Magnetic resonance imaging of canine mast cell tumors

Mast cell tumors (MCT) are the most common cutaneous tumors in dogs. Our purpose was to describe the magnetic resonance (MR) imaging characteristics of cutaneous MCT and to identify imaging characteristics that allow differentiation of metastatic from normal lymph nodes. Eight dogs with a total of nineMCT were imaged as were their presumed draining and associated contralateral lymph nodes. The signal intensity of tumors and lymph nodes was compared to adjacent musculature. On T2-W images, 7/9 MCT were hyperintense to muscle and 2/9 were isointense. On T1-W images, 8/9 MCT were isointense and 1/9 were mildly hypointense. All tumors were strongly contrast enhancing; 5/9 were homogeneous and 4/9 heterogeneous in their enhancement patterns. Six lymph node pairs were included in the evaluation (five sentinel lymph nodes with metastases, one without, and six contralateral lymph nodes). Metastatic lymph nodes were significantly larger than their contralateral lymph nodes (P = 0.039). All lymph nodes were isointense on T1-W images and hyperintense on T2-W images. 5/5 metastatic and 2/7 normal lymph nodes were heterogeneously T2-hyperintense. All lymph nodes were moderately to strongly contrast enhancing. 4/5 metastatic and 2/7 normal lymph nodes had heterogeneous enhancement patterns. While heterogeneity was more common in metastatic than in normal lymph nodes, this difference was not significant (P = 0.058 for T2-W images; P = 0.234 for postcontrast images). MR imaging may be useful in the presurgical evaluation and clinical staging of cutaneous MCT.     

Primary conjunctival mast cell tumor in a Labrador Retriever

A 4-year-old, intact male Labrador Retriever with a rapidly progressive conjunctival mass was evaluated. Ocular examination showed a 2-cm elongated mass arising from the superior bulbar conjunctiva of the left eye. The mass resulted in distortion of the palpebral fissure and contacted the superior aspect of the cornea without modifying its structure; no adhesion to the sclera was detected. The superior palpebral conjunctiva was unaffected, and the remaining ocular examination was normal. The initial diagnostic work-up included CBC, serum biochemical analysis, urinalysis, and fine needle biopsy of the mass. A poorly differentiated mast cell tumor was diagnosed by cytology. Immunocytochemistry was performed to evaluate Ki-67 proliferation index, and 54/1000 tumoral nuclei showed a dark red staining. After a complete clinical staging, the mass was excised and identified histologically as a grade-II mast cell tumor. An adjuvant treatment with prednisone and vinblastine was instituted because of the limited excisional margins. No evidence of local recurrence or metastasis has been apparent during the 29-month follow-up period. This report contributes to the current literature pertaining to canine conjunctival mast cell tumors; unfortunately, the paucity of case reports and the absence of large studies regarding this tumor make conclusions regarding its biologic behavior impossible.     

Phosphorylated KIT as a predictor of outcome in canine mast cell tumours treated with toceranib phosphate or vinblastine

Canine cutaneous mast cell tumour (MCT) is the most common malignant skin tumour in dogs and can exhibit variable biologic behaviour. Dysregulated signalling through the receptor tyrosine kinase (RTK) KIT can promote cell proliferation and survival, and assessment of its dysregulation via detection of activating c‐kit gene mutations or assessment of KIT protein localization is associated with multiple features of malignancy. The aim of the current study was to use a previously validated immunohistochemical (IHC) assay to directly measure phosphorylated KIT (pKIT) in order to investigate its association with other established prognostic markers, response to therapy, progression free interval (PFI) and overall survival time (OST) in dogs treated medically for measurable MCT. Tumour tissue from 74 dogs enrolled in a prospective study comparing toceranib and vinblastine for MCT treatment were evaluated for pKIT immunoreactivity. pKIT was variably expressed, with some degree of positivity observed in 49/74 cases (66%). pKIT immunoreactivity was significantly associated with aberrant KIT localization, high mitotic index and high histologic grade. On univariate analysis, pKIT immunoreactivity predicted shorter PFI and OST in the entire patient population as well as shorter PFI in the toceranib treated group, and was the sole predictive factor for OST upon multivariate analysis, while mitotic index was the sole independent predictive factor for PFI. These results demonstrate that IHC detection of pKIT correlates with several features of aggressive behaviour, and may confer information that is complementary to other prognostic factors. However, the role of pKIT in predicting outcome needs to be studied further before recommendations can be made for its routine use.     

