Ki-67 Score Research Articles

Comparison of Different Molecular Subtypes with 14% Ki-67 Cut-off Threshold in Breast Cancer Patients of Pakistan- An Indication of Poor Prognosis

Ki-67 is a proliferation marker that is used not only to categorize patients in luminal A and B subtypes of breast cancers, but also to determine the aggressiveness of the disease in triple negative and human epidermal growth factor 2 (HER2) over expressed molecular subtypes. The present study was designed to evaluate the role of Ki-67 with cut off value of 14% in molecular subgroups and its association with patient prognosis. Immunostaining was performed on histopathologically confirmed sections (n = 278) to assess expression of Ki-67, estrogen receptor (ER), progesterone receptor (PR) and HER2. Immunoreactivity of molecules was recorded as percentage scoring. Adopting a cut off value of 14%, Ki-67 was high in 88%of the cases included in the study. High Ki-67 was significantly associated with pathological parameters including histological grade, advanced stage and nodal/distant metastasis. Immunoexpression of ER, PR and HER2 also showed strong correlation with high expression of Ki-67. Based on the St. Gallen classification, the cases were categorized into luminal A (10%) and luminal B (51%), triple negative (20%) and HER2 enriched (18%). Ki-67 index was also significantly high in 98% of HER2 enriched and 95% of TNBC patients. Interestingly, Ki-67 score with cut off value of 14% proved to be significant in deciphering prognosis in luminal patients. Moreover, high expression of Ki-67 also proved to be a marker of poor prognosis, especially in triple negative patients. We suggest that utilization of IHC4 status i.e. ER, PR, HER2 and Ki-67 along with pathological findings and molecular subtyping can considerably affect clinical as well as therapeutic decisions.

Independent Clinical Validation of the Automated Ki67 Scoring Guideline from the International Ki67 in Breast Cancer Working Group.

Ki67 is an important biomarker with prognostic and potential predictive value in breast cancer. However, the lack of standardization hinders its clinical applicability. In this study, we aimed to investigate the reproducibility among pathologists following the guidelines of the International Ki67 in Breast Cancer Working Group (IKWG) for Ki67 scoring and to evaluate the prognostic potential of this platform in an independent cohort. Four algorithms were independently built by four pathologists based on our study cohort using an open-source digital image analysis (DIA) platform (QuPath) following the detailed guideline of the IKWG. The algorithms were applied on an ER+ breast cancer study cohort of 157 patients with 15 years of follow-up. The reference Ki67 score was obtained by a DIA algorithm trained on a subset of the study cohort. Intraclass correlation coefficient (ICC) was used to measure reproducibility. High interobserver reliability was reached with an ICC of 0.938 (CI: 0.920–0.952) among the algorithms and the reference standard. Comparing each machine-read score against relapse-free survival, the hazard ratios were similar (2.593–4.165) and showed independent prognostic potential (p ≤ 0.018, for all comparisons). In conclusion, we demonstrate high reproducibility and independent prognostic potential using the IKWG DIA instructions to score Ki67 in breast cancer. A prospective study is needed to assess the clinical utility of the IKWG DIA Ki67 instructions.

IHC-based Ki67 as response biomarker to tamoxifen in breast cancer window trials enrolling premenopausal women

Window studies are gaining traction to assess (molecular) changes in short timeframes. Decreased tumor cell positivity for the proliferation marker Ki67 is often used as a proxy for treatment response. Immunohistochemistry (IHC)-based Ki67 on tissue from neo-adjuvant trials was previously reported to be predictive for long-term response to endocrine therapy for breast cancer in postmenopausal women, but none of these trials enrolled premenopausal women. Nonetheless, the marker is being used on this subpopulation. We compared pathologist assessed IHC-based Ki67 in samples from pre- and postmenopausal women in a neo-adjuvant, endocrine therapy focused trial (NCT00738777), randomized between tamoxifen, anastrozole, or fulvestrant. These results were compared with (1) IHC-based Ki67 scoring by AI, (2) mitotic figures, (3) mRNA-based Ki67, (4) five independent gene expression signatures capturing proliferation, and (5) blood levels for tamoxifen and its metabolites as well as estradiol. Upon tamoxifen, IHC-based Ki67 levels were decreased in both pre- and postmenopausal breast cancer patients, which was confirmed using mRNA-based cell proliferation markers. The magnitude of decrease of Ki67 IHC was smaller in pre- versus postmenopausal women. We found a direct relationship between post-treatment estradiol levels and the magnitude of the Ki67 decrease in tumors. These data suggest IHC-based Ki67 may be an appropriate biomarker for tamoxifen response in premenopausal breast cancer patients, but anti-proliferative effect size depends on estradiol levels.

CDK14 expression is elevated in patients with non-small cell lung cancer and correlated with poor prognosis.

