Fish Oncology: Diseases, Diagnostics, and Therapeutics.

The scientific literature contains a wealth of information concerning spontaneous fish neoplasms, although ornamental fish oncology is still in its infancy. The occurrence of fish neoplasms has often been associated with oncogenic viruses and environmental insults, making them useful markers for environmental contaminants. The use of fish, including zebrafish, as models of human carcinogenesis has been developed and knowledge gained from these models may also be applied to ornamental fish, although more studies are required. This review summarizes information available about fish oncology pertaining to veterinary clinicians.

Pharmacological targeting of valosin containing protein (VCP) induces DNA damage and selectively kills canine lymphoma cells.

BACKGROUND: Valosin containing protein (VCP) is a critical mediator of protein homeostasis and may represent a valuable therapeutic target for several forms of cancer. Overexpression of VCP occurs in many cancers, and often in a manner correlating with malignancy and poor outcome. Here, we analyzed VCP expression in canine lymphoma and assessed its potential as a therapeutic target for this disease. METHODS: VCP expression in canine lymphomas was evaluated by immunoblotting and immunohistochemistry. The canine lymphoma cell lines CLBL-1, 17-71 and CL-1 were treated with the VCP inhibitor Eeyarestatin 1 (EER-1) at varying concentrations and times and were assessed for viability by trypan blue exclusion, apoptosis by TUNEL and caspase activity assays, and proliferation by propidium iodide incorporation and FACS. The mechanism of EER-1 action was determined by immunoblotting and immunofluorescence analyses of Lys48 ubiquitin and markers of ER stress (DDIT3), autophagy (SQSTM1, MAP1LC3A) and DNA damage (γH2AFX). TRP53/ATM-dependent signaling pathway activity was assessed by immunoblotting for TRP53 and phospho-TRP53 and real-time RT-PCR measurement of Cdkn1a mRNA. RESULTS: VCP expression levels in canine B cell lymphomas were found to increase with grade. EER-1 treatment killed canine lymphoma cells preferentially over control peripheral blood mononuclear cells. EER-1 treatment of CLBL-1 cells was found to both induce apoptosis and cell cycle arrest in G1. Unexpectedly, EER-1 did not appear to act either by inducing ER stress or inhibiting the aggresome-autophagy pathway. Rather, a rapid and dramatic increase in γH2AFX expression was noted, indicating that EER-1 may act by promoting DNA damage accumulation. Increased TRP53 phosphorylation and Cdkn1a mRNA levels indicated an activation of the TRP53/ATM DNA damage response pathway in response to EER-1, likely contributing to the induction of apoptosis and cell cycle arrest. CONCLUSIONS: These results correlate VCP expression with malignancy in canine B cell lymphoma. The selective activity of EER-1 against lymphoma cells suggests that VCP will represent a clinically useful therapeutic target for the treatment of lymphoma. We further suggest a mechanism of EER-1 action centered on the DNA repair response that may be of central importance for the design and characterization of VCP inhibitory compounds for therapeutic use.

Metastatic malignant pilomatrixoma, acanthomatous ameloblastoma, and liver tumor in a dog with polyphagia, polyuria, polydipsia, and weight loss.

A 12-year-old, spayed female, Labrador dog was presented for evaluation of polyphagia, polyuria, polydipsia, weight loss of 2 months duration, and multiple cutaneous and subcutaneous masses. The dog was diagnosed with malignant pilomatrixoma with renal, lung, and lumbar metastases. This report describes an atypical presentation of malignant pilomatrixoma.

Pilomatrixoma malin avec métastases, améloblastome acanthomateux et tumeur hépatique chez une chienne avec polyphagie, polyurie, polydipsie et amaigrissement. Une chienne Labrador, âgée de 12 ans, était présentée pour l'évaluation d'une polyphagie, polyurie, polydipsie et d'un amaigrissement durant depuis 2 mois, ainsi que de multiples masses cutanées et sous-cutanées. Elle présentait un pilomatrixoma avec métastases aux reins, aux poumons et à une vertèbre lombaire. Ce rapport de cas décrit une présentation atypique de pilomatrixoma malin.(Traduit par les auteurs).

