Percentage Of Ki-67 Research Articles

Association between Radiological and First-Order Statistical Features of the Mammogram, and the Tumor Phenotype in Breast Cancer Patients

Background: Breast cancer is among the most prevalent cancers which can effectively be screened by mammography. The spatial distribution of grey levels of the mammogram known as first-order statistical features (FOSFs) contain higher dimensional data which describe the breast composition. We aim to test these basic measures to differentiate density categories in breast cancer patients, and use them as covariates to investigate the relationship between radiologic and pathologic features of the tumor. Methods: Data from Eighty-five breast cancer subjects, their BI-RADS breast density category (a to d), percentage density (PD), and FOSFs of the mammogram including median, mean, interquartile range, kurtosis, maximum, minimum, standard deviation, skewness, and energy, were extracted. The tumor grade and the percentage of Ki67, ER, PR, and Her2 status were recorded. A linear discriminant analysis, and a support vector machine (SVM) were used to discriminate each density category from others. Then, the relation between variables were investigated using ANCOVA and regression analysis. Results: Density categories a and d were classified by SVM with high accuracy. The key feature of a and d were interquartile range and maximum intensities respectively. Reported tumor margins were related to Her2 overexpression and PR positivity. Spiculated tumor margin predicted percentage of PR expression, with a cumulative odds ratio of 7.85 (CI 2.5- 24.78), when adjusted for age, area of breast, density, and FOSFs, (p=0.0004). Conclusion: The findings of this study suggest that FOSFs can be incorporated in computer-aided systems to adjust for differences in breast composition and to refine risk profiles.

RE-VISITING BIOPSIES FOR SUSPECTED CNS LYMPHOMAS: TO BIOPSY OR NOT TO BIOPSY?

Abstract AIMS It is a common practice to get a histological diagnosis before treatment of suspected CNS (central nervous system) lymphoma. Should CNS lesions be routinely biopsied in patients with background systemic lymphoma? METHOD A 10-year retrospective review of biopsy-proven CNS lymphoma was conducted. Data was analyzed using SPSS for Windows, version 25 (SPSS, Inc., Chicago, Illinois, USA), and a p-value < 0.05 was considered significant for the appropriate test statistic. RESULTS Patients with background systemic lymphoma whose brain MRI appearances were in favour of lymphoma after MDT discussion all had histologically-proven lymphoma after brain biopsy. Pre-biopsy MDT diagnosis was congruent with post-biopsy histology in 87.6% of cranial lesions (p-value 0.003) and 40% of spinal lymphomas (p-value 0.002). Secondary CNS lymphoma was seen in 21.2%. Stereotactic brain biopsy was the most common biopsy technique for cranial lesions (79.8%). Histology was Diffuse B Cell Lymphoma in 93% of cases. The mean Ki67 percentage for cranial lymphomas was higher than that of spine (p-value 0.028) The complication rate for cranial biopsy was 8.9%. Transient motor deficits (4.4%) and post-operative haematoma (2.2%) were the most common. The 24-hour post-brain biopsy mortality rate was 1.1%, while overall 30-day post-op mortality rate was 4.4%. The mean percentage of Ki67 was higher among patients who had complications (p-value 0.001). CONCLUSION In a setting of properly constituted neurooncology MDT, a sub-set of patients with suspected brain lymphoma and background systemic lymphoma might not require brain biopsy. KEY WORDS Lymphoma, Brain, Spine, Biopsy, Complication

Abstract C064: Mapping cellular plasticity through the lens of FRA1 during Kras G12D mediated pancreatic acinar to ductal metaplasia

Abstract Introduction: Acinar cells have been proposed as a cell-of-origin for pancreatic ductal adenocarcinoma (PDAC) after undergoing acinar to ductal metaplasia (ADM). ADM can be triggered by pancreatitis causing acinar cells to de-differentiate to a ductal-like state. We identify FRA1 (gene name Fosl1) as the most active transcription factor during Kras G12D acute pancreatitis-mediated injury. Herein, we explore the role of the FRA1 in the onset of ADM and neoplastic transformation. Methods: We generated tamoxifen (TAM)-inducible Ptf1aCre ERT ;Kras G12D ; Rosa26 YFP/YFP (TAM-KCY Fosl1 WT ) mice and bred in the Fosl1 fl/fl allele to create TAM- KCY Fosl1 KO mice and subjected them to caerulin (CCK analog) induced acute pancreatitis. First, we performed a time course of hematoxylin and eosin (H&E) images from mice either with Fosl1 WT or Fosl1 KO after induction of acute pancreatitis to understand the functional role of FRA1 during acinar de-differentiation. Next, after snap-freezing pancreata at the Day 5 post-caerulein timepoint and extracting nuclei, we sorted YFP+ nuclei for snATAC sequencing and bulk-RNA sequencing. We developed a pseudotime model and gene-regulatory network (GRN) to understand the dynamic landscape as cells transitioned from normal acinar to late ADM. Finally, to determine the functional role of in tumorigenesis, we bred an inducible mutant p53 R172H gain- of-function allele into TAM-KCY Fosl1 WT/KO mice to create TAM-KPCY Fosl1 WT/KO mice. These mice were subjected to the same tamoxifen and acute pancreatitis protocol as TAM-KCY Fosl1 WT/KO mice and monitored for tumor growth and overall survival. Results: A time course of H&E images demonstrated that FRA1 depletion slowed progression to maximum ADM in the first 5 days after caerulein treatment. We additionally observed a dramatic reduction in the amount of PanIN lesions formed between TAM-KCY Fosl1 WT and TAM-KCY Fosl KO mice, which occurred on day 7 in this caerulein model. Pseudotime analysis proposed a time point when FRA1 becomes active to prime ADM cells for recovery from inflammation and simultaneously undergo PanIN transformation. Furthermore, GRN analysis validated the differentially active transcription factors identified by pseudotime analysis of snATAC-seq. To elaborate on further timepoints, we observed a delay in the progression of TAM-KCY Fosl KO PanIN lesions at later Day 21, 8-week and 12-week timepoints. In the survival analysis, TAM-KPCY Fosl1 WT mice had a median survival time of 25.86 weeks, while TAM-KPCY Fosl1 KO mice survived for almost twice as long with a median survival time of 40.57 weeks. Histologically, the percentage of Ki-67- positive cells in TAM-KPCY Fosl1 WT tumors was significantly higher than in TAM-KPCY Fosl1 KO tumors, suggesting the loss of FRA1 reduce tumor aggressiveness. Conclusions: Our findings reveal that FRA1 is a critical mediator of ADM progression to PanIN and PDAC. Fosl1 genetic depletion can alter the kinetics of ADM formation and lead to a decrease in PanIN and eventual malignant transformation. Citation Format: Alina Li, Noriyuki Nishiwaki, Kensuke Sugiura, Kensuke Suzuki, Constanza Nicole Tapia Contreras, Dorsay Sadeghian, Anirban Maitra, Jason R Pitarresi, Rohit Chandwani, Peter A Sims, Anil K Rustgi. Mapping cellular plasticity through the lens of FRA1 during Kras G12D mediated pancreatic acinar to ductal metaplasia [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C064.