ObjectiveTo investigate the clinical significance of cyclin-dependent kinase 14 (CDK14) expression in patients with non-small cell lung cancer (NSCLC).MethodsThe present prospective observational study included 193 patients diagnosed with NSCLC between January 2010 and December 2014. NSCLC tumor tissues and paired paracancerous normal tissues were obtained from all patients. CDK14, thyroid transcription factor 1 (TTF-1), cytokeratin 5/6 (CK5/6), and Ki67 expression was measured via immunohistochemistry (IHC)ResultsCDK14 staining was strong (>3) in 129 patients (66.49%) and weak (≤3) in 64 patients (33.16%). The mean IHC scores were markedly higher in tumor tissues than in paracancerous tissues. Pearson’s analysis demonstrated that the IHC scores of CDK14 expression were positively correlated with TTF-1, CK5/6, and Ki67 scores. Kaplan–Meier analysis illustrated that 5-year overall survival was markedly longer in patients with weak CDK14 staining. TNM stage, pleural invasion, lymph node metastasis, CDK14 expression, and Ki67 expression were risk factors for 5-year overall survival in patients with NSCLC.ConclusionCDK14 overexpression portended poor outcomes in patients with NSCLC, and CDK14 expression was correlated with TTF-1, CK5/5, and Ki67 expression.

The Oncotype DX recurrence score impact on the management of ER-positive, HER2-negative, node-negative breast cancer

Introduction: Oncotype DX recurrence score is used to categorize estrogen-receptor-positive, human epidermal growth factor receptor-2 negative, lymph-node negative early breast cancer in high- or low-recurrence risk groups. It has a guiding significance for whether post-operative chemotherapy or only hormonal manipulation is selected as an adjuvant treatment. Aims: Assess the impact of the Oncotype DX recurrence score on adjuvant chemotherapy-related management decision-making in the cases of ER+ve, HER2–ve, LN–ve early breast cancer in our local unit. Material and methods: A cohort of 76 patients with early breast cancer were included, two had bilateral disease. All were operated for estrogen-receptor-positive, human epidermal growth factor receptor-2 negative, lymph-node negative early breast cancer. Tumor grade, Ki67 proliferative index, PREDICT and Oncotype DX recurrence score results were obtained in addition to the offered treatment information for each case. Results: After the primary tumor surgery and an Oncotype DX recurrence score assessment, 18 patients (24%) were eligible for adjuvant chemotherapy; out of them, 10 patients (56%) had a chemotherapy absolute survival benefit of > 15% at 10 years, where 5 patients (30%) had a chemotherapy relative survival benefit of ~ 6.5%, 3 patients (17%) had a chemotherapy relative survival benefit of ~1.6%. In this cohort, 10 patients (13%) had a low Oncotype DX recurrence score; however, they received adjuvant chemotherapy based on other clinico-biological parameters. The other 48 patients (63%) with a low recurrence risk were spared potential adverse events related to the systemic therapy. Based on the menopausal status, every patient had received suitable hormonal manipulation therapy. The data also revealed the absence of a relationship between the Ki67 proliferative index and the Oncotype DX recurrence score (p = 0.06); moreover, the size of the tumor did not correlate with the Oncotype DX recurrence score (p = 0.5). Conclusion: The Oncotype DX recurrence score provides a credible prediction of distant disease recurrence risk in early breast cancer; however, it does not correlate with other prognostic markers, such as the Ki67 proliferative index as well as the tumor size. In this cohort, the use of the Oncotype DX recurrence score led to a 24% rate of treatment recommendations in the direction of adjuvant chemotherapy in addition to anti-hormonal therapy for estrogen-receptor-positive, human epidermal growth factor receptor-2 negative, lymph-node negative early breast cancer.

Improving Prognosis of Surrogate Assay for Breast Cancer Patients by Absolute Quantitation of Ki67 Protein Levels Using Quantitative Dot Blot (QDB) Method

BackgroundImmunohistochemistry (IHC)-based surrogate assay is the prevailing method in daily clinical practice to determine the necessity of chemotherapy for Luminal-like breast cancer patients worldwide. It relies on Ki67 scores to separate Luminal A-like from Luminal B-like breast cancer subtypes. Yet, IHC-based Ki67 assessment is known to be plagued with subjectivity and inconsistency to undermine the performance of the surrogate assay. A novel method needs to be explored to improve the clinical utility of Ki67 in daily clinical practice.Materials and MethodsThe Ki67 protein levels in a cohort of 253 specimens were assessed with IHC and quantitative dot blot (QDB) methods, respectively, and used to assign these specimens into Luminal A-like and Luminal B-like subtypes accordingly. Their performances were compared with the Kaplan–Meier, univariate, and multivariate survival analyses of the overall survival (OS) of Luminal-like patients.ResultsThe surrogate assay based on absolutely quantitated Ki67 levels (cutoff at 2.31 nmol/g) subtyped the Luminal-like patients more effectively than that based on Ki67 scores (cutoff at 14%) (Log rank test, p = 0.00052 vs. p = 0.031). It is also correlated better with OS in multivariate survival analysis [hazard ratio (HR) at 6.89 (95% CI: 2.66–17.84, p = 0.0001) vs. 2.14 (95% CI: 0.89–5.11, p = 0.087)].ConclusionsOur study showed that the performance of the surrogate assay may be improved significantly by measuring Ki67 levels absolutely, quantitatively, and objectively using the QDB method.