Long-term postsurgical outcomes of mast cell tumors resected with a margin proportional to the tumor diameter in 23 dogs

We evaluated the postsurgical outcomes of cutaneous or subcutaneous mast cell tumors (MCTs, n=25) in 23 dogs, resected with lateral surgical margins proportional to the widest tumor diameter, including at least one facial plane. The tumor diameter range was 0.3–2.6 cm (median: 0.9 cm), and all were histologically diagnosed as Kiupel’s low-grade MCT. Resection was histologically complete in 20, close (deep margin) in three, and incomplete (deep margin) in two. No dogs developed local recurrence at the site of initial surgery during follow-up of 161–2,219 days (median: 976 days). These results suggest that resection of low-grade, relatively small MCTs with surgical margins proportional to the tumor diameter is a practical procedure with high success rates.     

Apoptotic intrinsic pathway proteins predict survival in canine cutaneous mast cell tumours

Mast cell tumours (MCTs) are the most frequent canine round cell neoplasms and show variable biological behaviours with high metastatic and recurrence rates. The disease is treated surgically and wide margins are recommended. Adjuvant chemotherapy and radiotherapy used in this disease cause DNA damage in neoplastic cells, which is aimed to induce apoptotic cell death. Resisting cell death is a hallmark of cancer, which contributes to the development and progression of tumours. The aim of this study was to investigate the expression of the proteins involved in the apoptotic intrinsic pathway and to evaluate their potential use as prognostic markers for canine cutaneous MCTs. Immunohistochemistry for BAX, BCL2, APAF1, Caspase‐9, and Caspase‐3 was performed in 50 canine cases of MCTs. High BAX expression was associated with higher mortality rate and shorter survival. BCL2 and APAF1 expressions offered additional prognostic information to the histopathological grading systems. The present results indicate that variations in the expression of apoptotic proteins are related to malignancy of cutaneous MCTs in dogs.     

Disseminated mast cell tumor infiltrating the sphenoid bone and causing blindness in a dog

Mast cell tumors are found in most organs and tissues with variable biologic behavior in dogs. This case illustrates the clinical and magnetic resonance imaging (MRI) findings in a dog with disseminated mast cell tumor infiltrating the sphenoid bones. A 6‐year‐old male neutered Greyhound presented with a 3‐day history of acute onset of blindness. General physical examination was normal. Neurological examination revealed mildly disorientated mental status, absent menace response in both eyes, bilaterally decreased vestibulo–oculocephalic reflexes and absent direct and consensual pupillary light reflex in both eyes. An electroretinogram indicated normal retinal function in both eyes. A lesion involving the middle and rostral cranial fossa was suspected. Hematology and serum biochemistry were normal except decreased urea (1.2 mmol/L). MRI of the head revealed heterogeneous signal intensity of the sphenoid bones on T2‐weighted images and loss of their normal internal architecture. Cerebrospinal fluid analysis was normal. Abdominal ultrasound revealed hepatosplenomegaly and mesenteric lymphadenopathy. Fine needle aspirates were taken from the jejunal lymph nodes and the spleen. Results were consistent with disseminated mast cell tumor. The owner declined any treatment and the dog was euthanatized. Postmortem examination confirmed disseminated mast cell tumor affecting multiple organs, including the sphenoid bones. To our knowledge, this is the first case describing MRI features of disseminated mast cell tumor affecting the sphenoid bones and causing acute onset of blindness in a dog.     

