Natasha J. Olby

Genome-wide association analysis reveals a SOD1 mutation in canine degenerative myelopathy that resembles amyotrophic lateral sclerosis

Canine degenerative myelopathy (DM) is a fatal neurodegenerative disease prevalent in several dog breeds. Typically, the initial progressive upper motor neuron spastic and general proprioceptive ataxia in the pelvic limbs occurs at 8 years of age or older. If euthanasia is delayed, the clinical signs will ascend, causing flaccid tetraparesis and other lower motor neuron signs. DNA samples from 38 DM-affected Pembroke Welsh corgi cases and 17 related clinically normal controls were used for genome-wide association mapping, which produced the strongest associations with markers on CFA31 in a region containing the canine SOD1 gene. SOD1 was considered a regional candidate gene because mutations in human SOD1 can cause amyotrophic lateral sclerosis (ALS), an adult-onset fatal paralytic neurodegenerative disease with both upper and lower motor neuron involvement. The resequencing of SOD1 in normal and affected dogs revealed a G to A transition, resulting in an E40K missense mutation. Homozygosity for the A allele was associated with DM in 5 dog breeds: Pembroke Welsh corgi, Boxer, Rhodesian ridgeback, German Shepherd dog, and Chesapeake Bay retriever. Microscopic examination of spinal cords from affected dogs revealed myelin and axon loss affecting the lateral white matter and neuronal cytoplasmic inclusions that bind anti-superoxide dismutase 1 antibodies. These inclusions are similar to those seen in spinal cord sections from ALS patients with SOD1 mutations. Our findings identify canine DM to be the first recognized spontaneously occurring animal model for ALS.

Molecular analysis of a spontaneous dystrophin `knockout' dog

We have determined the molecular basis for skeletal myopathy and dilated cardiomyopathy in two male German short-haired pointer (GSHP) littermates. Analysis of skeletal muscle demonstrated a complete absence of dystrophin on Western blot analysis. PCR analysis of genomic DNA revealed a deletion encompassing the entire dystrophin gene. Molecular cytogenetic analysis of lymphocytes from the dam and both dystrophic pups confirmed a visible deletion in the p21 region of the affected canine X chromosome. Utrophin is up-regulated in the skeletal muscle, but does not appear to ameliorate the dystrophic canine phenotype. This new canine model should further our understanding of the physiological and biochemical processes in Duchenne muscular dystrophy.

Recovery of pelvic limb function in dogs following acute intervertebral disc herniations.

Chondrodystrophoid breeds of dog are prone to explosive herniation of mineralized disc material into the thoracolumbar spinal canal. The resulting acute spinal cord injury may represent an excellent spontaneous model of acute traumatic spinal cord injury. The aims of this study were to quantify the recovery of dogs following acute disc herniations, to evaluate external factors that influence recovery, and to identify a group of dogs suitable for use in clinical trials on neuroprotective drugs. The gait of 88 dogs with thoracolumbar disc herniations was scored at the time of injury and 2, 4, and 12 weeks after surgical decompression. Dogs were placed into four groups dependent on the severity of presenting signs; dogs in group 1 had the most severe injury severity. Group 1 dogs showed a variable but incomplete recovery by 12 weeks. Dogs in groups 2 and 3 recovered uniformly but more completely, while dogs in group 4 made a rapid and excellent recovery and were deemed unsuitable for clinical trials. Combining dogs in groups 1, 2 and 3 produced a population of dogs with incomplete recovery by 12 weeks. Power analysis revealed that 87 such dogs would be needed per treatment group to detect a 20% change in function with a power of 95%. The number needed reduced drastically to 19 by eliminating dogs in group 1, but this produced less room for functional improvement. External factors did not appear to influence outcome. We conclude that dogs with spontaneous disc herniations provide a useful model of acute spinal cord injury for clinical trials.