Bortezomib enhances the anti-cancer effect of the novel Bruton's tyrosine kinase inhibitor (BGB-3111) in mantle cell lymphoma expressing BTK.

BGB-3111, a novel Bruton’s tyrosine kinase (BTK) inhibitor, shows promising anti-cancer effects in chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), and Waldenstrom macroglobulinemia (WM). This study aimed to investigate the anti-cancer effects of BGB-3111 combined with bortezomib (BTZ) against the BTK-expressing MCL. We found that BTK, which was overexpressed in 59.4% of patients with MCL, was mainly characterized by high Ki67 and elevated MIPI scores. BGB-3111 strongly inhibited cell proliferation, induced cell cycle arrest in the G1/G0-phase, and promoted cell apoptosis in the MCL cells expressing BTK. BGB-3111 provides better safety than another BTK inhibitor, ibrutinib as ibrutinib inhibits the inducible T-cell kinase (ITK) as an off-target effect but BGB-3111 does not inhibit ITK. Low doses of BTZ enhanced the anti-cancer effect induced by the low dose of BGB-3111 by downregulating the expression levels of PARP and Bcl-2 and increasing the expression levels of cleaved PARP and cleaved caspase-9. In addition, low doses of BGB-3111, but not of BTZ, inhibited BTK phosphorylation. However, low-doses of BTZ strengthened the anti-cancer effect induced by the low-doses of BGB-3111 via synergistically suppressing the IκBα and P65 phosphorylation. Taken together, our findings validate that BGB-3111 is a novel and effective BTK inhibitor for MCL-expressing BTK. Hence, it can be harnessed as a potential therapeutic strategy through a combinatorial treatment comprising low-dose BGB-3111 and low-dose BTZ to gain strong anti-cancer effects and better safety for MCL patients.

Quantification of Global DNA Methylation in Canine Melanotic and Amelanotic Oral Mucosal Melanomas and Peripheral Blood Leukocytes From the Same Patients With OMM: First Study.

Oral mucosal melanomas (OMMs) are aggressive and resistant cancers of high importance in veterinary oncology. Amelanotic OMM produces comparatively less melanin and is considered to be more aggressive than melanotic OMM. Global DNA methylation profiles with hypomethylated or hypermethylated patterns have both been associated with aggressive neoplasms; however, global DNA hypomethylation seems to correlate to higher aggressiveness. Accordingly, global DNA methylation in peripheral blood leukocytes has been investigated to understand the role of systemic or environmental factors in cancer development. This study aimed to quantify global DNA methylation in canine melanotic and amelanotic OMM samples and in the peripheral blood leukocytes of the same dogs. Tumor tissue samples were collected from 38 dogs, of which 19 were melanotic and 19 were amelanotic OMM. These were submitted to immunohistochemistry (IHC) with anti-5-methylcytosine (5mC) and anti-Ki67 primary antibodies. Ki67- and 5mC-positive nuclei were manually scored with the help of an image analysis system. Peripheral blood samples were collected from 18 among the 38 OMM-bearing dogs and from 7 additional healthy control dogs. Peripheral blood leukocytes were isolated from the 25 dogs, and DNA was extracted and analyzed by high-performance liquid chromatography (HPLC) for global DNA methylation. The pattern of global DNA methylation in both canine melanotic and amelanotic OMM indicated higher percentages of weakly or negatively stained nuclei in most of the OMM cells, presuming predominant global DNA hypomethylation. In addition, Ki67 counts in amelanotic OMM were significantly higher than those in melanotic OMM (p < 0.001). Global DNA methylation different immunostaining patterns (strong, weak or negative) correlated with Ki67 scores. Global DNA methylation in circulating leukocytes did not differ between the 9 melanotic and 9 amelanotic OMM or between the 18 OMM-bearing dogs and the 7 healthy dogs. This study provides new information on canine melanotic and amelanotic OMM based on global DNA methylation and cell proliferation.

Prognostic significance of Ki67 during neoadjuvant chemotherapy in primary unresectable ovarian cancer.

