Feline HER2 Protein Expression Levels and Gene Status in Feline Mammary Carcinoma: Optimization of Immunohistochemistry (IHC) and<i>In Situ</i>Hybridization (ISH) Techniques

Soares, Maria Carolina S.; Correia, Jorge; Rodrigues, Pedro; Simões, Margarida; Matos, A. de O.; Ferreira, Fernando

doi:10.1017/s1431927613001529

Cited by 38 publications

(59 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Hormonal regulation of feline mammary carcinoma (FMC) is evident because tumor incidence is drastically reduced by ovariohysterectomy [50;52]. Disease progression and histopathology of FMC are similar to that of human breast cancer [51;53–60]. FMC express EGFR, HER2 and HER3 as we reported with two feline mammary tumor lines K12 [61] and K248 [62] as well as primary tumor tissue [63].…”

Section: Canine and Feline Mammary Tumor Modelsmentioning

confidence: 77%

Evolution of animal models in cancer vaccine development

Wei

Jones

Juhász

et al. 2015

Vaccine

View full text Add to dashboard Cite

Advances in cancer vaccine development are facilitated by animal models reflecting key features of human cancer and its interface with host immunity. Several series of transplantable preneoplastic and neoplastic mouse mammary lesions have been used to delineate mechanisms of anti-tumor immunity. Mimicking immune tolerance to tumor-associated antigens (TAA) such as HER2/neu, transgenic mice developing spontaneous mammary tumors are strong model systems for pre-clinical vaccine testing. In these models, HER2 DNA vaccines are easily administered, well-tolerated, and induce both humoral and cellular immunity. Although engineered mouse strains have advanced cancer immunotherapy, basic shortcomings remain. For example, multiple mouse strains have to be tested to recapitulate genetic regulation of immune tolerance in humans. Outbred domestic felines more closely parallel humans in the natural development of HER2 positive breast cancer and their varying genetic background. Electrovaccination with heterologous HER2 DNA induces robust adaptive immune responses in cats. Importantly, homologous feline HER2 DNA with a single amino acid substitution elicits unique antibodies to feline mammary tumor cells, unlocking a new vaccine principle. As an alternative approach to targeted vaccination, non-surgical tumor ablation such as cryoablation induces anti-tumor immunity via in situ immunization, particularly when combined with toll-like receptor (TLR) agonist. As strategies for vaccination advance, non-invasive monitoring of host response becomes imperative. As an example, magnetic resonance imaging (MRI) and positron emission tomography (PET) scanning following administration of tryptophan metabolism tracer [11C]-alpha-methyl-tryptophan (AMT) provides non-invasive imaging of both tumor growth and metabolic activities. Because AMT is a substrate of indoleamine-pyrrole 2,3-dioxygenase (IDO), an enzyme that produces the immune regulatory molecule kynurenine, AMT imaging can provide novel insight of host response. In conclusion, new feline models improve the predictive power of cancer immunotherapy and real-time PET imaging enables mechanistic monitoring of host immunity. Strategic utilization of these new tools will expedite cancer vaccine development.

show abstract

Section: Canine and Feline Mammary Tumor Modelsmentioning

confidence: 77%

Evolution of animal models in cancer vaccine development

Wei

Jones

Juhász

et al. 2015

Vaccine

View full text Add to dashboard Cite

show abstract

“…However these authors demonstrated that neither TAB250 nor SP3 antibodies showed reactivity with the extracellular regions of the cat erbB-2 protein [62]. The majority of works, with respect to erbB-2 IHC analysis in cat mammary tissues, uses antibodies with affinity for the intracellular region of the erbB-2 protein, which include: A0485 [4], [38], [43]; CB11 [41]; A0485 and CB11 [42]; and, 4B5, A0485 and CB11 [62], [63].…”

Section: Discussionmentioning

confidence: 99%

ERBB2 in Cat Mammary Neoplasias Disclosed a Positive Correlation between RNA and Protein Low Expression Levels: A Model for erbB-2 Negative Human Breast Cancer

et al. 2013

View full text Add to dashboard Cite

Human ERBB2 is a proto-oncogene that codes for the erbB-2 epithelial growth factor receptor. In human breast cancer (HBC), erbB-2 protein overexpression has been repeatedly correlated with poor prognosis. In more recent works, underexpression of this gene has been described in HBC. Moreover, it is also recognised that oncogenes that are commonly amplified or deleted encompass point mutations, and some of these are associated with HBC. In cat mammary lesions (CMLs), the overexpression of ERBB2 (27%–59.6%) has also been described, mostly at the protein level and although cat mammary neoplasias are considered to be a natural model of HBC, molecular information is still scarce. In the present work, a cat ERBB2 fragment, comprising exons 10 to 15 (ERBB2_10–15) was achieved for the first time. Allelic variants and genomic haplotype analyses were also performed, and differences between normal and CML populations were observed. Three amino acid changes, corresponding to 3 non-synonymous genomic sequence variants that were only detected in CMLs, were proposed to damage the 3D structure of the protein. We analysed the cat ERBB2 gene at the DNA (copy number determination), mRNA (expression levels assessment) and protein levels (in extra- and intra protein domains) in CML samples and correlated the last two evaluations with clinicopathological features. We found a positive correlation between the expression levels of the ERBB2 RNA and erbB-2 protein, corresponding to the intracellular region. Additionally, we detected a positive correlation between higher mRNA expression and better clinical outcome. Our results suggest that the ERBB2 gene is post-transcriptionally regulated and that proteins with truncations and single point mutations are present in cat mammary neoplastic lesions. We would like to emphasise that the recurrent occurrence of low erbB-2 expression levels in cat mammary tumours, suggests the cat mammary neoplasias as a valuable model for erbB-2 negative HBC.

