to a non-prescription diet causes an increase in urine cystine concentrations. While there has not yet been a formal diet study, the preliminary results on ten dogs from various breeds show little or no change in the nitroprusside test result after a diet change from a prescription diet that supports urine health to a “typical” kibble. More data, as well as affordable access to standard urine amino-acid analysis, will be necessary to produce more definitive results.
Looking ahead, there are several areas where we want to learn more by performing additional studies:
- Broaden the scope of the search beyond the regions surrounding the two cystinuria genes by performing additional SNP-chip analysis to identify the locations of DNA variations associated with cystinuria.
- Perform total urine amino acid analysis (at UC-Davis or elsewhere) of normal and cystinuric Deerhounds to allow us to analyze all four amino acids involved in cystinuria to continue to improve our understanding of the amino acid changes associated with Type-3 cystinuria and to develop better assays for detecting the disease.
- Conduct a short survey to collect urinary health data on Deerhounds that have been NP tested. This will not only allow us to explore the potential uses of our improved nitroprusside test but will also provide the needed information to allow us to use the banked Deerhound DNA samples in our ongoing studies.
We will be submitting a grant to support these studies in 2022.
As always, we plan on offering free cystinuria testing of urine samples at the 2022 National Specialty. See below for details.
Samples Needed: Dr. Henthorn still needs DNA and urine samples from affected dogs and their relatives. In addition, additional samples were provided last year for the neutering study. These samples support the previous findings that neutering prevents stone formation.
However, they still need more samples, 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. Directions for sample collection can be found here.
Dilated Cardiomyopathy (DCM)
As you may remember, before COVID struck the SDCA Board approved a project to Holter monitor 100 Deerhounds to learn more about how heart arrhythmias relate to DCM. COVID created some logistical problems, and there were other logistical problems not caused by COVID, but, as of this writing, it looks like we will be ready to go early next year. YAY!
In the meantime, because it’s always a good idea to add to a project with which you are having problems, we added a component to the project where we are going to compare the results we get from monitoring with an Alivecor device, which allows you to track heartbeats using a smart phone, to the Holter results.
You can see how the project is doing by going to the DCM Project page on the Deerhound Health web site.
WE NEED DOGS FOR THE HOLTER STUDY. We are looking for dogs age four and older. Please contact us if you are interested.
Necropsy Study: The final component of the study is to collect tissue samples from dogs diagnosed with DCM that have died. (Please note: the dog does not need to die from the DCM.) If your dog has a confirmed case of DCM and passes away, we hope you will have your vet take some small tissue samples so we can learn about this awful disease. Sample collection information can be found at https://deerhoundhealth.org/samples-needed/cardiomyopathy-samples/
DNA: Also, we have agreed to provide samples of dogs affected with DCM to the U.K. Deerhound Club, as they are continuing work on the genetics of DCM. We have arranged that CHIC will send them the DNA so all samples can go into our CHIC DNA bank. We are asking that a little more blood than usual (7–10ccs) be sent, so we can send the British researchers all the DNA they need and still have some left for other projects. We can reimburse the $20 CHIC fee for samples submitted for this project—once the sample has been sent, just let Miranda know if you would like this reimbursement. Here are the directions for sending a sample to CHIC.
Testing: Testing to identify heart defects and DCM are recommended health tests for Deerhounds. Breeding stock should have a cardiac ultrasound (echocardiogram) before breeding, and all puppies should have their hearts listened to (auscultation) by a veterinarian before they go to their homes. The club is hoping to once again offer cardiac ultrasounds at the 2022 specialty; see below.
To date, four studies have been funded by the Canine Health Foundation’s (CHF) Bloat Initiative, which the SDCA has supported with our CHF Fund monies. All four studies have been completed, and the final reports have been summarized in previous years’ research wrap-up articles. We will continue to stay in touch with the CHF about this initiative.
It has been suggested that we look into sponsoring some research on chronic/recurrent pneumonia in Deerhounds. To do that, we need samples—lots and lots of samples, along with any associated diagnostic tests that may have been run on these affected dogs. For now, we are asking that you put DNA from affected dogs in CHIC.
The sooner we start actively collecting samples, the sooner we will find a researcher interested in taking this on.
CHIC DNA Bank
Banking DNA samples from our dogs—and keeping the health records updated—continues to be the single most important thing we can do to help our research projects, as currently all of our researchers have access to our CHIC DNA bank and all have used our CHIC DNA Bank samples as well as shared samples with each other. We couldn’t have made the progress that we have without all of our banked samples and updated health information. We currently have 363 samples in our DNA Bank and several hundred other samples with our researchers.
As always, we will be offering free CHIC DNA bank sample collection at the 2022 specialty; see below for details.
One way to follow the SDCA’s research efforts through the year is through the SDCA’s Deerhound Health web site and affiliated Facebook page, which usually post research updates, interesting studies, clinical trials, health clinic information, and other health tidbits. Please note that the site was redesigned last year and now has a new web address.
If you want to receive blog posts automatically, just click the “Subscribe to Blog via Email” button in the right sidebar of the site.
2022 National Specialty Sample Collection
We are planning on having a health clinic and sample collection at the 2022 National Specialty in Frankenmuth, Michigan, May 31–June 4; it’s a great opportunity to catch up on health testing and DNA collection on your dogs at the best price, and we do the shipping. 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. We haven’t confirmed pricing for this year yet, but testing has been FREE (Penn and the SDCA underwrite the processing costs) as long as a blood sample for research 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 discounted Factor VII testing (the SDCA underwrites part of the processing cost).
Cardiac Testing: We are holding a cardiac clinic.
As always, you can bring blood and urine samples to the show from dogs that you are leaving at home.
Here is more information on the health clinics.
You can always make a donation to the Bunnie Austin Health Fund: donate when you renew your membership or subscription or go to the Deerhound Store.
We can’t do all this work without your support. Thanks to everyone who made generous donations of samples and funds last year. And thanks to everyone for their patience during these very frustrating last two years.
If you have any questions about any of these projects, please don’t hesitate to ask.