Ki-67 Score Research Articles

AI-Powered Segmentation of Invasive Carcinoma Regions in Breast Cancer Immunohistochemical Whole-Slide Images.

The automation of quantitative evaluation for breast immunohistochemistry (IHC) plays a crucial role in reducing the workload of pathologists and enhancing the objectivity of diagnoses. However, current methods face challenges in achieving fully automated immunohistochemistry quantification due to the complexity of segmenting the tumor area into distinct ductal carcinoma in situ (DCIS) and invasive carcinoma (IC) regions. Moreover, the quantitative analysis of immunohistochemistry requires a specific focus on invasive carcinoma regions. In this study, we propose an innovative approach to automatically identify invasive carcinoma regions in breast cancer immunohistochemistry whole-slide images (WSIs). Our method leverages a neural network that combines multi-scale morphological features with boundary features, enabling precise segmentation of invasive carcinoma regions without the need for additional H&E and P63 staining slides. In addition, we introduced an advanced semi-supervised learning algorithm, allowing efficient training of the model using unlabeled data. To evaluate the effectiveness of our approach, we constructed a dataset consisting of 618 IHC-stained WSIs from 170 cases, including four types of staining (ER, PR, HER2, and Ki-67). Notably, the model demonstrated an impressive intersection over union (IoU) score exceeding 80% on the test set. Furthermore, to ascertain the practical utility of our model in IHC quantitative evaluation, we constructed a fully automated Ki-67 scoring system based on the model's predictions. Comparative experiments convincingly demonstrated that our system exhibited high consistency with the scores given by experienced pathologists. Our developed model excels in accurately distinguishing between DCIS and invasive carcinoma regions in breast cancer immunohistochemistry WSIs. This method paves the way for a clinically available, fully automated immunohistochemistry quantitative scoring system.

Artificial Intelligence-Based Mitosis Scoring in Breast Cancer: Clinical Application

In recent years, artificial intelligence (AI) has demonstrated exceptional performance in mitosis identification and quantification. However, the implementation of AI in clinical practice needs to be evaluated against the existing methods. This study is aimed at assessing the optimal method of using AI-based mitotic figure scoring in breast cancer (BC). We utilized whole slide images from a large cohort of BC with extended follow-up comprising a discovery (n = 1715) and a validation (n = 859) set (Nottingham cohort). The Cancer Genome Atlas of breast invasive carcinoma (TCGA-BRCA) cohort (n = 757) was used as an external test set. Employing automated mitosis detection, the mitotic count was assessed using 3 different methods, the mitotic count per tumor area (MCT; calculated by dividing the number of mitotic figures by the total tumor area), the mitotic index (MI; defined as the average number of mitotic figures per 1000 malignant cells), and the mitotic activity index (MAI; defined as the number of mitotic figures in 3 mm2 area within the mitotic hotspot). These automated metrics were evaluated and compared based on their correlation with the well-established visual scoring method of the Nottingham grading system and Ki67 score, clinicopathologic parameters, and patient outcomes. AI-based mitotic scores derived from the 3 methods (MCT, MI, and MAI) were significantly correlated with the clinicopathologic characteristics and patient survival (P < .001). However, the mitotic counts and the derived cutoffs varied significantly between the 3 methods. Only MAI and MCT were positively correlated with the gold standard visual scoring method used in Nottingham grading system (r = 0.8 and r = 0.7, respectively) and Ki67 scores (r = 0.69 and r = 0.55, respectively), and MAI was the only independent predictor of survival (P < .05) in multivariate Cox regression analysis. For clinical applications, the optimum method of scoring mitosis using AI needs to be considered. MAI can provide reliable and reproducible results and can accurately quantify mitotic figures in BC.

