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OBJECTIVES: The standard treatment for canine and feline meningiomas includes radiotherapy, surgical excision or combined therapy. However, new therapeutic approaches are required due to the possible recurrence or progression of meningiomas despite initial therapy. Adjunctive therapy with synthetic long-acting somatostatin (SST) analogues has been described in humans with SST-expressing tumours. The expression of SST receptors (SSTRs) by feline meningiomas is currently unknown, and there are little data about canine meningiomas. We hypothesized that SSTR is expressed by canine and feline meningiomas (S1). METHODS: Seven canines and 11 felines with histologically confirmed meningiomas underwent STTR screening. RNA expressions of SSTR1, SSTR2, SSTR3 and SSTR5 (canine) and SSTR1-SSTR 5 (feline) in fresh frozen and formalin-fixed and paraffin-embedded (FFPE) samples were investigated using real-time (RT)-qPCR. The expression of SSTR1 and SSTR2 in FFPE samples was evaluated using immunohistochemistry (IHC). The specificity of applied antibodies for canine and feline species was confirmed by western blotting. RESULTS: In canine meningiomas (n = 7), RNA expression of SSTR1, SSTR2 and SSTR5 was detected in all samples; SSTR3 RNA expression was detected in only 33% of samples. In feline meningiomas (n = 12), RNA expression of SSTR1, SSTR4, SSTR5 and SSTR2 was detected in 91%, 46%, 46% and 36% of samples, respectively; SSTR3 was not expressed. Overall, the detection rate was lower in FFPE samples. IHC revealed the expression of SSTR1 and SSTR2 in all samples from both species. However, it is important to exercise caution when interpreting IHC results due to the presence of diffuse background staining. CONCLUSIONS: SSTRs are widely expressed in canine and feline meningiomas, thereby encouraging further studies investigating SSTR expression to conduct trials about the effect of adjunctive therapy with long-acting SST-analogues.
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Enfermedades de los Gatos , Enfermedades de los Perros , Meningioma , Receptores de Somatostatina , Receptores de Somatostatina/metabolismo , Receptores de Somatostatina/genética , Animales , Perros , Gatos , Enfermedades de los Gatos/metabolismo , Enfermedades de los Gatos/genética , Meningioma/veterinaria , Meningioma/metabolismo , Meningioma/genética , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/genética , Neoplasias Meníngeas/veterinaria , Neoplasias Meníngeas/metabolismo , Neoplasias Meníngeas/genética , Femenino , MasculinoRESUMEN
Gastrointestinal lymphoma is the most common form of lymphoma in domestic cats. Aggressive phenotypes are much less common but do bear and unfavorable prognosis. Immunophenotyping by flow cytometry (FCM) is not systematically performed in these patients, because of difficulties in the acquisition of suitable sample material from the gastrointestinal tract. A multimodal diagnostic approach is recommended to improve identification of subtypes targeting patient tailored therapeutic strategies. The aim of this prospective study was to present results of multicolor FCM immunophenotyping in surgically removed gastrointestinal mass and relate them with histopathology using the World Health Organization (WHO) classification and clonality PCR testing. Thirty-two patients were included. Eight cats (25%) had gastric, 23 (72%) had intestinal lymphoma and 1 (3%) had gastric/jejunal lymphoma. Intestinal lymphoma sites were represented by 18 small intestinal, 4 ileocaecal, 1 large intestinal. All gastric lymphomas were diffuse large B-cell lymphoma (DLBCL). Small intestinal lymphomas were 10 enteropathy associated T-cell lymphoma type I (EATL I), 2 enteropathy associated T-cell lymphoma type II (EATL II), 2 peripheral T-cell lymphoma (PTCL), 3 DLBCL and one DLBCL+EATL II. The most common small intestinal FCM T-cell phenotype was CD3+CD21- CD4-CD8-CD18+ CD5-CD79- in 7/10 EATL I and one EATL II. The most frequent FCM B-cell phenotype was CD3-CD21+ CD4-CD8-CD18+ CD5-CD79+ in 13/17 DLBCL and the DLBCL+EATL II. Clonality PCR results were positive in 87.5% (28/32) of all cases. No cross-lineage rearrangement was observed. IHC and FCM results agreed in 87.5% (28/32) of all cases. When all 3 methods were combined, consistent results were seen in 75% (24/32). This is the first demonstration of a multicolor FCM approach set in context to the gold standard histopathology and clonality testing results.