The purpose of this study was to investigate whether the Ki67 values were associated with survival for predicting prognosis in patients with advanced ovarian cancer receiving neoadjuvant chemotherapy (NACT). Among 17 patients treated with NACT, 13 patients were available for tissue samples from matched pre- and post-therapy tissues. Ki67 scores were transformed to a logarithmic scale for the statistical analyses. The optimal cutoff values of the log-phase Ki67 were assessed by receiver operating characteristic (ROC) analysis. Kaplan-Meier analysis, the log-rank test, and Cox regression analysis were carried out to analyze survival. The Ki67-decrease and post-NACT Ki67 were the independent factors associated with relapse-free survival (RFS) (p < 0.001 and p=0.003). No association was observed on overall survival. The optimal cutoff values for the Ki67-decrease and the post-NACT Ki67 were 6.67% and 5.46 based on ROC where the area under ROC curves (AUC) were 1.00 (p < 0.001) with the 100% sensitivity and specificity. The median RFS was 537 days in patients showing Ki67-decrease >6.66% or post-NACT Ki67 level <5.46, while it was 224 days in those with Ki67 decrease ≤6.66% or post-NACT Ki67 level ≥5.46 (p=0.001). The Ki67-decrease and the lower post-NACT Ki67 are independent factors associated with favorable RFS, indicating that they could be precise biomarker candidates for prognosis in NACT-administered patients with advanced ovarian cancer.

Ki-67 immunoexpression and radiological assessment of necrosis improves accuracy of conventional and modified core biopsy systems in predicting the final grade assigned to adult-soft tissue sarcomas. An international collaborative study

Based on the French Federation Nationale des Centers de Lutte Contre le Cancer (FNCLCC) grading system, this study assesses the accuracy of conventional and modified core biopsy (CB) systems in predicting the final grade (low vs high) assigned to the resected specimen. Substituting Ki-67 immunoexpression for mitotic count, and radiological for histological assessment of necrosis, we used two modified FNCLCC CB grading systems: (1) Ki-67 immunoexpression alone, and (2) Ki-67 plus radiological assessment of necrosis. We graded 199 soft tissue sarcomas (STS) from nine centers, and compared the results for the conventional (obtained from local histopathology reports) and modified CB systems with the final FNCLCC grading of the corresponding resected specimens. Due to insufficient sample quality or lack of available radiologic data, five cases were not evaluated for Ki67 or radiological assessment of necrosis. The conventional FNCLCC CB grading system accurately identified 109 of the 130 high-grade cases (83.8%). The CB grading matched the final FNCLCC grading (low vs high) in 175 (87.9%) of the 199 resected tumors; overestimating the final grade in three cases and underestimating in 21 cases. Modified system 1 (Ki-67) accurately identified 117 of the 130 high-grade cases (90.0%). The CB grading matched the final FNCLCC grading (low vs high) in 175 (89.7%) of the 195 evaluated cases; overestimating seven and underestimating 13 cases. Modified system 2 (Ki-67 plus radiological necrosis) accurately identified 120 of the 130 high-grade cases (92.3%). This last matched the final FNCLCC grading (low vs high) in 177 (91.2%) of the 194 evaluated cases; overestimating seven and underestimating 10 cases. Modified system 2 obtained highest area under ROC curves, although not statistically significant. Underestimated CB grades did not correlate with histological subtypes, although many of the discrepant cases were myxoid tumors (myxofibrosarcomas or myxoid liposarcomas), leiomyosarcomas or undifferentiated pleomorphic/spindle cell sarcomas. Using modified FNCLCC CB grading systems to replace conventional mitotic count and histologic assessment of necrosis may improve the distinction between low and high-grade STS on CB. Our study confirms that classifying grade 1 as low grade and grades 2 and 3 as high grade improves correlation between CB and final grade by up to 21%, irrespective of CB system used. A higher than expected Ki-67 score in a low-grade sarcoma diagnosed on CB should raise concern that a higher-grade component may not have been sampled. Furthermore, correlation of all clinicopathological and radiological findings at multidisciplinary meetings is essential to assess the histological grade on CB as accurately as possible.

English

BACKGROUND Breast cancer is the most common malignancy in females. Triple negative breast carcinomas (TNBC), one of the molecular subtypes of breast carcinoma, are not sensitive to hormonal therapy and are reported to have aggressive behaviour. The present study was done to evaluate the expression of Ki67 in triple negative breast cancer using immunohistochemistry (IHC) and correlate the Ki67 expression with other clinicopathological variables. METHODS Based on the IHC status (ER, PR, HER neu), 50 triple negative breast carcinoma cases were selected from January 2019 to June 2020 for a cross-sectional study in Department of Pathology, Government medical college Thrissur. Ki67 immunohistochemical staining was done on the tissue sections and the Ki67 score was correlated with clinicopathological variables. RESULTS Of the 50 cases, majority (74 % cases) had high Ki67 expression (score of more than 50 %). High Ki67 score was strongly associated with presence of lymphovascular emboli (LVE) (P value &lt; 0.05). Even though most of the patients were above 50 years (72 %), no significant correlation was seen between age and Ki67 score. The association with tumour size, histopathological type, tumour grade and lymph node status were not statistically significant. CONCLUSIONS Ki67 expression was high in triple negative breast cancer with mean score of 62 %. High Ki67 score correlated with presence of LVE. High Ki67 would predict increased proliferation of breast cancer cells and could be considered as a prognostic marker. KEYWORDS TNBC- Triple Negative Breast Cancer, Ki 67 Score, Clinicopathological Variables