Treatment Outcomes of Limited Stage Grade 3A Follicular Lymphoma

Introduction While radiotherapy is the established standard of care for limited stage, grade 1-2 follicular lymphoma (FL), the management of localized grade 3A (FL3A) disease remains controversial. We reviewed the outcomes of grade 3A patients treated with definitive RT, systemic therapy (ST) alone or combined modality therapy (CMT) of RT plus ST. Methods We analyzed a single institution database to identify newly-diagnosed, FL3A patients with Ann Arbor stage I/II disease treated between 2000-2021. All cases were reviewed by the Hematopathology staff at the time of diagnosis. Patients were stratified by definitive treatment modality including RT monotherapy, CMT, or ST. Best post-RT imaging response was evaluated using Lugano criteria for all by CT, and by PET if available. Rate of complete response (CR), overall survival (OS), progression-free survival (PFS) and time to out-of-field progression were compared between the three treatment modalities. Progression-free survival (PFS) and overall survival (OS) were calculated using Kaplan Meier from first day of treatment. Time to out-of-field progression was estimated via cumulative incidence function from treatment start date with competing events death without out-of-field progression of disease (PD) and in-field PD. Univariable Cox regression was used to assess possible clinicodemographic associations with PFS. Results 84 patients (median age 62, 38% male) were analyzed with median follow-up of 6.9 years from treatment. Patients were stage I (73%) or II (27%) and 92% were initially PET staged at diagnosis. 29 (35%) had extranodal disease. The most common site of extranodal disease was the head and neck (13% of patients). The median (IQR) baseline maximum SUV and maximum lesion size prior to treatment were 9.8 (5.4, 14.0) and 3.2 (2.0, 4.5) cm respectively, and did not differ between treatment groups. RT was the most commonly utilized treatment modality (n=48, 57%), followed by CMT (n=21, 25%), then ST (n=15, 18%). For RT/CMT patients the planned RT dose was most commonly 36 Gy (32%, range 4-40Gy). 2 patients (3%) received a very low dose program beginning with 4 Gy. Of the CMT/ST patients the most common regimen was 3-4 cycles of R-CHOP (53%). 3 patients (8%) received Rituximab alone. All ST patients were diagnosed after 2010. The ST subset had the highest proportion of patients with stage II disease (p&amp;lt;0.001), while CMT patients had the highest proportion of female patients (p=0.03) and grade 3 disease not further characterized (p=0.002). The rates of CR post-treatment were 88% for RT, 100% for CMT, and 87% for ST. Of the 22 patients who relapsed, only 1 suffered an infield relapse alone (4.5%) with the rest experiencing distant relapse (95.5%). The 5-year cumulative incidence of out-of-field progression was 32% (95%CI 18-46%) for RT, 11% (95%CI 1.7-30%) for CMT, and 17% (95%CI 2.3-43%) for ST. There were factors significantly associated with PFS univariably including the treatment modality (CMT vs. RT HR 0.30, 95% CI 0.10-0.91, p=0.03), and presence of low-grade FL component in the biopsy (HR 3.33, 95% CI 1.43-7.78, p=0.005). Age, sex, stage, extranodal status, Ki67 score, max pre-treatment size, and max pre-treatment SUV were not associated with PFS. The probability of transformation to DLBCL was 4.2% (95% CI 1.1-11%) at 5 years after treatment. Overall survival was excellent for all treatment modalities with median OS of 18 years for CMT, and not reached for RT or ST. Conclusion To our knowledge this is the largest study of the outcomes of early stage FL3A after therapy. CMT had the highest rate of CR in our cohort, and was associated with a statistically significantly lower hazard of PFS compared to RT. However, OS is excellent regardless of treatment modality suggesting all may be appropriate options.

Long-Term Remissions after First-Line Autologous Stem Cell Transplantation for Mantle Cell Lymphoma

Introduction: The role for autologous stem cell transplantation (ASCT) in mantle cell lymphoma (MCL) has been recently called into question by the TRIANGLE trial, which demonstrated similar 3-year outcomes with or without ASCT among patients treated with R-CHOP/R-DHAP and 2 years of maintenance ibrutinib +/- rituximab. However, long-term follow-up is necessary to determine the durability of remissions and ascertain the true value of first-line ASCT for MCL. Methods: This retrospective population-based study included all patients ≥18 years old who received ASCT as part of first-line therapy for MCL in the province of Alberta, Canada between 2000-2022. The primary objective was to determine freedom from progression (FFP), which was measured from the time of ASCT to disease progression. Secondary objectives were to determine progression-free survival (PFS) and overall survival (OS). FFP, PFS, and OS were estimated using the Kaplan-Meier method. Cumulative incidences of relapse and non-relapse mortality (NRM) were determined using competing risks analysis. Univariable and multivariable Cox regression was performed to identify risk factors for FFP. Results: The study population included 141 patients with median age at ASCT of 60 years (range 34-71). At diagnosis, 130 (92%) patients had advanced stage disease, 52 (37%) had intermediate/high risk MIPI score, 35 (25%) had Ki67 &amp;gt;30%, 32 (23%) had B symptoms, and 17 (12%) had blastoid/pleomorphic histology. Induction protocols included R-CHOP/cytarabine combinations (n=93), R-bendamustine/cytarabine combinations (n=36), or alternative regimens (n=12). PET-defined pre-transplant disease status was complete metabolic response in 75 (82%) and partial metabolic response in 17 (18%) patients. ASCT conditioning consisted of melphalan plus total body irradiation (n=93), BEAM (n=40), or other regimens (n=8). Maintenance rituximab was received by 96 (68%) patients. With median follow-up time 7.6 years (range 0.4-23.2), the median FFP was not reached (Figure 1) while the median PFS was 11.3 years and median OS was 14.0 years. At 8 years after ASCT, FFP was 67% (95% CI 56-75%), PFS was 57% (95% CI 46-66%), and OS was 70% (95% CI 60-78%). FFP at 8 years was 76% (95% CI 59-86%) with maintenance rituximab versus 49% (95% CI 33-63%) without maintenance rituximab (p&amp;lt;0.001). Multivariable analysis revealed that maintenance rituximab was associated with improved FFP (HR 0.26, 95% CI 0.10-0.72) and B symptoms were associated with inferior FFP (HR 8.45, 95% CI 2.86-25.0), whereas age, Ki67, blastoid/pleomorphic histology, and MIPI score were not significantly associated with FFP. The cumulative incidence of relapse at 8 years was 32% (95% CI 23-41%) and the mean relapse rate appeared to decrease over time, from 4.4%/year during years 0-5 to 2.5%/year during years 6-12 after ASCT. In a subgroup analysis to evaluate the very long-term outcomes of 49 patients who underwent ASCT &amp;gt;10 years ago, median follow-up time was 12.5 years (range 0.4-23.2) and 13-year FFP was 40% (95% CI 26-54%), PFS was 32% (95% CI 19-46%), and OS was 54% (95% CI 35-68%). No relapse occurred &amp;gt;11.2 years after ASCT. Twenty patients remain alive and in remission for &amp;gt;10 years, 5 patients for &amp;gt;15 years, and 1 patient for &amp;gt;20 years after ASCT. There were no cases of NRM within 1 year of ASCT. Therapy-related myeloid neoplasms occurred in 3 (2%) patients. Conclusions: This long-term follow-up study demonstrates that first-line ASCT frequently achieves durable remissions in MCL, with median PFS &amp;gt;11 years, a declining relapse rate over time, and notably high FFP rates among those who receive maintenance rituximab. A subset of patients experienced remissions lasting &amp;gt;10-20 years and may be functionally cured of their lymphoma, although additional follow-up is warranted to confirm the emergence of a plateau on the FFP curve. Given the durability of remissions and possibility of cure in some patients, longer follow-up of the TRIANGLE trial should be awaited before first-line ASCT is prematurely abandoned for MCL.