Anti-VEGFA Therapy Reduces Tumor Growth and Extends Survival in a Murine Model of Ovarian Granulosa Cell Tumor.

Although angiogenesis has been proposed as a therapeutic target for the treatment of ovarian granulosa cell tumor (GCT), its potential has not been evaluated in controlled studies. To do so, we used the Pten (tm1Hwu/tm1Hwu); Ctnnb1 (tm1Mmt/+);Amhr2 (tm3(cre)Bhr/+) (PCA) mouse model, which develops GCTs that mimic the advanced disease in women. A monoclonal anti-vascular endothelial growth factor A (VEGFA) antibody was administered weekly to PCA mice beginning at 3 weeks of age. By 6 weeks of age, anti-VEGFA therapy significantly decreased tumor weights relative to controls (P < .05) and increased survival, with all treated animals but none of the controls surviving to 8 weeks of age. Analyses of PCA tumors showed that anti-VEGFA treatment resulted in significant decreases in tumor cell proliferation and microvessel density relative to controls (P < .05). However, treatment did not have a significant effect on apoptosis or tumor necrosis. The VEGFA receptor 2 (VEGFR2) signaling effector p44/p42 mitogen-activated protein kinase (MAPK), whose activity is associated with cell proliferation, was significantly less phosphorylated (i.e., activated) in tumors from the treated group (P < .05). Conversely, no significant difference was found in the activation of protein kinase B, a VEGFR2 signaling effector associated with cell survival. Together, these results suggest that anti-VEGFA therapy is effective at inhibiting GCT growth in the PCA model and acts by reducing microvascular density and cell proliferation through inhibition of the VEGFR2-MAPK pathway. Findings from this preclinical model therefore support the investigation of targeting VEGFA for the adjuvant treatment of GCT in women.

Retrospective evaluation of potential causes associated with clinically relevant hyperlactatemia in dogs with lymphoma.

The aim of this study was to determine whether or not canine lymphoma could be associated with a clinically relevant type B hyperlactatemia (> 2.5 mmol/L). The medical database from the University of Montreal Veterinary Medical Teaching Hospital was searched for confirmed cases of canine lymphoma with a blood lactate measurement. Information retrieved included stage, clinical observations compatible with causes of type A and B hyperlactatemia other than cancer, hepatic involvement, and drugs administered. Twenty (40%) dogs were hyperlactatemic. Five dogs (10%) were classified as having cancer-related hyperlactatemia. Seventy-five percent of hyperlactatemic dogs had clinical evidence of type A hyperlactatemia. In addition to lymphoma, 70% of hyperlactatemic dogs had evidence of an additional cause of type B hyperlactatemia. A significant association (P = 0.01) was identified between corticosteroid administration and hyperlactatemia. Cytological, echographic, and/or biochemical tests revealed hepatic changes in all hyperlactatemic dogs. Lymphoma alone may not be sufficient to explain clinically relevant hyperlactatemia in dogs.

Proteomic profiling of a mouse model for ovarian granulosa cell tumor identifies VCP as a highly sensitive serum tumor marker in several human cancers.

The initial aim of this study was to identify novel serum diagnostic markers for the human ovarian granulosa cell tumor (GCT), a tumor that represents up to 5% of all ovarian cancers. To circumvent the paucity of human tissues available for analyses, we used the Ctnnb1(tm1Mmt/+);Pten(tm1Hwu/tmiHwu);Amhr2(tm3(cre)Bhr/+) transgenic mouse model, which features the constitutive activation of CTNNB1 signaling combined with the loss of Pten in granulosa cells and develops GCTs that mimic aggressive forms of the human disease. Proteomic profiling by mass spectrometry showed that vinculin, enolase 1, several heat shock proteins, and valosin containing protein (VCP) were more abundantly secreted by cultured mouse GCT cells compared to primary cultured GC. Among these proteins, only VCP was present in significantly increased levels in the preoperative serum of GCT cancer patients compared to normal subjects. To determine the specificity of VCP, serum levels were also measured in ovarian carcinoma, non-Hodgkin's lymphoma and breast, colon, pancreatic, lung, and prostate cancer patients. Increased serum VCP levels were observed in the majority of cancer cases, with the exception of patients with lung or prostate cancer. Moreover, serum VCP levels were increased in some GCT, ovarian carcinoma, breast cancer, and colon cancer patients who did not otherwise display increased levels of widely used serum tumor markers for their cancer type (e.g. inhibin A, inhibin B, CA125, CEA, or CA15.3). These results demonstrate the potential use of VCP as highly sensitive serum marker for GCT as well as several other human cancers.

