Recurrence rates and sites for grade II canine cutaneous mast cell tumors following complete surgical excision.

A retrospective study was performed on 31 dogs with completely excised, grade II, cutaneous mast cell tumors in order to determine recurrence rates and sites. Distant tumor recurrence developed in 22% of dogs, and local tumor recurrence developed in 11% of dogs; however, the vast majority of these animals were incompletely staged initially. Complete surgical excision of grade II mast cell tumors was associated with effective local control in 89% of these dogs. Therefore, adjuvant radiation therapy might not be indicated in the majority of dogs with complete surgical excision.     

Evaluation of surgical margins required for complete excision of cutaneous mast cell tumors in dogs

OBJECTIVE: To determine whether neoplastic mast cells extended into tissue 1, 2, or 3 cm laterally or deeper than 1 fascial plane from the visible edge of cutaneous mast cell tumors (MCTs) in dogs. DESIGN: Prospective study. ANIMALS: 21 client-owned dogs with > or = 1 cutaneous MCT PROCEDURES: After preparation for surgery, each dog's skin was marked 1, 2, and 3 cm from the tumor edge at 0 degrees, 90 degrees, 180 degrees, and 270 degrees. At each 3-cm mark, deep fascia was exposed and sutured to the skin; the tumor was excised in routine fashion and fixed in formalin. Tumors were graded; margins were examined histologically for neoplastic mast cells. RESULTS: 23 cutaneous MCTs in 21 dogs were included in this study. Fifteen (65%) tumors were located on the trunk, 5 (22%) on the hind limbs, and 3 (13%) on the head and neck. There were 3 (13%) grade-I and 20 (87%) grade-II tumors. All grade-I tumors were completely excised at all margins. Seventy-five percent of the grade-II tumors were completely excised at the 1-cm margin, and 100% were completely excised at the 2-cm margin. Two grade-II MCTs located on the hind limbs of dogs were excised with a complete but close (within 1 mm) deep margin. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a 2-cm lateral margin and a deep margin of 1 fascial plane appear to be adequate for complete excision of grade-I and -II MCTs in dogs.     

Periocular cutaneous mast cell tumors in cats: evaluation of surgical excision (33 cases)

Objective  To describe feline periocular cutaneous mast cell tumor (CMCT) clinical features, rates of local tumor recurrence and metastases, and cat survival time following surgical excision.
Animals studied  Thirty-three cats with periocular CMCTs.
Procedures  Medical records of cats diagnosed with periocular CMCTs were reviewed; cats were included if CMCTs were surgically excised and the diagnosis confirmed by histopathology. The appearance, size, location and histopathology findings of CMCTs were recorded. Rates of local recurrence, metastasis, and survival time following surgical excision were collected when available.
Results  All periocular CMCTs were restricted to the eyelids. In addition to surgical excision, three cats were treated with adjunctive therapy (strontium-90 irradiation or cryotherapy) intraoperatively. Local tumor control was achieved in 22/23 cats with a minimum follow-up of 30 days (median follow-up time of 711 days); one cat developed disseminated CMCTs but no local recurrence. Cats with periocular CMCTs had a median survival time of 945 days. Metastatic disease involving peripheral lymph nodes or abdominal viscera was not detected in any cat at any time during the study. All periocular CMCTs were classified as low-grade based on histopathology, and complete excision was achieved in approximately 50% of cases.
Conclusions  Surgical excision of periocular CMCTs in cats is an effective treatment option with rare local recurrence and metastases, even following incomplete surgical excision.     

Morphometry of canine cutaneous mast cell tumors

Twenty-four canine cutaneous nodules, diagnosed as mast cell tumors by fine-needle aspiration biopsy and confirmed by histopathologic analysis by staining with hematoxylin and eosin (HE) and toluidine blue, were analyzed by computerized nuclear morphometry on panoptic- and HE-stained cytopathology slides. Two hundred nuclei per lesion were examined. The morphometric parameters investigated were nuclear area, mean diameter, perimeter, regularity factor, and ellipticity factor. Lesions were graded as I (well differentiated), II (intermediate differentiation), or III (poorly differentiated) according to the following morphologic features: invasiveness, cellularity and cellular morphology, mitotic index, and stromal reaction. Nuclear morphometric results were then compared with histopathologic grades. Values of nuclear area, mean diameter, and perimeter increased with increase in histopathologic grade, but statistical analysis revealed significant differences only between grades II and III and between grades I and III when HE was used (P < 0.01) and between grades I and III with panoptic stain (P < 0.05). The ellipticity factor and regularity factor did not reveal significant differences between histopathologic grades. The results indicate that nuclear morphometric analysis, in combination with the rapid and inexpensive cytopathology technique, can help in mast cell tumor grading, thus contributing to the establishment of a more precise prognosis and treatment.     