Abstract 887: Clinicopathological and genetic features of breast cancer in Algerian population: A multicenter study

Abstract Background: Breast cancer is the most commonly diagnosed cancer and the leading cause of morbidity and mortality among Algerian women.The objective of this retrospective cohort study was to analyze some clinicopathological and molecular characteristics of breast cancer diagnosed between 2011 and 2019. Here we also report the experience of our research laboratory for the screening of BRCA1 and BRCA2 genes in hereditary breast and ovarian cancer (HBOC) patients. Materials and Methods: The population study included 7655 consecutive breast cancer patients. Data were collected from cancer registries of three medical oncology services and two anti-cancer centers located in north central region and eastern region of Algeria, respectively. Breast cancers were diagnosed between 2011 and 2019. Patient and tumor information included: age at diagnosis, menopausal status, receptor status, Ki-67 score, histological type,TNM stage and histological grade. Recurrent germline mutations on BRCA1 and BRCA2 genes previously found in Algerian patients were screened using PCR-Sanger sequencing in 230 HBOC patients. In addition, 18 HBOC patients were analyzed by NGS using a cancer panel of 30 hereditary cancer genes or BRCA1/2 genetic test. Results: The median age at diagnosis of the patients was 49.52 years. Data for age at diagnosis were available for 7389 patients. 55.29% patients were diagnosed under the age of 50 years and 55.83% had a premenopausal status. Data for receptor status were available for a total of 5599 patients. Of these, 55.75% patients were luminal A, 19.79% were TNBC, 17.39% were luminal B and 7.05% were HER2+. Data for Ki-67 score were available for 3357 cases. We noticed that 62.73% patients had a Ki-67 score ≥ 20% and 37.27% had a score &amp;lt;20%. Data for histological tumor type were available for 6145 cases. 84.28% patients had invasive ductal carcinoma followed by 8.84% with invasive lobular carcinoma. Data for TNM stage were available for 3524 cases. We identified 59.3% patients with stage III, 22.3% with stage II and 9.5% with stage IV. Data for histological grade were available for 6693 patients. We found 67.45% patients with grade II and 25.51% with grade III, respectively. 10 distinct BRCA1 germline mutations have been detected in 18 families and 5 distinct BRCA2 germline mutations have been identified in 5 families. The BRCA1 recurrent mutation c.83_84delTG has been detected in 5 unrelated families.The BRCA1 recurrent mutation c.2125_2126insA has been identified in 3 unrelated families. In addition, the BRCA1 c.181T&amp;gt;G has been found in two unrelated families. Interestingly, our results also showed differences in the distribution of the mutation spectrum of BRCA1 and BRCA2 genes between the eastern region and the north central region of Algeria. Conclusion: Our current study will contribute in developing optimal clinical trial protocols and personalized management strategies for breast cancer patients. Citation Format: Farid Cherbal, Leticia Ledmila Saada, Massila Nehar, Kamelia Yatta, Lydia Souha Zerrouki, Wafa Abdou, Chiraz Mehemmai, Hadjer Gaceb, Wassila Benbrahim, Hassen Mahfouf, Kada Boualga. Clinicopathological and genetic features of breast cancer in Algerian population: A multicenter study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 887.

Application Test of the AI-Automatic Diagnostic System for Ki-67 in Breast Cancer

To study the different methods of artificial intelligence (AI)-assisted Ki-67 scoring of clinical invasive ductal carcinoma (IDC) of the breast and to compare the results. A total of 100 diagnosed IDC cases were collected, including slides of HE staining and immunohistochemical Ki-67 staining and diagnosis results. The slides were scanned and turned into whole slide image (WSI), which were then scored with AI. There were two AI scoring methods. One was fully automatic counting by AI, which used the scoring system of Ki-67 automatic diagnosis to do counting with the whole image of WSI. The second method was semi-automatic AI counting, which required manual selection of areas for counting, and then relied on an intelligent microscope to conduct automatic counting. The diagnostic results of pathologists were taken as the results of pure manual counting. Then the Ki-67 scores obtained by manual counting, semi-automatic AI counting and automatic AI counting were pairwise compared. The Ki-67 scores obtained from the manual counting (pathological diagnosis results), semi-automatic AI and automatic AI counts were pair-wise compared and classified according to three levels of difference: difference ≤10%, difference of >10%-<30% and difference ≥30%. Intra-class correlation coefficient ( ICC) was used to evaluate the correlation. The automatic AI counting of Ki-67 takes 5-8 minutes per case, the semi-automatic AI counting takes 2-3 minutes per case, and the manual counting takes 1-3 minutes per case. When results of the two AI counting methods were compared, the difference in Ki-67 scores was all within 10% (100% of the total), and the ICC index being 0.992. The difference between manual counting and semi-automatic AI was less than 10% in 60 cases (60% of the total), between 10% and 30% in 37 cases (37% of the total), and more than 30% in only 3 cases (3% of the total), ICC index being 0.724. When comparing automatic AI with manual counting, 78 cases (78% of the total) had a difference of ≤10%, 17 cases (17% of the total) had a difference of between 10% and 30%, and 5 cases (5%) had a difference of ≥30%, the ICC index being 0.720. The ICC values showed that there was little difference between the results of the two AI counting methods, indicating good repeatability, but the repeatability between AI counting and manual counting was not particularly ideal. AI automatic counting has the advantage of requiring less manpower, for the pathologist is involved only for the verification of the diagnosis results at the end. However, the semi-automatic method is better suited to the diagnostic habits of pathologists and has a shorter turn-over time compared with that of the fully automatic AI counting method. Furthermore, in spite of its higher repeatability, AI counting, cannot serve as a full substitute for pathologists, but should instead be viewed as a powerful auxiliary tool.