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Lymphoma is the most common tumour of domestic cats, developing most frequently in the small intestine. Feline small intestinal lymphoma predominantly demonstrates a T-cell immunophenotype identified by standard immunopositivity for T cells with CD3 or immunopositivity for B cells with CD20. In contrast, a wide spectrum of immunohistochemical antibodies are applied in humans to diagnose the various specific lymphoma subtypes according to the WHO classification. Our aim was to augment our knowledge of immunophenotypes in feline non-B-cell lymphomas forming macroscopic masses in the intestinal tract. We evaluated the combined immunohistochemistry and flow cytometry findings from 15 cases. Neoplastic lymphoid cells were immunopositive for CD3 in 93% (14/15), granzyme B in 87% (13/15), CD5 in 20% (3/15), CD8 in 13% (2/15), CD4 in 7% (1/15) and CD56 in 7% (1/15) of cases. Cytotoxic granules indicating a cytotoxic origin of the neoplastic cells were identified by histopathology only in 13% (2/15) and by cytology in 47% (7/15) of the cases. Without immunohistochemical labelling of the cytotoxic protein granzyme B, the cytotoxic status would have been missed in 46% (6/13) of the cytological and in 85% (11/13) of the histopathological slides. These findings suggest that more complex immunophenotyping may advance our understanding and help prognosticate small intestinal T-cell lymphoma in cats.
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Enfermedades de los Gatos , Inmunofenotipificación , Neoplasias Intestinales , Gatos , Animales , Inmunofenotipificación/veterinaria , Enfermedades de los Gatos/patología , Enfermedades de los Gatos/inmunología , Neoplasias Intestinales/veterinaria , Neoplasias Intestinales/patología , Inmunohistoquímica , Masculino , FemeninoRESUMEN
OBJECTIVES: Flow cytometric (FCM) immunophenotyping of lymphoid tissue aspirates is an available adjunct for feline lymphoma diagnostics. Reference data have only been established for feline peripheral blood. Studies investigating the composition of normal and mildly reactive feline lymph nodes (LNs) are lacking. The aim of this prospective study was to establish reference data for lymphocyte subpopulations in normal and mildly reactive feline peripheral LNs using a standardised multicolour panel of antibodies. METHODS: Macroscopically inconspicuous mandibular and/or popliteal LNs from 31 adult cats, which were euthanased for reasons other than haematological diseases, were excised and processed within 5 h after death. Multicolour flow cytometry using eight different feline-specific, anti-canine and human cross-reactive monoclonal antibodies used in current diagnostic marker panels was performed after cytological exclusion of pathological states and complemented by lymphocyte clonality testing, histopathology and immunohistochemistry (IHC) to ensure the absence of lymphoid disease. RESULTS: Of 31 cats, the immunophenotyping data of 24 individuals could be included as histopathology and clonality testing excluded a pathological condition. Lymphocyte populations showed the following positive antibody reactions: CD18+ 86.3% ± 13.86%, CD3+ 54.81% ± 11.10%, CD5+ 57.39% ± 12.66%, CD21+ 40.42% ± 12.40%, CD79alphacy+ (CD79αcy) 30.41% ± 13.49% and CD14+ 0.75% ± 1.35%. There were 30.88% ± 13.48% CD4+ and 12.91% ± 6.68% CD8+ cells. Cytology revealed a mixed population of mostly lymphoid cells in all samples. The absence of a monoclonal/oligoclonal neoplastic population was confirmed by lymphocyte clonality testing. Histopathology and IHC showed a normal or mildly reactive pattern in all cases. CONCLUSIONS AND RELEVANCE: This study establishes FCM immunophenotyping data of lymphocyte populations of normal and mildly reactive feline peripheral LNs. For the first time, anti-CD5, CD4, CD8 and CD21 reference data in normal and mildly reactive feline peripheral LNs are presented. CD18, CD3, CD14 and CD79αcy have been used to establish reference data for the first time in any feline material.