Comparative study of droplet-digital PCR and absolute Q digital PCR for ctDNA detection in early-stage breast cancer patients

BackgroundAnalysis of circulating tumor DNA (ctDNA) is increasingly used for clinical decision-making in oncology. However, ctDNA could represent ≤ 0.1 % of cell-free DNA in early-stage tumors and its detection requires high-sensitive techniques such as digital PCR (dPCR). MethodsIn 46 samples from patients with early-stage breast cancer, we compared two leading dPCR assays for ctDNA analysis: QX200 droplet digital PCR (ddPCR) system from Bio-Rad which is the gold-standard in the field, and Absolute Q plate-based digital PCR (pdPCR) system from Thermo Fisher Scientific which has not been reported before. We analyzed 5 mL of baseline plasma samples prior to any treatment. ResultsBoth systems displayed a comparable sensitivity with no significant differences observed in mutant allele frequency. In fact, ddPCR and pdPCR possessed a concordance > 90 % in ctDNA positivity. Nevertheless, ddPCR exhibited higher variability and a longer workflow. Finally, we explored the association between ctDNA levels and clinicopathological features. Significantly higher ctDNA levels were present in patients with a Ki67 score > 20 % or with estrogen receptor-negative or triple-negative breast cancer subtypes. ConclusionBoth ddPCR and pdPCR may constitute sensitive and reliable tools for ctDNA analysis with an adequate agreement in early-stage breast cancer patients.

Augmented anti-cancer effect of hypoxia-inducible factor 2α (HIF2α)- targeting PT2385 delivered in α-synuclein-AuNP microcapsules for clear cell renal carcinoma mouse model

Development of anti-cancer drug carrier system capable of delivering chemotherapeutic and immunotherapeutic agents to cancerous lesions is essential to overcome severe side effects and reduced therapeutic efficacy of the drugs. The microcapsules of gold nanoparticles (AuNPs) fabricated with a self-assembly protein of alpha-synuclein (αS) have been employed to carry the hypoxia-inducible factor 2α (HIF2α)-targeting PT2385 in a mouse model xenografted with clear cell renal cell carcinoma (ccRCC). Hydrophobic cargo like rhodamine 6G entrapped within the microcapsules was demonstrated to be selectively released under cancer-related stimuli including acidic pH, local hyperthermia, and proteases such as matrix metalloproteinases. The αS-AuNP microcapsules containing the hydrophobic drug of PT2385 suppressed the expression of HIF2α and its downstream genes in ccRCC A498 cells in vitro to the level comparable to that obtained with direct PT2385 treatment, indicating that the PT2385 remained active inside the capsules. In the A498 xenograft mouse model, intraperitoneal administration of the αS-AuNP-PT2385 microcapsules significantly reduced the tumor volume and Ki-67 scores from those monitored in the PT2385-treated group. Therefore, the αS-AuNP microcapsule is suggested to be a promising cancer-targeting drug delivery system which exhibits several merits such as improved loading efficiency for hydrophobic drugs, their protection from metabolism, and selective cargo release in tumor microenvironments in addition to its physicotherapeutic potential due to the photothermal effect of AuNPs.

PATH-48. KI-67 INDEX AS A PREDICTIVE MARKER OF MENINGIOMA CLINICO-PATHOLOGIC COURSE AND RECURRENCE FOLLOWING SURGICAL RESECTION: A RETROSPECTIVE STUDY

Abstract BACKGROUND/OBJECTIVE Meningiomas are the most common primary intracranial tumor in adults. Although usually benign, they have a variable clinical course. Ki-67, a nuclear protein involved in cell cycle regulation, has been widely studied as a marker of cellular proliferation in various cancers. However, the prognostic significance of Ki-67 in meningiomas remains controversial. Here, we investigate the Ki-67 index, as a predictive marker of meningioma recurrence following surgical resection and compare it to established prognostic markers such as WHO grade and degree of resection. METHODS The medical records of 451 consecutive patients with previously untreated cranial meningiomas who underwent resections from January 2011 to January 2021 at North Shore University Hospital (NSUH) were reviewed. Collected data included WHO grade, Ki-67 proliferative index, degree of resection, and meningioma recurrence. RESULTS There were 290 patients with grade I, 154 with grade II, and 7 with grade III meningiomas. The average post-resection follow-up period was 4 years, and 82 tumors (18%) recurred. Recurrence occurred significantly more frequently in tumors with a Ki67 index of 5 or more (31%) compared to those with a Ki67 index less than 5 (10.4%). Likewise, higher WHO grades were associated with a higher rates of recurrence, with rates of 11.3%, 27.9%, and 71.4% for grades 1, 2, and 3, respectively. Higher WHO grades also correlated with higher Ki67 scores (2.59, 10.01, and 20.71) for grades 1, 2, and 3, respectively. Additionally, a multivariate logistic regression model identified ki67, WHO grade and degree of resection as independent predictive variables for meningioma recurrence. CONCLUSION Our 10-year retrospective study suggests that the Ki67 index is a promising predictive marker for recurrence of intracranial meningioma following surgical resection. Our findings add to a growing body of data which supports inclusion of ki67 index in the WHO grading criteria.