Clinical Significance of Low Grade Follicular Lymphoma with High Proliferative Index

Introduction: Follicular lymphoma (FL) accounts for about 20% of lymphomas. The time to treatment, time to progression and overall survival are widely variable and additional prognostic factors are needed to identify patients who might benefit from upfront therapy. The association of high Ki67 proliferative index in FL with recurrence, progression, and decreased survival in unclear. Furthermore, the contribution of Ki67 proliferative index score in follicular versus interfollicular area for prognosis has not been reported.Methods: We performed a retrospective observational analysis of newly diagnosed patients with FL evaluated at University of Minnesota, from 2003 to April 2016. All cases were graded using WHO Classification histologic grading criteria. We excluded patients with no follow-up and grade 3B. We examined medical records and performed Ki67 immunohistochemical nuclear stains. Two pathologists reviewed Ki67 staining in follicular and interfollicular area and selected areas with maximum staining. Ten consecutive follicular and interfollicular areas were scored separately for Ki67 nuclear staining. Association of Ki67 with overall survival (OS) and progression-free survival (PFS) from date of diagnosis was analyzed using Cox regression. We defined PFS as time from diagnosis to initiation of first treatment or death, whichever occurred first.Results: We examined 69 cases, 63 were grade 1 and 2 follicular lymphoma and 6 were grade 3A. Median age at diagnosis was 59 years. Gender was distributed equally. At time of diagnosis FLIPI score was 0 in 9 patients (13%), 1 in 15 patients (21.7%), 2 in 19 patients (27.5%), 3 in 15 patients (21.7%), 4 in 3 patients (4.3%), and unknown in 8 patients (11.6%). 27.5% patients (27 cases) showed bone marrow involvement at the time of diagnosis. Median (interquartile range) Ki67 was 25% (15%, 40%) in follicular area and 5% (5%, 15%) in interfollicular area.Median follow-up time was 4.0 years (range 0.1 to 11.7 years). 37.7% underwent upfront observation, 14.5% were treated with RCHOP as first line of therapy, 23.2% received rituximab as maintenance therapy, few (13%) cases transformed into diffuse large B-cell lymphoma, and 20% of the patients died. OS at 5-years was 86% (95% CI: 76%, 97%), and PFS at 5-years was 25% (95% CI: 16%, 40%). Median PFS was 1.6 months. In univariate analysis, follicular Ki67 was not associated with PFS (p > 0.40) or OS. Interfollicular Ki67 was highly associated with shorter PFS with hazard ratio 1.32 per 10 units of Ki67 (95% CI: 1.08, 1.63). Results were similar adjusting for FLIPI score.Conclusions: Our data suggests that interfollicular area Ki67 score at diagnosis is prognostic to predict PFS in patients with FL. The relationship appears to be linear and doesn't change when adjusting for FLIPI score. Follicular area Ki67 has no impact on clinical outcomes. We propose that the Ki67 proliferative index should be scored in the interfollicular areas to assess the prognosis in FL. DisclosuresBachanova:Novartis Pharmaceuticals Corporation: Membership on an entity's Board of Directors or advisory committees, Research Funding; Zymogen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Oxis: Membership on an entity's Board of Directors or advisory committees, Research Funding; Seattle-Genetics: Consultancy, Membership on an entity's Board of Directors or advisory committees; Juno: Membership on an entity's Board of Directors or advisory committees.