The oculomotor cistern and pituitary adenomas: anatomical and clinical study.

The oculomotor cistern (OMC) is a meningeal cuff filled with CSF that contains the oculomotor nerve (cranial nerve [CN] III) at the level of the lateral wall of the cavernous sinus. Only a few studies have investigated the involvement of the OMC by pituitary adenomas (pituitary neuroendocrine tumors [PitNETs]), mainly with relatively small case series. The aim of this study was to perform a histomorphological description of the OMC and systematically analyze its involvement by PitNETs from radiological, clinical, and surgical perspectives. Ten hemisellae from formalin-fixed specimens were studied with 3-µm sections. Digital image analysis software was used for morphological and quantitative assessments. Clinical, radiological, surgical, and histological data of patients undergoing endoscopic transsphenoidal surgery for PitNETs at the University of Brescia, Italy, between 2014 and 2021 were recorded. OMC involvement was graded as not compressed, compressed, and invaded. The same surgical team operated on all patients. The OMC had an elliptical shape with an average area of 3.1 mm2 and a length of 5.5 mm. No cisternal points of weakness were recognized in the histomorphological study. Of 315 patients, 246 had complete data: apoplexy and CN III palsy were documented in 6.9% and 8.5%, respectively. OMC compression and invasion were recorded in 106 (43.1%) and 23 (9.3%) patients. Significant associations between OMC involvement and PitNET dimensions (p < 0.001), Knosp grade (p < 0.001), preoperative oculomotor palsy (p < 0.001), Ki-67 percentage (p = 0.009), and recurrence/progression of residual tumor (p = 0.008) were found. A new postoperative CN III palsy was evident in 2%: transient in 4 cases, and persistent in 1 patient treated for a recurrent PitNET who experienced a local infection complication. Preoperative CN III palsy improved in 10 cases. Significant OMC involvement by PitNETs might be underrecognized, but it can be treated using the endoscopic transsphenoidal approach, and it affects patient outcomes.

Immuno-hormonal regulation of tumor proliferation in breast cancer patients

It is well known that tumor cells proliferation is regulated by sex steroid hormones, estradiol and progesterone (E2 and Pg) in breast cancer patients (BCP). Moreover, specific auto-antibodies to estrogen receptor (ERα) were detected in blood serum of BCP. Their levels positively correlated with the percentage of Ki-67 expressing breast cancer cells. We proposed that antiidiotypic auto-antibodies to E2 and Pg (IgG2-E2 and IgG2-Pg) could act as antibodies against membrane ER and progesterone receptor (PR). Our study aimed for research of IgG2-E2 and IgG2-Pg according to E2 and Pg serum levels in association with Ki-67 positive tumors in BCP. Antiidiotypic antibodies were studied in ER-positive patients with breast cancer (stage I, n = 374, and stage II-IV, n = 379,) using ELISA technique with monoclonal antibodies against E2 and Pg as adsorbed antigens. Blood serum concentrations of E2 and Pg were measured using “ImmunoEA-Estradiol” and “ImmunoEA-Progesterone” test-systems (“Immunotech”, Russia). Tumor Ki-67 was studied by standard immunohistochemical technique at the Kemerovo Oncological Hospital. Higher percentage of Ki-67 positive breast cancer cells (Ki-67 30) was increased in BCP II-IV stages compared with stage I patients (50.7% vs 29.8%, p 0,001). Such increased values were detected for the BCP with low levels of both IgG2-E2 and IgG2-Pg antibodies in the following subgroups: 1) at low serum E2 concentration of ≤ 200 pmol/L (50.9% vs 21.3%, р 0.001); 2) at the E2 exceeding 200 pmol/L (74.2% vs 34.1%, р = 0.003); 3) at the Pg levels under 600 pmol/L (60.6% vs 22.2%, р 0.001); 4) at Pg values exceeding 600 pmol/L (58.5% vs 30.8%, р = 0.02). Similar differences were not revealed between stage II-IV and stage I BCP with low levels of both IgG2-E2 and IgG2-Pg. Corresponding Ki-67 30 indices were as follows: 1) 36.9% vs 22.0% (р = 0.14); 2) 48.4% vs 34.5% (р = 0.08); 3) 34.0% vs 27.7% (р = 0.75). Significant differences were detected in BCP with Pg 600 pmol/L and high IgG2-E2 and IgG2-Pg levels only: 48.1% vs 26.6%, (р = 0.01). Hence, antiidiotypic auto-antibodies to steroid hormones may participate in regulation of tumor proliferation in BCP. Immunoassay of IgG2-E2 and IgG2-Pg may be used for prognosis of tumor proliferation upon breast cancer progression.

