Margaret C. McEntee

Feline Vaccine‐Associated Sarcomas

Feline vaccine-associated sarcomas have presented many challenges. Initially, the etiopathogenesis and biological behavior of these tumors had to be characterized, and strategies implemented to move tumors away from problematic sites. Next, diagnostic and treatment recommendations evolved as the biologic behavior of vaccine-associated sarcomas forced early and aggressive intervention. Current therapeutic strategies are expensive, at times debilitating, and frequently fail to effect tumor control. This review summarizes the known history, epidemiology, etiology, and clinical management of vaccine-associated sarcomas after a decade of work. The next challenges must be to find more practical and effective solutions, and to eliminate the cause of vaccine-associated sarcomas.

Production of donor-derived sperm after spermatogonial stem cell transplantation in the dog.

Spermatogonial stem cell transplantation (SSCT) offers unique approaches to investigate SSC and to manipulate the male germline. We report here the first successful performance of this technique in the dog, which is an important model of human diseases. First, we investigated an irradiation protocol to deplete endogenous male germ cells in recipient testes. Histologic examination confirmed >95% depletion of endogenous spermatogenesis, but retention of normal testis architecture. Then, 5-month-old recipient dogs (n=5) were focally irradiated on their testes prior to transplantation with mixed seminiferous tubule cells (fresh (n=2) or after 2 weeks of culture (n=3)). The dogs receiving cultured cells showed an immediate allergic response, which subsided quickly with palliative treatment. No such response was seen in the dogs receiving fresh cells, for which a different injection medium was used. Twelve months post-injection recipients were castrated and sperm was collected from epididymides. We performed microsatellite analysis comparing DNA from the epididymal sperm with genomic DNA from both the recipients and the donors. We used six markers to demonstrate the presence of donor alleles in the sperm from one recipient of fresh mixed tubule cells. No evidence of donor alleles was detected in sperm from the other recipients. Using quantitative PCR based on single nucleotide polymorphisms (SNPs), about 19.5% of sperm were shown to be donor derived in the recipient. Our results demonstrate the first successful completion of SSCT in the dog, an important step toward transgenesis through the male germline in this valuable biomedical model.

Enhanced bioavailability of oral docetaxel by co-administration of cyclosporin A in dogs and rats

Pharmacokinetic and pharmacodynamic endpoints of intravenously and orally administered docetaxel (DT) with or without oral cyclosporine were characterized in rats and dogs. Plasma samples were analysed for DT using liquid chromatography-mass spectrometry. DT area-under-the-concentration-time curve, plasma clearance and maximum serum clearance were significantly affected by cyclosporine in rats (P <or= 0.0005). Bioavailability of DT alone and DT combined with cyclosporine was 1.3 and 82%, respectively, in rats. In dogs, cyclosporine resulted in a 14-17-fold increase in peak DT concentrations (P < 0.0005) and area-under-the-curve (P < 0.0005), and a 17-fold reduction in DT clearance (P < 0.005). Bioavailability of DT alone was 18 +/- 16% and statistically exceeded 100% (251 +/- 104%) when combined with cyclosporine in dogs. Two of the six dogs receiving DT plus cyclosporine developed neutropenia, and one of the six dogs experienced vomiting and diarrhoea. Cyclosporine significantly alters the disposition of oral DT in dogs and rats, such that clinically relevant serum concentrations are achievable.

Completeness of reporting of radiation therapy planning, dose, and delivery in veterinary radiation oncology manuscripts from 2005 to 2010.

Surrounding a shift toward evidence-based medicine and widespread adoption of reporting guidelines such as the Consolidated Standards of Reporting Trials (CONSORT) statement, there has been a growing body of literature evaluating the quality of reporting in human and veterinary medicine. These reviews have consistently demonstrated the presence of substantive deficiencies in completeness of reporting. The purpose of this study was to assess the current status of reporting in veterinary radiation oncology manuscripts in regards to treatment planning methods, dose, and delivery and to introduce a set of reporting guidelines to serve as a standard for future reporting. Forty-six veterinary radiation oncology manuscripts published between 2005 and 2010 were evaluated for reporting of 50 items pertaining to patient data, treatment planning, radiation dose, delivery of therapy, quality assurance, and adjunctive therapy. A mean of 40% of checklist items were reported in a given manuscript (range = 8-75%). Only 9/50 (18%) checklist items were reported in > or = 80% manuscripts. The completeness of reporting was best in regards to a statement of prescription radiation protocol (91-98% reported) and worst in regards to specification of absorbed dose within target volumes and surrounding normal tissues (0-6% reported). No manuscripts met the current International Commission of Radiation Units and Measurements (ICRU) dose specification recommendations. Incomplete reporting may stem from the predominance of retrospective manuscripts and the variability of protocols and equipment in veterinary radiation oncology. Adoption of reporting guidelines as outlined in this study is recommended to improve the quality of reporting in veterinary radiation oncology.

Three‐dimensional conformal radiation therapy alone or in combination with surgery for treatment of canine intracranial meningiomas

Treatment protocols, treatment planning methods and tumour types in studies evaluating radiotherapy for canine brain tumours have been varied. This case series retrospectively evaluated the outcome of definitive, three-dimensional conformal radiation therapy (3D-CRT) as either a sole modality or as an adjuvant to surgery in 31 dogs diagnosed with meningioma by histopathology (n = 10) or cross-sectional imaging of the head (n = 21, assessed independently by two board certified radiologists). Prescribed dose ranged from 45 to 54 Gy in 2.5 to 3 Gy fractions. Median overall survival was 577 days (interquartile range = 272-829 days; range = 30-1942 days) when all deaths were considered and 906 days (interquartile range = 336-912 days; range = 10 -1942 days) when only dogs dying due to meningioma were considered. No significant difference in survival time was detected for the defined clinical or imaging findings or between treatment with radiotherapy alone versus adjuvant radiotherapy, suggesting that 3D-CRT may be a viable alternative to surgery.