KI67/BCL2 index in invasive breast carcinoma (NST)

Background: B-cell lymphoma-2 (BCL2) is an anti-apoptotic protein that has an anti-proliferative effect and is a favourable prognostic marker in breast carcinoma. Ki67/BCL2 ratio is emerging as a superior prognostic marker than either marker alone. Aim: This study investigated the association of Ki67/Bcl2 index with hormonal receptor status and other clinicopathological parameters in premenopausal and postmenopausal primary invasive breast carcinomas (NST). Method: Clinicopathological data were obtained from 63 (42%) premenopausal and 87 (58%) postmenopausal carcinomas from a prospective study. Immunohistochemical assessment of ER, PR (Allred score), HER2 (ASCO/CAP 2013), Ki67 and BCL2 in the tumours were performed using routine protocols. BCL2 and Ki67 scores were obtained based on the percentage of positively stained cells (0=0–10%, 1=11–32%, 2=>33%). BCL2 staining score was subtracted from the Ki67 score to produce five categories (−2, −1, 0, 1, 2) and further categorised into two groups, low-index (-2, -1, 0) and high-index (1, 2). Association of Ki67/Bcl2 index with tumour grade, tumour size, T and N stage, nodal status, node ratio (<0.15), ER, PR and HER2 status and molecular subtype, were evaluated using the Chi-square test (SPSS-20). Associations were considered significant when p≤0.05. Results and conclusion: High Ki67/BCL2 index was significantly associated with negative ER (premenopausal χ2=4.701(1), p=0.030; postmenopausal χ2=23.104(1), p≤0.001) and negative PR (premenopausal χ2=12.106(1), p=0.001; postmenopausal χ2=12.956(1), p≤0.001) expression in both groups. It was also significantly associated with premenopausal grade ΙΙΙ tumours (χ2=8.168(2), p=0.017) and postmenopausal triple negative breast cancers (χ2=19.510(4), p=0.001). It is evident that high Ki67/BCL2 index, indicating a high proliferation rate, is associated with poor prognosticators in breast carcinoma and is suggestive of a poor clinical outcome.

Primary small cell carcinoma of the breast

Small cell carcinoma arising in breast is very rare. The diagnosis of primary small cell carcinoma can only be made if a non-mammary site is excluded clinically or if in situ component can be demonstrated histologically as in this case. We reoprt a case of a 68-year-old female patient who presented with a left breast lump, which was diagnosed as a triple negative high grade cancer on core biopsy. The patient received neoadjuvant chemotherapy followed by a left breast lump removal (lumpectomy) with sentinel lymph node biopsy. Macroscopically, there is a well circumscribed tan to pink mass up to 24 mm. Microscopically, the breast shows nests of tumour cells. The tumour cells are small with scant cytoplasm, salt and pepper chromatin, nuclear moulding. Mitotic and apoptotic figures are plentiful. There is ductal carcinoma in situ (DCIS) with similar cytological features. Immunohistochemistry was performed. The tumour cells were positive for E-Cadherin, p53 and CK7, they were weakly positive for NSE and INSM1 and they were negative for CK5/6, S100 and TTF1. Ki67 score was 90%. Features are compatible with primary small cell carcinoma. Small cell carcinoma has a very aggressive behaviour. The management of the primary tumour has not been standardised, due to the rarity of this entity.

Clinical and Histopathological Variables and Prognostic Factors of Adrenocortical Carcinoma.

Adrenocortical carcinoma (ACC) is a rare and highly aggressive tumor with a poor prognosis. The literature on prognosis from low-income or low-middle-income countries is limited and scarce. This study aimed to determine the clinical and histopathological characteristics, recurrence-free survival (RFS), overall survival (OS), and the factors affecting ACC's prognosis.This was a retrospective study of patients that presented with ACC to the Shaukat Khanum Memorial Cancer & Research Center, Lahore, Pakistan, between January 2011 and May 2018. Information regarding demographics and clinical and histopathological variables were extracted and analyzed. Of the 25 subjects, 16 (64%) were female. The median age of the sample was 35 years (range; 21 - 72 years).Statistically significant associations were found between RFS and functional status of the tumor (p = 0.014), cortisol overproduction (p = 0.02), androgen excess (testosterone [p = 0.03] and dehydroepiandrosterone sulfate [DHEA SO4] [p = 0.004]), Ki-67 score (p = 0.03), mitotic rate (p = 0.02), stratified mitotic rate (p = 0.01), and composite variable of disease (p = 0.004). The OS was found to have statistical associations with cortisol hypersecretion (p = 0.02), DHEA SO4 excess (p = 0.01), Modified Weis Score (p < 0.001), mitotic rate (p = 0.02), stratified mitotic rate (p = 0.003), and composite variable of disease (p = 0.001). Linear regression (forward-type) analysis suggested that the functional status of the tumor and the disease recurrence index statistically predicted the variance in RFS and OS, respectively.Multiple clinical and histopathological variables appear to affect the prognosis of ACC. However, based on multivariable analysis, it appears that the functional status of the tumor and the composite variable of disease recurrence are predictors of RFS and OS, respectively.

Immunohistochemical expression of substance P in breast cancer and its association with prognostic parameters and Ki-67 index.