Identification of matrix metalloproteinases in canine cutaneous mast cell tumors

Presence of matrix metalloproteinases has been associated with tumor invasion and metastasis in human neoplasia. The presence of matrix metalloproteinase 2 and matrix metalloproteinase 9 was determined in canine mast cell tumor tissue and normal stromal tissue from 24 dogs with spontaneously occurring cutaneous mast cell tumors. Seventeen of the mast cell tumors were of histologic grade 2, and 7 were of histologic grade 3. Gelatin zymography and computer assisted densitometry image analysis were used to quantify matrix metalloproteinase concentration. Bands from canine tissues migrated in the same location as human proenzyme and active enzyme matrix metalloproteinase 2 and matrix metalloproteinase 9 standards. A semiquantitative value for each patient sample was obtained by comparing the optical assessment density of each unknown band to the optical density of the human standard. The presence of matrix metalloproteinase 2 and matrix metalloproteinase 9 in histologic grade 2 mast cell tumors and histologic grade 3 mast cell tumors was compared, as was presence of matrix metalloproteinases in tumor and stromal tissue. There was dramatically more proenzyme matrix metalloproteinase 9 activity in histologic grade 3 mast cell tumors when compared to grade 2 tumors (P = .03). There was also dramatically more active enzyme matrix metalloproteinase 2 and active enzyme matrix metalloproteinase 9 activity in tumor tissue compared to stromal tissue (P = .02, P < .0001). This study demonstrates that the proenzyme and active enzyme forms of matrix metalloproteinase 2 and matrix metalloproteinase 9 are present in canine mast cell tumors. This appears to be related to the degree of histologic malignancy, although histologic grade 1 tumors were not evaluated.     

The treatment of canine mast cell tumours with electrochemotherapy with or without surgical excision

To describe the results of electrochemotherapy (ECT) in dogs with mast cell tumours (MCTs) either as first line therapy or as an adjuvant to surgery. The treatment combines administration of low dose chemotherapeutic drugs with the application of microsecond electric pulses, which cause the temporary permeabilization and increased porosity of the tumour cell membranes. The design of this study is a retrospective case series. A total of 51 dogs with MCTs were included and classified according to ECT procedure into 4 groups (ECT only, 15 cases, intra‐surgery ECT, 11, ECT Adjuvant to surgery, 14, Surgery followed by ECT, 11). The four groups (staged with location, size and grade) were evaluated to assess complete or partial remission, disease free interval, overall survival time and local toxicity. In this case series, Boxers, mixed breed and Labrador Retrievers, male dogs, between 4 and 9 years old were more represented. MCTs were predominantly grade 2 (Patnaik) and T stage 0–1, I‐1 (World Health Organization). Treated lesions were most commonly identified on the hindlimb and head where curative surgery would involve cosmetic or functional compromise. The intra‐surgery group of dogs showed the best disease free interval with Kaplan–Meyer analysis. Local toxicity induced by ECT ranged mostly from 1 to 4 in a 5‐point arbitrary scale with 0 – no toxicity to 5 – highest toxicity. In this study, ECT can be applied successfully as an exclusive therapy in smaller MCTs as an alternative to surgery. ECT can be combined with surgery either intra‐operatively or post operatively for larger lesions without significant toxicity.     