The Pathogenesis and Treatment of Acute Spinal Cord Injuries in Dogs

Mechanical damage to the spinal cord initiates a secondary injury cascade that results in ischemia, disturbances in ion concentrations, excitotoxicity, production of reactive oxygen species, inflammation, and apoptosis. Over weeks to months a glial scar forms, and scarring can result in the development of syringomyelia. In the early phase of the disease, treatment should focus on maintaining systemic blood pressure and oxygenation, decompression of the spinal cord, and stabilization, if indicated. There are currently no proven neuroprotective drug therapies for limiting secondary damage, but blinded clinical trials are underway. Transplantation with olfactory ensheathing cells and mesenchymal stem cells show promise, as do potassium channel antagonists. Canine clinical trials of these therapies are ongoing.

‘Putting our heads together’: insights into genomic conservation between human and canine intracranial tumors

Numerous attributes render the domestic dog a highly pertinent model for cancer-associated gene discovery. We performed microarray-based comparative genomic hybridization analysis of 60 spontaneous canine intracranial tumors to examine the degree to which dog and human patients exhibit aberrations of ancestrally related chromosome regions, consistent with a shared pathogenesis. Canine gliomas and meningiomas both demonstrated chromosome copy number aberrations (CNAs) that share evolutionarily conserved synteny with those previously reported in their human counterpart. Interestingly, however, genomic imbalances orthologous to some of the hallmark aberrations of human intracranial tumors, including chromosome 22/NF2 deletions in meningiomas and chromosome 1p/19q deletions in oligodendrogliomas, were not major events in the dog. Furthermore, and perhaps most significantly, we identified highly recurrent CNAs in canine intracranial tumors for which the human orthologue has been reported previously at low frequency but which have not, thus far, been associated intimately with the pathogenesis of the tumor. The presence of orthologous CNAs in canine and human intracranial cancers is strongly suggestive of their biological significance in tumor development and/or progression. Moreover, the limited genetic heterogenity within purebred dog populations, coupled with the contrasting organization of the dog and human karyotypes, offers tremendous opportunities for refining evolutionarily conserved regions of tumor-associated genomic imbalance that may harbor novel candidate genes involved in their pathogenesis. A comparative approach to the study of canine and human intracranial tumors may therefore provide new insights into their genetic etiology, towards development of more sophisticated molecular subclassification and tailored therapies in both species.

Intervertebral Disk Degeneration in Dogs: Consequences, Diagnosis, Treatment, and Future Directions

Evidence of intervertebral disk degeneration (IVDD) is extremely common in dogs, and its prevalence increases with age. It has many important consequences because degeneration of the intervertebral disks often is a prelude to disk herniation, which can injure the spinal cord, spinal nerves, or both. This review summarizes the advances in diagnosis and treatment of IVDD that have been made since the 1950s when the first detailed description of the degenerative changes was published. It also discusses new approaches to treatment of the associated spinal cord injury and new methods by which to classify injury severity that are currently under development.

Evaluation of levetiracetam as adjunctive treatment for refractory canine epilepsy: a randomized, placebo-controlled, crossover trial

BACKGROUND There is little evidence-based information available to guide treatment of refractory epilepsy in dogs. The antiepileptic drug levetiracetam (LEV) is administered to dogs, although its safety and efficacy are unknown. OBJECTIVE To evaluate the safety and efficacy of LEV as adjunctive therapy for refractory epilepsy in dogs. ANIMALS Thirty-four client-owned dogs with idiopathic epilepsy. METHODS Randomized, blinded trial involving dogs resistant to phenobarbital and bromide. Dogs received LEV (20 mg/kg PO q8h) or placebo for 16 weeks, and after a 4-week washout were crossed over to the alternate treatment for 16 weeks. Owners kept records on seizure frequency and adverse events. Hemogram, chemistry profile, urinalysis, and serum antiepileptic drug concentrations were evaluated at established intervals. RESULTS Twenty-two (65%) dogs completed the study. Weekly seizure frequency during the 1st treatment period decreased significantly during LEV administration relative to baseline (1.9 ± 1.9 to 1.1 ± 1.3, P = .015). The reduction in seizures with LEV was not significant when compared to placebo (1.1 ± 1.3 versus 1.5 ± 1.7, P = .310). The most common adverse event was ataxia, with no difference in incidence between LEV and placebo (45 versus 18%, P = .090). No changes in laboratory parameters were identified and owners reported an improved quality of life (QOL) with LEV compared to placebo (QOL score 32.7 ± 4.3 versus 29.4 ± 4.5, P = .028). CONCLUSIONS AND CLINICAL IMPORTANCE Adjunctive treatment with LEV appears safe in epileptic dogs. Efficacy of LEV over placebo was not demonstrated, although the power of the study was limited. Further evaluation of LEV as treatment for epilepsy in dogs is warranted.