Implementation of Digital Image Analysis in Assessment of Ki67 Index in Breast Cancer.

The clinical utility of the proliferation marker Ki67 in breast cancer treatment and prognosis is an active area of research. Studies have suggested that differences in pre-analytic and analytic factors contribute to low analytical validity of the assay, with scoring methods accounting for a large proportion of this variability. Use of standard scoring methods is limited, in part due to the time intensive nature of such reporting protocols. Therefore, use of digital image analysis tools may help to both standardize reporting and improve workflow. In this study, digital image analysis was utilized to quantify Ki67 indices in 280 breast biopsy and resection specimens during routine clinical practice. The supervised Ki67 indices were then assessed for agreement with a manual count of 500 tumor cells. Agreement was excellent, with an intraclass correlation coefficient of 0.96 for the pathologist-supervised analysis. This study illustrates an example of a rapid, accurate workflow for implementation of digital image analysis in Ki67 scoring in breast cancer.

High-risk histological subtype-related FAM83A hijacked FOXM1 transcriptional regulation to promote malignant progression in lung adenocarcinoma.

According to the histopathology, lung adenocarcinoma (LUAD) could be divided into five distinct pathological subtypes, categorized as high-risk (micropapillary and solid) group, intermediate-risk (acinar and papillary) group, and low-risk (lepidic) group. Despite this classification, there is limited knowledge regarding the role of transcription factors (TFs) in the molecular regulation of LUAD histology patterns. Publish data was mined to explore the candidate TFs associated with high-risk histopathology in LUAD, which was validated in tissue samples. Colony formation, CCK8, EdU, transwell, and matrigel assays were performed to determine the biological function of FAM83A in vitro. Subcutaneous tumor-bearing in BALB/c nude mice and xenograft perivitelline injection in zebrafish were utilized to unreal the function of FAM83A in vivo. We also performed chromatin immunoprecipitation (ChIP), dual-luciferase reporter, and rescue assays to uncover the underline mechanism of FAM83A. Immunohistochemistry (IHC) was performed to confirm the oncogenic role of FAM83A in clinical LUAD tissues. Screening the transcriptional expression data from TCGA-LUAD, we focus on the differentially expressed TFs across the divergent pathological subtypes, and identified that the expression of FAM83A is higher in patients with high-risk groups compared with those with intermediate or low-risk groups. The FAM83A expression is positively correlated with worse overall survival, progression-free survival, and advanced stages. Gain- and loss-of-function assays revealed that FAM83A promoted cell proliferation, invasion, and migration of tumor cell lines both in vivo and in vitro. Pathway enrichment analysis shows that FAM83A expression is significantly enriched in cell cycle-related pathways. The ChIP and luciferase reporter assays revealed that FAM83A hijacks the promoter of FOXM1 to progress the malignant LUAD, and the rescue assay uncovered that the function of FAM83A is partly dependent on FOXM1 regulation. Additionally, patients with high FAM83A expression positively correlated with higher IHC scores of Ki-67 and FOXM1, and patients with active FAM83A/FOXM1 axis had poor prognoses in LUAD. Taken together, our study revealed that the high-risk histological subtype-related FAM83A hijacks FOXM1 transcriptional regulation to promote malignant progression in lung adenocarcinoma, which implies targeting FAM83A/FOXM1 is the therapeutic vulnerability.

Evaluation of the relationship between Ki67 expression level and neoadjuvant treatment response and prognosis in breast cancer based on the Neo-Bioscore staging system

BackgroundNeoadjuvant chemotherapy (NAC) is widely used in the treatment of primary breast cancer. Different staging systems have been developed to evaluate the residual tumor after NAC and classify patients into different prognostic groups. Ki67, a proliferation marker, has been shown to be useful in predicting treatment response and prognosis. We aimed to investigate the prognostic importance Neo-Bioscore stage and pretreatment and posttreatment Ki67 levels in breast cancer patients who received NAC and correlations between Neo-Bioscore stage and pretreatment and posttreatment Ki67 levels.MethodsA total of 176 invasive breast carcinoma patients who underwent NAC were included in the study. Ki67 levels were evaluated by immunohistochemical methods in Trucut biopsy and surgical excision specimens. Patients were classified into prognostic groups using the Neo-Bioscore staging system.ResultsPatients with high pretreatment Ki67 score were more likely to be in the higher Neo-Bioscore risk group (p < 0.001). Patients with a high posttreatment Ki67 score were more likely to be in the higher Neo-Bioscore prognostic risk group (p < 0.001). Overall survival (OS) and disease-free survival (DFS) were shorter in patients with high posttreatment Ki67 scores and in patients in the higher Neo-Bioscore risk group. We also determined a cutoff 37% for pathological complete response.ConclusionNeo-Bioscore staging system is found to be important in predicting survival. The posttreatment Ki67 level is more important than pretreatment Ki67 level in predicting survival.