NCAPD2 augments the tumorigenesis and progression of human liver cancer via the PI3K‑Akt‑mTOR signaling pathway.

Non‑SMC condensin I complex subunit D2 (NCAPD2) is a newly identified oncogene; however, the specific biological function and molecular mechanism of NCAPD2 in liver cancer progression remain unknown. In the present study, the aberrant expression of NCAPD2 in liver cancer was investigated using public tumor databases, including TNMplot, The Cancer Genome Atlas and the International Cancer Genome Consortium based on bioinformatics analyses, and it was validated using a clinical cohort. It was revealed that NCAPD2 was significantly upregulated in liver cancer tissues compared with in control liver tissues, and NCAPD2 served as an independent prognostic factor and predicted poor prognosis in liver cancer. In addition, the expression of NCAPD2 was positively correlated with the percentage of Ki67+ cells. Finally, single‑cell sequencing data, gene‑set enrichment analyses and in vitro investigations, including cell proliferation assay, Transwell assay, wound healing assay, cell cycle experiments, cell apoptosis assay and western blotting, were carried out in human liver cancer cell lines to assess the biological mechanisms of NCAPD2 in patients with liver cancer. The results revealed that the upregulation of NCAPD2 enhanced tumor cell proliferation, invasion and cell cycle progression at the G2/M‑phase transition, and inhibited apoptosis in liver cancer cells. Furthermore, NCAPD2 overexpression was closely associated with the phosphatidylinositol 3‑kinase (PI3K)‑Akt‑mammalian target of rapamycin (mTOR)/c‑Myc signaling pathway and epithelial‑mesenchymal transition (EMT) progression in HepG2 and Huh7 cells. In addition, upregulated NCAPD2 was shown to have adverse effects on overall survival and disease‑specific survival in liver cancer. In conclusion, the overexpression of NCAPD2 was shown to lead to cell cycle progression at the G2/M‑phase transition, activation of the PI3K‑Akt‑mTOR/c‑Myc signaling pathway and EMT progression in human liver cancer cells.

Ki67 is a better marker than PRAME in risk stratification of BAP1-positive and BAP1-loss uveal melanomas

BackgroundAccurate risk stratification of uveal melanoma (UM) patients is important for determining the interval and frequency of surveillance. Loss of BAP1 expression has been shown to be strongly associated with...

Screening optimal candidates with operable, early-stage triple-negative breast cancer benefitting from capecitabine maintenance: A post-hoc analysis of the SYSUCC-001 study

BackgroundTo explore whether specific clinicopathological covariates are predictive for a benefit from capecitabine maintenance in early-stage triple-negative breast cancer (TNBC) in the SYSUCC-001 phase III clinical trial. MethodsCandidate covariates included age, menstrual status, type of surgery, postoperative chemotherapy regimen, Ki-67 percentage, histologic grade, primary tumor size, lymphovascular invasion, node status, and capecitabine medication. Their nonlinear effects were modeled by restricted cubic spline. The primary endpoint was disease-free survival (DFS). A survival prediction model was constructed using Cox proportional hazards regression analysis. ResultsAll 434 participants (306 in development cohort and 128 in validation cohort) were analyzed. The estimated 5-year DFS in development and validation cohorts were 77.8 % (95 % CI, 72.9%–82.7 %) and 78.2 % (95 % CI, 70.9%–85.5 %), respectively. Age and node status had significant nonlinear effects on DFS. The prediction model constructed using four covariates (node status, lymphovascular invasion, capecitabine maintenance, and age) demonstrated satisfactory calibration and fair discrimination ability, with C-index of 0.722 (95 % CI, 0.662–0.781) and 0.764 (95 % CI, 0.668–0.859) in development and validation cohorts, respectively. Moreover, patient classification was conducted according to their risk scores calculated using our model, in which, notable survival benefits were reported in low-risk subpopulations. An easy-to-use online calculator for predicting benefit of capecitabine maintenance was also designed. ConclusionsThe evidence-based prediction model can be readily assessed at baseline, which might help decision making in clinical practice and optimize patient stratification, especially for those with low-risk, capecitabine maintenance might be a potential strategy in the early-disease setting.

Abstract PO1-14-12: Quantitative Biomarkers and 21-Gene Assay Results Predict Breast-Conserving Surgery Following Neoadjuvant Therapy in Early-Stage, Hormone Receptor-Positive, HER2-Negative Breast Cancer