show abstract

“…The antigen retrieval was performed by boiling samples for 2 minute in a sodium citrate buffer solution (0.01M NaCH 3 COO, pH 6.0) using a pressure cooker (2 atm). 3,38 For PR immunodetection, antigen retrieval was achieved with the same sodium citrate buffer in a water bath for 60 minutes at 95 C. For CK 5/6, samples were microwaved at 900 W for 15 minutes in Tris-EDTA buffer (pH 9.0). 2 Immunohistochemical analysis was performed with the primary antibodies summarized in Supplemental Table 1, and staining was achieved with a modified streptavidin-peroxidase conjugate method (Novolink MaxPolymer Detection System, Leica Biosystems, Wetzlar, Germany).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Ki-67 as a Prognostic Factor in Feline Mammary Carcinoma

et al. 2015

View full text Add to dashboard Cite

Ki-67 is a nuclear protein and a proliferation marker frequently used in establishing the prognosis for breast cancer patients. To investigate the prognostic value of the Ki-67 proliferation index in female cats with mammary carcinoma, a prospective study was conducted with 96 animals. The Ki-67 index of primary tumors (n ¼ 96) was initially determined, and whenever possible, the Ki-67 index of regional lymph node metastasis (n ¼ 38) and distant metastasis (n ¼ 16) was also estimated. The optimal cutoff value for the Ki-67 index was determined by univariate and multivariate analysis. Ki-67 indices 14% were detected in 72.9% (70 of 96) of the tumors. Tumors with a Ki-67 index 14% were significantly associated with large size (P ¼ .022), poor differentiation (P ¼ .009), presence of necrotic areas (P ¼ .008), estrogen receptor-negative status (P < .0001), fHER2-negative status (P ¼ .003), and shorter overall survival (P ¼ .012). Moreover, Ki-67 expression in the primary tumor was strongly and positively correlated with both regional metastasis (P < .0001; r ¼ 0.83) and distant metastasis (P < .0001; r ¼ 0.83), and was significantly higher in distant metastases when compared with the primary tumor (P ¼ .0009). A similar correlation was also observed between regional and distant metastasis (P < .0001; r ¼ 0.75). On the basis of the above results, the authors propose the adoption of the 14% value as the optimal cutoff for Ki-67 to identify tumors with high risk of disease progression.Keywords feline mammary carcinoma, Ki-67 proliferation index, prognostic factor, cutoff value Feline mammary carcinomas (FMCs) are among the most prevalent tumors in cats, with an incidence that can reach 40% of all tumor cases in this species. 23,39,42 Contrary to those of humans and dogs, 85% to 93% of feline mammary tumors are considered malignant, and the most common histotypes are the tubular, papillary, solid, and cribriform carcinomas. 39 Feline mammary tumors may metastasize to the regional lymph nodes, lungs, and liver, as well as other organs. Although some studies correlate age, tumor size, presence of regional metastasis, and malignancy grade with overall survival (OS), 19,37,43 still scarce information is available regarding the proliferation index of FMC. Based on this scenario, additional prognostic and predictive factors may provide useful insights into tumor biology to achieve a more efficient therapy and a better follow-up of the affected female cats.Ki-67 is a protein found only in growing, dividing cells. It is expressed in all cell cycle phases, except in the resting phase (G0). With an intranuclear localization, the expression levels of Ki-67 are low during the G1 and S phases, rapidly increase during the G2 phase, and reach a peak in mitosis. This well-defined expression pattern makes the Ki-67 antigen a good proliferation marker, very useful and reliable in the prognosis of human breast cancer. 9,45 Several studies of women demonstrated that high Ki-67 expression is a poor prognostic indicator of 5-year rec...

show abstract

Feline HER2 Protein Expression Levels and Gene Status in Feline Mammary Carcinoma: Optimization of Immunohistochemistry (IHC) andIn SituHybridization (ISH) Techniques

Cited by 38 publications

References 28 publications

Evolution of animal models in cancer vaccine development

Evolution of animal models in cancer vaccine development

ERBB2 in Cat Mammary Neoplasias Disclosed a Positive Correlation between RNA and Protein Low Expression Levels: A Model for erbB-2 Negative Human Breast Cancer

Ki-67 as a Prognostic Factor in Feline Mammary Carcinoma

Contact Info

Product

Resources

About