Germline gene polymorphisms predisposing domestic mammals to carcinogenesis

Cancer is a complex disease caused in part by predisposing germline gene polymorphisms. Knowledge of carcinogenesis in companion mammals (dog and cat) and some livestock species (pig and horse) is quite advanced. The prevalence of certain cancers varies by breed in these species, suggesting the presence of predisposing genetic variants in susceptible breeds. This review summarizes the present understanding of germline gene polymorphisms, including BRCA1, BRCA2, MC1R, KIT, NRAS and RAD51, associated with predisposition to melanoma, mammary cancer, osteosarcoma and histiocytic sarcoma in dogs, cats, pigs and horses. The predisposing variants in these species are discussed in the context of human germline gene polymorphisms associated with the same types of cancer.     

Timely adjuvant chemotherapy improves outcome in dogs with non-metastatic splenic hemangiosarcoma undergoing splenectomy

Timely delivery of adjuvant chemotherapy has been shown to be advantageous in many human cancers and canine osteosarcoma. Adjuvant chemotherapy has been shown to improve outcome for canine splenic hemangiosarcoma. The aim of this retrospective study was to investigate whether timely adjuvant chemotherapy administration resulted in better outcome in dogs with non-metastatic splenic hemangiosarcoma undergoing splenectomy. Medical records were searched for dogs with non-metastatic, splenic hemangiosarcoma that received splenectomy and adjuvant chemotherapy. The number of days from surgery to the first chemotherapy dose (StoC) was evaluated to identify the cut-off value associated with the best survival advantage. StoC and other possible prognostic factors were tested for influence on time to metastasis (TTM) and overall survival (OS). Seventy dogs were included. Median StoC was 20 days (range: 4–70). The time interval associated with the greatest survival benefit was 21 days. Median TTM and OS of dogs with StoC ≤ 21 days were significantly longer than those with StoC >21 days (TTM: 163 vs. 118 days, p = .001; OS: 238 vs. 146 days, p < .001). On multivariable analysis, StoC >21 days was the only variable significantly associated with increased risk of tumour progression (HR 2.1, p = .010) and death (HR 2.3; p = .008). Starting adjuvant chemotherapy within 21 days of surgery may be associated with a survival benefit in dogs with non-metastatic splenic hemangiosarcoma, possibly due to the early targeting of newly recruited metastatic cells after surgery.     

Adjuvant anthracycline‐based vs metronomic chemotherapy vs no medical treatment for dogs with metastatic splenic hemangiosarcoma: A multi‐institutional retrospective study of the Italian Society of Veterinary Oncology

Treatment options for dogs with metastatic (stage III) splenic hemangiosarcoma are limited. A doxorubicin‐based chemotherapy regimen is commonly administered; however, there are no published data to support this practice. The aim of this study was to investigate the impact of maximum‐tolerated‐dose chemotherapy (MTD), metronomic chemotherapy (MC) and no adjuvant treatment on outcome in dogs with stage III splenic hemangiosarcoma undergoing splenectomy. Medical records of dogs with stage III splenic hemangiosarcoma that underwent splenectomy followed by MTD chemotherapy, MC or no adjuvant treatment were retrieved. Time to progression (TTP), survival time (ST) and toxicity were evaluated. One hundred three dogs were identified: 23 received adjuvant MTD, 38 MC and 42 were not medically treated. Overall median TTP and ST were 50 (95% confidence interval [CI], 39‐61) and 55 days (95% CI, 43‐66), respectively. Dogs treated with adjuvant MTD had a significantly longer TTP and ST compared with dogs receiving MC (median TTP, 134 vs 52 days, P = .025; median ST, 140 vs 58 days, P = .023, respectively). Dogs treated by splenectomy only had the shortest median TTP (28 days) and ST (40 days). However, treatment‐related adverse events (AEs) were significantly more frequent in the MTD group (P = .017). The outcome for dogs with metastatic splenic hemangiosarcoma is poor. While MTD showed greater efficacy compared to MC, toxicity was higher in this group. Treatment‐related AEs need to be carefully balanced against this modest survival prolongation when offering adjuvant MTD to dogs with advanced stage hemangiosarcoma.     