Abstract Background: Neoadjuvant chemotherapy (NACT) and neoadjuvant endocrine therapy (NET) are often used to downstage tumors and/or enable breast-conserving surgery (BCS) in patients (pts) with early-stage, hormone receptor-positive/HER2-negative (HR+/HER2-) breast cancer. Given the increasing use of neoadjuvant therapy for HR+ breast cancer, studies are needed to better quantify the success of NACT/NET in downstaging tumors as a function of estrogen (ER), progesterone (PR), and Ki-67 expression, and 21-gene recurrence score (RS) in this patient population. Methods: Data were from pts with stage I-III, HR+/HER2- breast cancer diagnosed from 2010-2020 in the National Cancer Database (NCDB). Quantitative ER, PR, and Ki-67 data became available in 2018. We included pts who received either NACT or NET, with surgical plan categorized as mastectomy or BCS. Pts who received NACT for 12–30 weeks or NET for 4–36 months prior to surgery were eligible. Predicted rates of BCS by quantitative biomarkers (ER, PR, Ki-67 percentage, and RS) were estimated using restricted cubic spline logistic regression. Multivariable logistic regression models were fit to examine the associations between surgical plan and quantitative biomarkers for the two neoadjuvant cohorts. Adjusted odds ratios (aOR) and 95% confidence intervals (CI) were calculated per 10% increase in percentages of ER, PR, and Ki-67 expression. Results: Of 44,589 pts treated with NACT, the mean age was 53 years, the median amount of time they were on NACT before surgery was 21 weeks, and 65% had cT1-2 disease. Of 10,466 pts treated with NET, the mean age was 68 years, the median amount of time they were on NET before surgery was 6 months, and 76% had cT1-2 disease. Overall, 42% of pts underwent BCS after NACT; while 64% did after NET. In the NACT cohort, 13,752 of the pts with available quantitative biomarker data were included in regression analyses; in the NET cohort, 4,003 were included. After adjusting for demographic and clinicopathologic factors, increasing ER% (aOR=0.96, 95% CI: 0.94–0.97) and PR% (aOR=0.98, 95% CI: 0.96–0.99) were associated with lower odds of BCS after NACT. Increasing Ki-67% was associated with greater odds of BCS after NACT (aOR=1.07, 95% CI: 1.04–1.10). Pts with a low (aOR=0.50, 95% CI: 0.29–0.88) or intermediate (aOR=0.58, 95% CI: 0.41–0.81) RS were significantly less likely than pts with a high RS to undergo BCS after NACT. Asian pts were less likely than White pts to have BCS (aOR=0.75, 95% CI: 0.56–0.99), while rates of BCS after NACT were similar to pts from other racial and ethnic groups. In the NET cohort, increasing ER% was associated with greater odds of BCS (aOR=1.09, 95% CI: 1.01–1.17). There was no significant association between baseline PR% or Ki-67% and BCS after NET. Pts with a low (aOR=0.92, 95% CI: 0.60–1.43) or intermediate (aOR=0.96, 95% CI: 0.65–1.41) RS were numerically less likely than pts with a high RS to have BCS after NET, though not statistically significant. Compared with White pts, Asian pts were less likely to have BCS after NET (aOR=0.60, 95% CI: 0.40–0.91). Conclusions: In this analysis of pts from the NCDB who received neoadjuvant therapy for early-stage, HR+/HER2- breast cancer, we found that BCS after NACT was higher for patients with a high RS or high Ki-67, suggesting that NACT is unlikely to downstage tumors with a low/intermediate RS or low Ki-67. Most pts receiving NET underwent BCS (likely due to a smaller clinical tumor size); BCT after NET was most dependent on ER expression. Asian pts were less likely to undergo BCS after either NACT or NET, which is consistent with previous reports. These data could facilitate appropriate neoadjuvant treatment selection based on tumor biology and improve patient counseling on the likelihood of successful BCS. Citation Format: Jincong Freeman, Sarah Shubeck, Nan Chen, Rita Nanda, Dezheng Huo, Frederick Howard. Quantitative Biomarkers and 21-Gene Assay Results Predict Breast-Conserving Surgery Following Neoadjuvant Therapy in Early-Stage, Hormone Receptor-Positive, HER2-Negative Breast Cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-14-12.

Abstract PO5-06-05: DISCORDANCE BETWEEN HORMONE RECEPTORS AND HER2 STATUS IN BREAST CANCER PATIENTS RELAPSE

Abstract Introduction: Breast cancer (BC) is a heterogeneous disease, and during its progression, the tumor phenotype can undergo changes that are often associated with a poor prognosis. In this study, we aim to compare the discordance in estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER-2), and Ki-67 expression between primary and recurrent/metastatic lesions. Methods: This study is a retrospective analysis of 132 non-advanced breast cancer patients who experienced confirmed recurrence/metastasis at the Instituto Nacional de Enfermedades Neoplasicas between 2011 and 2022. Descriptive analysis was conducted to assess the expressions of ER, PgR, HER-2, and Ki-67 in primary and metastatic breast cancer, as well as clinical variables. Normality tests, including Kolmogorov-Smirnov and Shapiro-Wilk, were performed. The paired samples Wilcoxon test was used to determine significant differences between primary and recurrent/metastatic tumors. Additionally, Spearman's rank correlation coefficient was employed to evaluate the relationships of expression discordance. Results: Fifty-nine percent of patients presented locoregional recurrence, while 41% presented visceral metastasis. In the primary tumor, ER and PR expression were found in 63.6% and 55.3% of cases, respectively. HER2 status was determined as 0 (36.3%), +1 (22%), +2 (14.4%), +3 (26.5%), and not determined (0.8%) by IHC. For the recurrent/metastatic tumor, ER and PR expression were found in 59% and 34% of cases, respectively. HER2 status was determined as 0 (27.3%), +1 (19.7%), +2 (25%), +3 (25%), and not determined (3%). The median Ki-67 levels were 37% in the primary tumor and 40% in the recurrent/metastatic tumor. Significant differences were found for ER (p=0.014), PR (p&amp;lt; 0.001), and HER2 (p=0.019), but not for Ki-67 percentage (p=0.139). The discordance rates of ER, PR, HER2, and Ki-67 expression were 27.9%, 47.7%, and 26.3%, respectively. Conclussions: Discordance in hormone receptor expression between primary and recurrent/metastatic tumors is a common phenomenon, with ER showing a discordance rate of 27.9% and PR showing a higher rate of 47.7%. HER2 status also displayed significant discordance, with a 26.3% rate of discordant expression between primary and recurrent/metastatic tumors. The findings from this study emphasize the dynamic nature of breast cancer and the importance of comprehensive evaluation of biomarker expression in both primary and recurrent/metastatic tumors, to choose the best treatment options. Citation Format: Katia Roque, Iris Otoya, Natalia Valdivieso, Zaida Morante, Hugo Fuentes, Silvia Neciosup, Henry Gómez, Jorge Cotito, Gonzalo Ziegler, Estefania Peña, Tatiana Vidaurre, Ramon Andrade de Mello, Carlos Castañeda. DISCORDANCE BETWEEN HORMONE RECEPTORS AND HER2 STATUS IN BREAST CANCER PATIENTS RELAPSE [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-06-05.

