Feline mammary

Question 1

Maniscalco et al. 2012. Activation of AKT in feline mammary carcinoma: a new prognostic factor for feline mammary tumors

Prognostic value of PI3K/AKT/PTEN pathway dysregulation in feline mammary tumors

p-AKT expression significantly correlated with?

Tumors expressing p-AKT had ____ disease free period than those with negative tumors

AKT activation was associated with ___ and ____

No AKT activation was observed in relation to ___

Answer 1

Tumor malignancy, histological differentiation, and clinical recurrence

Shorter

HER-2 expression and PTEN downregulation

ER or PR

Question 2

Baptista et al. 2012. Sequence variation and mRNA expression of the TWIST1 gene in cats with mammary hyperplasia

Humans have germline mutation in TWIST1 oncogene may predispose to breast cancer and expression has been correlated with tumor progression and metastasis

Feline mammary CA had ___ of expression of TWIST1 mRNA than benign mammary tumors

Variations in TWIST1 coding region in feline mammary tumors

Answer 2

Lower levels

None, and no mutationa as people have

Question 3

Penafiel-Verdu et al. 2012. Reduced expression of E-cadherin and B-catenin and high expression of basal cytokeratins in feline mammary carcinomas with regional metastasis

21 adenomas and 139 mammary carcinomas

66 of the carcinomas had metastasized, while 73 had regional LN mets at diagnosis

Preservation of E-cadherin and B-catenin is a significant feature of carcinomas without metastasis, wheras metastasis reveal the loss of one or both adhesion molecules

Basal cytokeratin was statistically associated with presence of regional metastasis

Expression of E-cadherin-B-catenin was correlated with high expression of CK18 and low expression of CK5/6

Question 4

Ferrari et al. 2012. CXCR4 expression in feline mammary carcinoma cells: evidence of a proliferative role for the SDF-1/CXCR4 axis

Malignant feline mammary tumors express CXCR4 with higher level in malignant tumors and metastatic cells have stronger reactivity for CXCR4 than primary tumors

CXCR4 activation in primary cultures of feline mammary carcinoma causes increase in proliferation rate

SDF-1/CXCR4 system seems to play a tumorigneic role in feline mammary gland malignancy

Question 5

Cherrington et al. 2012. Comparative analysis of peptidylarginine deiminase-2 expression in canine, feline, human mammary tumors

PAD converts arginine residues in proteins to citrulline

Canine mammary gland, PAD2 is detected in epithelial cells in estrus and becomes widely expresses in diestrus

PAD2 appears to modify nuclear histone, suggesting a role for the enzyme in chromatin remodelling and gene regulation

Normal human and canine mammary epithelium showed strong cytoplasmic and nuclear expression of PAD2, but reduced expression in mammary carcinomas from both species

Feline mammary Ca had complete loss of nuclear PAD2 expression

Loss of PAD2 may represent marker of progression towards more aggressive neoplasia

Question 6

Zappulli et al. 2012. Immunohistochemical expression of E-cadherin and B-catenin in feline mammary tumors

IHC expression of E-cadherin and B-catenin in 53 mammary tumors and 48 hyperplastic or dysplastic lesions from 57 queens

E-cadherin and B-catenin expression was membranous in all samples and significant decrease in malignant tumors and metastases

Cytoplasmic expression of both markers was inversely correlated to membrane localization

B-catenin nuclear labeling detected in 1 LN (60% positive cells) and in basal/myoepithelial cells of 6/7 ductal tumors

No correlation with survival was found with either marker

Question 7

Pang et al. 2013. Feline mammary carcinoma stem cells are tumorigenic, radioresistant, chemoresistant, and defective in activation of ATM/p53 DNA damage pathway

Feline mammary carcinoma stem cells were resisatnt to chemotherapy and radiation, possibly due to aberrant activation of the ATM/p53 DNA damage pathway.

EMT was feature of invasive phenotype

Results demonstrate CSC are a feature of mammary cancer in cats

Question 8

Maniscalco et al. Activation of mTOR in triple negative feline mammary carcinomas

Triple negative breast cancer (TNBC) - absent ER, PR, and HER2 overexpression. mTOR is overexpressed in TNBC and represents a target

Study investigated mTOR and p-mTOR expression in FMC in relation to TN phenotype

53.3% of FMC were ER, PR, HER2 negative while 56.9% and 55.2% expressed mTOR and p-mTOR

Question 9

Wiesse et al. 2013. Feline mammary basal-like ACAs: a potential model for human triple-negative breast cancer (TNBC) with basal-like subtype

TNBC - negative for ER, PR, HER2

Subpopulation of TNBC have basal like morphology - positive for CK5/6 and/or EGFR and high incidence of BRCA mutations

24 FMAs were classified

14 of 24 (58%) FMA were triple negative, 11/14 (79%) had basal like subtype with no genetic abnormality in BRAC1 or BRAC2

Question 10

Michishita et al. 2013. Flow cytometric analysis for detection of tumor-initiating cells in feline mammary carcinoma cell lines

(TICs) - have the capacity to self-renew, differentiate, and form tumors at high frequency, has a potential role in tumor initiation, aggression, and recurrence.

