Negative Human Epidermal Growth Factor Research Articles

High Expression of CLDN 18.2 is Associated with Poor Disease-Free Survival of HER-2 Positive Gastric Cancer.

Anti-claudin (CLDN) 18.2 therapy has been proven to be effective in treating advanced gastric cancer with negative human epidermal growth factor receptor 2 (HER-2). This study purposed to investigate the relationship of CLDN 18.2 expression with prognosis of HER-2-positive gastric cancer patients. To investigate the expression of claudin (CLDN) 18.2 in Human epidermal growth factor receptor 2 (HER-2) positive gastric cancer patients after radical resection and its relationship with gastric cancer prognosis. A total of 55 postoperative HER-2-positive gastric cancer patients were included in this study. CLDN 18.2 protein was detected by immunohistochemistry, and detailed clinical and pathological information was collected. Factors considered potentially important in the univariate analysis were included in the multivariate analysis, which involved COX regression to find the independent prognostic factors affecting disease-free survival (DFS). Immunohistochemistry showed that different levels of CLDN 18.2 protein were expressed in HER-2 positive gastric cancer tissues, and the Chi-square analysis showed that the expression level of CLDN 18.2 was significantly correlated with the lymph node stage. Higher expression levels of CLDN 18.2 were found in patients with lymph node positivity and were associated with poor prognosis in HER-2-positive gastric cancer patients. Gastric cancer patients with low and high expressions of CLDN 18.2 had postoperative median DFS of 38.5 months (95% confidence interval (CI) 28.8-48.2 months) and 12.1 months (95% CI, 11.7-41.0 months), respectively. High expression of CLDN 18.2 in HER-2 positive gastric cancer is associated with poor prognosis, and the optimal treatment mode for this population is worth exploring after the approval of anti-CLDN 18.2 drugs.

Efficacy of nivolumab in combination with chemotherapy after radiotherapy for meningeal carcinomatosis in patients with Epstein-Barr virus-associated gastric cancer:a case report

A 62-year-old man presented with fever and anorexia since July X. Initial treatments were rendered ineffective, and due to altered consciousness and vomiting, he was referred to our hospital. On admission, he manifested delirium, drowsiness, and disorientation. While blood tests were normal, gastroscopy identified a type 3 tumor in his lower gastric body, later diagnosed as a poorly differentiated adenocarcinoma. Immunohistochemistry demonstrated negative human epidermal growth factor receptor 2 and positive programmed death-ligand 1 expression with a combined positive score ≥5. Furthermore, a positive Epstein-Barr virus-encoded small RNA in situ hybridization result was noted. Abdominal contrast-enhanced CT and PET-CT scans demonstrated multiple lymph node metastases around the stomach and liver, establishing the diagnosis of stage IVB gastric cancer (T4aN2M1). Brain magnetic resonance imaging (MRI) demonstrated enhanced lesions in the brainstem, cerebellar sulci, and right occipital lobe. Although cerebrospinal fluid cytology was negative for malignancy, the clinical symptoms and MRI findings confirmed leptomeningeal carcinomatosis (LMC). The patient underwent radiotherapy for LMC (total of 30Gy in 10 fractions), followed by combination therapy with a nivolumab and SOX regimen. Posttreatment, the LMC symptoms resolved;however, he experienced grade 3 immune-related adverse events related to liver dysfunction. Nivolumab was discontinued, and with steroid administration, the adverse events improved. Imaging evaluations posttreatment showed gastric tumor reduction and the absence of LMC. After 7 cycles, nivolumab was reintroduced, with no liver dysfunction recurrence noted through 15 cycles. Endoscopic examination 1 year postonset demonstrated that the gastric tumor had scarred, and MRI showed no signs of LMC recurrence. In 5-8% of solid tumors, LMC complications are present, resulting in limited treatment options and poor prognosis. Recent reports suggest the potential of immune checkpoint inhibitors in treating intracranial metastasis from solid tumors. In Japan, nivolumab was approved for gastric cancer treatment in 2017 and for first-line therapy in combination with chemotherapy since 2021. We report a case in which radiotherapy and chemotherapy combined with nivolumab provided durable control of LMC originating from gastric cancer for more than 1 year.

Comparison of Mast Cell Density and Prognostic Factors in Invasive Breast Carcinoma: A Single-Centre Study in Malaysia.