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Ganglios Linfáticos , Subgrupos Linfocitarios , Animales , Anticuerpos Monoclonales , Gatos , Citometría de Flujo/veterinaria , Inmunofenotipificación/veterinaria , Ganglios Linfáticos/citología , Estudios ProspectivosRESUMEN
Gastrointestinal lymphomas are uncommon in dogs and little is known about their distinct subtypes or proliferation rate. The aim of this study was to stratify 33 canine gastrointestinal lymphoma samples according to the latest World Health Organization classification and to determine the Ki67 proliferation index by manual counting, digital image analysis and visual estimation. The Ki67 index was then correlated with subtype, immunophenotype, mitotic index, grade and tumour location. The mitotic index correlated positively with the Ki67 index. A significantly higher number of Ki67-positive cells was found in enteropathy-associated T-cell lymphoma type I and in diffuse large B-cell lymphoma compared with enteropathy-associated T-cell lymphoma type II. There was also a significant difference in Ki67 immunolabelled cells between grade 1 and grade 2 lymphomas. Moderate agreement was found between the Ki67 index as obtained by manual counting and visual estimation, but there was strong agreement between manual counting and digital image analysis. The user-friendly digital imaging system used in this study could have potential for future determination of the Ki67 index in lymphoid neoplasms.
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Enfermedades de los Perros , Neoplasias Gastrointestinales , Linfoma de Células B Grandes Difuso , Animales , Proliferación Celular , Perros , Neoplasias Gastrointestinales/veterinaria , Antígeno Ki-67 , Linfoma de Células B Grandes Difuso/veterinaria , Índice Mitótico/veterinariaRESUMEN
Differentiation between resident mature lymphocyte populations and small-cell lymphoma cannot be made by cytological review alone and remains challenging in histopathological review. These cases warrant application of complementary tools like PCR-based immunoglobulin (IG) and T-cell receptor (TCR) clonality testing for confirmation. Although primer coverage of potential IG gene rearrangements in feline B-cell neoplasms constantly improves, the possibility of false negative and false positive test results still poses a problem. In this retrospective study, we assessed diagnostic sensitivity and specificity of a novel developed multiplex PCR assay for routine diagnosis of B-cell clonality. Therefore, 24 feline patients were subjected to comparative clonality testing by using different primer sets. Feline lymphoma cell lines and confirmed patient material served as positive control. Compared to previous studies, this novel developed multiplex PCR assay showed positive effects on diagnostic sensitivity, specificity, accuracy, and positive predictive value accompanied by a slight impairment of negative predictive value. Notably, none of the primer sets was superior; hence, we recommend the combined application of the herein tested primer sets in routine diagnostics. However, a more in-depth-evaluation of the dynamic of assay specific parameters in dependency on primer set usage requires prospective studies on larger cohorts of feline patients.
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Linfocitos B , Enfermedades de los Gatos , Linfoma de Células B , Animales , Enfermedades de los Gatos/diagnóstico , Gatos , Linfoma de Células B/genética , Linfoma de Células B/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Estudios RetrospectivosRESUMEN
Alimentary lymphomas arising from T cells are rare and aggressive malignancies in humans. In comparison, they represent the most common anatomical form of lymphoma in cats. Due to the low prevalence in humans, the underlying pathomechanism for these diseases is poorly characterised, limiting experimental analysis and therapeutic exploration. To date, activating mutations of the JAK/STAT core cancer pathway and particularly the STAT5B oncoprotein have been identified in human enteropathy-associated T cell lymphoma. Here, we describe a high homology of human and feline STAT3 and STAT5B proteins and strong conservation at the genomic level. Analysis of 42 samples of feline T cell alimentary lymphoma reveals broad activation of STAT3 and STAT5B. Screening for known activating mutations in STAT3 or STAT5B identifies the presence of the STAT5BN642H driver mutation in feline enteropathy-associated T cell lymphoma in 7 out of 42 (16.67%) samples in total. Regarding lymphoma subtypes, the majority of mutations with 5 out of 17 (29.41%) cases were found in feline enteropathy-associated lymphoma type II (EATL II). This identification of an oncogenic STAT5B driver mutation in felines recapitulates the genetic situation in the corresponding human disease, thereby establishing the cat as a potential new model for a rare and incurable human T cell disease.
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Corticosteroid administration prior to the application of chemotherapy in small animal lymphoma patients is a concern, as it is discussed to negatively influence the therapeutic outcome due to corticosteroid-induced drug resistance. Using feline lymphoma cell lines FT-1 and MS4 we have shown, that prednisolone pre-treatment alters the susceptibility of these cells towards doxorubicin or vincristine treatment in vitro. The observed effect was negative as for the killing potential and it was cell line and drug (doxorubicin or vincristine) dependent. Furthermore, increase in mRNA expression of selected proteins with multidrug resistance potential (MDR1, BCRP, LRP, MT) was observed after prednisolone pre-treatment. Administration of chemical inhibitors of these proteins did not lead to reversal in sensitivity of tested cell lines to doxorubicin or vincristine.
