High mitotic index and high nuclear grade are part of representative hallmarks of adrenocortical cancer (ACC), but the analysis of these parameters is known to be operator-dependent. A characteristic neutrophil/T-lymphocytes infiltrate ratio has been often implicated in carcinogenesis, progression and clinical outcome of several cancer types. However, its role in adrenal cortical tumors is unclear. Aim: to assess by computerized morphometry morphological features, vascular and inflammatory pattern in adrenocortical adenomas (ACAs) and carcinomas. Methods: A single Institution series of 11 ACAs and 18 ACCs samples was analyzed using a Kontron-Zeiss KS400 image analyzer. Four consecutive sections 4 µm thick were obtained with a total of 250–300 HPF examined for each case. Immunohistochemistry for Ki67 and CD8/CD15 was obtained to assess proliferation index and inflammatory infiltrate respectively. To minimize subjectivity, particularly relevant when quantitative results are expected, we generated a morphometric model based on analysis of volume fractions occupied by Ki67 positive and negative cells (nuclei, cytoplasm) and inflammatory compartments (CD15+ granulocytes, CD8+ lymphocytes). Lastly, the assessment of Ki-67 by computerized morphometry was compared with pathologist’s evaluation. Results: volume fraction of Ki-67+ cells was higher in ACCs (ACC .11951, ACA .06637; p<0.001); nuclei volume fraction resulted higher in ACCs, in both Ki-67- (ACC .11951, ACA .06637; p<0.001) and Ki-67+ cells (ACC .01293, ACA .00104; p<0.001). Nuclear/cytoplasmic ratio was higher in ACCs, both Ki-67- (ACC .20535, ACA .09260; p<0.001) and Ki-67+ cells (ACC .66141, ACA .27281; p<0.001). Volume fractions of CD15+ (ACC .00312, ACA .00098; p<0.001) and CD8+ cells (ACC .00731, ACA .00356; p<0.05) were also significantly higher in ACCs. Moreover, when comparing morphometric analysis of Ki67+ cells to pathologist’s scores, the data of the point grid analysis revealed significantly lower values compared to conventional histopathology. Conclusions: Our computerized morphometric model is simple, repeatable (lacking observer bias) and flexible, as it can be upgraded to include newly described histological or immunohistochemical features. This method could be integrated into a classification tool to complement conventional histological analysis to achieve quantification of morphological characteristics and histological biomarkers of adrenocortical tumors. In our experience, nuclear/cytoplasmic ratio differs mostly between ACCs and ACAs, both in Ki-67+ and Ki-67- cells. We speculate that neutrophils may play a role in ACC milieu and that the quantitative assessment of inflammatory infiltrate may find a place in the diagnostic algorithm of adrenal benign and malignant tumors.

Quantitative Assessment By Computerized Morphometry of Ki-67 and Intratumoral Inflammatory Infiltrate in Benign and Malignant Adrenocortical Tumors / P. Dalino Ciaramella, M. Vertemati, D. Petrella, E. Grossrubatscher, B. Zampetti, M. Boniardi, R. Pugliese, P. Loli. - In: ENDOCRINE REVIEWS. - ISSN 1945-7189. - 35:3 suppl.(2014 Jun). (Intervento presentato al 96. convegno Endocrine Society's Annual Meeting and Expo tenutosi a Chicago nel 2014).

Quantitative Assessment By Computerized Morphometry of Ki-67 and Intratumoral Inflammatory Infiltrate in Benign and Malignant Adrenocortical Tumors

P. Dalino Ciaramella
Primo
;
M. Vertemati
Secondo
;
2014

Abstract

High mitotic index and high nuclear grade are part of representative hallmarks of adrenocortical cancer (ACC), but the analysis of these parameters is known to be operator-dependent. A characteristic neutrophil/T-lymphocytes infiltrate ratio has been often implicated in carcinogenesis, progression and clinical outcome of several cancer types. However, its role in adrenal cortical tumors is unclear. Aim: to assess by computerized morphometry morphological features, vascular and inflammatory pattern in adrenocortical adenomas (ACAs) and carcinomas. Methods: A single Institution series of 11 ACAs and 18 ACCs samples was analyzed using a Kontron-Zeiss KS400 image analyzer. Four consecutive sections 4 µm thick were obtained with a total of 250–300 HPF examined for each case. Immunohistochemistry for Ki67 and CD8/CD15 was obtained to assess proliferation index and inflammatory infiltrate respectively. To minimize subjectivity, particularly relevant when quantitative results are expected, we generated a morphometric model based on analysis of volume fractions occupied by Ki67 positive and negative cells (nuclei, cytoplasm) and inflammatory compartments (CD15+ granulocytes, CD8+ lymphocytes). Lastly, the assessment of Ki-67 by computerized morphometry was compared with pathologist’s evaluation. Results: volume fraction of Ki-67+ cells was higher in ACCs (ACC .11951, ACA .06637; p<0.001); nuclei volume fraction resulted higher in ACCs, in both Ki-67- (ACC .11951, ACA .06637; p<0.001) and Ki-67+ cells (ACC .01293, ACA .00104; p<0.001). Nuclear/cytoplasmic ratio was higher in ACCs, both Ki-67- (ACC .20535, ACA .09260; p<0.001) and Ki-67+ cells (ACC .66141, ACA .27281; p<0.001). Volume fractions of CD15+ (ACC .00312, ACA .00098; p<0.001) and CD8+ cells (ACC .00731, ACA .00356; p<0.05) were also significantly higher in ACCs. Moreover, when comparing morphometric analysis of Ki67+ cells to pathologist’s scores, the data of the point grid analysis revealed significantly lower values compared to conventional histopathology. Conclusions: Our computerized morphometric model is simple, repeatable (lacking observer bias) and flexible, as it can be upgraded to include newly described histological or immunohistochemical features. This method could be integrated into a classification tool to complement conventional histological analysis to achieve quantification of morphological characteristics and histological biomarkers of adrenocortical tumors. In our experience, nuclear/cytoplasmic ratio differs mostly between ACCs and ACAs, both in Ki-67+ and Ki-67- cells. We speculate that neutrophils may play a role in ACC milieu and that the quantitative assessment of inflammatory infiltrate may find a place in the diagnostic algorithm of adrenal benign and malignant tumors.
computerized morphometry ; adrenocortical carcinomas ; Ki67
Settore BIO/16 - Anatomia Umana
Settore MED/08 - Anatomia Patologica
Settore MED/13 - Endocrinologia
giu-2014
Endocrine Society
https://endo.confex.com/endo/2014endo/webprogram/Paper15214.html
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/244011
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