Unraveling the chaotic genomic landscape of primary and metastatic canine appendicular osteosarcoma with current sequencing technologies and bioinformatic approaches.

Osteosarcoma is a rare disease in children but is one of the most common cancers in adult large breed dogs. The mutational landscape of both the primary and pulmonary metastatic tumor in two dogs with appendicular osteosarcoma (OSA) was comprehensively evaluated using an automated whole genome sequencing, exome and RNA-seq pipeline that was adapted for this study for use in dogs. Chromosomal lesions were the most common type of mutation. The mutational landscape varied substantially between dogs but the lesions within the same patient were similar. Copy number neutral loss of heterozygosity in mutant TP53 was the most significant driver mutation and involved a large region in the middle of chromosome 5. Canine and human OSA is characterized by loss of cell cycle checkpoint integrity and DNA damage response pathways. Mutational profiling of individual patients with canine OSA would be recommended prior to targeted therapy, given the heterogeneity seen in our study and previous studies.

A case of feline injection-site sarcoma at the site of cisplatin injections.

A spayed 14-year-old female domestic shorthair cat presented with a squamous cell carcinoma of the nasal planum and was treated with intralesional chemotherapy. During nasal infiltrations with cisplatin mixed with the cat's own serum, a new carcinomatous lesion developed at the medial canthus of the right eye, which was also treated using intralesional chemotherapy. Two months after the treatment course, the cat developed a new mass at the site of the eyelid chemotherapy, which was diagnosed as a soft tissue sarcoma. At the owner's request, the tumour was marginally excised, but it recurred after 10 months. No lung or lymph node metastases were evident at the time of euthanasia. The histotype of the tumour, the coincidence with injections and the histological description make the hypothesis of an injection-site sarcoma likely. To the authors' knowledge, this is the first case of an injection-site sarcoma at the site of a cisplatin injection.

Localised radiotherapy for a ferret with possible anal sac apocrine adenocarcinoma.

A seven-year-old, neutered male ferret was referred to our hospital with two perianal masses (2.4x3.0 and 2.4x3.5 cm, respectively) that had recurred after initial surgical excision. Complete resection of the masses was impossible as there was deep invasion along the rectum. On histopathology, the masses were diagnosed as apocrine adenocarcinoma possibly of anal gland origin based on tumour location. There was marked response to localised radiotherapy using an orthovoltage unit at 4 Gy, twice weekly. No visible mass was detectable after six doses of radiation. However, at that time, pleural effusion was diagnosed and radiotherapy was discontinued. Cytology of a sample of the pleural effusion suggested mesothelioma, and no obvious pulmonary metastasis of anal sac adenocarcinoma were identified on thoracic radiography. The ferret died at home on day 71 after the first admission.

Lack of guanylyl cyclase C, the receptor for Escherichia coli heat-stable enterotoxin, results in reduced polyp formation and increased apoptosis in the multiple intestinal neoplasia (Min) mouse model.

Guanylyl cyclase C (GC-C), a transmembrane receptor for bacterial heat-stable enterotoxin and the mammalian peptides guanylin and uroguanylin, mediates intestinal ion secretion and affects intestinal cell growth via cyclic GMP signaling. In intestinal tumors, GC-C expression is maintained while guanylin and uroguanylin expression is lost, suggesting a role for GC-C activation in tumor formation or growth. We show by in situ hybridization that GC-C expression is retained in adenomas from multiple intestinal neoplasia (Apc(Min/+)) mice. In order to determine the in vivo role of GC-C in intestinal tumorigenesis, we generated Apc(Min/+) mice homozygous for a targeted deletion of the gene encoding GC-C and hypothesized that these mice would have increased tumor multiplicity and size compared to wild-type Apc(Min/+) mice on the same genetic background. In contrast, the absence of GC-C resulted in a reduction of median polyp number by 55%. There was no change in the median diameter of polyps, suggesting no effect on tumor growth. Somatic loss of the wild-type Apc allele, an initiating event in intestinal tumorigenesis, also occurred in polyps from GC-C-deficient Apc(Min/+) mice. We have found increased levels of apoptosis as well as increased caspase-3 and caspase-7 gene expression in the intestines of GC-C-deficient Apc(Min/+) mice compared with Apc(Min/+) mice. We propose that these alterations are a possible compensatory mechanism by which loss of GC-C signaling also affects tumorigenesis.

Effect of ovariohysterectomy in bitches with mammary neoplasms.

Ninety bitches with mammary tumours were studied for two years after the surgical removal of the primary tumour(s). Twenty-nine of the bitches had been spayed before the development of the mammary tumour, 22 were spayed when the tumours were removed and 39 were left entire. Fifty-eight of the bitches (64 per cent) had benign tumours and, of these, 15 (26 per cent) developed a new mammary tumour within two years, irrespective of whether the bitch was spayed. The other 32 bitches had malignant tumours which were grouped into 'invasive' and 'well defined' carcinomas. Sixty-three per cent of the spayed bitches and 57 per cent of the entire bitches, with invasive carcinoma were dead within two years of surgery as a result of their mammary tumours. For those with well defined carcinomas the tumour-related death rates were 18 per cent and 33 per cent respectively for the spayed and entire bitches. These findings suggest that ovariohysterectomy when mammary tumours are removed does not have a significant effect on the progression of malignant disease, and that about one in four bitches with a benign mammary tumour is likely to develop a further tumour in another gland.

Multiple digital tumours in a rottweiler.

