Recurrence rates and clinical outcome for dogs with grade II mast cell tumours with a low AgNOR count and Ki67 index treated with surgery alone

Grade II mast cell tumours (MCT) are tumours with variable biologic behaviour. Multiple factors have been associated with outcome, including proliferation markers. The purpose of this study was to determine if extent of surgical excision affects recurrence rate in dogs with grade II MCT with low proliferation activity, determined by Ki67 and argyrophilic nucleolar organising regions (AgNOR). Eighty‐six dogs with cutaneous MCT were evaluated. All dogs had surgical excision of their MCT with a low Ki67 index and combined AgNORxKi67 (Ag67) values. Twenty‐three (27%) dogs developed local or distant recurrence during the median follow‐up time. Of these dogs, six (7%) had local recurrence, one had complete and five had incomplete histologic margins. This difference in recurrence rates between dogs with complete and incomplete histologic margins was not significant. On the basis of this study, ancillary therapy may not be necessary for patients with incompletely excised grade II MCT with low proliferation activity.     

Hypotonic water as adjuvant therapy for incompletely resected canine mast cell tumors: a randomized, double-blind, placebo-controlled study

Objective— To evaluate efficacy of hypotonic water as adjuvant therapy after marginal resection of canine mast cell tumors (MCT).
Study Design— Double-blinded, placebo-controlled, prospective, randomized study.
Animals— Dogs (n=30) with spontaneous, cutaneous, solitary MCT.
Methods— The wound bed of MCT, resected with margins <0.5 cm, was injected with either hypotonic or isotonic water according to a standardized protocol. Follow-up was obtained by clinical examination at 1, 2, 3, 6, and 12 months and annual telephone contact with the owner.
Results— Eighteen dogs were treated with isotonic lactated Ringer's solution and 12 dogs with hypotonic distilled water. All MCT were stage 0 tumors and most grade II. Six tumors (4 isotonic, 2 hypotonic) recurred locally, 3 of these dogs died from disease-related reasons within 4 months. The surviving 3 dogs were alive with a median survival time (ST) of 1092 days. The calculated 2-year recurrence-free rate was 92.7%; the 2-year disease-free rate 79.1%; and the 2-year survival rate 89.5%. No significant differences in local recurrence and ST were observed between treatment groups. Histologic grading was the only significant prognosticator for ST and recurrence-free periods.
Conclusion— No significant differences in local recurrence and ST were observed between adjunctive hypotonic water and placebo treatment after marginal resection of solitary MCT.
Clinical Relevance— Hypotonic water does not decrease the rate of local recurrence in dogs with solitary MCT after marginal surgical excision.     

Recurrence rate, clinical outcome, and cellular proliferation indices as prognostic indicators after incomplete surgical excision of cutaneous grade II mast cell tumors: 28 dogs (1994-2002)

The objectives of this study were to determine local recurrence rate, clinical outcome, and prognostic value of the number of argyrophylic nucleolar organizer regions (AgNORs), presence of proliferating cell nuclear antigen (PCNA), and number of Ki-67-positive nuclei after incomplete surgical excision of canine cutaneous grade II mast cell tumors (MCTs). This retrospective study included 30 MCTs in 28 dogs. Medical records were examined and follow-up information was obtained from owners and referring veterinarians. Only cases in which excision was incomplete and no anvcillary therapy (other than prednisone) for MCT was given were included. Paraffin-embedded tumor tissues were retrieved for AgNORs, PCNA, and Ki-67 staining. Median follow-up time was 811.5 days. Seven (23.3%) tumors recurred locally. Median time to local recurrence was not reached with a mean of 1,713 days. The estimated proportions of tumors that recurred locally at 1, 2, and 5 years were 17.3, 22.1, and 33.3%, respectively. Eleven (39.3%) dogs developed MCTs at other cutaneous locations. Median progression-free survival was 1,044 days. Median overall survival was 1,426 days. The combination of Ki-67 and PCNA scores was prognostic for local recurrence (P = .03) and development of local recurrence was prognostic for decreased overall survival (P = .04). Results suggest that a minority of incompletely excised MCTs recur. Therefore, ancillary local therapies may not always be necessary. However, local recurrence can negatively affect survival of the affected dogs. Cellular proliferation indices may indicate the likelihood of MCT recurrence after incomplete excision.     

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.     

Vinblastine and prednisolone as adjunctive therapy for canine cutaneous mast cell tumors

Twenty-seven dogs with inadequately excised, cutaneous mast cell tumors (MCT; 20 residual microscopic disease, seven marginal excision) were treated with a vinblastine and prednisolone chemotherapeutic protocol. Twenty dogs were available for follow-up examination after 12 months. One dog suffered local recurrence of the tumor, four dogs developed new cutaneous tumors, and one dog had both events. Fourteen dogs were free of MCT. There was no confirmed tumor-related mortality. Although toxicity from the chemotherapy was generally mild, one dog died of sepsis during treatment.     