Evaluation of neoadjuvant prednisone administration and surgical excision in treatment of cutaneous mast cell tumors in dogs

OBJECTIVE: To determine response rate and reduction in tumor burden and effect of dose on tumor response in dogs treated with neoadjuvant prednisone for cutaneous mast cell tumors (MCTs). DESIGN: Combined prospective clinical study and retrospective case series. ANIMALS: 49 dogs with MCT. PROCEDURES: Medical records were retrospectively reviewed for dogs with primary untreated cutaneous MCT managed with neoadjuvant prednisone administration and surgery. Tumor characteristics and response to treatment were recorded. A subset of dogs assigned to low-dose (LD) treatment with neoadjuvant prednisone (1.0 mg/kg [0.45 mg/lb], PO, q 24 h) or high-dose (HD) treatment (2.2 mg/kg [1.0 mg/lb], PO, q 24 h) was used to determine the effects of dose. RESULTS: The overall objective response rate was 70% for dogs treated with neoadjuvant prednisone; prednisone dose was not significantly associated with response. Prospectively, the median sum maximal diameter (MaxD) reduction was 45.2%, and reduction in tumor volume was 80.6%. In both treatment groups, the mean percentage MaxD reduction and tumor volume reduction were significant. The difference in response between the LD and HD groups was not significant. The LD group had mean MaxD and tumor volume reductions of 35.4% and 52.5%, respectively, compared with mean reductions of 48.8% in MaxD and 78% in tumor volume in the HD group. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with neoadjuvant prednisone appears to be useful for inducing reduction of MCTs and may facilitate resection when adequate surgical margins cannot be confidently attained because of mass location or size or both.     

Factors influencing complete tumor excision of mast cell tumors and soft tissue sarcomas: a retrospective study in 100 dogs

The recommended treatment for soft tissue sarcomas (STS) and mast cell tumors (MCT) is complete surgical removal, provided that the tumor is amenable to surgical excision. The objective of this study was to evaluate possible risk factors for incomplete surgical excision of skin and subcutaneous STS and MCT in 100 dogs treated with wide excision with curative intent. Decreased body weight was a risk factor (P = 0.03, odd's ratio = 0.96) as well as increased tumor size (1.4% increase in risk of incomplete excision per cm(2); P = 0.02). Gender, age, breed, location, grade, tumor type, re-excision, and level of surgeon's training (P = 0.0711) were not significant. Veterinary surgery residents were at increased risk of incompleteness of excision compared with ACVS surgeons and ACVS surgeons with additional training in surgical oncology.     

Evaluation of a two-centimeter lateral surgical margin for excision of grade I and grade II cutaneous mast cell tumors in dogs

OBJECTIVE: To evaluate completeness of excision and clinical outcome in dogs with cutaneous mast cell tumors (MCTs) excised with a lateral margin of 2 cm and a deep margin of 1 fascial plane. DESIGN: Prospective study. ANIMALS: 16 client-owned dogs with 1 or more cutaneous MCTs. PROCEDURE: Excision of MCTs was performed with a 2-cm lateral margin and a deep margin of 1 fascial plane. Histologic tumor grading was performed; surgical margins were categorized as complete or incomplete. Follow-up information was obtained via repeat examination of the dogs by veterinarians or client-completed questionnaires. RESULTS: 4 grade I and 19 grade II cutaneous MCTs were evaluated. Overall, 21 (91%) MCTs were completely excised; 2 grade II tumors had foci of mast cells at the 2-cm margin. Two dogs received adjunctive treatments following surgery. Follow-up information was available for all dogs (median follow-up period, 379 days; range, 51 to 538 days); no local recurrence was detected during this time. De novo MCTs were detected in 3 of 16 dogs at 37, 54, and 154 days after surgery. Via Kaplan-Meier analysis, median survival time and disease-free interval were both > 538 days (medians not yet reached). No prognostic variables were identified. CONCLUSIONS AND CLINICAL RELEVANCE: Excision with a 2-cm lateral margin and a deep margin of 1 fascial plane may result in satisfactory excision of grades I and II MCTs in dogs, with recurrence rates similar to those reported previously. Use of these margins may minimize complications associated with larger local tumor resection.     