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Antineoplásicos/administración & dosificación , Enfermedades de los Gatos/tratamiento farmacológico , Doxorrubicina/administración & dosificación , Expresión Génica , Linfoma/veterinaria , Prednisolona/administración & dosificación , Vincristina/administración & dosificación , Animales , Gatos , Línea Celular Tumoral , Resistencia a Múltiples Medicamentos , Linfoma/tratamiento farmacológico , ARN Mensajero/metabolismoRESUMEN
In humans B-symptoms refer to systemic symptoms of lymphoma such as fever, weight loss, and night sweats and influence the prognosis of patients. In canine lymphoma, substage B is used to describe any clinical sign observed. Aim of the retrospective study was to compare the prognostic value of substage B with B-symptoms to predict treatment response and survival in canine nodal diffuse large B-cell lymphoma. Affected dogs treated with CHOP chemotherapy between 2008 and 2019 were included. B-symptoms were defined by weight loss greater than 10% of normal weight, fever and the occurrence of unexplained resting tachypnoea substituted human night sweats. Substage B was defined as any symptoms but lymphadenopathy. Fifty-five cases were included. B-symptoms were present in 20/55 (36%) and substage B in 40/55 (74%) patients. No significant associations between B-symptoms or substage B and weight, sex, breed, WHO stage and lymphoma grade were found. Treatment response was negatively associated with both substage B (P = .02) and B-symptoms (P = .001). B-symptoms significantly decreased progression free survival (PFS) (95 vs 330 days, P = .001) and lymphoma specific survival (LSS) (160 vs 462 days, P = .001). Data showed that B-symptoms might be a more reliable prognostic indicator than substage B in canine nodal diffuse large B-cell lymphoma. Prospective studies assessing B-symptoms in a larger cohort of patients and in other common lymphoma types are warranted. The abstract was presented at the fourth meeting of the European Canine Lymphoma Network Group in Lugano, 22 June 2019 and published in the proceeding of the meeting on the page 26.
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Enfermedades de los Perros/patología , Linfoma de Células B Grandes Difuso/veterinaria , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Ciclofosfamida/uso terapéutico , Enfermedades de los Perros/tratamiento farmacológico , Perros , Doxorrubicina/uso terapéutico , Femenino , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Linfoma de Células B Grandes Difuso/patología , Masculino , Prednisona/uso terapéutico , Pronóstico , Estudios Retrospectivos , Vincristina/uso terapéuticoRESUMEN
The use of transgenic mouse models has revolutionized the study of many human diseases. However, murine models are limited in their representation of spontaneously arising tumors and often lack key clinical signs and pathological changes. Thus, a closer representation of complex human diseases is of high therapeutic relevance. Given the high failure rate of drugs at the clinical trial phase (i.e., around 90%), there is a critical need for additional clinically relevant animal models. Companion animals like cats and dogs display chronic inflammatory or neoplastic diseases that closely resemble the human counterpart. Cat and dog patients can also be treated with clinically approved inhibitors or, if ethics and drug safety studies allow, pilot studies can be conducted using, e.g., inhibitors of the evolutionary conserved JAK-STAT pathway. The incidence by which different types of cancers occur in companion animals as well as mechanisms of disease are unique between humans and companion animals, where one can learn from each other. Taking advantage of this situation, existing inhibitors of known oncogenic STAT3/5 or JAK kinase signaling pathways can be studied in the context of rare human diseases, benefitting both, the development of drugs for human use and their application in veterinary medicine.