Abstract PO4-03-10: HER2 Expression in Triple-Negative Breast Cancer: Clinicopathologic Features and Response to Neoadjuvant Chemotherapy

Abstract BACKGROUND: Half of the primary breast cancers reported as HER-2 negative have low HER-2 expression, defined as an immunohistochemical (IHC) score of 1+ or 2+, with negative in situ hybridization. Among these cases, 37% correspond to the triple-negative immunophenotype. The primary objective of this study was to evaluate HER-2 expression in patients with triple-negative breast cancer and compare the clinicopathologic features and response to neoadjuvant chemotherapy between HER-2 negative (0) and HER-2 low (1+/2+) subtypes. METHODS: We retrospectively analyzed 157 patients with early-stage triple-negative breast cancer who underwent neoadjuvant chemotherapy followed by surgery with curative intent and met the study's inclusion criteria. These patients were treated at the National Institute of Neoplastic Diseases between 2010 and 2021. HER-2 analysis was performed using immunohistochemistry (IHC), and in cases of HER-2 (2+) amplification, in situ hybridization was performed. RESULTS: Of all the included patients, 107 were classified in the HER-2 negative group (68%), and 50 were classified in the HER-2 low group (32%). Seventy-four percent of patients with HER-2 negative were in the age range of 40-64 years, compared to 68% in the HER-2 low group. A significant difference was found between the HER2-negative and HER2-low groups regarding age and Ki67. The median age of HER-2 low patients is significantly higher than the median age of HER-2 negative patients (52 years vs. 45 years, respectively). No significant association was found between HER-2 status and clinical stages or histological grade. However, the median Ki67 percentage of HER-2 negative patients is significantly higher than the median Ki67 of HER-2 low patients (70% vs. 50%, respectively (p=0.035)). Thirty-two percent of patients with HER-2 negative achieved a pathological complete response (pCR) compared to 18% of patients with HER-2 low. There were no differences in the pCR rate in the HER-2 1+ and 2+ subgroups. Among the patients included in both groups, who achieved pCR, no disease progression was observed at the time of analysis. CONCLUSIONS: Patients in the HER-2 negative group had a higher rate of complete pathological response to neoadjuvant chemotherapy compared to those in the HER-2 low group. No differences were observed in the pCR rate between HER-2 1+ and 2+ subgroups, suggesting that HER2 expression levels within the low range may not significantly impact treatment response. Table. Table. Citation Format: Zaida Morante, Iris Otoya, Natalia Valdivieso, Cindy Calle, Hugo Fuentes, Silvia Neciosup, Carlos Castañeda, Tatiana Vidaurre, Martin Falla, Valeria Colomo, Henry Gómez. HER2 Expression in Triple-Negative Breast Cancer: Clinicopathologic Features and Response to Neoadjuvant Chemotherapy [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-03-10.

Pathological proliferation: a potential mechanism for poor CD4+ T cell recovery in people living with HIV.

People living with HIV (PLWH) fail to achieve normalization of CD4+ T cell counts and function, especially in immunological non-responders (INRs). The frequencies of Ki67+CD4+ T cells were inversely associated with CD4+ T cell counts in HIV infected patients. Early ART did not normalize CD4+ T cell proliferation. However, the features of the abnormal proliferation CD4+ T cell in INRs are far from known. PLWH were divided into INRs (n= 16) and immunological responders (IRs, n= 53) groups. Mass cytometry was applied to peripheral blood T cells to profile the immune cells and liquid chip technique was used to measure plasma levels of cytokines and chemokines. Correlation analyses were conducted to evaluate associations between the degree of CD4+ T cell proliferation and immune function. The percentage of Ki67+ CD4+ T cells were significant higher in INRs, and we defined these cells with significant higher level of Ki67, as over-proliferating cells. No significant difference of markers' expression (HLA-DR, CD38, CD57, PD-1, PD-L1, CD107a, perforin) was found between INRs and IRs. Compared with naïve CD4+ T cells in INRs, Ki67+ CD4+ T cells exhibited lower levels of CD57 and CD38. Whereas Ki67+ T cells exhibited higher levels of CD38 and CD57 and activation compared with differentiated mature central memory CD4+ T cells and effector memory CD4+ T cells. Ki67+ cells did not show higher levels of senescence and activation compared to certain Ki67- CD4+ central memory T cells in IRs. Furthermore, Ki67+ CD4+ Tcm cells exhibited positive correlations with pro-inflammatory cytokines. We proposed and validated the hypothesis of "pathological proliferation" in INRs: excessive proliferation of CD4+ T cells in INRs may be accompanied by aberrant activation, senescence and loss of immune function. Eventually, such over-proliferating but poor-quality cells in INRs result in incomplete recovery of both CD4+ T cell counts and function. An intervention that enhancing the proliferative capacity or functional ability or both of CD4+ T cell in INRs might therefore be beneficial.

Abstract 4771: Assessment of Ki67 proliferation index in pre-malignant colonic adenomas: Implication for new biomarker testing to predict metachronous disease (The INCISE project)