Pharmacological targeting of mammalian target of rapamycin inhibits ovarian granulosa cell tumor growth.

Few targeted therapies have been developed for ovarian granulosa cell tumor (GCT), even though it represents 5% of all malignant ovarian tumors in women. As misregulation of PI3K/AKT signaling has been implicated in GCT development, we hypothesized that the AKT signaling effector mammalian target of rapamycin (mTOR) may play a role in the pathogenesis of GCT and could represent a therapeutic target. Analyses of human GCT samples showed an increase in protein levels of mTOR and its downstream effectors RPS6KB1, RPS6, eIF4B and PPARG relative to normal granulosa cells, suggestive of an increase in mTOR pathway activity and increased translational activity and/or protein stability. We next sought to evaluate mTOR as a GCT therapeutic target using the Pten (tm1Hwu/tmiHwu);Ctnnb1 (tm1Mmt/+);Amhr2 (tm3(cre)Bhr/+) (PCA) mouse model, in which mTOR, RPS6KB1, eIF4B and PPARG are upregulated in tumor cells in a manner similar to human GCT. Treatment of PCA mice with the mTOR-specific inhibitor everolimus reduced tumor growth rate (1.5-fold; P < 0.05) and also reduced total tumor burden (4.7-fold; P < 0.05) and increased survival rate (78 versus 44% in the vehicle group) in a PCA surgical model of GCT peritoneal carcinomatosis. Everolimus decreased tumor cell proliferation and tumor cell volume relative to controls (P < 0.05), whereas apoptosis was unaffected. Phosphorylation of RPS6KB1 and RPS6 were decreased (P < 0.05) by everolimus, but RPS6KB1, RPS6, eIF4B and PPARG expressions were not affected. These results suggest that mTOR is a valid and clinically useful pharmacological target for the treatment of GCT, although its inhibition does not reverse all consequences of aberrant PI3K/AKT signaling in the PCA model.

A comparison of computed tomography, computed radiography, and film-screen radiography for the detection of canine pulmonary nodules.

Computed tomography (CT) has become more widely available and computed radiography (CR) has replaced film-screen radiography for canine thoracic imaging in many veterinary practices. There are limited data comparing these modalities in a veterinary clinical setting to detect pulmonary nodules. We compared CT, CR, and film-screen radiography for detecting the presence, number, and characteristics of pulmonary nodules in dogs. Observer performance for a variety of experience levels was also evaluated. Twenty-one client-owned dogs with a primary neoplastic process underwent CT and CR; nine also received film-screen radiographs. Positive/negative classification by consensus agreed between the three modalities in 8/9 dogs and between CR and CT in the remaining 12. CT detected the greatest (P = 0.002) total number of nodules and no difference was seen between CR and films. The greatest number of nodules was seen in the right middle and both caudal regions, but only using CT (P < 0.0001). Significantly smaller nodules were detected with CT (P = 0.0007) and no difference in minimum size was detected between CR and films. Observer accuracy was high for all modalities; particularly for CT (90.5-100%) and for the senior radiologist (90.5-100%). CT was also characterized by the least interobserver variability. Although CT, CR, and film-screen performed similarly in determining the presence or absence of pulmonary nodules, a greater number of smaller nodules was detected with CT, and CT was associated with greater diagnostic confidence and observer accuracy and agreement.

Clinical and computed tomography features of secondary renal hyperparathyroidism.

An atypical case of secondary renal hyperparathyroidism was diagnosed in a 9-year-old miniature schnauzer after a skull computed tomography (CT) showed the presence of 2 bilateral and symmetrical soft tissue maxillary masses, and osteopenia of the skull.