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Objectives The purpose of this study was to specify lymphoma subtypes according to the World Health Organization (WHO) classification in a group of cats and to investigate their potential prognostic value. Methods Records of cats from the University of Veterinary Medicine Vienna suffering from lymphoma were reviewed in this retrospective study. To diagnose various subtypes specified in the WHO classification, histopathological and immunohistochemical examinations, as well as clonality assays in some cases, were performed. Results Of the 30 cats included in this study and classified according to the WHO guidelines, peripheral T-cell lymphoma was the most prevalent lymphoma subtype (37% of cases; n = 11), followed by diffuse large B-cell (23%; n = 7), intestinal T-cell (10%; n = 3), T-cell-rich B-cell (10%; n = 3), large granular lymphocytic (7%; n = 2), anaplastic large T-cell (7%; n = 2), B-cell small lymphocytic (3%; n = 1) and T-cell angiotropic lymphoma (3%; n = 1). The median survival time (MST) was 5.4 months (range 6 days to 2.2 years), with two cats still alive after 1.7 and 2.0 years, respectively. Treating cats prior to chemotherapy with glucocorticoids did not worsen their prognosis. Adding to chemotherapy, radiotherapy or surgery did not improve the clinical outcome. We observed that patients with intestinal T-cell lymphoma lived significantly longer (MST 1.7 years) than those with a diffuse large B-cell (MST 4.5 months) or peripheral T-cell lymphoma (MST 6.1 months). Cats with T-cell-rich B-cell lymphoma survived significantly longer (MST 1.2 years) than those with a diffuse large B-cell lymphoma. Conclusions and relevance A detailed diagnosis of feline lymphoma can be obtained by allocating different subtypes according to the WHO classification. From the eight detected lymphoma subtypes, two, intestinal T-cell lymphoma and T-cell-rich B-cell lymphoma, showed promising survival times in cats.
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Enfermedades de los Gatos/mortalidad , Linfoma/veterinaria , Estadificación de Neoplasias , Animales , Austria/epidemiología , Enfermedades de los Gatos/clasificación , Enfermedades de los Gatos/patología , Gatos , Femenino , Linfoma/clasificación , Linfoma/mortalidad , Masculino , Prevalencia , Pronóstico , Estudios Retrospectivos , Análisis de Supervivencia , Organización Mundial de la SaludRESUMEN
Biological features of canine osteosarcomas (OS) differ markedly from those found in feline and resemble more human osteosarcomas, in particular for their high rate of metastasis and poor prognosis. Ezrin, radixin and moesin are members of the ERM protein family and link the actin cytoskeleton with the cell membrane. Ezrin and moesin have been shown to be of prognostic significance in tumor progression due to their role in the metastatic process. The objective of this study was to analyze ezrin and moesin protein expression in a series of dog (n = 16) and cat (n = 8) osteosarcoma samples using immunohistochemistry and western blot techniques. We found that cat OS have a higher moesin expression compared to dog OS, however, the active phosphorylated forms of moesin and ezrin Tyr353 were more abundant in the dog samples. A statistically significant difference was found for the low and high immunohistochemical scores of ezrin and pan-phospho-ERM proteins between cat and dog. Although phospho-ezrin Thr567 was higher in feline OS, the membranous localization in dog OS samples indicates the presence of the biologically active form. Therefore, the observed differences in phosphorylated forms of ezrin and moesin status should be further studied to demonstrate if they are relevant for different biological behavior between dog and cat OS.
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Neoplasias Óseas/metabolismo , Proteínas del Citoesqueleto/metabolismo , Regulación Neoplásica de la Expresión Génica , Proteínas de Microfilamentos/metabolismo , Osteosarcoma/metabolismo , Citoesqueleto de Actina/metabolismo , Animales , Enfermedades de los Gatos/metabolismo , Gatos , Línea Celular Tumoral , Membrana Celular/metabolismo , Citoesqueleto/metabolismo , Progresión de la Enfermedad , Enfermedades de los Perros/metabolismo , Perros , Femenino , Perfilación de la Expresión Génica , Masculino , Proteínas de la Membrana/metabolismo , Metástasis de la Neoplasia , Fenotipo , FosforilaciónRESUMEN
Osteosarcoma is considered the most common bone cancer in cats and dogs, with cats having a much better prognosis than dogs, since the great majority of dogs with osteosarcoma develop distant metastases. In search of a factor possibly contributing to this disparity, the stem cell growth factor receptor KIT was targeted, and the messenger (m)RNA and protein expression levels of KIT were compared in canine vs. feline osteosarcomas, as well as in normal bone. The mRNA expression of KIT was quantified by reverse transcription-quantitative polymerase chain reaction, and was observed to be significantly higher in canine (n=14) than in feline (n=5) osteosarcoma samples (P<0.001). KIT protein expression was evaluated by immunohistochemistry, which revealed that 21% of canine osteosarcoma samples did not exhibit KIT staining in their neoplastic cells, while in 14% of samples, a score of 1 (<10% positive tumour cells) was observed, and in 50% and 14% of samples, a score of 2 (10-50% positivity) and 3 (>50% positivity), respectively, was observed. By contrast, the cancer cells of all the feline bone tumour samples analysed were entirely negative for KIT. Notably, canine and feline osteocytes of healthy bone tissue lacked any KIT expression. These results could be the first evidence that KIT may be involved in the higher aggressiveness of canine osteosarcoma compared with feline osteosarcoma.