Treatment of vascular and soft-tissue sarcomas in dogs using an alternating protocol of ifosfamide and doxorubicin

A retrospective analysis was done to assess the toxicity and efficacy associated with an alternating chemotherapy protocol of ifosfamide (375 mg m^?2) and doxorubicin (30 mg m^?2) for adjuvant treatment of 39 dogs with sarcomas. Twelve dogs had various soft‐tissue sarcomas and 27 dogs had hemangiosarcoma (HSA). Complete blood counts were evaluated 7 days after the first dose of ifosfamide and doxorubicin. One dog had grade 4 neutropenia (<500 µL^?1) after treatment with ifosfamide and one dog had grade 3 neutropenia (500–1000 µL^?1) after treatment with doxorubicin. One dog treated with doxorubicin was hospitalized for 24 h due to vomiting. The median survival time (ST) for the 27 dogs with HSA treated by surgery and with doxorubicin/ifosfamide was 149 days (mean 366 days). Although the protocol of alternating ifosfamide and doxorubicin was well tolerated, it failed to result in a statistically significant improvement in the ST when compared to a historical population of dogs with stage 2 splenic HSA treated by surgery alone.     

Evaluation of optimal water fluoridation on the incidence and skeletal distribution of naturally arising osteosarcoma in pet dogs

Experimental toxicological studies in laboratory animals and epidemiological human studies have reported a possible association between water fluoridation and osteosarcoma (OSA). To further explore this possibility, a case‐control study of individual dogs evaluated by the UC Davis Veterinary Medical Teaching Hospital was conducted using ecologic data on water fluoridation based on the owner's residence. The case group included 161 dogs with OSA diagnosed between 2008–2012. Two cancer control groups included dogs diagnosed with lymphoma (LSA) or hemangiosarcoma (HSA) during the same period (n = 134 and n = 145, respectively). Dogs with OSA were not significantly more likely to live in an area with optimized fluoride in the water than dogs with LSA or HSA. Additional analyses within OSA patients also revealed no significant differences in age, or skeletal distribution of OSA cases relative to fluoride status. Taken together, these analyses do not support the hypothesis that optimal fluoridation of drinking water contributes to naturally occurring OSA in dogs.     

Comparison of doxorubicin–cyclophosphamide with doxorubicin–dacarbazine for the adjuvant treatment of canine hemangiosarcoma

Canine hemangiosarcoma (HSA) is a neoplasm of vascular endothelial origin that has an aggressive biological behaviour, with less than 10% of dogs alive at 12‐months postdiagnosis. Treatment of choice consists of surgery followed by adjuvant doxorubicin‐based chemotherapy. We prospectively compared adjuvant doxorubicin and dacarbazine (ADTIC) to a traditional doxorubicin and cyclophosphamide (AC) treatment, aiming at determining safety and assessing whether this regimen prolongs survival and time to metastasis (TTM). Twenty‐seven dogs were enrolled; following staging work‐up, 18 were treated with AC and 9 with ADTIC. Median TTM and survival time were longer for dogs treated with ADTIC compared with those receiving AC (>550 versus 112 days, P = 0.021 and >550 versus 142 days, P = 0.011, respectively). Both protocols were well tolerated, without need for dose reduction or increased interval between treatments. A protocol consisting of combined doxorubicin and dacarbazine is safe in dogs with HSA and prolongs TTM and survival time.     

Feline cutaneous hemangiosarcoma: a retrospective study of 18 cases (1998-2003)

Cutaneous hemangiosarcoma (HSA) has been infrequently reported in dogs and cats. Medical records of 18 cats diagnosed with cutaneous HSA were reviewed. Age at the time of diagnosis, breed, sex, tumor location, tumor size, treatment type, survival time, disease-free interval, and cause of death were evaluated. Aggressive surgical excision of the tumor was attempted in 10 cats. A complete surgical excision was achieved in five of the 10 cats. Median survival times were statistically longer in cats that underwent surgery versus cats that did not. Cats with cutaneous HSA treated with aggressive surgical excision of their tumors may have a good long-term prognosis.     