Deionised water as an adjunct to surgery for the treatment of canine cutaneous mast cell tumours

Medical records of 55 dogs with a diagnosis of cutaneous mast cell tumour were reviewed. Twenty-seven of the dogs were treated with surgery plus deionised water and the remaining 28 with surgery alone. A survival analysis was performed to determine whether deionised water, as an adjunct to surgery for cutaneous mast cell tumour, affected survival time or time to tumour recurrence. Dogs in which mast cell tumour recurred had a significantly shorter survival time compared with dogs with no recurrence (P=0.05), regardless of the method of treatment. A significant negative association between tumour recurrence and method of treatment (P=0.0097) and clinical stage (P=0.0223) was observed. Dogs treated with surgery and deionised water had a significantly shorter time to recurrence of their mast cell tumour (P=0.0113). Based on these results, deionised water does not appear to be beneficial in prolonging survival time or time to tumour recurrence for dogs with cutaneous mast cell tumours.     

Radiation therapy for incompletely excised grade II canine mast cell tumors

Forty-five dogs with incompletely excised grade II mast cell tumors were treated with radiation using a cobalt 60 teletherapy unit (15 fractions of 3.2 Gy for a total of 48 Gy). Twenty-four of the dogs underwent prophylactic regional lymph node irradiation. Three (6.7%) dogs had tumor recurrence, two (4.4%) dogs developed metastasis, and 14 (31%) dogs developed a second cutaneous mast cell tumor. No difference in overall survival rate was observed between the dogs receiving and not receiving prophylactic irradiation of the regional lymph node.     

Mast cell tumour destruction in dogs by hypotonic solution

Distilled water was studied as an hypoosmotic oncolysis agent to prevent mast cell tumour (MCT) recurrence following surgical excision. The results from 118 MCTS excised in 74 dogs were: surgery alone, a recurrence rate of 52·6 per cent (10/19); and surgery followed by distilled water injections into the wound, a 26·2 per cent (26/99) recurrence rate. All of 10 small MCTS (0·03 to 0·25 cm³) injected with distilled water in situ with no surgery, regressed. The present data suggest that distilled water should be considered as an adjunct to surgery for canine MCTS.     

Evaluation of Cisplatin as an electrochemotherapy agent for the treatment of incompletely excised mast cell tumors in dogs

Background: Electrochemotherapy (ECT) couples the administration of anticancer drugs with the delivery of electric pulses that increase the drug uptake through the cell membranes, resulting in an improved efficacy. Hypothesis: To evaluate the tolerability and efficacy of cisplatin (CDDP) as an ECT agent to prevent recurrence of incompletely resected mast cell tumors (MCTs). Animals: Thirty‐seven dogs. Methods: Prospective study recruiting dogs with incompletely excised MCTs as confirmed by surgeon and pathology reports. After debulking, the tumor bed and margins were infiltrated with CDDP, and then exposed to trains of biphasic electrical pulses under sedation. Five minutes after the injection of the chemotherapy agent, sequences of 8 biphasic pulses lasting 50 + 50 μs each, were delivered in bursts of 1,300 V/cm for sclerosed and of 800 V/cm for exposed lesions, with caliper or needle array electrodes, respectively. A second session was performed 1 or 2 weeks later based on clinical considerations. Results: The treatment was well tolerated with minimal adverse effects. Twenty‐nine dogs had no evidence of recurrence over the 6‐year study period, 6 had tumor recurrence, 1 died of multiple cutaneous MCTs, and 1 died of unrelated causes. The estimated median time to recurrence was 1,200 days. Recurrence was not observed among the long‐term (>1 year) treated dogs. Conclusions and Clinical Importance: ECT with CDDP appears effective in the treatment of incompletely resected MCT in dogs and could be a useful addition to the current options based on its low cost, limited toxicity, and ease of administration.     

Canine subcutaneous mast cell tumors: cellular proliferation and KIT expression as prognostic indices

Molecular assays are widely used to prognosticate canine cutaneous mast cell tumors (MCT). There is limited information about these prognostic assays used on MCT that arise in the subcutis. The aims of this study were to evaluate the utility of KIT immunohistochemical labeling pattern, c-KIT mutational status (presence of internal tandem duplications in exon 11), and proliferation markers--including mitotic index, Ki67, and argyrophilic nucleolar organizing regions (AgNOR)--as independent prognostic markers for local recurrence and/or metastasis in canine subcutaneous MCT. A case-control design was used to analyze 60 subcutaneous MCT from 60 dogs, consisting of 24 dogs with subsequent local recurrence and 12 dogs with metastasis, as compared to dogs matched by breed, age, and sex with subcutaneous MCT that did not experience these events. Mitotic index, Ki67, the combination of Ki67 and AgNOR, and KIT cellular localization pattern were significantly associated with local recurrence and metastasis, thereby demonstrating their prognostic value for subcutaneous MCT. No internal tandem duplication mutations were detected in exon 11 of c-KIT in any tumors. Because c-KIT mutations have been demonstrated in only 20 to 30% of cutaneous MCT and primarily in tumors of higher grade, the number of subcutaneous MCT analyzed in this study may be insufficient to draw conclusions on the role c-KIT mutations in these tumors.     