Clinical Characterization of a Familial Degenerative Myelopathy in Pembroke Welsh Corgi Dogs

BACKGROUND Adult dogs with degenerative myelopathy (DM) have progressive ataxia and paresis of the pelvic limbs, leading to paraplegia and euthanasia. Although most commonly reported in German Shepherd dogs, high disease prevalence exists in other breeds. OBJECTIVE Our aim was the clinical and histopathologic characterization of familial degenerative myelopathy (FDM) in Pembroke Welsh Corgi (PWC) dogs. ANIMALS Twenty-one PWCs were prospectively studied from initial diagnosis until euthanasia. METHODS Neurologic examination, blood tests, cerebrospinal fluid (CSF) analysis, electrodiagnostic testing, and spinal imaging were performed. Concentrations of 8-iso-prostaglandin F2alpha (8-isoprostane) were measured in CSF. Routine histochemistry was used for neuropathology. Deoxyribonucleic acid and pedigrees were collected from 110 dogs. RESULTS Median duration of clinical signs before euthanasia was 19 months. Median age at euthanasia was 13 years. All dogs were nonambulatory paraparetic or paraplegic, and 15 dogs had thoracic limb weakness at euthanasia. Electrodiagnostic testing and spinal imaging were consistent with noncompressive myelopathy. No significant difference was detected in 8-isoprostane concentrations between normal and FDM-affected dogs. Axonal and myelin degeneration of the spinal cord was most severe in the dorsal portion of the lateral funiculus. Pedigree analysis suggested a familial disease. CONCLUSIONS AND CLINICAL IMPORTANCE Clinical progression of FDM in PWC dogs was similar to that observed in other breeds but characterized by a longer duration. Spinal cord pathology predominates as noninflammatory axonal degeneration. Oxidative stress injury associated with 8-isoprostane production is not involved in the pathogenesis of FDM-affected PWC dogs. A familial disease is suspected.

Intervertebral disk disease in 10 cats.

The medical records of 10 cats diagnosed with intervertebral disk disease were reviewed. No apparent sex or breed predilection was found. The mean age of cats in the study was 9.8 years. Clinical signs included back pain, difficulty ambulating, and incontinence. Radiographs revealed narrowed disk spaces, mineralized intervertebral disks, and evidence of extradural compression on myelography or computed tomography. All intervertebral disk herniations occurred in the thoracolumbar spine, with a peak incidence at the fourth to fifth lumbar (L4-L5) intervertebral disk space. Eight cats had Hansens type I intervertebral disk herniation. Surgery was performed in seven cats. All cats judged to have an excellent outcome had undergone surgical decompression.

Microarray Analysis of Differentially Expressed Genes of Primary Tumors in the Canine Central Nervous System

The pathophysiologic similarities of many human and canine cancers support the role of the domestic dog as a model for brain tumor research. Here we report the construction of a custom canine brain-specific cDNA microarray and the analysis of gene expression patterns of several different types of canine brain tumor The microarray contained 4000 clones from a canine brain specific cDNA library including 2161 clones that matched known genes or expressed sequence tags (ESTs) and 25 cancer-related genes. Our study included 16 brain tumors (seven meningiomas, five glial tumors, two ependymomas, and two choroid plexus papillomas) from a variety of different dog breeds. We identified several genes previously found to be differentially expressed in human brain tumors. This suggests that human and canine brain tumors share a common pathogenesis. In addition, we also found differentially expressed genes unique to either meningiomas or the glial tumors. This report represents the first global gene expression analysis of different types of canine brain tumors by cDNA microarrays and might aid in the identification of potential candidate genes involved in tumor formation and progression.