Epidemiologic, clinical, pathologic, and prognostic characteristics of splenic hemangiosarcoma and splenic hematoma in dogs: 217 cases (1985)

Data on age, sex, and breed were obtained from surgical pathologic records of 92 dogs with splenic hemangiosarcoma (SHS) and for 125 dogs with splenic hematoma (SHA) diagnosed in 1985 at the University of Pennsylvania School of Veterinary Medicine. Further information on body weight, clinical and surgical findings, and survival time was obtained for 59 dogs (64.1%) with SHS and 91 dogs (72.8%) with SHA. Splenic hemangiosarcoma was markedly more common in dogs 8 to 13 years old, and SHA was appreciably more common in dogs greater than or equal to 8 years old, compared with dogs 1 to 7 years old. Compared with sexually intact females, only spayed females were at significantly (odds ratio [or], 2.2; 95% confidence interval [CI], 1.2 to 4.1) increased risk for developing SHS; sex predisposition was not found for dogs with SHA. The German Shepherd Dog was the only breed with increased risk for development of either SHS (OR, 4.7; 95% CI, 2.7 to 7.8) or SHA (OR, 2.8; 95% CI, 1.7 to 4.9), compared with all other purebred dogs. Association of tumor type for 7 commonly reported clinical signs with observance of hemoperitoneum at surgery was determined; anorexia (P = 0.01), collapse (P = 0.01), and hemoperitoneum (P less than 0.001) were significantly more common in dogs with SHS. The median survival time for dogs with SHS was 19 days, compared with 338 days for dogs with SHA (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Concurrent splenic and right atrial mass at presentation in dogs with HSA: a retrospective study

The objective of this retrospective study was to evaluate the presence of concurrent splenic and cardiac hemangiosarcoma (HSA). Dogs were divided into two groups: group 1 included 23 dogs with splenic HSA, and group 2 included 31 dogs with a cardiac HSA. All dogs were fully assessed for metastasis with thoracic radiography, abdominal and/or cardiac ultrasound, and/or postmortem examination. Two dogs (8.7%) in group 1 had a concurrent cardiac mass. Neither of these dogs had pericardial effusion, and both were golden retrievers. Thirteen of the dogs in group 1 presented with a hemoabdomen. Concurrent intra-abdominal metastasis was noted in seven dogs. In group 2, 9/31 (29%) of the dogs had a concurrent splenic HSA, and 13/31 (42%) of the dogs had evidence of metastasis to another site. There was a significant association between age and the presence of nonsplenic metastasis (odds ratio, 0.457). The rate of concurrent right atrial mass detected by cardiac ultrasound in dogs with splenic HSA was 8.7%, which is less than previously reported. For dogs with right atrial HSA, the risk of metastasis to nonsplenic sites decreases with age.     

Metastatic pattern in dogs with splenic haemangiosarcoma: Clinical implications

A retrospective study was undertaken to characterise the biological behaviour of splenic haemangiosarcoma (HSA) in dogs. Metastatic pattern data for 25 dogs with splenic HSA that were presented for clinical signs relating to splenic lesions (eg, abdominal mass and, or, haemoperitoneum) and had undergone necropsy were analysed. Six of 25 dogs with splenic HSA that were presented for abdominal mass/haemoperitoneum had right atrial HSA. Fifteen of 19 (79 per cent dogs) with splenic HSA associated with abdominal mass/hemoperitoneum without right atrial involvement had disease confined to the peritoneal cavity. The most common metastatic sites in these dogs were liver, omentum and mesentery. Extraperitoneal metastases were seen in four of 19 (21 per cent) dogs without right atrial involvement. Analysis of signalment data of dogs in this series and the literature revealed no differences between dogs with disease confined to the peritoneal cavity and dogs with extraperitoneal metastases. The subjectivity of primary site designation, importance of ante mortem identification of individuals with concurrent right atrial involvement, and need for more aggressive therapy directed at intraperitoneal metastases are discussed.     