Mast cell tumor invading the cornea in a horse

A 3‐year‐old Marwari mare was presented for evaluation of an irregular, reddish mass protruding from behind the right third eyelid. The mass appeared to arise at the ventral limbal area, involved the perilimbal bulbar conjunctiva and widely extended into corneal tissue. No other ocular or systemic abnormalities were detected at the time of presentation. The mass was surgically removed by lamellar keratectomy, with defocused CO₂ laser used as adjunctive therapy to treat the surgical exposed area and its surroundings. Histopathologic evaluation showed sheets of densely packed, well‐differentiated neoplastic mast cells separated by fibrovascular connective tissue. Nuclear staining for Ki‐67 was performed, and an average of 370 cells were positive per 1000 counted cells. Two months postoperatively, the surgical site was filled with flat fibrovascular and pigmented tissue, while the surrounding cornea was transparent with no superficial vascularization around the fibrotic scar. Thirty‐two months after treatment, no recurrence of the neoplasia was reported.     

Treatment of canine mast cell tumours with prednisolone and radiotherapy

This retrospective study describes 35 dogs with non‐resectable, grade I–III mast cell tumours on the head or limb treated with prednisolone (40 mg m^?2 daily) for 10–14 days prior to radiotherapy (4 × 800 cGy fractions at 7‐day intervals) from a 4 MV linear accelerator. Prednisolone was continued at a reduced dose rate (20 mg m^?2) during radiotherapy and for 2 months or longer afterwards. Eighteen of 24 tumours (75%) decreased in size in response to prednisolone treatment. By 6–8 weeks following radiotherapy, 12 dogs had achieved a complete remission and 19 a partial response. Two tumours remained static and two progressed during the course of treatment. The overall response rate was 88.5%. With long‐term follow‐up, 11 dogs experienced local recurrence (n = 4), metastasis (n = 5) or both (n = 2). The median progression‐free interval was 1031 days (95% CI 277.44–1784.56, Kaplan–Meier), with 1‐ and 2‐year progression‐free rates of 60 and 52%, respectively. Tumour grade did not predict the prognosis for this group of dogs, but tumour location did affect the outcome. Dogs with tumours located on the limb survived longer than those with tumours on the head. The combination of prednisolone with radiotherapy appears to have a useful role in the management of measurable mast cell tumours sited on the head and distal extremities.     

Moh’s micrographic surgery for the management of a periocular mast cell tumor in a dog

A 3‐year‐old neutered male boxer dog presented with a 6‐month history of a waxing and waning mass of the left dorsotemporal eyelid margin. Cytology and biopsy confirmed a diagnosis of mast cell neoplasia. Systemic staging of the dog failed to reveal any evidence of metastatic neoplasia. Owing to the location of the tumor within the eyelid margin and the wide surgical margins recommended for excision of mast cell tumors, Mohs micrographic surgery (MMS) was chosen for its potential to conserve tissues while providing intraoperative confirmation the tumor was completely excised. Utilizing MMS horizontal sectioning technique, 100% of the surgical margins were assessed prior to closure of the surgical wound. This represents the first time a comprehensive MMS protocol was used in a veterinary patient under general anesthesia.     

Marked subcutaneous mast cell and eosinophilic infiltration associated with the presence of multiple Dirofilaria repens microfilariae in 4 dogs

Dirofilaria repens is a parasitic nematode in the subcutaneous tissue of carnivores, including dogs and cats, transmitted by mosquitoes. Human beings may be accidental hosts. Infection of a dog with D repens was first reported in Palestine in 1934, and 2 additional cases were reported in dogs in Israel to date. This report describes D repens infection in 4 dogs in Israel that presented with subcutaneous masses, which were cytologically characterized by marked mast cell and eosinophil infiltration. In 3 cases, multiple microfilariae were present in the lesions; rare microfilariae were present in the 4^th case. In all 4 dogs, PCR of fine‐needle aspirates from the lesions were positive for D repens. The mast cells observed in all lesions were uniform and highly granulated, and with the presence of the microfilariae, a mast cell tumor was considered less likely. This report suggests that D repens infection‐associated subcutaneous lesions, characterized cytologically by massive mast cell and eosinophil infiltration, should be considered a differential diagnosis for mast cell tumor, especially in geographic locations endemic for this nematode. Notably, all 4 dogs were infected with D repens despite a routine prophylactic doramectin therapy administered every 3 months, probably due to the relatively long time interval between treatments.     

