Automated Analysis of Proliferating Cells Spatial Organisation Predicts Prognosis in Lung Neuroendocrine Neoplasms

Lung neuroendocrine neoplasms (lung NENs) are categorised by morphology, defining a classification sometimes unable to reflect ultimate clinical outcome. Subjectivity and poor reproducibility characterise diagnosis and prognosis assessment of all NENs. Here, we propose a machine learning framework for tumour prognosis assessment based on a quantitative, automated and repeatable evaluation of the spatial distribution of cells immunohistochemically positive for the proliferation marker Ki-67, performed on the entire extent of high-resolution whole slide images. Combining features from the fields of graph theory, fractality analysis, stochastic geometry and information theory, we describe the topology of replicating cells and predict prognosis in a histology-independent way. We demonstrate how our approach outperforms the well-recognised prognostic role of Ki-67 Labelling Index on a multi-centre dataset comprising the most controversial lung NENs. Moreover, we show that our system identifies arrangement patterns in the cells positive for Ki-67 that appear independently of tumour subtyping. Strikingly, the subset of these features whose presence is also independent of the value of the Labelling Index and the density of Ki-67-positive cells prove to be especially relevant in discerning prognostic classes. These findings disclose a possible path for the future of grading and classification of NENs.

Evaluation of Ki67 Expression in Relation to Tumor Stage and Fuhrman Nuclear Grade of Renal Cell Carcinoma and MUC1 Expression in Clear Cell Renal Cell Carcinoma

BACKGROUND Renal cell carcinoma (RCC) is the most common malignant renal tumour in adults. Prognosis of RCC depends on various factors like tumour stage, nuclear grade and histological type. For planning adjuvant therapy, accurate prediction of prognosis is mandatory. In many studies, ki67 and MUC1 has shown to be of prognostic significance and immunohistochemical expression of these two markers plays an important role in determining the prognosis of RCC. The purpose of this study was to evaluate the Ki67 expression in histologically confirmed cases of RCC and MUC1 expression in clear cell renal cell carcinoma, and to correlate them with the stage and Fuhrman nuclear grade of the tumour, in order to determine their role as prognostic markers in RCC. METHODS This study was a retrospective study. A total of 50 specimens of renal cell carcinoma were studied. The specimens were total and partial nephrectomy done in the department of urology for a period of 3 years. Expression of Ki67 and MUC1 in RCC were studied by immunohistochemistry (IHC). Statistical analysis was performed and P value < 0.05 was considered significant. RESULTS Out of 50 RCC studied, Ki67 labelling index ≥ 15 % was found in 35 cases. For MUC1, immunoreactivity of more than 10 % of tumor cell was found in 28/34 of clear cell RCC. In this study, Ki67 labelling index showed statistically significant expression with the stage of tumor and the nuclear grade. MUC1 expression also showed significant correlation with nuclear grade and stage of clear cell RCC. CONCLUSIONS High Ki67 labelling index in renal cell carcinoma is seen to correlate with higher nuclear grade and stage of tumor. High level expression with circumferential staining pattern of MUC1 is seen in high grade tumours with increased risk of metastasis. So MUC1 and Ki67 can be considered as a marker of prognosis of RCC. KEY WORDS Renal Cell Carcinoma, Immunohistochemistry, Ki67, MUC1

Ki67 Protein expression and correlation with the histological grade and pTNM stage of colorectal carcinoma

: Colorectal cancer, is third most frequently encountered malignancy worldwide. Histological grade, stage, and proliferative index act as vital prognostic markers, playing a decisive role in patient care and prognosis. While histopathological examination can determine grade and stage, Ki67 protein expression by immunohistochemistry represents the proliferative capacity of the malignant cell population. : This Study was conducted to immunohistochemically analyze expression of Ki67 protein and its change with respect to different grades and stage of colorectal adenocarcinoma. : A total of 66 histologically diagnosed cases of colorectal carcinoma underwent histopathological examination followed by immunohistochemistry for Ki67 protein. : A statistically significant (p=0.04) correlation was obtained between Ki67 labelling index and histopathological grade, with higher values of Ki67 labelling index in poorly differentiated carcinomas (43.2±1.7). The Ki67 labelling index value was lowest in stage IV disease (11.4±2.4) with metastatic burden, with higher values in lower stage diseases, however this correlation was not found to be significant statistically (p=0.07). : The rate of cell division and proliferation measured by Ki-67 antibody is related to histological grade of the malignancy, demonstrating higher mitotic activity with loss of differentiation and anaplasia. Stage IV disease have lower mitotic activity, thus may be less amenable to cytostatic chemotherapy drugs, and require multimodality treatment with addition of radiotherapy and other drug regimes.

Development of a Standardized Semantic Feature-Based Reporting Proforma for Intraoperative Ultrasound Findings in Brain Tumors and Application in High-Grade Gliomas – A Preliminary Study

Purpose A semantic feature-based reporting proforma for intraoperative ultrasound findings in brain tumors was devised to standardize reporting. It was applied as a pilot study on a cohort of histologically confirmed high-grade supratentorial gliomas (Grade 3 and 4) for internal validation. Materials and Methods This intraoperative semantic ultrasound proforma was used to evaluate 3D ultrasound volumes using Radiant DICOM software by 3 surgeons. The ultrasound semantic features were correlated with histological features like tumor grade, IDH status, and MIB index. Results 68 patients were analyzed using the semantic proforma. Irregular crenated was the most common margin (63.2%) and lesions were heterogeneously hyperechoic (95.6%). Necrosis was commonly seen and noted as single (67.6%) or multiple (13.2%) in over 80% cases. A separate perilesional zone, which was predominantly hyperechoic in 41.8% and both hypo and hyperechoic in 12.7%, could be identified in 54.5% of cases. Grade 4 tumors were more likely to have an irregular crenated margin (71.2%) with a single large area of necrosis, while Grade 3 tumors were likely to have smooth (31.3%) or non-characterizable margins (31.2%) with no or multiple areas of necrosis. IDH-negative tumors were more likely to have a single large focus of necrosis. Among the GBMs (52 cases), MIB labelling index of>15% was associated with poorly delineated, uncharacterizable margins, when compared with MIB labelling index<15% (23.5 vs. 0%), (p=0.046). Conclusion A detailed semantic proforma was developed for brain tumors and was internally validated. A few ultrasound sematic features were identified correlating with histological features in high-grade gliomas. It will require further external validation for refinement and acceptability.

Challenges in High-grade Neuroendocrine Neoplasms and Mixed Neuroendocrine/Non-neuroendocrine Neoplasms

The growth in knowledge of the pathogenesis, molecular background, and immunohistochemical profile of neuroendocrine neoplasms (NENs) has led not only to an increased awareness of these diseases but also to several changes of the nomenclature. In particular, the concept and terminology of high-grade (grade 3) NENs and mixed neoplasms have changed considerably over the last 20 years, creating some confusion among pathologists and clinicians. The aim of this review is to elucidate the diagnostic criteria, including the most important differential diagnoses of high-grade NENs and mixed neuroendocrine/non-neuroendocrine neoplasms (MiNENs). The role of the Ki67 labelling index and morphology, used to define grade 3 NENs of the digestive system and lungs, is also discussed. The evolution of the concepts and terminology of MiNENs is revised, including the most important differential diagnoses.

Author Correction: Validation of Clinical Treatment Score post-5 years (CTS5) risk stratification in premenopausal breast cancer patients and Ki-67 labelling index

An amendment to this paper has been published and can be accessed via a link at the top of the paper.