Immunohistochemical and clinical evaluation of p53 in canine cutaneous mast cell tumors

One hundred twenty-six cutaneous mast cell tumors obtained by excisional biopsy from 106 dogs were evaluated using immunohistochemical staining for the presence of p53 protein. A standard avidin-biotin immunohistochemical protocol was used incorporating a polyclonal antibody of rabbit origin (CM-1) as the primary antibody. Histopathologic grading of tumors was performed on hemotoxylin and eosin-stained samples. There was a significant difference in the percentage of cells staining positive for p53 for the histopathologic grades (P = 0.0005). Grade III tumors had a significantly greater p53 content than did grade I or II tumors (P < 0.05). Clinical data obtained retrospectively was available for 54 dogs. Tumor recurred in 19 of 54 (35.2%) dogs. Twenty-nine dogs died by the end of the study; 9 of 29 (31.0%) died of mast cell tumor disease. Histopathologic grade showed a significant negative association with survival time. Both clinical stage and histopathologic grade showed a significant negative association with time to recurrence. The percentage of cells staining positive for p53 did not significantly improve the forward analysis. Immunohistochemical detection of p53 did not appear useful in characterizing the clinical association between cutaneous mast cell tumor cellular features and survival time or time to tumor recurrence in dogs.     

Variation among pathologists in histologic grading of canine cutaneous mast cell tumors.

Ten veterinary pathologists at 1 veterinary institution independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors (MCTs). There was significant variation among pathologists in grading the MCTs (P < 0.001). The probability of assigning a low grade was significantly higher for the pathologists in this study who use a published reference for histologic grading of canine cutaneous MCTs that allows subcutaneous MCTs or MCTs with mitotic figures to be included in the low-grade category (P < 0.0001 and P < 0.0001, respectively).     

Cellular proliferation in canine cutaneous mast cell tumors: associations with c-KIT and its role in prognostication

Canine cutaneous mast cell tumor (MCT) is a common neoplastic disease in dogs. Due to the prevalence of canine MCTs and the variable biologic behavior of this disease, accurate prognostication and a thorough understanding of MCT biology are critical for the treatment of this disease. The goals of this study were to evaluate and compare the utility of the proliferation markers Ki67, proliferating cell nuclear antigen (PCNA), and argyrophilic nucleolar organizing region (AgNOR) as independent prognostic markers for canine MCTs and to evaluate the use of these markers in combination, as each marker assesses different aspects of cellular proliferation. An additional goal of this study was to evaluate the associations between cellular proliferation and c-KIT mutations and between cellular proliferation and aberrant KIT protein localization in canine MCTs. Fifty-six MCTs treated with surgical excision alone were included in this study. Each MCT was evaluated for Ki67 expression, PCNA expression, and KIT protein localization using immunohistochemistry; for AgNOR counts using histochemical staining; and for the presence of internal tandem duplication c-KIT mutations using polymerase chain reaction amplification. In this study, increased Ki67 and AgNOR counts were both associated with significantly decreased survival. On the basis of these results, we recommend that the evaluation of cellular proliferation, including evaluations of both Ki67 expression and AgNORs, should be routinely used in the prognostication of canine MCTs. Additionally, the results of this study show that MCTs with aberrant KIT protein localization or internal tandem duplication c-KIT mutations are associated with increased cellular proliferation, further suggesting a role for c-KIT in the progression of canine MCTs.     

Prognosis of canine patients with nasal tumors according to modified clinical stages based on computed tomography: a retrospective study

To evaluate the efficacy of clinical staging based on computed tomography (CT) imaging over the World Health Organization (WHO) staging system based on radiography for nasal tumors in dogs, a retrospective study was conducted. This study used 112 dogs that had nasal tumors; they had undergone radiography and CT and had been histologically confirmed as having nasal tumors. Among 112 dogs, 85 (75.9%) were diagnosed as adenocarcinoma. Then they were analyzed for survival time according to each staging system. More than 70% of the patients with adenocarcinoma were classified as having WHO stage III. The patients classified under WHO stage II tended to survive longer than those classified under WHO stage III. Dogs classified under WHO stage III were further grouped into CT stages III and IV, and CT stage III patients had a significantly longer survival time than CT stage IV patients. In addition, patients treated with a combination of surgery and radiation had a significantly longer survival time than the patients who did not receive any treatment in CT stage III. On the other hand, different treatment modalities did not show a significant difference in the survival time of CT stage IV dogs. The results suggest that WHO stage III dogs may have various levels of tumor progression, indicating that the CT staging system may be more accurate than the WHO staging system.     