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Feline alimentary lymphoma is the most common hematopoietic neoplasia in cats. It affects mainly the small intestines and is most frequently of T-cell origin. Evaluation of a fine needle aspirate is often the first step in the diagnostic work-up. Differentiation between a resident mature lymphocyte population as encountered in inflammatory bowel disease and small cell lymphoma cannot be achieved by cytology alone. Even full thickness biopsies evaluated by histopathology can be inconclusive. These cases warrant the application of complementary tools like PCR-based T-cell receptor (TCR) clonality testing for confirmation. The aim of this study was to optimize the DNA extraction protocol for formalin fixed and paraffin embedded tissues (FFPE) and to establish a heteroduplex analysis to enhance resolution of the PCR fragments of the T-cell receptor gamma (TCRG) V-J gene. The new protocols resulted in improved quantity and quality of the extracted DNA. Heteroduplex analysis of the samples improved the resolution of the electrophoresis results so that rules for interpretation of the different patterns could be established. Application of this improved setup detected clonal rearrangements in at least one TCRG primer reaction in 31 of 36 of our feline intestinal lymphoma samples after DNA quality testing.
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Enfermedades de los Gatos/diagnóstico , Neoplasias Gastrointestinales/veterinaria , Linfoma de Células T/veterinaria , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Animales , Enfermedades de los Gatos/patología , Gatos , Neoplasias Gastrointestinales/diagnóstico , Neoplasias Gastrointestinales/patología , Regulación Neoplásica de la Expresión Génica , Linfoma/genética , Linfoma de Células T/metabolismo , Linfoma de Células T/patología , Mutación , Reacción en Cadena de la Polimerasa/métodos , Estudios RetrospectivosRESUMEN
A 7-year-old male Boxer with a 3.5-year history of atopy and food hypersensitivity was presented with multiple poorly circumscribed nodules and maculae of the skin and tongue, and jaundiced mucosal membranes. Cytologic and histopathologic examination of the skin lesions revealed cutaneous epitheliotropic lymphoma. Cells were CD3(+) and CD8(+) in flow cytometry. The CBC showed a moderate leukocytosis with 16% atypical lymphocytes with irregularly cleaved nuclei. Flow cytometric phenotyping of peripheral blood showed an elevated proportion of the CD8(+) T-lymphocyte subpopulation, indicating a malignant population of T-cell origin, and the electropherogram of the PCR antigen receptor rearrangement produced a monoclonal peak for TCRγ. Liver enzyme activities were markedly increased and abdominal ultrasound examination showed increased echogenicity of the liver and enlarged abdominal lymph nodes. Fine-needle aspirates of the liver confirmed infiltration with lymphocytes exhibiting the same morphology as the cells detected in skin and peripheral blood. Treatment was induced with L-asparaginase, lomustine, and prednisone. Partial clinical remission of the skin and tongue lesions was achieved within 10 days, and hematologic abnormalities resolved. Despite further treatment with L-asparaginase and lomustine, the dog relapsed within one month and was euthanized. Presence of malignant lymphocytes in skin, peripheral blood, and liver indicate a rare variant of leukemic cutaneous T-cell lymphoma, equivalent of Sézary syndrome in a dog. This case report describes the use of flow cytometry as a complementary tool for lymphocyte characterization of skin lesions for the first time.