Evaluation of the use of chemotherapy and other prognostic variables for surgically excised canine thyroid carcinoma with and without metastasis.

This retrospective study compared the efficacy of surgery alone versus surgery in combination with chemotherapy in the treatment of canine thyroid carcinoma; potential prognostic factors were evaluated. Forty-four dogs with biopsy-confirmed thyroid carcinoma met the inclusion criteria. Twenty-eight dogs were treated with surgery alone and 16 with surgery and chemotherapy. The median survival of dogs treated with surgery and chemotherapy was 518 d, which was not statistically different from that of the dogs treated with surgery alone. The number of thyroid lobes removed at surgery was prognostic with respect to survival. Despite an overall metastatic rate of 48%, the addition of chemotherapy to surgical excision did not improve survival; however, this finding may be due to inadequate power to demonstrate a difference.

Laparoscopic splenectomy for treatment of splenic hemangiosarcoma in a dog.

OBJECTIVE: To report laparoscopic splenectomy in a dog. STUDY DESIGN: Clinical report. ANIMALS: Mixed breed dog (n=1). METHODS: Hemangiosarcoma was diagnosed by ultrasound-guided fine-needle aspiration of a splenic mass in an 11-year-old, 30 kg, mixed breed dog. No metastatic disease was identified during complete staging (chest radiographs, echocardiogram, and abdominal ultrasonography); however, cystic calculi were identified. Laparoscopic splenectomy using Ligasure V was performed through 3 portals and the calculi were removed by laparoscopic-assisted cystoscopy. RESULTS: Total surgical time was 2 hours and for laparoscopic splenectomy, 65 minutes. The celiotomy incision for splenic removal was 7 cm. The dog recovered uneventfully and was ambulatory 2 hours postoperatively. CONCLUSION: Laparoscopy with Ligasure V facilitated successful removal of a spleen with a 3 cm mass. CLINICAL RELEVANCE: Laparoscopic splenectomy in dogs is feasible for removal of a normal-sized spleen with a moderate-sized mass.

A mouse surgical model for metastatic ovarian granulosa cell tumor.

We recently described a genetically engineered mouse model that develops ovarian granulosa cell tumors (GCTs) that mimic many aspects of the advanced human disease, including distant dissemination. However, because the primary tumors killed their hosts before metastases were able to form, the use of these mice to study metastatic disease required the development of a simple, reliable, and humane surgical protocol for the excision of large GCTs from debilitated mice. Here we describe a protocol involving multimodal anesthesia, tumor removal through ventral midline celiotomy and perioperative fluid therapy, and analgesia that led to the postoperative survival of more than 90% of mice, despite the removal of tumors representing as much as 10% of the animal's body weight. Intraabdominal recurrence of the GCT did not occur in surviving animals, but most developed pulmonary or adrenal metastases (or both) by 12 wk after surgery. We propose that this mouse model of metastatic GCT will serve as a useful preclinical model for the development of novel treatment modalities and diagnostic techniques. Furthermore, our results delineate anesthetic and surgical principles for the removal of large abdominal tumors from mice that will be applicable to other models of human cancers.

Cobalt radiation with or without low-dose cisplatin for treatment of canine naso-sinus carcinomas.

The objective of this study was to determine if low-dose cisplatin could be added safely to radiation therapy for the treatment of naso-sinus carcinomas in dogs. Thirty-one dogs were evaluated; 18 of these dogs received cobalt radiation in combination with low-dose cisplatin while 13 dogs received radiation alone. No difference was observed for acute or late radiation effects. Cisplatin was administered at a dosage of 7.5 mg/m2 20 min prior to every other radiation treatment. An initial dose of 10 mg/m2 was intended but toxicity (primarily azotemia) was unacceptable. Cisplatin was administered as prescribed in 12 of 18 dogs. Cisplatin was discontinued in 2 dogs because of azotemia. In the other 4 dogs cisplatin was not administered as prescribed because the dogs were withdrawn from treatment due to disease progression or radiation effects. There was no long-term renal disease in patients who developed azotemia. The overall median survival was 433 days with 4 (12.9%) dogs still alive at the completion of the study.