Efficacy and Toxicity of VAC Chemotherapy (Vincristine, Doxorubicin, and Cyclophosphamide) in Dogs with Hemangiosarcoma

Fifteen dogs with hemangiosarcoma were treated with a combination of vincristine, doxorubicin, and cyclophosphamide after incisional or excisional biopsy. The median survival for all fifteen dogs was 172 days (mean survival = 316 days). The median survival for those dogs with splenic hemangiosarcoma was 145 days (mean survival = 271 days) as compared with previously published median survival times in dogs with splenic hemangiosarcoma treated with surgery alone of 19 to 65 days. Toxicities included neutropenia (11/15), severe gastroenteritis (4/15), cardiotoxicity (3/15), and sepsis (2/15). The authors conclude that vincristine, doxorubicin, and cyclophosphamide chemotherapy may be an efficacious treatment modality in dogs with hemangiosarcoma and is associated with acceptable toxicity.     

Clinical and pharmacokinetic characteristics of intracavitary administration of pegylated liposomal encapsulated doxorubicin in dogs with splenic hemangiosarcoma

BACKGROUND: Canine splenic hemangiosarcoma (HSA) is a fatal malignancy, and most affected dogs die within a few months of diagnosis. Most dogs present with signs from tumor rupture, resulting in hemoabdomen and intra-abdominal dissemination. The abdomen is also the main site of disease recurrence. HYPOTHESIS: Intraperitoneal (IP) administration of doxorubicin will delay or prevent intra-abdominal tumor recurrence and prolong survival in dogs with HSA. ANIMALS: Fourteen dogs with splenic HSA. METHODS: A prospective, unmasked, uncontrolled clinical trial. After staging of disease status and splenectomy, pegylated liposomal encapsulated doxorubicin was administered intraperitoneally (1 mg/kg body weight) every 3 weeks for 4 cycles. All dogs were monitored for recurrence of HSA. Samples of plasma and abdominal fluid were collected for measurement of doxorubicin concentration and pharmacokinetic analysis. Nonlinear mixed-effect modeling was used to describe the pharmacokinetics of liposomal doxorubicin administered IP. RESULTS: All 14 dogs died, 12 because of HSA and 2 from other causes. Postmortem examination was performed on 12 dogs. All 12 dogs died because of HSA-related causes and had hepatic metastases and hemoabdomen. The IP-treated dogs had fewer serosal, mesenteric, and omental metastases than historical controls treated with systemic doxorubicin. Results of the postmortem examination and pharmacokinetic analysis confirmed that IP delivery of doxorubicin resulted in an effective drug concentration with a clearance comparable with that after i.v. delivery. CONCLUSIONS AND CLINICAL IMPORTANCE: IP pegylated liposomal encapsulated doxorubicin administration did not prevent intraabdominal recurrence of HSA in dogs.     

Elevated serum thymidine kinase activity in canine splenic hemangiosarcoma*

Thymidine kinase 1 (TK1) is a soluble biomarker associated with DNA synthesis. This prospective study evaluated serum TK1 activity in dogs presenting with hemoabdomen and a splenic mass. An ELISA using azidothymidine as a substrate was used to evaluate TK1 activity. Sixty‐two dogs with hemoabdomen and 15 normal controls were studied. Serum TK1 activity was significantly higher in dogs with hemangiosarcoma (HSA) than in normal dogs (mean ± SEM = 17.0 ± 5.0 and 2.01 ± 0.6, respectively), but not dogs with benign disease (mean ± SEM = 10.0 ± 3.3). Using a cut‐off of 6.55 U/L, TK activity demonstrated a sensitivity of 0.52, specificity of 0.93, positive predictive value of 0.94 and negative predictive value of 0.48 for distinguishing HSA versus normal. When interval thresholds of <1.55 and >7.95 U/L were used together, diagnostic utility was increased. Serum TK1 evaluation may help to discriminate between benign disease and HSA in dogs with hemoabdomen and a splenic mass.     

Prevalence of hemangiosarcoma in anemic dogs with a splenic mass and hemoperitoneum requiring a transfusion: 71 cases (2003-2005)