An eight-and-a-half-year-old rottweiler bitch was examined on four separate occasions over a period of 32 months for lameness caused by a mass on a digit. On each occasion the mass and affected digit were amputated. Three different neoplasms were diagnosed: two squamous cell carcinomas, a melanocytoma and an intracutaneous cornifying epithelioma. There was no recurrence following surgical excision.

Radiation therapy for incompletely resected canine mast cell tumors.

The records of 56 dogs treated with megavoltage radiation for mast cell neoplasia were reviewed to determine the efficacy of this treatment modality. Total radiation dose ranged from 45 to 57 Gray (Gy), dose per fraction ranged from 3.0 to 4.0 Gy, and radiation treatment time ranged from 14-28 days. Median disease free interval (95% CI) was 32.7 (19-70) months. Median disease free interval for dogs older than 7.5 years was 15 (lower limit 7) months as compared to 62 (lower limit 20) for dogs younger than 7.5 years of age (p = 0.006). Median disease free interval for dogs with measurable disease was 12 (lower limit 5) months as compared to 54 (32-70) months for dogs with microscopic disease (p = 0.006). Radiation treatment time was also significantly related to disease free interval. Median disease free interval for dogs treated longer than 22 days was 12 (7-19) months as compared to greater than 50 (lower limit 20) months for dogs treated in 22 or fewer days (p < 0.001). This appeared to be due to more recurrences in dogs treated with 3-per-week fractionation and suggests that tumor proliferation in the interfraction interval may be important. Sex, tumor location, histologic grade, WHO clinical stage, number of radiation fractions, total radiation dose, and dose-per-fraction, as well as the following "yes/no" variables: steroids given, surgery prior to radiation, lymph nodes irradiated, and development of another mast cell tumor did not appear to influence median disease free interval or survival. Data presented herein support megavoltage radiation as an effective treatment for canine mast cell neoplasia, and suggest that disease free interval in dogs treated with daily fractions may be longer than that achieved with alternating day fractions.

Mutations of the p53 gene in canine lymphoma and evidence for germ line p53 mutations in the dog.

Mutations of the p53 gene are associated with a number of non-lymphoid cancers of the dog. The present study investigates the p53 gene status within canine patients treated for primary and secondary lymphoma. Three out of eight patients exhibited p53 gene mutations. These included one patient with a germ line mutation and two patients with de novo p53 mutations associated with the secondary lymphoma. Allelic loss of the p53 gene was also observed within primary and secondary tumours of the three canine patients. The results indicate that germ line p53 mutations exist in dogs and may be involved in the known predisposition of some breeds to cancer. The presence of therapy-related p53 point mutations was found to be associated with chemoresistant secondary lymphomas. A causative role for DNA-damaging chemotherapy in de novo mutation of the p53 gene is discussed. Characterization of p53 inactivation in canine tumorigenesis may provide a valuable clinical model for assessing the efficacy and optimal therapeutic regimens of anti-cancer agents.

Multiple malignant meningiomas in a young cat.

A 2-year-old cat was presented with generalised muscle tremors and progressive fore- and hindlimb ataxia, 5 months after the initiation of chemotherapy for thymic lymphoma. The lymphoma was treated with combination chemotherapy (cyclophosphamide, vincristine and prednisolone), which resulted in remission. The neurological signs progressed to paralysis and the cat subsequently died. On autopsy, multiple meningiomas were diagnosed, which is an unusual finding. It is possible that the lymphoma chemotherapy resulted in the development of the multiple meningiomas as secondary malignancies.

Pathology of canine bladder and urethral cancer and correlation with tumour progression and survival.

Biopsy and necropsy specimens, comprising 107 primary carcinomas and three mesenchymal tumours, were reviewed from 110 dogs with cancer of the bladder, urethra, or both. Histological classifications developed for the assessment of human bladder cancer were found to be readily applicable to the dog. These classifications are based on histological features, including the pattern of growth, the cell type, the grade of transitional tumour and the depth of invasion of the bladder wall. Features associated with localized disease in canine transitional cell carcinoma included papillary architecture, "in-situ" tumour, low tumour grade and a strong peritumoral lymphoid cell reaction. Features of tumours with metastasis included infiltrating and non-papillary architecture, increasing tumour grade, depth of invasion, vascular invasion and presence of peritumoral fibrosing reaction. Wide variability was found within single tissue samples, indicating that multiple sample sites are necessary for the adequate characterization of a given lesion. Statistically significant correlations were found between: tumour grade and depth of invasion (P < 0.0001); tumour grade and presence of metastases (P < 0.029); and peritumoral desmoplasia and metastases (P < 0.029). It was concluded that canine bladder cancer could be classified for the purpose of clinical management with a modified World Health Organization system as developed for human tumours.

Two haemangioendotheliomas in a bovine brain.

The microscopical, ultrastructural and immunohistochemical features of an unusual type of haemangioendothelioma in the brain of a calf are described. The salient features of this tumour were an exuberant proliferation of vasoformative elements whose endothelial cells exhibited epithelioid and spindle cell features. Intracytoplasmic miniature lumen formation in endothelial cells was characteristic of this vascular neoplasm.

Pathological and radiographical features of multicentric malignant fibrous histiocytoma in two dogs.

Widespread organ distribution of malignant fibrous histiocytoma, including osseous involvement, was demonstrated in two dogs. Both cases had a storiform-pleomorphic pattern histologically and immunohistochemical stains were used to differentiate this from other types of neoplasms with the same histological pattern. Radiographically the lesions were predominantly lytic in the metaphysis of long bones, although periosteal proliferation and axial skeletal involvement were seen in one dog.