Use of the Gene Expression Test Prosigna® in Premenopausal Patients with HR+, HER2− Early Breast Cancer: Correlation of the Results with the Proliferation Marker Ki-67

Introduction: In hormone receptor-positive (ER+/PR+) and human epidermal growth factor receptor 2-negative (HER2−) early-stage breast cancer (EBC), gene expression tests such as the Prosigna are increasingly used since classic clinicopathological parameters and the proliferation factor Ki-67 often do not allow a definite therapy decision regarding an adjuvant chemotherapy. While the Prosigna test has been validated for postmenopausal patients, few data are available regarding its use in premenopausal patients. The present study compared the Prosigna test with the Ki-67 index in premenopausal patients. Materials and Methods: Premenopausal patients with HR+ HER2−, pN0-1, G1-2 EBC were retrospectively enrolled (n = 55). The Prosigna assay was performed in formalin-fixed paraffin-embedded tumor samples of surgical resection specimens. Ki-67 was reassessed in original diagnostic core needle biopsy specimens and defined as low, intermediate, or high with the threshold of <10%, 10–24%, ≥25%. Results: According to Ki-67, patients were in the low (LR)-, intermediate (IR)-, and high-risk (HR) groups in 40%, 36%, and 24% of the cases. The Prosigna gene signature assay assessed the risk of recurrence as LR for 45% of the patients, IR for 35%, and HR for 20%. The most frequent intrinsic subtypes were luminal A in 73% and luminal B in 24% of the patients. A moderate correlation was found between Prosigna and Ki-67 scores with a Pearson correlation coefficient of 0.51. In the overall cohort, 47% of the Ki-67-based therapy decision would correspond to those based on the Prosigna score. After exclusion of IR patients, matching of low/low or high/high results was observed in 57% of the cases. Conclusion: According to the present study, there is only limited concordance regarding the risk group stratification between Ki-67 and Prosigna-based risk assessment. The relevance and frequency of premenopausal breast cancer emphasizes the need for further evaluation of gene expression analyses in this setting and the correlation with classic clinicopathological parameters regarding therapy decision-making.

Evaluation of the international Ki67 working group cut point recommendations for early breast cancer: comparison with 21-gene assay results in a large integrated health care system

PurposeThe International Ki67 Working Group (IKWG) has developed training for immunohistochemistry (IHC) scoring reproducibility and recommends cut points of ≤ 5% and ≥ 30% for prognosis in ER+, HER2−, stage I/II breast cancer. We examined scoring reproducibility following IKWG training and evaluated these cut points for selecting patients for further testing with the 21-gene Recurrence Score (RS) assay.MethodsWe included 307 women aged 50+ years with node-negative, ER+PR+HER2− breast cancer and with available RS results. Slides from the diagnostic biopsy were stained for Ki67 and scored using digital image analysis (IA). Two IHC pathologists underwent IKWG training and visually scored slides, blinded to each other and IA readings. Interobserver reproducibility was examined using intraclass correlation (ICC) and Kappa statistics.ResultsDepending on reader, 8.8–16.0% of our cohort had Ki67 ≤ 5% and 11.4–22.5% had scores ≥ 30%. The ICC for Ki67 scores by the two pathologists was 0.82 (95% CI 0.78–0.85); it was 0.79 (95% CI 0.74–0.83) for pathologist 1 and IA and 0.76 (95% CI 0.71–0.80) for pathologist 2 and IA. For Ki67 scores ≤ 5%, the percentages with RS < 26 were 92.6%, 91.8%, and 90.9% for pathologist 1, pathologist 2, and IA, respectively. For Ki67 scores ≥ 30%, the percentages with RS ≥ 26 were 41.5%, 51.4%, and 27.5%, respectively.ConclusionThe IKWG’s Ki67 training resulted in moderate to strong reproducibility across readers but cut points had only moderate overlap with RS cut points, especially for Ki67 ≥ 30% and RS ≥ 26; thus, their clinical utility for a 21-gene assay testing pathway remains unclear.

Is Ki-67 Really Useful as a Predictor for Response to Neoadjuvant Chemotherapy in Locally Advanced Breast Cancer?

Neoadjuvant chemotherapy (NACT) is routinely offered to operable locally advanced breast cancer (LABC) patients desirous of breast conservation surgery and inoperable LABC patients. Pathological complete response (pCR) following chemotherapy is recognized as a surrogate for survival outcomes in high grade tumour subtypes. Many biological and tumor characters have been shown to predict pCR. The current study was performed with the aim of investigating the ability of Ki-67 in predicting pCR with NACT in breast cancer patients. A total of 105 patients with locally advanced breast cancer who completed NACT followed by surgery were included in this study from January 2020 till December 2022. Patients with advanced metastatic breast carcinoma, who did not give consent for NACT, who did not complete NACT and who did not undergo surgery were excluded. All patients were assessed for Ki-67 score on core-needle biopsy samples and response rate was assessed clinically and by histopathological examination of resected specimen. Quantitative variables were compared using unpaired t-test or Mann-Whitney 'U' test and for categorical variables Chi-square or Fisher's exact test were used. Receiver operating characteristic (ROC) curve analysis was performed to assess the predictive potential of Ki-67 expression levels in predicting pCR. To identify the predictive factors associated with pCR, univariate analysis was performed. The P value < 0.05 was considered as statistically significant. Mean age was 51.57 ± 10.8years. 51 patients achieved clinical complete response (cCR) and 33 achieved pCR after NACT. Mean Ki-67 index in overall study population, in pCR group and no pCR group was 46.44 ± 22.92%, 51.60 ± 22.3% and 44.06 ± 22.7%, respectively. On univariate analysis, ER negativity, PR negativity and Her 2neu positivity were found predictive of pCR. On subgroup analysis, TNBC and Her 2neu positive sub groups were associated with higher cCR and pCR rate. We found no significant association between Ki-67 and pCR. This result may be confounded by the fact that a significant duration of the study was in the COVID-19 pandemic. Validation of this data is required in a large prospective study.