Evaluation of anti-human leukocyte antigen-DR monoclonal antibody therapy in spontaneous canine lymphoma

A pilot study of anti-human leukocyte antigen (HLA)-DR monoclonal antibody (mAb) in dogs with lymphoma was undertaken to verify the suitability of a canine model to address therapeutically relevant endpoints prior to a full trial in dogs, and ultimately human investigation. In vitro studies demonstrated that L243, a murine IgG1 anti-HLA-DR, binds to normal and malignant canine lymphocytes and induces apoptosis in canine lymphoma cells. Moreover, L243 was administered safely to normal dogs and dogs with lymphoma, and bound to malignant cells in nodal tissue. Preliminary evidence of transient disease stabilization was observed in a subset of dogs with advanced-stage lymphoma following L243 immunotherapy. hL243γ4P (IMMU-114), a humanized IgG4 anti-HLA-DR, currently under evaluation preclinically for human trials, was also shown to bind malignant canine lymphocytes, and safety and pharmacokinetic data from the administration of IMMU-114 to normal dogs indicate similar behavior to L243 in these assessments. These findings provide a rationale for the use of dogs with lymphoma in safety and efficacy evaluations of anti-HLA-DR mAbs for both veterinary and human applications.

A SURVEY OF VETERINARY RADIATION FACILITIES IN 2010

A survey of veterinary radiation therapy facilities in the United States, Canada, and Europe was done in 2010, using an online survey tool, to determine the type of equipment available, radiation protocols used, caseload, tumor types irradiated, as well as other details of the practice of veterinary radiation oncology. The results of this survey were compared to a similar survey performed in 2001. A total of 76 facilities were identified including 24 (32%) academic institutions and 52 (68%) private practice external beam radiation therapy facilities. The overall response rate was 51% (39/76 responded). Based on this survey, there is substantial variation among facilities in all aspects ranging from equipment and personnel to radiation protocols and caseloads. American College of Veterinary Radiology boarded radiation oncologists direct 90% of the radiation facilities, which was increased slightly compared to 2001. All facilities surveyed in 2010 had a linear accelerator. More facilities reported having electron capability (79%) compared to the 2001 survey. Eight facilities had a radiation oncology resident, and academic facilities were more likely to have residents. Patient caseload information was available from 28 sites (37% of radiation facilities), and based on the responses 1376 dogs and 352 cats were irradiated in 2010. The most frequently irradiated tumors were soft tissue sarcomas in dogs, and oral squamous cell carcinoma in cats.

Phase I and Pharmacokinetic Evaluation of the Combination of Orally Administered Docetaxel and Cyclosporin A in Tumor-Bearing Cats

Intravenously administered docetaxel (DT) is problematic in cats because of the requirement for premedication to ameliorate acute vehicle-induced hypersensitivity reactions. Previously we have revealed that therapeutic plasma concentrations of DT can be achieved in normal and tumor-bearing dogs when DT is administered PO in combination with oral cyclosporin A (CSA). The purpose of this study was to identify the maximally tolerated dosage and characterize the pharmacokinetic disposition of oral DT combined with CSA in cats with tumors. Eighteen tumor-bearing cats were enrolled in this phase I dose escalation and pharmacokinetic study. DT was administered by gavage with CSA (5 mg/kg) twice over a 3-week period. The starting dose of DT was 1.0 mg/kg. Based on the clinical toxicity profile, with gastrointestinal adverse effects and hematologic toxicity the maximal tolerated dose of oral DT was 1.75 mg/kg in combination with 5 mg/kg CSA. Additional studies are necessary to determine the efficacy of DT/CSA in cats with epithelial tumors.

Three‐dimensional conformal versus non‐graphic radiation treatment planning for apocrine gland adenocarcinoma of the anal sac in 18 dogs (2002–2007)

Differences in dose homogeneity and irradiated volumes of target and surrounding normal tissues between 3D conformal radiation treatment planning and simulated non-graphic manual treatment planning were evaluated in 18 dogs with apocrine gland adenocarcinoma of the anal sac. Overall, 3D conformal treatment planning resulted in more homogenous dose distribution to target tissues with lower hot spots and dose ranges. Dose homogeneity and guarantee of not under-dosing target tissues with 3D conformal planning came at the cost, however, of delivering greater mean doses of radiation and of irradiating greater volumes of surrounding normal tissue structures.

Treatment of advanced canine anal sac adenocarcinoma with hypofractionated radiation therapy: 77 cases (1999-2013).

Currently no standard of care exists for advanced, inoperable or metastatic anal sac adenocarcinoma (ASAC). The objective of this retrospective study was to assess the role of hypofractionated radiation therapy (RT) in 77 dogs with measurable ASAC. A total of 38% of dogs experienced a partial response to RT. For dogs presenting with clinical signs related to the tumour, improvement or resolution of signs was noted in 63%. For dogs presenting with hypercalcemia of malignancy, resolution was noted in 31% with RT alone and an additional 46% with radiation, prednisone, and/or bisphosphonates. Median overall survival was 329 days (range: 252-448 days). Median progression free survival was 289 days (range: 224-469). There was no difference in survival based on radiation protocol, use of chemotherapy, previous surgery or advanced stage. Radiation toxicities were mild and infrequent. Hypofractionated RT is well tolerated and is applicable in the treatment of advanced primary, locoregional or metastatic ASAC.