Diagnostic value of serum survivin, Ki-67 and thymidine kinase in dogs with nasal cavity disease

Abstract

Background: The most common nasal cavity disease (ND) in dogs is the malignant nasal cavity tumor. Prognosis and survival after radiation therapy are reported to correlate with tumor size, and therefore indirectly with the time to diagnosis. Diagnosis of a nasal tumor requires imaging procedures under anesthesia. Thus, diagnostic serum markers are urgently needed for early detection and for therapeutic monitoring.

Materials and methods: This prospective, blinded study included dogs with nasal discharge that completed a comprehensive diagnostic workup for ND. Dogs were evaluated by blood testing and whole-body CT and those with concomitant diseases or with steroid pre-treatment were excluded. Serum survivin, Ki-67, and thymidine kinase 1 (TK1) concentrations were determined, and the survivin-lymphocyte ratio (SLR) calculated. Results were compared between groups of dogs with different NDs and to ten healthy controls.

Results: A total of 55 dogs were included, consisting of 25 with malignant ND (12 sarcomas, 13 carcinomas) and 30 with benign ND (7 benign tumors, 13 dogs with idiopathic rhinitis (IR), 10 others including dogs with dental diseases and sinonasal aspergillosis). Survivin and SLR were significantly increased in dogs with malignant ND and in subgroup comparison in sarcomas compared to controls. In addition, the SLR was significantly increased in carcinomas and IR compared to controls. In dogs with IR, no differences were observed in survivin concentrations or SLR based on microbiological or histopathological findings. Survivin concentrations or SLR in dogs with nasal tumors were not significantly different between T-categories. No significant differences were detected in TK1 concentrations among the groups, nor in Ki-67, except for significantly lower Ki-67 concentrations in benign tumors compared to carcinomas and the group others including, e.g., dental diseases.

Conclusion: Although not diagnostic for ND, increased survivin serum concentrations or SLR can be detected in dogs with malignant nasal tumors and IR. In malignant nasal tumors, survivin and SLR did not correlate with tumor size and therefore may be useful in the detection of even small nasal tumors. Therefore, in dogs with nasal tumors and IR, survivin and SLR could serve as a target for disease monitoring or as therapeutic target.

Keywords: blood marker; carcinoma; lymphoplasmacytic rhinitis; nasal discharge dog; nasal neoplasia; sarcoma.

Figure 1

Serum survivin, survivin-lymphocyte ratio (SLR) and serum Ki-67 in dogs with chronic nasal discharge. (A) Compared to controls (CG), survivin was significantly increased in dogs with malignant ND (p = 0.028) and (B) in subgroup comparison significantly increased in dogs with sarcomas (S; p = 0.009). Additionally, survivin concentrations of dogs with S were significantly increased compared to dogs of the group others (O; p = 0.003). (C) SLR in malignant ND was significantly increased compared to controls (p = 0.002), (D) as well as in subgroup comparison in dogs with S (p = 0.001), carcinomas (C; p = 0.030) and idiopathic rhinitis (IR; p = 0.014). Additionally, the SLR in S was significantly increased compared to that of O (p = 0.001). (E, F) Only Ki-67 concentrations in dogs with C (p = 0.040) and O (p = 0.026) were significantly increased compared to concentrations in benign tumors. Data are shown in box and whisker plots. Upper and lower boxes represent the 25th and 75th percentiles (lower whiskers = minimum, upper whiskers = maximum values) and the line represents the median. In the graphs, the in the present study calculated cut-off values (88.5 pg/mL for survivin and 44.55 for SLR) are marked by a dotted horizontal line.

Figure 2

Evaluation of the influence of various factors on survivin serum concentrations in dogs with idiopathic rhinitis (IR) or malignant nasal tumors. (A,B) show the survivin concentrations or SLR in the different groups in relation to a positive or negative result of the microbiological examination (ME) of a nasal swab. No significant differences in sense of influence of bacterial colonization on marker concentrations were detected. n.a. = ME was not available in 4 dogs with nasal tumor due to financial constraints of the owners (n = 3) or because it was performed at the regular veterinarian (n = 1) before presentation. (C,D) Additionally, no statistically significant difference in survivin concentrations or SLR was detected in the different T-categories [T-category according to Adams et al. (14)] with T4 indicating cribriform plate lysis. Due to the low number of dogs with T1-T3 category in different tumor groups, these categories were grouped together (6 carcinomas, 8 sarcomas, and 7 benign tumors) against T4 (7 carcinomas and 3 sarcomas). (E,F) In dogs with IR, the result of the histopathological examination in regard of the predominant inflammatory cell type is shown. No statistically significant differences in serum survivin concentrations or SLR were detected. C = carcinoma, S = sarcoma, BT = benign tumor, IR = idiopathic rhinitis, O = others. In the graphs, the in the present study calculated cut-off values (88.5 pg/mL for survivin and 44.55 for SLR) are marked by a dotted horizontal line.

Figure 3

Survivin serum concentrations and survivin-lymphocyte ratio (SLR) in dogs detected in control examinations (EX). The dogs had not received any steroids between examinations. In panel (A–D) survivin concentrations and SLR 6–8 weeks after diagnosis and palliative endoscopic interventional cytoreduction through the nasal entrance are illustrated in malignant (A,B) and benign (C,D) nasal tumors. In some dogs, re-examinations were performed on owner’s request. In panel (A,B) 7 sarcomas and 3 carcinomas were evaluated (with only 4 concentrations in EX2 and 2 concentrations in EX3 available). In panel (C,D) values of 5 benign tumors are illustrated at two different time points, 6–8 weeks apart. Despite tumor volume reduction, survivin concentrations and SLR values did not show a statistically significant decrease. (C,D) However, in dogs with benign tumors, concentrations fell below the cut-off values set in the present study. In the graphs illustrating survivin concentrations and SLR respectively, the previously calculated cut-off values (88.5 pg/mL for survivin and 44.55 for SLR) are marked by a dotted horizontal line. Data are shown in box and whisker plots. Upper and lower boxes represent the 25th and 75th percentiles (lower whiskers = minimum, upper whiskers = maximum values) and the line represents the median.

Figure 4

Comparison of (A) survivin concentrations and (B) survivin-lymphocyte ratio (SLR) in included dogs of the groups others, the control group and dogs with idiopathic rhinitis (IR) versus excluded IR dogs. If evaluating only dogs without a detectable nasal mass, including the control dogs, survivin concentrations and SLR were significantly increased in dogs with IR, in contrast to the other group with dental diseases or sinonasal aspergillosis (Kruskal-Wallis test: survivin p = 0.049 and SLR p = 0.029). In excluded dogs, survivin and SLR were suggested to be decreased due to steroid pre-treatment or pneumonia, as in human medicine both variables were reported to negatively influence survivin values. In the graphs, the in the present study calculated cut-off values (88.5 pg/mL for survivin and 44.55 for SLR) are marked by a dotted horizontal line. Data are shown in box and whisker plots. Upper and lower boxes represent the 25th and 75th percentiles (lower whiskers = minimum, upper whiskers = maximum values) and the line represents the median.