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Enfermedades de los Perros/patología , Citometría de Flujo/veterinaria , Linfoma Cutáneo de Células T/veterinaria , Síndrome de Sézary/veterinaria , Enfermedades de la Piel/veterinaria , Neoplasias Cutáneas/veterinaria , Animales , Biopsia con Aguja Fina/veterinaria , Perros , Hígado/enzimología , Hígado/patología , Ganglios Linfáticos/patología , Linfocitos/patología , Linfoma Cutáneo de Células T/patología , Masculino , Síndrome de Sézary/patología , Piel/patología , Enfermedades de la Piel/patología , Neoplasias Cutáneas/patologíaRESUMEN
Osteosarcoma is a rare but aggressive bone neoplasm in humans, which is commonly treated with surgery, classical chemotherapy and radiation. Sorafenib, an inhibitor of a number of kinases targeting the Raf/MEK/ERK pathway, is a promising new chemotherapeutic agent in human medicine that has been approved since 2006 for the therapy of renal cell carcinoma and since 2007 for the treatment of hepatocellular carcinoma. Here, we studied the antimetastatic potential of 4 µM of this multikinase inhibitor in a human osteosarcoma cell line. DNA microarray-based gene expression profiling detected 297 and 232 genes upregulated or downregulated at a threshold of >2-fold expression alteration (P<0.05) in the sorafenib-treated cells. Three genes (CXCR4, FOS and S100A4) that are involved in tumor progression were chosen for validation by quantitative PCR (qPCR) and protein expression analysis. The decrease in RNA expression detected by microarray profiling was confirmed by qPCR for all three genes (P<0.01). On the protein level, sorafenib-induced reduction of S100A4 was verified both by western blotting and immunohistochemistry. For CXCR4 and c-Fos, a reduced protein expression was shown by immunohistochemistry, for c-Fos also by immunoblotting. We conclude that sorafenib could serve as a potent chemotherapeutical agent by which to inhibit the metastatic progression of osteosarcomas.
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Antineoplásicos/farmacología , Niacinamida/análogos & derivados , Proteínas Oncogénicas v-fos/metabolismo , Compuestos de Fenilurea/farmacología , Receptores CXCR4/metabolismo , Proteínas S100/metabolismo , Neoplasias Óseas , Línea Celular Tumoral , Regulación hacia Abajo , Ensayos de Selección de Medicamentos Antitumorales , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Niacinamida/farmacología , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteínas Oncogénicas v-fos/genética , Osteosarcoma , Receptores CXCR4/genética , Proteína de Unión al Calcio S100A4 , Proteínas S100/genética , Sorafenib , Transcriptoma/efectos de los fármacosRESUMEN
The role of VEGF and its receptors has extensively been studied in tumours. In contrast, the presence and function of VEGF in normal tissues like the lymph node has not been given much attention until now. To study the expression of VEGF, VEGFR-1, VEGFR-2 and VEGFR-3 in the heterogenous cell population of the canine lymph node, laser capture microdissection was used to isolate pure cell fractions of macrophages, lymphocytes, endothelial cells, and capsule cells of the canine lymph node. To clarify if macrophages take up VEGF from the environment or express VEGF, VEGFR-1, VEGFR-2 or VEGFR-3 themselves, the mRNA expression was studied by real-time RT-PCR. After RNA isolation and subsequent analysis with the Agilent 2100 Bioanalyzer only RNA samples with appropriate RNA integrity were used for real-time PCR. For the accurate relative quantification of mRNA expression levels several reference genes were evaluated. It was shown that the reference genes HPRT1 and B2M serve as reliable reference genes for gene expression studies in the canine lymph node. Expression data analysis revealed no significant difference in VEGF expression levels between endothelial cells and the other investigated cells. VEGFR-1 expression was significantly lower in lymphocytes. Also macrophages showed a highly significant lower expression of VEGFR-1 compared to endothelial cells. In addition, the VEGFR-2 expression in lymphocytes and macrophages was significantly lower in comparison to endothelial cells. We were not able to detect VEGFR-3 mRNA in the lymphocyte cell population, in macrophages and cells of the lymph node capsule VEGFR-3 was expressed at very low levels. It was shown that laser capture microdissection in combination with quantitative real-time PCR is a valuable tool for studying the expression patterns of specific cells in their microenvironment. Our results support the hypothesis that VEGF and its receptors have other biological roles besides stimulating angiogenesis in the normal lymph node. These biological functions need to be clarified in further studies.
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Perros/genética , Ganglios Linfáticos/metabolismo , Receptores de Factores de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/genética , Animales , Secuencia de Bases , Cartilla de ADN/genética , Perros/metabolismo , Perfilación de la Expresión Génica , Rayos Láser , Ganglios Linfáticos/citología , Microdisección/métodos , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genética , Receptor 3 de Factores de Crecimiento Endotelial Vascular/genéticaRESUMEN
An 8-year-old male Austrian Pinscher and a 14-year-old male Golden Retriever were presented for evaluation due to unexplainable high fructosamine values despite euglycemia and epistaxis in combination with polydipsia/polyuria, respectively. Blood analysis revealed severe hyperglobulinemia, hypoalbuminemia and markedly elevated fructosamine concentrations in both dogs. Multiple myeloma with IgA-monoclonal gammopathy was diagnosed by serum and urine electrophoresis including immunodetection with an anti-dog IgA antibody and bone marrow aspirations. Diabetes mellitus was excluded by repeated plasma and urine glucose measurements. Fructosamine values were positively correlated with globulin, but negatively correlated with albumin concentrations. These cases suggest that, as in human patients, monoclonal IgA gammopathy should be considered as a possible differential diagnosis for dogs with high fructosamine concentrations.