Treatment of canine mast cell tumours with vinblastine, cyclophosphamide and prednisone: 35 cases (1997–2004)

The purpose of this retrospective study was to determine the efficacy and toxicity of a combined protocol of vinblastine, cyclophosphamide and prednisone (VCP) in 35 dogs with mast cell tumours (MCTs). Eleven dogs had measurable disease (group 1) and 24 dogs had incompletely excised MCT or were at high risk for metastasis (group 2). Five patients in group 1 achieved complete response, two partial responses, two stable diseases and two progressive diseases. The median progression‐free survival time (PFST) for group 1 and 2 dogs was 74 and 865 days, respectively. The median overall survival time (OST) for group 1 and 2 dogs was 145 and >2092 days, respectively. Significant negative multivariate prognostic factors included macroscopic disease and reduced vinblastine (VBL) treatments for PFST, and presence of MCT in bone marrow analysis, Patnaik grade III MCT and reduced VBL treatments for OST. Toxicity was infrequent and self‐limiting. This study suggests that the VCP protocol should be considered as an option in the treatment of MCT in dogs.     

Frequency and severity of mastocytemia in dogs with and without mast cell tumors: 120 cases (1995-1997).

OBJECTIVE: To elucidate frequency of detection on blood smears and severity on quantitative buffy coat evaluation of mastocytemia between dogs without mast cell tumors (MCT) and dogs that had MCT, and to expand the list of diseases associated with mastocytemia in dogs without MCT. DESIGN: Retrospective study. ANIMALS: 94 dogs without MCT and 26 dogs with MCT. PROCEDURE: Medical records of all dogs with mast cells detected on blood or buffy coat smears during a 2-year period were reviewed. Dogs with mastocytemia were grouped by disease into dogs with MCT and dogs without MCT. Twenty-five of the dogs without MCT that had mast cells detected on blood smears also had evaluations of buffy coat smears. Quantitative buffy coat results of the 25 dogs without MCT were compared with those of the 26 dogs with MCT. RESULTS: 95.5% of blood smears with mast cells detected during CBC determination were from dogs without MCT. For these dogs, diagnoses included inflammatory disease (28.2%), regenerative anemia (27%), neoplasia other than MCT (25.9%), and trauma (11.8%). Dogs with MCT had a mean of 71.4 mast cells/buffy coat smear, whereas dogs without MCT had a mean of 276.2 mast cells/buffy coat smear. The 2 highest counts of mast cells/buffy coat smear were for dogs without MCT. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of results of quantitative buffy coat evaluations, severity of mastocytemia in dogs without MCT often exceeds that detected during tumor staging in dogs with MCT. Random detection of mast cells in blood smears during CBC determination in dogs is usually not secondary to MCT.     

Aggressive local therapy combined with systemic chemotherapy provides long‐term control in grade II stage 2 canine mast cell tumour: 21 cases (1999–2012)

This retrospective case series evaluates the outcome of 21 dogs with grade II stage 2 mast cell tumour (MCT) treated with adequate local therapy and adjuvant systemic chemotherapy (prednisone, vinblastine and CCNU). The median survival for all dogs was 1359 days (range, 188–2340). Median disease‐free interval was 2120 days (149–2325 days). Dogs treated with surgery and chemotherapy had shorter survival (median, 1103 days; 188–2010 days) than those that underwent surgery, radiation therapy and chemotherapy as part of their treatment (median, 2056 days; 300–2340 days). Two patients had local recurrence in the radiation field and four patients had de novo MCT. Distant metastasis was not observed in any dogs. The results of this study suggest that, in the presence of loco‐regional lymph node metastasis in grade II MCT, the use of prednisone, vinblastine and CCNU after adequate local‐regional therapy can provide a median survival in excess of 40 months.     

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.     

Incorporation of sentinel lymph node mapping in dogs with mast cell tumours: 20 consecutive procedures

The study hypothesis is that incorporation of sentinel lymph node (SLN) mapping in dogs presenting for mast cell tumour (MCT) removal would impact the recommended adjuvant therapy offered. Nineteen dogs were enrolled having either spontaneously occurring or incompletely excised MCTs. Staging included regional lymph node aspiration. SLN mapping was done with regional lymphoscintigraphy combined with intra‐operative lymphoscintigraphy and blue dye. Twenty MCTs in 19 dogs were excised with SLN mapping. Eight dogs had SLNs different from the closest node. Twelve dogs had metastasis in extirpated SLNs, seven occurred in MCTs with a MI ≤ 5. No correlation was noted between patient stage and the c‐KIT proto‐oncogene. Because of SLN staging, 8 of 19 dogs were offered additional therapy that would have otherwise been excluded. Anatomic sampling of lymph nodes in dogs with MCTs does not accurately reflect which lymph nodes are most likely to be receiving the draining tumour lymph.     