OBJECTIVE: To determine prevalence of splenic hemangiosarcoma in anemic dogs with a splenic mass and hemoperitoneum requiring a transfusion and to identify factors that could differentiate between dogs with hemangiosarcoma and dogs with other splenic masses at the time of hospital admission. DESIGN: Retrospective case series. ANIMALS: 71 dogs. PROCEDURES: Medical records, blood bank logs, and histologic reports of dogs with a splenic mass and hemoperitoneum that required a transfusion between 2003 and 2005 were reviewed. Dogs that received a transfusion of packed RBCs, were splenectomized, and had a definitive histologic diagnosis were included. RESULTS: Signalment of dogs was similar to that in other reports. Malignant splenic neoplasia was identified in 54 of 71 (76.1%) dogs, whereas 17 of 71 (23.9%) dogs had a benign splenic lesion. Of 54 dogs with malignant splenic neoplasia, 50 (92.6% [70.4% of all dogs]) had splenic hemangiosarcoma. In addition, dogs with splenic hemangiosarcoma had significantly lower total solids (TS) concentrations and platelet counts at admission. Finally, hemoperitoneum was strongly associated with a diagnosis of splenic hemangiosarcoma. CONCLUSIONS AND CLINICAL RELEVANCE: In this clinical population of dogs, prevalence of hemangiosarcoma was higher than in other studies. Dogs with hemangiosarcoma in this study had significantly lower TS concentrations and platelet counts at the time of admission, compared with values for dogs with other splenic masses. No other markers were useful in differentiating dogs with hemangiosarcoma. It is important to discuss the prevalence of and poor prognosis associated with hemangiosarcoma with owners when they are contemplating whether to proceed with treatment.     

Clinical outcome in 20 cases of lingual hemangiosarcoma in dogs: 1996–2011

With the exception of solar‐induced dermal hemangiosarcoma (HSA), the biologic behaviour of canine HSA is characterised by rapid tumour growth, a high metastatic rate and short survival times. Outcome of dogs with HSA of the tongue has not been previously reported. The purpose of this study was to assess outcome and prognostic factors in dogs with lingual HSA. Clinical data was collected retrospectively and histopathology was reviewed for 20 dogs. Median progression free survival was 524 days and the median overall survival time was 553 days. All dogs had low or intermediate grade tumours; most tumours were small and located on the ventral surface of the tongue. Prognostic factors significantly associated with increased survival included small tumour size and absence of clinical signs of an oral mass at the time of diagnosis. Dogs with HSA confined to the tongue may have a better prognosis compared with HSA in other organs.     

Non-coagulopathic spontaneous hemothorax in dogs

Objective: To determine the history, clinicopathologic findings, underlying causes, and outcomes for dogs with non‐coagulopathic spontaneous hemothorax. Design: Retrospective case series. Setting: University referral hospital. Animals: Sixteen client‐owned dogs. Interventions: The medical records database was searched for dogs with hemothorax. Dogs with trauma, secondary coagulopathy, recent thoracic surgery, or pericardial intervention were excluded. For the remaining dogs, signalment, clinical signs, clinicopathologic findings, radiographic findings, histopathologic findings, interventions, and outcome were recorded. Measurements and main results: The most common presenting signs were tachypnea (n=9) and lethargy (n=5), typically of <1‐week duration. The most common cause of non‐coagulopathic spontaneous hemothorax in dogs was neoplasia, which was diagnosed in 14 patients (88%). Identified malignancies included hemangiosarcoma (n=1), malignant mesothelioma (n=1), metastatic ovarian carcinoma (n=1), osteosarcoma (n=2), and pulmonary carcinoma (n=2). An intrathoracic mass was visualized in 7 other dogs; however, histopathology was not obtained. Pancreatitis and lung lobe torsion were each diagnosed in 1 dog, and survival was prolonged with both surviving at least 1 year post discharge. Only 6 of 14 dogs that were diagnosed with neoplasia were discharged from the hospital. For the 4 dogs with cancer with available outcome data, median survival time was 16 days (range 1–70 days). Two dogs were lost to follow‐up and had unknown survival times. Conclusions: The development of non‐coagulopathic spontaneous hemothorax warrants a high‐index suspicion for neoplasia, in particular thoracic wall neoplasia.     