In human breast cancers, TICs are identified by surface markers, such as CD44 and CD24, and an aldefluor assay based on aldehyde dehydrogenase activity (ALDH(+)) using flow cytometry.

We attempted to identify CD44(+)CD24(-) and ALDH(+) cells using 8 feline mammary carcinoma cell lines, including FKNp, which was obtained from a primary lesion, and the capacity to generate tumor nodules was analyzed in immunodeficient mice injected with ALDH(+) FKNp-derived cells.

The CD44(+)CD24(-) and ALDH(+) cells were detected in all cell lines derived from feline mammary carcinomas. Xenograft transplantation into immunodeficient mice demonstrated that as few as 1 × 10(2) ALDH(+) cells could initiate tumor growth in 1 out of 4 mice, while 1 × 10(3) ALDH(+) cells initiated tumor growth in 5 out of 6 mice. However, 1 × 10(3) ALDH(-) cells failed to initiate tumors in all the tested mice. ALDH(+)-derived tumors contained both ALDH(+) and ALDH(-) cells, indicating that ALDH(+) FKNp-derived cells had higher tumorigenicity than ALDH(-) cells.

These results suggest that TICs may exist in feline mammary carcinomas, and further characterization of CD44(+)CD24(-) and ALDH(+) cells is needed to define novel therapies targeted against TICs. This study provides the foundation for elucidating the contribution of TICs in tumorigenesis.

Question 11

Buendia et al. 2013. N-cadherin expression in feline mammary tumors is associated with reduced E-cadherin expression and the presebce if regional metastasis

N-cadherin is expressed by neural and fibroblast cells but not epithelial cells

Some studies in human suggest switch from N-cad to E-cad as a step in malignant progression

21 adenomas and 139 carcinomas

Results showed statistically significant relation between expression of N-cad and 2 prognostic factors and also reduced expression of E-cad in tumors that expressed N-cad

Question 12

Flores et al. 2014. Reduced ezpression of claudin-2 is associated with high histological grade and metastasis of feline mammary carcinomas

Claudins are family of tight junction proteins and play role in cell polarity, controlling paracellular ion flux and regulating proliferation and differentiation

Evidence suggets changes in caludin expression and development of human breast cancer

CLDN expression in normal feline mammary tissue (5), mammary carcinomas (52), metastatic lesions (29)

77% of carcinomas had reduced CLDN expression compared to normal mammary gland

Reduced expression of CLDN-2 was significantly associated with high histologic grade of carcinoma, with 88.6% of grade II/III carcinomas showing decreased expression

CLDN-2 downregulation was associated with metastatic disease, with 93% have decreased protein expression

Question 13

Santos et al. 2013. ERB2 in cat mammary neoplasias disclosed a positive correlation between RNA an dprotein low expression levels: a model for erbB2 negative human breast cancer

Question 14

Millanta et al. 2016. COX-2, mPGES-1 and EP2 receptor immunohistochemical expression in canine and feline malignant tumors

PGE2 is controlled by COX-2 and microsomal PGE synthase-1 (mPGES-1). PG singlaing is involved in human and animal cancer development

COX-2 positivity occured in 83% canine and 81% feline mammary tumors

mPGES-1 in 75% of canine and 66% feline mammary carcinomas and the EP2 receptor expression was observed in 89% canine and 54% feline carcinomas

Frequency of COX-2, EP2 receptor and mPGES-1 expression was significantly higher in carcinomas than non-neoplastic tisssues and adenomas

Question 15

Mills et al. 2015. Prognostic value of histologic grading for feline mammary carcinoma: a retrospective survival analysis

Evaluate the Elston and Ellis grading system (originally developed for human breast cancer) for feline mammary carcinoma

Elston and Ellis grading failed to correlate with overall survival

Using multivariable analysis, lymphovascular invasion, nuclear form, and mitotic counr each demonstrated independentd prognostic significance

Modification of the Elston and Ellis grading system - correlation with overall survival

MST 27, 29, 31 months for grade 1, 14, 12, or 14 months for grade 2, 13, 5, 8 for grade 3 using modified Ellis system

Question 16

Adelfinger et al. 2014. Evaluation of a new recombinant oncolytic vaccinia virus strain GLV-5b451 for feline mammary carcinoma therapy

Question 17

Flores et al. 2014. Clinicopathologic significance of immunoexpression of caludin-1 and claudin-7 in feline mammary CAs

Inverse association between CLDN-7 and histological grade of tumor

Reduced expression of CLDN-7 was significantly associated with decreased tubule formation, high proliferative activity and metastasis

No significant association was found between CLDN-1 expression

Question 18

Caliari et al. 2014. Triple-negative vimentin positive heterogeneous feline mammary carcinomas as a potential comparative model for breast cancer

A large group of hormone receptors (HRs)-negative aggressive carcinomas - that did not overexpress HER2 - could be distinguished from the less aggressive (10.8%) and benign (8%) HRs + tumors, that showed bilineage (luminal and myoepithelial) differentiation.