Pre-operative neoadjuvant vinblastine-prednisolone in canine mast cell tumours: A single-centre retrospective cohort study

Neoadjuvant chemotherapy can be used in canine mast cell tumours (MCTs) to optimise surgical margins or to enable marginal excision in challenging locations. The objective of this study was to describe the outcome of dogs with cutaneous and subcutaneous MCTs treated with neoadjuvant vinblastine-prednisolone (NA-VP). Records of treatment-naïve dogs with cutaneous/subcutaneous MCT that received NA-VP were reviewed including signalment, indication for NA-VP, staging results, clinical response, surgical data and histopathology reports. For dogs with post-operative follow-up ≥365 days, predictive factors for local recurrence (LR) were evaluated. Forty-four dogs were included. NA-VP was indicated to optimise surgical margins (group MARG) in 19 dogs (43.2%) and to enable surgery (group MORB) in 25 dogs (56.8%). Complete and partial response were documented in 40.9% of dogs and 30 dogs (68.2%) underwent surgery. The indication for NA-VP was significantly associated with undergoing surgery (p < .001) on multivariable analysis. Twelve (48%) and 18 dogs (94.7%) underwent surgery in the group MORB and MARG, respectively. Five dogs (16.7%) experienced wound dehiscence. Complete excision was achieved in 14 dogs (46.7%). In dogs undergoing surgery with ≥365 days of follow-up, LR was documented in five cases (20.8%). None of the factors analysed including mitotic count, completeness of excision and response to NA-VP were associated with LR; notably, LR occurred in 3/11 (27.2%) completely excised MCTs. In a pre-operative setting, NA-VP appears safe and could be beneficial in selected cases. Prognostic factors such as clinical response, mitotic count and completeness of excision should be interpreted with caution following NA-VP.     

Immunohistochemical expression of p27 and p21 in canine cutaneous mast cell tumors and histiocytomas

The objective of this study was to evaluate by immunohistochemical means the nuclear expression of p27 and p21 proteins in cutaneous mast cell tumors and histiocytomas of dogs. In mast cell tumors, nine of the 13 grade I tumors, 13 of the 19 grade II tumors, and 10 of the 15 grade III tumors showed no detectable or mild p27 immunoreactivity. In contrast, one of the 13 grade I tumors, 12 of the 19 grade II tumors, and 11 of the 15 grade III tumors showed moderate or marked p21 immunoreactivity. Nineteen of the 28 histiocytomas showed no detectable or mild p27 immunoreactivity, and 24 cases showed moderate or marked p21 immunoreactivity. These findings indicate that a loss or absence of p27 expression is an early pathogenic event in mast cell and histiocyte tumorigenesis and that p21 expression may be a marker of mast cell tumor progression and histiocytoma cell proliferation.     

Treatment of canine mast cell tumors with CCNU (lomustine)

The efficacy and toxicity of CCNU (1-[2-chloroethyl]3-cyclohexyl-1-nitrosourea) were evaluated in 23 dogs with measurable mast cell tumors (MCT). Twenty-two dogs had cutaneous MCT and 1 dog had an intranasal MCT Nineteen (83%) dogs had biopsy of their original mass performed and 4 (17%) had aspiration cytology of masses. Of the 19 tumors histologically graded, 1 (5%) neoplasm was classified as grade I, 10 (53%) were grade II, and the remaining 8 (42%) were grade III. Dogs were treated with CCNU at a dosage of 90 mg/m2 body surface area every 3 weeks. Response could be evaluated in 19 dogs. Eight of the 19 dogs (42%) had a measurable response to CCNU. One dog had a durable complete response for 440 days. Seven dogs (37%) had a partial response for a median and mean duration of 77 days and 109 days, respectively (range, 21-254 days). Treatment with CCNU resulted in stable disease in 6 dogs (32%) for a median and mean duration of 78 days and 122 days, respectively (range, 42-347 days). The acute dose-limiting toxicity was neutropenia 7 days after administration of CCNU. The median and mean neutrophil counts 7 days after CCNU were 1,452 cells/microL and 1,683 cells/microL, respectively (n = 17). Other toxicoses were uncommon. CCNU should be considered an active agent in the treatment of MCT in dogs.