The neuropeptide substance P is a potential biomarker and therapeutic target in cancer. The main objectives of this study were to investigate the expression level of substance P in different breast cancer molecular subtypes and identify its association with clinicopathological parameters of patients and with Ki-67 index. A retrospective analysis was performed for a total of 164 paraffin-embedded breast cancer tissue samples [42 Her2/neu-enriched, 40 luminal A, 42 luminal B (triple-positive) and 40 triple negative subtypes]. The tissue microarray slides containing specimens were used to determine the expression of substance p and Ki-67 by immunohistochemical staining. The mean age of the cohort was 51.35 years. Twenty two percent of cases had low substance P expression levels (TS ≤ 5), while 78% had high expression levels (TS > 5). A significant association was found between SP expression level and breast cancer molecular subtype (p = 0.002), TNM stage (p = 0.034), pN stage (p = 0.013), axillary lymph node metastasis (p = 0.004), ER and PR statuses (p<0.001) and history of DCIS (p = 0.009). The average percentage of Ki-67 expression was 27.05%. When analyzed as a continuous variable, significant differences were observed between the mean Ki-67 scores and molecular subtype (p = 0.001), grade (p = 0.003), pN stage (p = 0.007), axillary lymph node metastasis (p = 0.001), and ER and PR statuses (p <0.001). SP is overexpressed in most of the analyzed tissues and has a negative prognostic value in the breast cancer patients. Besides substance P is a potential therapeutic target in breast cancer.

Influence of Human Telomerase Reverse Transcriptase Mutation on the Aggressiveness and Recurrence in Meningiomas.

Background: Over 200 human telomerase reverse transcriptase (hTERT) polymorphism combinations have been implicated in the development of cancer. This study aimed to evaluate hTERT mutations in meningioma tissue and its association with meningioma.Material and Methods: A total of 90 patients who underwent surgery between 2006 and 2015 and were histopathologically diagnosed with meningioma (WHO 2016) were included.Results: Among the 90 participants included herein, 50 (55.5%) and 40 (44.5%) were female and male, respectively, with an average age of 56.2 ± 14 years. Mean Ki-67 values were 10.56% (SD 12.41, range 0-60), while the mean follow-up duration was 39.1 months (SD 26.3). Low- and high-grade patients had a mean Ki-67 score of 4.31% (SD 3.58, range 0-16) and 19.92% (SD 14.91, range 2-60) (p = 0.0001). Our results showed a moderate positive correlation between Ki-67 score and the presence of hTERT mutation (Pearson correlation test, r = 0.5161; p = 0.0001). Patients with an hTERT mutation > 30% had significantly higher risk for reoperation than those with lower levels of mutation (p = 0.016, chi square test). None of the patients requiring reoperation had an hTERT mutation < 10%. Moreover, high-grade patients had a 7.2 times higher risk of reoperation than those with an hTERT mutation > 30%.Conclusion: The presence of hTERT mutation, in addition to high Ki-67, indicated a more aggressive meningioma disease course and potentially increased risk of recurrence.

Prognostic parameter differences in breast cancer patients between luminal A and luminal B types after application of the new classification according to Ki67 score

BackgroundThe conventional histological classification of breast cancer doesn't provide sufficient information about the biological characteristics and treatment prediction for breast cancer subtypes. Luminal breast cancer has been classified recently according to the expression of ki67 which has a great impact of the biological activity of the tumor. Patients and methodsA total of 300 patients with breast cancer who underwent modified radical mastectomy were investigated. Patients were divided into 2 groups: Luminal A (101 patients) and B (199 patients) according to the new classification based on hormone receptor and Ki67 expressions and the prognostic parameters were investigated. ResultsThe mean age was 48.81 years with a mean BMI of 30.7, the age showed no significant correlation (P value 0.996), but it was significant with BMI (P value 0.045). Lobular breast cancer was commoner in luminal type-A (13.9%) comparing to type-B patients (5%) with a significant correlation with the histopathological type (P value 0.006). Most patients presented with clinical stage IIB, IIA, and IIIA respectively. The tumor size, axillary LN status, clinical stage, and recurrence or metastasis showed no statistical significance (P values 0.9, 0.078, 0.584, and 0.943) respectively, 66.3% of luminal type-A patients had intermediate grade and no tumor necrosis (72.5%), while 55.3% of luminal type-B patients had high grade and tumor necrosis (61.8%), the correlation was significant for tumor grade and necrosis (P values 0.000) for each parameter. Luminal type-B had higher breast cancer fatality rate compared to type-A (11.2% vs. 2%) with a significant correlation with survival status and duration of survival (P values 0.003 & 0.025) respectively. ConclusionLuminal type-A cancer is less common than type-B but has better survival and duration of survival than type-B. Ki67 biomarker affects tumor aggressiveness and invasiveness. More genetic analysis is required to detect the molecular diversity of both types.