Abstract Approximately 40% of patients are found to have polyps at bowel screening colonoscopy, which are associated with a higher risk of developing future polyps or colorectal cancer. The British Society of Gastroenterology 2020 (BSG20) guidelines stratify patients into high/low risk of metachronous disease, using the size, dysplasia, and number of polyps found at index colonoscopy. This study aims to explore the prognostic significance of Ki67 proliferation percentages in adenomatous polyps, with metachronous disease. Tissue microarray (TMA) was constructed from 1201 index adenomatous polyps with representative cores of basal and luminal adenomatous epithelia. 67% patients were identified as high risk at index colonoscopy. The median time to detect metachronous polyps was 55 months (range 6-72 months) with 56% of patients developing metachronous polyps by the end of follow-up. TMA was stained for Ki67 using immunohistochemistry. Ki67 proliferation percentages in adenomatous epithelia was quantified using QuPath. Data was split into training (n=830) and test (n=371) datasets. The Ki67 proliferation cutpoint was made using Survmine R package using the training dataset. X2 test was used to study associations with clinicopathological criteria and Kaplan survival analysis was used to assess Ki67 association with metachronous lesions. Ki67 percentages were higher in lumina than in basal adenomatous epithelium (median Ki67 were 63% &amp; 48%; respectively). Total Ki67 percentages were computed by averaging luminal and basal values. Thresholds of 70%, 64% and 59% were employed to classify patients into low/high categories based on luminal, basal or total percentages; respectively. In the training cohort, high luminal, basal or total Ki67 were associated with metachronous lesions (p values 0.032, &amp;lt;0.001 and 0.006; respectively). Other associations were significant between luminal or total Ki67 and location of index polyps (p=0.017 &amp; p=0.044; respectively), histology (p=0.019) and advanced future lesion status (p=0.002 &amp; p&amp;lt;0.001; respectively). In multivariate cox regression analysis, high Ki67 was an independent prognostic factor for metachronous polyps (luminal Ki67 HR=1.28, CI: 1.052-1.560, p=0.014; basal Ki67 HR=1.639, CI: 1.313-2.046, p&amp;lt;0.001; total Ki67 HR=1.344, CI: 1.105-1.635, p=0.003) when adjusted to number of index polyps and BSG20 risk score. BSG20 risk score stratification of patients in terms of metachronous disease was improved when combined with luminal, basal or total Ki67 in the training datasets (all p&amp;lt;0.05). Prediction of metachronous disease by the combined groups was validated in the test cohort for luminal, basal and total Ki67 (all p&amp;lt;0.05). High Ki67 proliferation rates in adenomatous polyps are independently associated with the development of metachronous polyps and may help improve the BSG20 criteria. Citation Format: Gerard P. Lynch, Aula Ammar, Christopher Bigley, Mark Johnstone, Jakub Jawny, Hannah Morgan, Jennifer Hay, Noori Maka, Stephen McSorley, Joanne Edwards. Assessment of Ki67 proliferation index in pre-malignant colonic adenomas: Implication for new biomarker testing to predict metachronous disease (The INCISE project) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4771.

Abstract 2606: Targeting mesothelin on tumor cells with a novel, completely human fusion construct delivering the serine protease granzyme B

Abstract Mesothelin is a cell-surface antigen identified as an excellent cancer-associated target upregulated on numerous tumors, including pancreatic and ovarian cancers, as well as mesotheliomas and lung cancers. Granzyme B is a highly cytotoxic serine protease nominally delivered by immune effector cells to target cells, resulting in induction of cell death pathways that are both caspase-dependent and -independent. We developed a fusion protein (GrB-Fc-SD1) composed of GrB and containing SD1, a novel human single-domain antibody that uniquely targets Region III of the mesothelin glycoprotein. The SD1 and GrB domains are tethered by a human IgG heavy chain fragment which enables dimerization. The fusion protein was transiently expressed in HEK-293E cells in serum-free conditions with a final yield of 5 mg/L. Western blot analysis confirmed the expression of this construct as a homodimer. The GrB enzymatic activity in GrB-Fc-SD1 was similar to that of commercially available human granzyme B. ELISA studies confirmed the specific recognition of the SD1 antibody to mesothelin extra-cellular domain. Biacore analysis confirmed binding of GrB-Fc-SD1 with a dissociation constant between 10−7 and 10−8 M. In vitro cytotoxicity against a limited panel of lung, pancreatic and ovarian cancer cell lines expressing surface mesothelin indicate cytotoxicity in the nanomolar range, while cytotoxicity against mesothelin-negative cells was at micromolar levels. Ex vivo serum stability studies showed excellent stability of GrB-Fc-SD1 at both 4°C and -20°C over ninety days and a half-life of approximately three days at elevated temperatures (37°C). In vivo efficacy studies against Capan-2 xenografts in nude mice using both weekly x 5 and QOD x 5 doses of GrB-Fc-SD1 (130 mg/mg total dose, delivered IP) indicated highly significant efficacy in both dose regimens, with p &amp;lt; 0.001 compared to vehicle-treated mice. Mouse body weights were not impacted at the efficacy doses used in this study, indicating that this dose was non-toxic. Histopathology studies revealed a significant decrease in the percentage of Ki-67+ nuclei in treated tumors compared to controls. These data warrant continued investigation of the protein as a therapeutic candidate. Research supported by H2Biologics. Research conducted, in part, by the Clayton Foundation for Research. Citation Format: Khalid A. Mohamedali, Lawrence H. Cheung, Mihai G. Iurascu, Louis R. DePalatis, Claire Thuning-Roberson, Michael G. Rosenblum. Targeting mesothelin on tumor cells with a novel, completely human fusion construct delivering the serine protease granzyme B [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2606.

Abstract 6675: Dissecting the relationship between mucin 5AC and the lung/gut microbiome in the pathogenesis of K-ras mutant lung cancer