In vitro effects of Yunnan Baiyao on canine hemangiosarcoma cell lines

Yunnan Baiyao is a Chinese herbal medicine that has been utilized for its anti‐inflammatory, haemostatic, wound healing and pain relieving properties in people. It has been utilized in the veterinary profession to control bleeding in dogs with hemangiosarcoma (HSA) and has been anecdotally reported to prolong survival times in dogs with this neoplasm. This study evaluated the in vitro activity of Yunnan Baiyao against three canine HSA cell lines after treatment with increasing concentrations of Yunnan Baiyao (50, 100, 200, 400, 600 and 800 µg mL^?1) at 24, 48 and 72 h. Mean half maximum inhibitory concentration (IC₅₀) at 72 h for DEN, Fitz, SB was 369.9, 275.9 and 325.3 µg mL^?1, respectively. Caspase‐3/7 activity increased in correlation with the IC₅₀ in each cell line which was confirmed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL, APO‐BRDU Kit; BD Biosciences, San Jose, CA, USA) assay. VEGF in cell supernatant was also quantified. Overall, the study found that Yunnan Baiyao causes dose and time dependent HSA cell death through initiation of caspase‐mediated apoptosis, which supports future studies involving Yunnan Baiyao.     

Ultra‐frequent HRAS p.Q61R somatic mutation in canine acanthomatous ameloblastoma reveals pathogenic similarities with human ameloblastoma

Ameloblastoma is a locally aggressive odontogenic tumour that occurs in humans and dogs. Most ameloblastomas (AM) in humans harbour mutually‐exclusive driving mutations in BRAF, HRAS, KRAS, NRAS or FGFR2 that activate MAPK signalling, and in SMO that activates Hedgehog signalling. The remarkable clinical and histological similarities between canine acanthomatous ameloblastoma (CAA) and AM suggest they may harbour similar driving mutations. In this study, aimed at characterizing the mutational status of SMO, BRAF, HRAS, KRAS, NRAS and FGFR2 in CAA, we used RNA sequencing, Sanger sequencing and restriction fragment length polymorphism assays to demonstrate that 94% of CAA (n = 16) harbour a somatic HRAS p.Q61R mutation. The similarities in MAPK‐activating mutational profiles between CAA and AM implicate conserved molecular mechanisms of tumorigenesis, thus, qualifying the dog as a potentially useful model of disease. Given the relevance of RAS mutations in the pathogenesis of odontogenic tumours and other types of cancer, the results of this study are of comparative, translational, and veterinary value.     

A Retrospective Study of Canine Hemangiosarcoma and its Association with Acanthocytosis

A retrospective study was conducted on cases of canine hemangiosarcoma diagnosed at the Western College of Veterinary Medicine between 1969 and 1979. A total of 33 dogs were assessed with respect to breed, age, sex and site of neoplasia. Hemangiosarcoma was most common in older dogs and there was no sex predilection. Incidence was highest in German Shepherds. The primary sites in order of most frequent occurrence were spleen, right atrium and liver. Acanthocytes were found in six of 12 dogs examined. There was no association between acanthocytosis and such factors as breed, primary site of the neoplasm and splenic or hepatic involvement.     

Retrospective comparison of first‐line adjuvant anthracycline vs metronomic‐based chemotherapy protocols in the treatment of stage I and II canine splenic haemangiosarcoma

Splenectomy followed by adjuvant chemotherapy is commonly used to treat canine splenic haemangiosarcoma (HSA), although it is unclear if different treatment protocols may have a similar efficacy. The objective of this retrospective study was to assess outcome in dogs with stage I and II splenic HSA treated with either first‐line adjuvant anthracycline (AC) or metronomic (MC)‐based chemotherapy protocols, by comparing median time to progression (TTP) and median survival time (MST). Medical records of nine institutions were searched for dogs diagnosed with stage I and II splenic HSA that underwent adjuvant treatment with AC‐ or MC‐based protocols following splenectomy. Patients treated with MC following AC were included in an additional group (AMC). Ninety‐three dogs were included: 50 in the AC group, 23 in the AMC group and 20 in the MC group. The overall MST was 200 days (range 47‐3352) and the overall median TTP was 185 days (range 37‐1236). The median TTP of stage I dogs was significantly longer compared to stage II dogs (338 vs 151 days, respectively, P = .028). When adjusting for treatment type, the MST was 154 days for the AC group (range 47‐3352 days), 338 days for the AMC group (range 79‐1623 days) and 225 days for the MC group (range 57‐911 days). The difference in MST and median TTP was not found to be statistically significant between treatment groups. This study suggests that adjuvant MC in canine splenic HSA may result in a similar outcome when compared to other treatment protocols. Further studies are warranted to confirm these findings.