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Canine mast cell tumors: utility of stereologic tools in cytology.

ABSTRACT: Quantitative morphologic parameters assessed in cytologic samples of canine cutaneous mast cell tumors (ccMCTs) may assist with surgical planning and prognostication. Robust cutoffs can be defined, with high reproducibility, for parameters such as the nuclear area (NA). The NA may be determined by morphometry (image analysis, NAI) or by stereology, such as the 2D-nucleator method (NAN); stereologic techniques have not been applied to cytologic specimens of ccMCT, to our knowledge. We retrospectively selected routine cytology smears from 51 ccMCT cases and screened them to determine the percentage of neoplastic mast cells with indistinct nuclear borders; this was repeated after the slides were restained with H&E. The NAI and the NAN were estimated in 100 mast cells per animal in H&E-stained slides. All nuclei were visible in H&E smears, and unbiased quantification was feasible. The NAN was similar to NAI, but less time-consuming. Both the NAN and NAI determined by cytology differed in histologic low- and high-grade ccMCTs, and in histologic grade I plus II versus grade III ccMCTs. Stereologic parameters such as the NAN could be considered as complementary techniques for the cytologic evaluation of ccMCTs.

SUBMITTER: Marcos R

PROVIDER: S-EPMC8921802 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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Canine mast cell tumors: utility of stereologic tools in cytology.

Marcos Ricardo R Almeida João J Marques Joana J Moreira Raquel R Dias-Pereira Patrícia P Correia-Gomes Carla C Santos Marta M

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 20211111 2

Quantitative morphologic parameters assessed in cytologic samples of canine cutaneous mast cell tumors (ccMCTs) may assist with surgical planning and prognostication. Robust cutoffs can be defined, with high reproducibility, for parameters such as the nuclear area (NA). The NA may be determined by morphometry (image analysis, NAI) or by stereology, such as the 2D-nucleator method (NAN); stereologic techniques have not been applied to cytologic specimens of ccMCT, to our knowledge. We retrospecti ...[more]

PMID: 34763591

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Project description:BackgroundYKL-40, a secretory glycoprotein, is involved in tumor cell proliferation, metastasis, and angiogenesis in human cancers. Its overexpression has been correlated with unfavorable prognosis in many human cancers. In veterinary medicine, elevated YKL-40 levels in the serum of canine cutaneous mast cell tumors (cMCTs) were observed in our previous study. However, the expression pattern of YKL-40 in canine cMCT tissues, along with its association with clinical and pathological features, is still unknown. This study aims to retrospectively investigate the expression level of YKL-40 in the tissues of canine cMCTs and its correlation with clinical features, pathological characteristics, and clinical outcomes. Forty formalin-fixed paraffin-embedded cMCT tissues collected from forty dogs were diagnosed as low-grade (n = 20) or high-grade s(n = 20) MCT according to the Kiupel grading system. The expression level of YKL-40 in cMCT tissues was investigated using immunohistochemical staining and immunoreactivity score (IRS).ResultsYKL-40 was expressed in all cMCTs at different levels, with significantly stronger expression in low-grade cMCTs compared to high-grade cMCTs. The expression level was also associated with tumor diameter, histological grade, mitotic counts, vessel density, and survival of cMCTs. The overall survival of cMCT dogs showed significant differences (p < 0.01) among mild (n = 15, MST 219 days), moderate (n = 19, MST not reached), and high (n = 6, MST not reached) YKL-40 expression groups. Among low-grade cMCTs, overall survival was significantly different between mild YKL-40 expression (MST 319 days) and moderate to high YKL-40 (MST not reached) expression (p < 0.01). In high-grade cMCTs, overall survival was not correlated with YKL-40 expression (p = 0.6589).ConclusionsThis study found that the YKL-40 expression level was significantly stronger in low-grade than in high-grade canine cutaneous mast cell tumors and was associated with various clinical and pathological features. Stronger YKL-40 expression level correlated with longer survival time, especially in low-grade cMCTs. Therefore, YKL-40 could serve as a prognostic marker for cMCTs.

Project description:BackgroundCanine cutaneous mast cell tumors (MCTs) are a common, yet clinically challenging tumor type given their variable biological behavior. Although patients with low grade MCTs can often be effectively managed with surgery alone, most dogs with high grade MCTs succumb to their disease despite multimodal therapy. An improved understanding of the immune tumor microenvironment (TME) may help identify novel prognostic and therapeutic targets.MethodsIn this study, we interrogated the immune transcriptional profiles of the TME in low and high grade MCTs, and quantified intratumoral T cells. Twelve client-owned dogs with MCTs (6 Kiupel low grade with clinically benign behavior and 6 Kiupel high grade with clinically aggressive behavior) that underwent curative-intent surgery were selected. Tumor grade was confirmed by a single veterinary pathologist. RNA was extracted from all tumors followed by immune transcriptional profiling utilizing the NanoString Canine IO panel and analysis using the ROSALIND platform. T cell density was determined by immunohistochemical staining for CD3 and quantified using ImageScope software (Leica Biosystems) following digital slide capture. Lymphocytic infiltrate was further characterized in the TME of one high grade MCT using co-immunofluorescence.ResultsImmune transcriptional profiling identified 9 differentially expressed genes between low and high grade MCTs (p-adj < 0.05). Programmed cell death protein 1 (PDCD1) and inducible T-cell costimulator ligand (ICOSLG) gene expression were significantly higher in a subset of high grade MCTs. ICOSLG expression positively correlated with T cell score (rs = 0.6434, p = 0.0278). Although the T cell density was not significantly different between low (mean of 76.42 CD3 + /mm2, SD 12 CD3 + /mm2) and high grade MCTs (mean of 129.1 CD3 + /mm2, SD 96.06 CD3 + /mm2), greater variation of T cell densities was observed across high grade MCTs compared to low grade (p = 0.0059). Immunofluorescence of one high grade MCT with marked T cell infiltration revealed organized aggregates of T and B cells consistent with tertiary lymphoid structures (TLS).ConclusionsOur data revealed significant differences in the immune TME of low and high grade MCTs and provides rationale to further investigate potential prognostic and therapeutic roles of immune checkpoints in canine MCTs.Supplementary informationThe online version contains supplementary material available at 10.1186/s44356-025-00020-9.

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Canine mast cell tumors: utility of stereologic tools in cytology.

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Canine mast cell tumors: utility of stereologic tools in cytology.

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