Immunohistochemical evaluations of cytoplasmic filaments indicated that HRs- FMCs are vimentin+, CK14+, and CK5_6+ carcinomas that may resemble the TNBCs (basal like/claudin low) described in women. The identification of luminal and myoepithelial progenitors within the mammary ductal system suggested potential cells/sites of origin of these tumors. A diffuse and never previously described CKs/vimentin luminal cell co-expression was detected in the non-neoplastic ducts, indicating a potential bilineage progenitor.

Question 19

Soares et al. 2016. Ki-67 as a Prognostic factor in feline mammary carcinoma: what is the optimal cutoff value?

Ki-67 indices of >14% were detected in 72.9% of the tumors.

Tumors with Ki-67 index>14% were signifcantly associated with large size, poor differentiation, and presence of necrotic areas, ER-negative status, fHER2-negative status, and shorter survival time

Ki-67 expression in primary tumor was strongly and positively correlated with both regional and distant metastasis

Cut-off value of 14% to identify high risk of disease progression

Question 20

Beffagna et al. 2016. A preliminary investigation of the role of transcription co-activators YAP/TAZ of the Hippo signaling pathway in canine and feline mammary tumors

YAP/TAZ transcription co-activators of the Hippo pathway sustain self-renewal and tumor initiation of CSCs

Increased cytoplasmic and nuclear expression of YAP/TAZ was observed in all carcinomas compared to normal tissues

Nuclear expression significantly increased in grade III compared to grade I - YAP/TAZ play a role increased aggressiveness

Question 21

Silva et al. 2017. Histologic evaluation of Ki-67 and cleaves caspace-3 expression in feline mammary carcinoma

Positive immunostaining was observed in all cancer samples for both nuclear Ki-67 and cleaved caspace-3 with mean positive staining percentage of 27.5% and 21.2%

No correlation between ki-67 and cleaved caspace-3 observe in feline mammary carcinoma

High proliferative index in FMC

Question 22

Graf et al. 2016. Swiss feline cancer registry 1965-2008: the influence of sex, breed and age on tumor types and tumor locations

Largest breed difference was found in development of FSA and SCC and tumors in skin/subcutis and mammary gland

Tumors mor frequent in middle-ages to older cats

Question 23

Gray et al. 2017. Dual targeting of EGFR and ERBB2 pathways produces a synergistic effect on cancer cell proliferation and migration in vitro

EGFR/ERBB are frequently disregulated in human cancers

EGFR and ERBB2 combines siRNA targeting produced synergistic effects in feline and canine cell lines similar to reported human cell lines

Dual EGFR and ERBB2 targeting urins TKIs should be evaluated

Question 24

Hassan et al. 2017. Feline Mammary Cancer

Question 25

Arbe et al. 2017. Inhibition of bioenergetic metabolism by the combination of metformin and 2-deoxyglucose highly decreased viability of feline mammary carcinoma cells

MET-antidiabetic drug 2DG-hexokinase inhibitor

Treatments significantly decreased both FMC cell viability up to 80%

AIRB (Her3+ and Ki67<5%) resulted more sensitive to 2DG than AIRATN (HER- and Ki67>15%)

The combination of MET/DG potentiated the effects of individually added drugs on FMC cells

Only MET significantly altered plasma membrane integrity, presented late apoptotic/necrotic cells and increased glucose consumption and lactate concentration

Question 26

Pisamai et al. 2017. Immunohistochemical expression profiles of cell adhesion molecules, MMP and their tissue inhibitors in central and peripheral neoplastic foci in FMC

Question 27

Gemignani et al. 2018. Association of surgical survival approach with complication rate, PFS time, and disease-specific survival time in cats with mammary ACA: 107 cases (1991-2014)

Complications occured in 19.7% cats treated with unilateral mastectomy, 35.7% treated with staged bilateral mastectomy and 40% with single-session bilateral mastectomy

MPFS for cats wuth bilateral mastectomy 542 days vs. 289 for unilateral mastectomy

Significant risk factors for disease progression - unilateral mastectomy, tumor ulceration, LN mets, tumors arising from fourth MG

Significant risk factors for disease specific death - LN mets and regional or distant metastasis

Question 28

Marques et al. 2018. CXCR4 and its ligand CXCL12 display opposite expression profiles in mammary metastatic disease, with the exception of HER-2 overexpressing cells

CXCR4 was more expressed in primary tumors than mets

CXCL12 was highly expressed in mets in liver and lung

Cats with CXCR4 positive primary tumors had significantly lower serum CXCL12 levels than cats with CXCR4- mammay tumors

At mets, HER2 overexpressing tumors presented higher CXCR4 expression than other tumor subtypes and significant difference in overall and disease free survival between the cats with CXCL12 positive and negative tumors

CXCL12- PT were associated with unfavorable prognosis incats with HER2-overexpressing tumors