Abstract Non-small cell lung cancer, including its most common subtype, lung adenocarcinoma (LUAD), is one of the most prevalent and lethal forms of cancer worldwide. This is largely due to patients having progressed to an advanced stage of disease by the time of diagnosis. Thus, better understanding the pathogenesis of early LUAD is paramount for the identification of key targets and development of preventive and therapeutic strategies for clinical management of the disease. KRAS mutations are the most common oncogenic driver alteration in human LUAD. Previously, we have found that KRAS mutant LUAD patients whose tumors harbor high expression of Mucin 5 AC (MUC5AC), a main airway secretory mucin, have significantly lower survival and poor prognosis. Mucins are membrane-bound and secreted hydrophilic proteins essential in clearance of allergens, pathogens, and debris in the lung airway. However, overproduction of mucins, such as MUC5AC, plays roles in the pathogenesis of airway diseases such as chronic obstructive pulmonary disease (COPD) and lung cancer. Given the known effect of mucin polymers on bacterial growth and the immune microenvironment in the context of other airway inflammatory diseases, in this study, our objective is to better understand the MUC5AC-to-microbiome relationship using a lung epithelial specific mouse model of KRAS-mutant LUAD (CC-LR) and test the combinatorial effects of genetic deletion of MUC5AC (CC-LR/Muc5ac−/−) and the pan-microbiome depletion on tumor progression, the lung immune contexture, and microbiome (lung and gut). Thus far, we found that genetic deletion of MUC5AC or pan-microbiome depletion significantly decreases the tumor burden (66% vs 50%, respectively). Additionally, both groups show to have a significantly decreased percentage of tumor-specific Ki67+ cells, a cell proliferation marker, and ERG+ cells, an angiogenesis marker, in comparison to the CC-LR controls. The combination of the lack of MUC5AC with pan-microbiome depletion has also shown to reduce tumor development that needs further investigation. These findings highlight the need to better understand the mechanistic interaction between mucins and the lung/gut microbiome in the context of KRAS-mutant lung cancer which we are currently exploring. It also suggests that their relationship could be a key target for modulation, potentially leading to the development of a minimally invasive alternative preventive and/or treatment modality against KRAS-mutant LUAD. Citation Format: Maria T. Grimaldo, Michael J. Clowers, Avantika Krishna, Katherine E. Larsen, Nastaran Karimi, Yasmina H. Rezai, Arnav Gaitonde, Farbod Khalaj, Haoyue Liu, Florencia McAllister, Seyed Javad Moghaddam. Dissecting the relationship between mucin 5AC and the lung/gut microbiome in the pathogenesis of K-ras mutant lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6675.

Abstract A021: Pilot study of daily exemestane in patients with Endometrial Intraepithelial Neoplasia (EIN) or grade 1 endometrial cancer

Abstract Introduction: The purpose of this study is to evaluate exemestane, an aromatase inhibitor, as a potential agent for the chemoprevention of endometrial cancer. Experimental procedures: This is a multi-center, phase IIA, single arm ‘window of opportunity’ pilot study of 25 mg per day of exemestane for 21-42 days in postmenopausal patients undergoing hysterectomy for endometrial intraepithelial neoplasia (EIN) or low-grade endometrial cancer. The primary objective is to determine if there is a decrease in proliferation index, measured by percent Ki-67 expression in endometrial tissue, from pre-treatment to post-exposure. Results of Ki-67 are compared with a historic control cohort of tumors from patients matched by preoperative histologic diagnosis, age, BMI category. Specimens from the historic control cohort included a preoperative endometrial biopsy as well as a surgical specimen, separated in time by less than 42 days. Secondary outcomes include comparison of Ki67 to historic controls, circulating serum estradiol and progesterone levels, pathologic response, tissue biomarkers, safety and adverse effects. Summary of Data: Forty patients accrued to the study between January of 2018 and March of 2021. The mean age was 63 (range 52-75). Thirty-nine patients (97.5%) self identifiedself-identified as white; 1 patient (2.5%) identified as Asian. The mean BMI was 40.3 (SD 9.75, range 22.8-60.5). Preoperative diagnoses included EIN, grade 1 endometrial adenocarcinoma, and grade 2 endometrial adenocarcinoma in 11 (27.5%), 26 (65%), and 3 (7.5%) patients respectively. Thirty-nine patients (97.5%) were compliant with exemestane dosing. All patients completed the study and no patients discontinued or interrupted therapy due to treatment related adverse events. Thirty-eight patients had evaluable specimens for both pre and post-treatment Ki-67 analysis. Median Ki-67 score decreased from 40.7% [IQR (33.9%, 50.3] at baseline to 18.1% [IQR (8.8%, 31.8%)]. This represents a median absolute change from baseline of -20.4% [IQR (-29.9, -6.7), p&amp;lt;0.001]. In total, percentage of Ki-67 in tumor cells declined in 31 patients and increased in 7 patients. In the historic control cohort, patients also had a decrease in Ki-67 score with a median absolute change from baseline of -6.7% [IQR (-12.7%, -1.3%), p&amp;lt;0.001]. However, the decrease in Ki-67 was greater in the study patients than the matched historic controls, with a median difference between the groups of -13.4% [IQR= (-23.3%, 6.9%), p=0.0095]. Serum estradiol levels did not change significantly between baseline and post-treatment (p=0.16). There was a small but significant increase from baseline to post-treatment in serum progesterone (0.14 ng/mL vs 0.37 ng/mL, p&amp;lt;0.001). Conclusion: In this pilot window of opportunity study, exemestane demonstrates anti-proliferative effects in endometrial intraepithelial neoplasia and low-grade endometrial cancer. This agent warrants further evaluation for the chemoprevention of endometrial cancer. Citation Format: Britt K. Erickson, Lisa Barrioullet, Rebecca C. Arend, Dina El-Rayes, Mahmoud Khalifa, Amy Skubitz, Kristin Boylan, Andrew Nelson, Bharat Thyagarajan, Thomas Havighurst, Richard Chappell, Eileen Dimond, Katina Deshong, Goli Samimi, Eva Szabo, Howard Bailey. Pilot study of daily exemestane in patients with Endometrial Intraepithelial Neoplasia (EIN) or grade 1 endometrial cancer [abstract]. In: Proceedings of the AACR Special Conference on Endometrial Cancer: Transforming Care through Science; 2023 Nov 16-18; Boston, Massachusetts. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(5_Suppl):Abstract nr A021.

Clinical and Pathological Predictors of Death for Adrenocortical Carcinoma.