Protective effect of paricalcitol in rat testicular damage induced by subchronic 1800 MHz radiofrequency radiation

In the present study, the possible protective effects of paricalcitol (P) were investigated in testicular damage because of 1800 MHz radiofrequency radiation (RFR) exposure. Male Sprague Dawley rats 8–10 weeks old (n = 28) were randomly divided into four groups as control (C) (n = 7), RFR (n = 7, 1800 MHz RFR 1 h/day for 30 days), P (n = 7, 0.2 μg/kg paricalcitol, 3 times a week for 30 days), and RFR + P (n = 7, 1800 MHz RFR 1 h/day for 30 days +0.2 μg/kg paricalcitol, 3 times a week for 30 days). Testicular tissue was evaluated with histological and biochemical methods. No statistically significant differences were detected between the groups in seminiferous tubule diameters and germinal epithelial thicknesses. While ultrastructural changes were observed in the seminiferous tubule and Leydig cells in the RFR group, these changes were decreased in the RFR + P group. It was found that the Johnsen Score, Ki67, and p63 immunoreactivity scores (IRS), superoxide dismutase (SOD), and catalase (CAT) activities in the RFR + P group were statistically increased as compared to the RFR group and the malondialdehyde (MDA) levels were decreased statistically and significantly. These results show that paricalcitol administration may have an ameliorative effect on testicular damage occurring because of 1800 MHz RFR exposure.

P11.55.B ROLE OF FRAILTY IN THE MANAGEMENT OF PATIENTS WITH GLIOBLASTOMA: A RETROSPECTIVE MULTICENTER STUDY

Abstract BACKGROUND Frailty is a state characterized by reduced physiological reserve and loss of resistance to stressors caused by accumulated age-related deficits. It has been associated with increased morbidity and is a strong predictor of mortality. Few data show the relationship of frailty with adverse outcomes and survival after elective craniotomy. Identification of risk factors that distinguish surgical candidates at elevated risk of peri-operative morbidity or mortality is imperative for informed surgical decision-making and post-operative radio and chemotherapy. MATERIAL AND METHODS This multicenter ongoing study is an analysis of a retrospective cohort, including 800 patients. We have currently analyzed the correlations in 364 patients underwent elective craniotomy for glioblastoma between July 2015 and October 2019. Frailty was assessed using validated scale, defined by the frailty index constructed according to the standard methodology, that includes age, diabetes, functional status, chronic obstructive pulmonary disease, congestive heart failure, hypertension, renal failure, antiplatelet drugs, liver diseases. Patients were classified as not frail (score of ≤1) and moderately-severely frail (score ≥2). Kaplan-Meier estimates and multivariable Cox proportional hazard models were used to evaluate the prognostic value adjusted for clinically relevant variables, including age, sex, average length of stay, pre and post-operative KPS, Ki-67, and ASA score. RESULTS The duration of follow-up from surgery was 50 months. The median age was 62 years (IQR, 54-71). 283 patients (80.63%) were not frail, and 68 patients (19.37%) were moderately-severely frail. From the overall survival curve, patients' 1-, 2-, and 3-year survival rates were about 51.4%, 19.0%, and 8.2%, respectively. Cox regression analysis revealed a significant association between age (HR, 1.02; 95% confidence interval [CI], 1.01-1.03; p = 0.003) and post KPS (HR, 0.98; 95% CI, 0.97-0.99; p = 0.001) with survival. No association between frailty, survival and length of stay, was found (HR, 1.19; 95% CI, 0.88-1.62; p = 0.26). CONCLUSION From preliminary statistical analysis, frailty is not associated with overall survival and length of stay and does not contraindicate neurosurgical treatment. From the statistical analyses we propose to define subgroups of patients who experienced complications related to surgery, characteristics of tumor (site, volume, extent of resection), or for cofactors related to hospitalization. We can improve “fragile” patient’s planning in pre-operative management and post-operative care protocols and determine «long-term consequences» of surgery. Updating this data could define a more effective clinical practice. Finally, we will be able to develop a novel frailty score capable of predicting mortality and severe morbidity outcomes after neurosurgical treatment.