Evaluation of bone marrow aspirates from multiple sites for staging of canine lymphoma and mast cell tumours

This prospective study evaluated the utility of bone marrow aspirates (BMAs) obtained from multiple sites for staging of canine lymphoma (LSA) and mast cell tumours (MCTs). Forty dogs (LSA, n = 24; MCTs, n = 16) were enrolled, but only 33 (82.5%) had diagnostic bone marrow (BM) aspirates obtained from two sites for inclusion in the study. Nineteen dogs with LSA were included, and 6 (31.6%) had BM involvement. Neoplastic lymphocytes were present in BM from both sites in all of these dogs. Fourteen dogs with MCTs were included, and 3 (21.4%) had BM involvement. Neoplastic mast cells were present at both sites in two dogs and at only one site in the third. These results indicate that BMAs from multiple sites may not be needed for accurate staging of canine LSA patients, but more studies evaluating the pattern of BM infiltration in dogs with high‐grade MCTs are warranted.     

Ultrasonographic findings in abdominal mast cell disease: a retrospective study of 19 patients

A retrospective survey from January 1989 to January 1999 of Tufts University Foster Hospital for Small Animals radiology records of 12 dogs and seven cats with cytologically or histopathologically confirmed abdominal mast cell disease was performed. Ultrasound changes in hepatic mast cell infiltration in dogs included a subjective increase in size, a diffuse increase in echogenicity, and one or more hypoechoic nodules. Ultrasound findings in the affected canine spleen included one or more hypoechoic nodules and a subjective increase in size. Two ultrasonographically unremarkable canine livers and one unremarkable spleen were found to be infiltrated by mast cells. The mast cell-infiltrated feline spleen was subjectively increased in size, mottled, irregular, or contained nodules. The affected lymph nodes in both dogs and cats were hypoechoic or inhomogeneous, subjectively increased in size, and rounded. Gastrointestinal involvement in cats was characterized by a thickened ileocecocolic junction or colon with loss of wall layering. Mast cells were not found in the gastrointestinal tract in any dog. One dog with mast cell infiltrate of the kidneys had multiple hypoechoic nodules in the cortex that distorted the outer contour of the kidney. Although these findings are not specific to the disease in either species, abdominal ultrasound is considered a useful tool for determining the extent of disease in small-animal patients with mast cell tumor if used in conjunction with histopathology or cytology.     

Eosinophilic conjunctivitis, herpes virus and mast cell tumor of the third eyelid in a cat

A 3-year-old Himalayan cat was diagnosed with concurrent eosinophilic conjunctivitis, herpes virus, and a conjunctival mast cell tumor. Eosinophilic conjunctivitis was verified via cytology from a conjunctival scraping, which revealed 50% eosinophils and 50% neutrophils. Herpes virus was verified via a positive polymerase chain reaction (PCR). Conjunctival scrapings for chlamydia immmunofluorescent antibody (IFA) and herpes IFA were negative. A mycoplasma was detected by a general mycoplasma PCR but the organism did not grow on the available mycoplasma media. The mass was excised and microscopic evaluation revealed a histiocytic mast cell tumor. The mast cell did not recur following local excision (at 1 year follow-up). The eosinophilic conjunctivitis was treated with both topical steroids and systemic megesterol acetate (Ovaban). When topical steroids were used, the herpes virus flared up and resulted in dendritic and geographic corneal ulceration. Therefore, the cat was treated with megesterol acetate and the eosinophilic conjunctivitis was well controlled. Treatment of eosinophilic conjunctivitis in the cat with megesterol acetate may be the treatement of choice due to the possibility of herpes virus.     

In situ c‐KIT mRNA quantification of canine cutaneous mast cell tumours and its relationship to prognostic factors