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Enfermedades de los Perros/sangre , Fructosamina/sangre , Inmunoglobulina A/metabolismo , Mieloma Múltiple/veterinaria , Paraproteinemias/veterinaria , Animales , Proteínas Sanguíneas/análisis , Enfermedades de los Perros/tratamiento farmacológico , Perros , Masculino , Melfalán/uso terapéutico , Mieloma Múltiple/complicaciones , Mieloma Múltiple/tratamiento farmacológico , Agonistas Mieloablativos/uso terapéutico , Paraproteinemias/sangre , Paraproteinemias/complicaciones , Paraproteinemias/tratamiento farmacológicoRESUMEN
Local recurrence of feline soft tissue sarcomas is common despite aggressive treatment. Liposomal doxorubicin might serve as a depot radiosensitizer if administered concomitantly with daily radiotherapy and thus improve tumor control. In this pilot study, the feasibility of concomitant liposomal radiochemotherapy was evaluated in a palliative setting in 10 cats with advanced soft tissue sarcomas. Cats were treated with median number of 5 (range 5-7) daily fractions of radiotherapy and a median total dose of 20 Gy (range 20-31.5 Gy). One dose of liposomal doxorubicin was administered at the beginning of radiotherapy. Seven cats received further free or liposomal doxorubicin after completion of the liposomal doxorubicin/radiation protocol. Seven of the treated 10 cats (70%) achieved a partial (n=5) or complete (n=2) response with a median response duration of 237 days. The median progression free interval in all 10 cats was 117 days and the median overall survival time was 324 days. Concomitant liposomal radiochemotherapy was tolerated well in nine cats, one cat experienced temporary anorexia. Although the number of patients is too small to make definitive conclusions, results appear promising enough to investigate the role of liposomal doxorubicin as a radiosensitizer further.
Asunto(s)
Enfermedades de los Gatos/tratamiento farmacológico , Enfermedades de los Gatos/radioterapia , Doxorrubicina/uso terapéutico , Cuidados Paliativos , Fármacos Sensibilizantes a Radiaciones/uso terapéutico , Sarcoma/veterinaria , Neoplasias de los Tejidos Blandos/veterinaria , Animales , Enfermedades de los Gatos/mortalidad , Gatos , Terapia Combinada/veterinaria , Fibrosarcoma/tratamiento farmacológico , Fibrosarcoma/radioterapia , Fibrosarcoma/veterinaria , Liposomas , Dosificación Radioterapéutica/veterinaria , Sarcoma/tratamiento farmacológico , Sarcoma/mortalidad , Sarcoma/radioterapia , Neoplasias de los Tejidos Blandos/tratamiento farmacológico , Neoplasias de los Tejidos Blandos/mortalidad , Neoplasias de los Tejidos Blandos/radioterapiaRESUMEN
Acute myeloid leukemia (AML) in dogs is a rare disease with poor prognosis. In most subjects, palliative treatment or euthanasia is performed. A 3.5-year-old male castrated labrador with AML-M7, which was treated with induction polychemotherapy (8 cycles) using vincristine (0.5 mg/m(2)/cycle), daunorubicin (20 mg/m(2)/cycle), cytosine arabinoside (ARA-C, 100 mg/m(2)/cycle) and prednisolone (1 mg/kg/day) is reported. Treatment was well tolerated and complete remission was achieved. Postinduction chemotherapy consisted of ARA-C, daunorubicin and prednisolone. After 3, 5 and 18 months, the subject relapsed. Each relapse was treated with ARA-C (up to 1,000 mg/m(2)) and etoposide or daunorubicin. Again, no severe side-effects occurred and the disease was controlled, with 37 chemotherapy-cycles (ARA-C, 3 x 1,000 mg/m(2)/cycle), for 24 months. Based on a literature-search, this is the first report documenting a long-term response of canine AML, probably resulting from the high-dose ARA-C. Clinical trials using high-dose ARA-C are now required to confirm antileukemic efficacy in canine leukemias.