Expression of programmed death-ligand 1, IRF1 and CD8 T lymphocyte infiltration in a primary subset of breast cancer patients in Sudan

ObjectivesThis study aimed to investigate the protein expression of programmed death ligand 1 (PD-L1) in breast cancer (BC) tissues and link this data with estrogen status, the expression of interferon regulatory factor1 (IRF-1), and CD8+T lymphocyte infiltration by immunohistochemistry (IHC). We also attempted to identify the association between PD-L1 expression, the cell proliferation index marker (Ki67), and lymph node involvement. MethodsOne hundred and fifty formalin-fixed and paraffin-embedded (FFPE) blocks of breast tissue were acquired from Sudanese females via The National Public Health Laboratory. FFPE blocks were subjected to antigen/antibody detection by IHC with antibodies raised against PD-L1, IRF1, and CD8. These data were analyzed alongside data extracted from medical records relating to estrogen receptor (ER) status, Ki67 index, and lymph node (LN) status. ResultsIHC analysis revealed a significant association between PD-L1 and CD8 (p = 0.010). In addition, regression analysis indicated the ability of IRF1 to induce PD-L1 expression levels in IRF1-positive cases that were two-fold higher than IRF1-deficient cases (odds ratio [OR]: 2.441 p = 0.035). Analysis also suggested that PD-L1 exerts impact on cell proliferation, as reflected by the Ki67 index. An independent t test showed that higher Ki67 scores were more frequent among PD-L1-positive patients than in PD-L1-negative patients (t = 2.608 p = 0.014). There was an inverse association between PD-L1 and ER status; ER-positive tumors exhibited negative PD-L1 expression and vice versa (p = 0.04). Furthermore, we investigated the prognostic value of PD-L1 by evaluating the association between PD-L1 and LNs dispersed variably with tumor cells; there was no statistically significant relationship between these factors (p > 0.05). ConclusionThe expression of PD-L1 and IRF-1, along with the infiltration of CD8, represents a potent panel of biomarkers with which to identify BC patients with the highest probabilities of achieving an excellent response to immune therapy, particularly when taking ER status into account, as ER expression levels are known to be high when immune checkpoint blockers (ICBs) generate a poor response.

Neoadjuvant palbociclib plus either giredestrant or anastrozole in oestrogen receptor-positive, HER2-negative, early breast cancer (coopERA Breast Cancer): an open-label, randomised, controlled, phase 2 study

The development of more potent selective oestrogen receptor antagonists and degraders (SERDs) that can be orally administered could help to address the limitations of current endocrine therapies. We report the primary and final analyses of the coopERA Breast Cancer study, designed to test whether giredestrant, a highly potent, non-steroidal, oral SERD, would show a stronger anti-proliferative effect than anastrozole after 2 weeks for oestrogen receptor-positive, HER2-negative, untreated early breast cancer. In this open-label, randomised, controlled, phase 2 study, postmenopausal women were eligible if they were aged 18 years or older; had clinical T stage (cT)1c to cT4a-c (≥1·5 cm within cT1c) oestrogen receptor-positive, HER2-negative, untreated early breast cancer; an Eastern Cooperative Oncology Group performance status of 0-1; and baseline Ki67 score of at least 5%. The study was conducted at 59 hospital or clinic sites in 11 countries globally. Participants were randomly assigned (1:1) to giredestrant 30 mg oral daily or anastrozole 1 mg oral daily on days 1-14 (window-of-opportunity phase) via an interactive web-based system with permuted-block randomisation with block size of four. Randomisation was stratified by cT stage, baseline Ki67 score, and progesterone receptor status. A 16-week neoadjuvant phase comprised the same regimen plus palbociclib 125 mg oral daily on days 1-21 of a 28-day cycle, for four cycles. The primary endpoint was geometric mean relative Ki67 score change from baseline to week 2 in patients with complete central Ki67 scores at baseline and week 2 (window-of-opportunity phase). Safety was assessed in all patients who received at least one dose of study drug. The study is registered with ClinicalTrials.gov (NCT04436744) and is complete. Between Sept 4, 2020, and June 22, 2021, 221 patients were enrolled and randomly assigned to the giredestrant plus palbociclib group (n=112; median age 62·0 years [IQR 57·0-68·5]) or anastrozole plus palbociclib group (n=109; median age 62·0 [57·0-67·0] years). 15 (7%) of 221 patients were Asian, three (1%) were Black or African American, 194 (88%) were White, and nine (4%) were unknown races. At data cutoff for the primary analysis (July 19, 2021), the geometric mean relative reduction of Ki67 from baseline to week 2 was -75% (95% CI -80 to -70) with giredestrant and -67% (-73 to -59) with anastrozole (p=0·043), meeting the primary endpoint. At the final analysis (data cutoff Nov 24, 2021), the most common grade 3-4 adverse events were neutropenia (29 [26%] of 112 in the giredestrant plus palbociclib group vs 29 [27%] of 109 in the anastrozole plus palbociclib group) and decreased neutrophil count (17 [15%] vs 16 [15%]). Serious adverse events occurred in five (4%) patients in the giredestrant plus palbociclib group and in two (2%) patients in the anastrozole plus palbociclib group. There were no treatment-related deaths. One patient died due to an adverse event in the giredestrant plus palbociclib group (myocardial infarction). Giredestrant offers encouraging anti-proliferative and anti-tumour activity and was well tolerated, both as a single agent and in combination with palbociclib. Results justify further investigation in ongoing trials. F Hoffmann-La Roche.

Whole transcriptome analysis of canine pheochromocytoma and paraganglioma.