Adrenocortical carcinoma (ACC) is a rare and lethal disease with a poor prognosis. This study aims to share our 41-year experience as a referral center, focusing on identifying risk factors associated with ACC mortality. Our retrospective analysis included a cohort of 150 adult patients with ACC in all stage categories, treated between 1981 and 2022. Tumor hormonal hypersecretion was observed in 78.6% of the patients, and the median age of diagnosis was 40 years. The majority presented as European Network for the Study of Adrenal Tumors (ENSAT) III or IV (22.9% and 31.2%, respectively), and the overall mortality rate was 54.6%. Independent predictors of death were elevated secretion of cortisol (HR = 2.0), androstenedione (HR = 2.2), estradiol (HR = 2.8), 17-OH progesterone (HR = 2.0), and 11-deoxycortisol (HR = 5.1), higher Weiss (HR = 4.3), modified Weiss (HR = 4.4), and Helsinki scores (HR = 12.0), advanced ENSAT stage (HR = 27.1), larger tumor size (HR = 2.7), higher Ki-67 percentage (HR = 2.3), and incomplete surgical resection (HR = 2.5). Mitosis greater than 5/50 high-power field (HR = 5.6), atypical mitosis (HR = 2.3), confluent necrosis (HR = 15.4), venous invasion (HR = 2.8), and capsular invasion (HR = 2.4) were also identified as independent predictors of death. Knowing the risk factors for ACC's mortality may help determine the best treatment option.

Neoadjuvant Chemotherapy and Pathologic Complete Response in HR+/HER2− Breast Cancer: Impact of Tumor Ki67 and ER Status

Introduction: Neoadjuvant chemotherapy (NAC) is extensively employed in breast cancer (BC), primarily for aggressive subtypes like triple-negative and human epidermal growth factor receptor 2 (HER2)-positive BC and in estrogen receptor-positive (ER+)/HER2− BC with high-risk features. In ER+/HER2− BC, pathological complete rates are much lower (Methods: Retrospective data analysis from Kartal Dr. Lütfi Kırdar City Hospital included HR+/HER2− BC patients who received NAC. Clinicopathological factors, NAC response, and surgical outcomes were assessed. Statistical analyses evaluated the association between Ki67, ER status, and pCR. Results: Of 203 patients, 11.8% achieved pCR. Ki67 (p p p = 0.003, HR: 1.02) and ER percentage (p = 0.019, HR: 0.97) as independent predictors of pCR probability. Conclusion: Consideration of Ki67 and ER percentage aids in NAC decisions for HR+/HER2− BC, identifying patients with high NAC response rates, facilitating axillary preservation, and potentially avoiding axillary dissection. The pCR rates in patients with Ki67 ≤24 are particularly low, especially in patients with a high ER percentage. In these cases, upfront surgery and adjuvant treatment should be considered instead of NAC.

Human Leukocyte Antigen Class I Expression and Natural Killer Cell Infiltration and Its Correlation with Prognostic Features in Luminal Breast Cancers.

The aim of this study was to determine whether low HLA-I expression and NK cells infiltration are related to prognostic features in breast cancer, as observed in cancers in other locations and non-hormone dependent breast cancers. Particularly, we explored their relation to infiltrated axillary lymph nodes (ALNs), with the aim of finding new predictors helping to decide the extent of axillary surgery. We conducted a retrospective correlational analysis of 35 breast cancers from 35 breast cancer patients showing axillary infiltration at diagnosis and with upfront surgery. HLA-I H-score and the number of NK cells x 50high power fields (HPF) in the biopsy specimen were correlated with pathological variables of the surgical specimen: number of infiltrated ALNs, tumor size, histological type, the presence of ductal carcinoma in situ, focality, histological grade, necrosis, lymphovascular and perineural invasion, Her2Neu status, and the percentages of tumor-infiltrating lymphocytes (TILs), estrogen receptor, progesterone receptor, ki67, and p53. All tumors showed hormone receptor expression and three of them Her2Neu positivity. A positive correlation (p=0.001**) was found between HLA-I H-score and TILs and Ki67 expression. HLA H-score increased with histological grade and was higher in unifocal than in multifocal disease (p=0.044 and p=0.011, respectively). No other correlations were found. High HLA-I H-score values correlated with features of poor prognosis in this cohort of luminal breast tumors, but not with infiltrated ALNs. This finding highlights the differences between luminal breast cancer, and cancers in other locations and non-hormone dependent breast cancers, in which low HLA-I expression tends to be associated with poor prognostic features.

RIPK4 downregulation impairs Wnt3A-stimulated invasiveness via Wnt/β-catenin signaling in melanoma cells and tumor growth in vivo.

The role of Wnt signaling in oncogenesis and drug resistance is well known. Receptor-interacting protein kinase (RIPK4) contributing to the increased activity of many signaling pathways, including Wnt/β-catenin, may be an important target for designing new drugs for metastatic melanoma, but its role in melanoma is not fully understood. We tested the effect of genetic manipulation of RIPK4 (CRISPR/Cas9) on xenograft growth. In addition, immunohistochemistry was used to detect active β-catenin, Ki67 and necrosis in xenografts. Wnt signaling pathway activity was examined using Western blot and Top-Flash. The effect of RIPK4 knockout on melanoma cells in vitro stimulated Wnt3A on wound overgrowth, migration and invasion ability was then evaluated. Our study showed that CRISPR/Cas9-mediated RIPK4 knockout (KO) significantly reduced tumor growth in a mouse model of melanoma, particularly of WM266.4 cells. RIPK4 KO tumors exhibited lower percentages of Ki67+ cells as well as reduced necrotic area and decreased levels of active β-catenin. In addition, we observed that RIPK4 knockout impaired Wnt3A-induced activation of LRP6 and β-catenin, as manifested by a decrease in the transcriptional activity of β-catenin in Top-Flash in both tested melanoma cell lines, A375 and WM266.4. Prolonged incubation (48h) with Wnt3A showed reduced level of MMP9, C-myc, and increased SOX10, proteins whose transcription is also dependent on β-catenin activity. Moreover, RIPK4 knockout led to the inhibition of scratch overgrowth, migration and invasion of these cells compared to their controls. RIPK4 knockdown inhibits melanoma tumor growth and Wnt3A stimulated migration and invasion indicating that RIPK4 might be a potential target for melanoma therapy.