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 detection of proliferating cell nuclear antigen and Ki-67 in mast cell tumors from dogs

OBJECTIVE: To determine whether immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and Ki-67 correlated with prognosis for dogs with cutaneous mast cell tumors (MCT). DESIGN: Case series. ANIMALS: 120 dogs with solitary cutaneous MCT that were excised. PROCEDURE: Information on signalment, history, and outcome was obtained by sending a questionnaire to referring veterinarians. Tumors were graded histologically, and immunohistochemical staining for Ki-67 and PCNA was performed. RESULTS: Survival rates 12, 18, and 24 months after surgery were significantly different among groups when dogs were grouped on the basis of histologic grade. Although mean number of PCNA-positive nuclei/1,000 tumor nuclei was significantly higher for dogs that died of MCT than for those that survived, there was great overlap in values. Mean number of Ki-67-positive nuclei/1,000 tumor nuclei was significantly higher for dogs that died of MCT than for those that survived, without any overlap in values between groups, and number of Ki-67-positive nuclei/1,000 tumor nuclei was significantly different among groups when tumors were grouped on the basis of histologic grades. For dogs with grade-II tumors, number of Ki-67-positive nuclei/1,000 tumor nuclei (< 93 vs > or = 93) was significantly associated with outcome (survived vs died). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for dogs with solitary cutaneous MCT, determining number of Ki-67-positive nuclei may be useful in predicting prognosis, particularly for dogs with grade-II tumors.     

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.     

Incomplete surgery, local immunostimulation, and recurrence of some tumour types in dogs and cats

Histologically confirmed inadequate treatment resulted in a lower than expected recurrence percentage in dogs with haemangiopericytoma (38%) and mastocytoma (30%). Clinical suspicion of inadequate tumour treatment did not always correlate with the histologically assessed inadequacy, nor with the appearance of local recurrence. Local recurrence did not seem to be correlated with histological grade of malignancy and tumour size. Local injection of C. parvum vaccine did not result in a lower percentage of local recurrence or longer recurrence-free intervals in any of the three tumour groups (canine haemangiopericytoma, canine mastocytoma, feline mammary carcinoma). Nor was palliative local adjuvant injection of Cp successful in dogs and cats with soft tissue sarcomas or in dogs with gingival melanoma. Re-operation of locally recurrent tumour was successful in some dogs with haemangiopericytoma, in a few with mastocytoma, but not in cats with mammary carcinoma. A trend toward histological progression of recurrences and metastases, when compared with the primary tumours, was not evident. The possible reasons for the relatively low recurrence rate of some tumour types and for the failure of Cp-treatment are discussed.     

Efficacy of radiation therapy for incompletely resected grade-III mast cell tumors in dogs: 31 cases (1987-1998)

OBJECTIVE: To determine the efficacy (durations of remission and survival) of an alternating-day radiation protocol for incompletely excised histologic grade-III solitary mast cell tumors (MCTs) in dogs. DESIGN: Retrospective study. ANIMALS: 31 dogs. PROCEDURE: Radiation (52 Gy in an 18-fraction alternating-day protocol) was delivered to an area bordered by margins > or = 3 cm around the surgical scar and to the associated local-regional lymph nodes. Dogs were not given chemotherapeutic agents concurrently or after radiation. Information on signalment, duration of remission, and survival time was obtained from medical records. RESULTS: Median and mean durations of remission were 27.7 and 17.0 months, respectively (range, 1 to 47 months). Median and mean durations of survival were 28 and 20 months, respectively (range, 3 to 52 months). Dogs with tumors located on the skin of the pinna, perineum, and prepuce had a median duration of remission greater than dogs with tumors located at other sites (27.7 and 14.4 months, respectively). Dogs with tumors < or = 3 cm in maximum diameter before surgery survived longer than dogs with tumors > 3 cm (31 and 24 months, respectively). The remission rate was 65% and survival rate was 71% at 1 year after treatment. Sixteen dogs that were euthanatized had complications associated with local-regional tumor progression. Systemic metastases to liver, spleen, intestine, and bone marrow were detected in 1 dog. CONCLUSIONS AND CLINICAL RELEVANCE: Without further treatment, incompletely excised grade-III mast cell tumors have high local-regional recurrence; local-regional treatment with radiation may effectively be used to manage many such tumors.