Pheochromocytomas and paragangliomas (PPGLs) are neuroendocrine tumors arising from the chromaffin cells in the adrenal medulla and extra-adrenal paraganglia, respectively. Local invasion, concurrent disorders, and metastases prevent surgical removal, which is the most effective treatment to date. Given the current lack of effective medical treatment, there is a need for novel therapeutic strategies. To identify druggable pathways driving PPGL development, we performed RNA sequencing on PPGLs (n = 19) and normal adrenal medullas (NAMs; n = 10) of dogs. Principal component analysis (PCA) revealed that PPGLs clearly clustered apart from NAMs. In total, 4,218 genes were differentially expressed between PPGLs and NAMs. Of these, 232 had a log2 fold change of >3 or < -3, of which 149 were upregulated in PPGLs, and 83 were downregulated. Compared with NAMs, PPGLs had increased expression of genes related to the cell cycle, tumor development, progression and metastasis, hypoxia and angiogenesis, and the Wnt signaling pathway, and decreased expression of genes related to adrenal steroidogenesis. Our data revealed several overexpressed genes that could provide targets for novel therapeutics, such as Ret Proto-Oncogene (RET), Dopamine Receptor D2 (DRD2), and Secreted Frizzled Related Protein 2 (SFRP2). Based on the PCA, PPGLs were classified into 2 groups, of which group 1 had significantly higher Ki67 scores (p = 0.035) and shorter survival times (p = 0.04) than group 2. Increased expression of 1 of the differentially expressed genes between group 1 and 2, pleiotrophin (PTN), appeared to correlate with a more aggressive tumor phenotype. This study has shed light on the transcriptomic profile of canine PPGL, yielding new insights into the pathogenesis of these tumors in dogs, and revealed potential novel targets for therapy. In addition, we identified 2 transcriptionally distinct groups of PPGLs that had significantly different survival times.

Relationship between the change in tumour-infiltrating lymphocyte level and residual tumour after neoadjuvant chemotherapy in patients with locally advanced breast cancer

To evaluate the tumour-infiltrating lymphocyte (TIL) rates in breast tissue before and after neoadjuvant chemotherapy (NAC) and their impact on survival, eighty-four patients with locally advanced breast cancer (LABC) were assessed. Pre- and post-NAC TIL levels were determined using biopsy and surgical specimens, respectively. The median TIL rate was significantly different before (17.5%) and after (5%) NAC. Pre- and postoperative Ki-67 index, molecular subtype, pre- and post-NAC TIL concentration, and preoperative residual-cancer-burden TIL were significantly associated with pathological complete response (pCR). Specifically, higher pre-NAC TIL levels were associated with higher pCR rates. Postoperative Ki-67 index and pCR, and postoperative Ki-67 index were significant predictors of disease-free (DFS) and overall survival, respectively. The independent prognostic factors for DFS were postoperative Ki-67 score (hazard ratio [HR]: 6.16; p = 0.012), post-NAC TIL score (HR: 0.42; P = 0.041), and pCR (HR: 0.10; P = 0.038). Our study confirms that higher pre-NAC and lower postoperative TIL levels may be surrogate factors for longer DFS, and postoperative TIL rate may predict post-NAC pCR in patients with LABC.

Diagnostic and prognostic values of flow cytometry in diffuse large B-cell lymphoma with bone marrow involvement

Objective: To analyze the diagnostic and prognostic values of flow cytometry (FC) in diffuse large B cell lymphoma (DLBCL) with bone marrow involvement (BMI). Methods: The clinical data of 412 patients with newly diagnosed DLBCL, including 243 males and 169 females, aged 64 (28-92) years old, in the Department of Hematology at Peking University Third Hospital from December 2012 to June 2022 were retrospectively analyzed. All patients underwent bone marrow biopsy (BMB) and bone marrow FC. The patients with BMI by FC were further detected by fluorescence in situ hybridization (FISH) for gene analysis. The positive rates and consistency of BMI detected by BMB and FC were evaluated. According to the results of BMB and FC examinations, all patients were divided into four groups: the BMB+FC+group (115 cases), the BMB-FC+group (50 cases), the BMB+FC-group (8 cases, the results did not include in statistical analysis because of small sample size), and the BMB-FC-group (239 cases). The clinical features, treatment response rates, 5-year survival rates, and immunophenotype characteristics by FC in different groups were analyzed. Results: Among the 412 patients with DLBCL, the positivity rates of BMB and FC for BMI detection were 29.9% (123/412) and 40.0% (165/412), respectively. Good consistency between BMB and FC was found (Kappa=0.841, P=0.001). The numbers of extranodal involvement≥2, splenomegaly, huge mass, higher Ki-67 score, higher international prognostic index (IPI) score, thrombocytopenia, and elevated lactate dehydrogenase level were more prevalent in the BMB+FC+group than those in the BMB-FC+group and the BMB-FC-group (all P<0.05). The treatment response rate in BMB+FC+group was 63.5% (73/115), which was lower than those in BMB-FC+group (88.0%, 44/50, P=0.048) and BMB-FC-group (90.0%, 215/239, P=0.032), respectively. The 5-year overall survival rates in three groups were (53.6±9.7) %, (72.5±8.6) %, and (75.2±7.6) %, respectively, with a statistically significant difference (P=0.037). According to the FISH results of bone marrow, 102 cases were diagnosed as not otherwise specified (NOS), 48 cases were diagnosed as double hit lymphoma (DHL), and 15 cases were diagnosed as triple hit lymphoma (THL). Compared with NOS subtypes, the tumor cells in DHL or THL subtypes had higher proportion of increased side scatter (SSC), higher positive rates of CD10 expression, CD38 strong expression and CD56 expression, and lower proportion of surface immunoglobulin light chain restriction (all P<0.05). Conclusions: FC is well consistent with BMB in diagnosing DLBCL with BMI. Combined with FISH detection, FC can contribute to the auxiliary diagnosis and risk stratification for DHL and THL, and provide reference for the prognostic evaluation in DLBCL with BMI.