Cutaneous mast cell tumours (MCTs) are the most frequent malignant skin tumours in dogs. Mutations in the c‐KIT proto‐oncogene are correlated with the pathogenesis and aggressiveness of MCTs. To date, studies have focused on c‐KIT mutations and KIT protein localization, with a general lack of mRNA‐level analyses. In this study, c‐KIT mRNA expression was investigated in canine MCTs by RNA in situ hybridization (RNA‐ISH). Furthermore, we evaluated associations between c‐KIT mRNA expression and the histological grade, KIT immunohistochemical staining pattern and other clinicopathological parameters. c‐KIT mRNA expression was observed in all MCT samples, appearing as clusters of dots in the cytoplasm of neoplastic cells. A significant correlation was detected between c‐KIT mRNA expression (quantified according to the H‐score and the percentage of positive cells) and the histological grade (determined using two‐and three‐tier grading systems; P < .05). We also found a significant positive correlation (all P < .05) between c‐KIT mRNA expression and the proliferation indices (mitotic index, Ki‐67, and Ag67). However, no significant associations with c‐KIT expression from RNA‐ISH were found with respect to different KIT staining patterns. Overall, these results demonstrate that c‐KIT mRNA expression might be an additional tool for measuring the c‐KIT status in canine cutaneous MCTs and could serve as a potential prognostic factor. Further studies should evaluate the prognostic significance of c‐KIT mRNA expression in a large and uniform cohort of canine MCTs.     

Mast cell and plasma cell collision tumor in the spleen of a dog

A 9‐year‐old spayed female English Mastiff was referred for outpatient ultrasound due to a 3‐week history of weight loss, vomiting, and decreased appetite. Abdominal ultrasound showed multiple splenic masses of varying sizes and serum chemistry panel showed hyperglobulinemia. Cytologic examination of fine‐needle aspirates of the splenic masses indicated a mast cell and plasma cell collision tumor. Results of serum and urine protein electrophoresis and immunofixation indicated the plasma cell neoplasia was producing IgA immunoglobulins.     

Detection of canine transitional cell carcinoma using a bladder tumor antigen urine dipstick test

Canine transitional cell carcinoma (TCC) carries a poor prognosis in part due to late disease detection. The measurement of specific tumor markers shed in the urine may aid in sensitive, early disease detection and therefore improved prognosis. A 1-year prospective clinical trial was designed to assess the efficacy, sensitivity and specificity of the first generation Bard BTA test to diagnose TCC in dogs. This test is a qualitative, rapid, latex agglutination, dipstick test run on voided urine, which measures a glycoprotein antigen complex associated with bladder cancer in human patients. Sixty-five dogs were entered in the study: 20 TCC confirmed patients, 19 healthy controls and 26 urologic controls with a variety of conditions including urinary tract infection, crystalluria and proteinuria. Overall test sensitivity was 90% and specificity was 78%. False positive test results were noted in the presence of significant glucosuria (4+), proteinuria (4+), and pyuria or hematuria (> 30-40 WBC or RBC per hpf). Urine parameters that had no effect on efficacy included collection method (cystocentesis or free catch), pH, specific gravity, crystalluria, bilirubinuria, bacteriuria and casts. These data indicated that the Bard BTA test was sensitive for the detection of the bladder tumor-associated antigen complex in canine TCC. As evaluated, this test may serve as a useful adjunct to diagnosis, especially when cytology or biopsy is questionable or impractical. Furthermore, because of the high sensitivity of the test, it may be a practical screening test to rule out TCC in geriatric patients or patients with clinical signs related to the lower urinary tract, particularly before pyuria and hematuria develop which may interfere with test results.     

Expression of receptor tyrosine kinase targets PDGFR‐β, VEGFR2 and KIT in canine transitional cell carcinoma

Transitional cell carcinoma (TCC) is the most common neoplasia of the canine urinary tract. It tends to be locally invasive and has a moderate metastatic rate. Receptor tyrosine kinases (RTKs) play an important role in promoting cell growth, differentiation and regulation of cell function. RTK inhibitor toceranib phosphate has been used anecdotally to treat TCC. The goal of this study was to evaluate archived normal urinary bladder, TCC and cystitis bladder samples for expression of toceranib phosphate targets: VEGFR2, PDGFR‐β and stem cell factor receptor (KIT). A significant number of TCC samples expressed PDGFR‐β compared with cystitis and normal bladder samples (P<.0001). While all the tumour samples stained positively for VEGFR2, there was no significant difference between tumour, cystitis and normal bladder samples in intensity scores or staining distribution. Minimal positive staining for KIT was noted in the tumour samples. Based on this proof of target study, further investigation is warranted to determine clinical response of TCC to toceranib phosphate.