Mast cells influence tumour growth, neo-angiogenesis and the propensity for metastasis by contributing to innate and adaptive immune responses in the tumour microenvironment. The number of mast cells has increased in various malignant tumours and their abundance has been associated with either a favourable or unfavourable prognosis. This study investigated the significant difference in stromal mast cell density among multiple prognostic factor groups in invasive breast carcinoma. CD117 (c-KIT) antibodies were used to stain 160 formalin-fixed and paraffin-embedded invasive breast carcinoma tissues to demonstrate the presence of mast cells. Then the labelled mast cells were counted in 10 fields at 400× magnification and the mean value was used to represent the mast cell density. The demographic distribution revealed that most patients were 40 years old or older (92.5%) and of Malay ethnicity (66.3%). With regard to prognostic factors, the most prevalent subtype was invasive carcinoma of no special type (80.6%), followed by tumour grade 3 (41.3%), T2 tumour size (63.1%), N0 lymph node stage (51.3%), presence of lymphovascular invasion (59.4%), positive oestrogen (64.4%) and progesterone receptors (53.1%), and negative human epidermal growth factor receptor 2 (HER2) expression (75.0%). However, there was no significant difference in stromal mast cell density among the different demographic and prognostic factor groups in invasive breast carcinoma. The findings from this study suggest that stromal mast cells do not play a significant role in preventing or promoting tumour growth in invasive breast carcinoma.

Omitting Radiotherapy after Breast-Conserving Surgery in Luminal A Breast Cancer

BackgroundAdjuvant radiotherapy is prescribed after breast-conserving surgery to reduce the risk of local recurrence. However, radiotherapy is inconvenient, costly, and associated with both short-term and long-term side effects. Clinicopathologic factors alone are of limited use in the identification of women at low risk for local recurrence in whom radiotherapy can be omitted. Molecularly defined intrinsic subtypes of breast cancer can provide additional prognostic information.MethodsWe performed a prospective cohort study involving women who were at least 55 years of age, had undergone breast-conserving surgery for T1N0 (tumor size <2 cm and node negative), grade 1 or 2, luminal A–subtype breast cancer (defined as estrogen receptor positivity of ≥1%, progesterone receptor positivity of >20%, negative human epidermal growth factor receptor 2, and Ki67 index of ≤13.25%), and had received adjuvant endocrine therapy. Patients who met the clinical eligibility criteria were registered, and Ki67 immunohistochemical analysis was performed centrally. Patients with a Ki67 index of 13.25% or less were enrolled and did not receive radiotherapy. The primary outcome was local recurrence in the ipsilateral breast. In consultation with radiation oncologists and patients with breast cancer, we determined that if the upper boundary of the two-sided 90% confidence interval for the cumulative incidence at 5 years was less than 5%, this would represent an acceptable risk of local recurrence at 5 years.ResultsOf 740 registered patients, 500 eligible patients were enrolled. At 5 years after enrollment, recurrence was reported in 2.3% of the patients (90% confidence interval [CI], 1.3 to 3.8; 95% CI, 1.2 to 4.1), a result that met the prespecified boundary. Breast cancer occurred in the contralateral breast in 1.9% of the patients (90% CI, 1.1 to 3.2), and recurrence of any type was observed in 2.7% (90% CI, 1.6 to 4.1).ConclusionsAmong women who were at least 55 years of age and had T1N0, grade 1 or 2, luminal A breast cancer that were treated with breast-conserving surgery and endocrine therapy alone, the incidence of local recurrence at 5 years was low with the omission of radiotherapy. (Funded by the Canadian Cancer Society and the Canadian Breast Cancer Foundation; LUMINA ClinicalTrials.gov number, NCT01791829.)

Breast cancer with scalp metastases: a case report

BackgroundWhile breast cancer is the most common cancer in women, cutaneous metastases are rare in breast cancer. Additionally, scalp involvement in breast cancer metastasis is extremely rare. That being said, scalp lesions should always be thoroughly investigated to distinguish metastatic lesions from other neoplasms.Case presentationA 47-year-old female Middle-Eastern patient presented with metastatic breast cancer in the lungs, bone, liver, and brain with no signs of multiple organ failure, in addition to cutaneous metastases, including the scalp. Between 2017 and 2022, she was managed through modified radical mastectomy, radiotherapy, and several lines of chemotherapy. She presented in September of 2022 with enlarging scalp nodules, which started developing 2 months prior to her presentation. Physical examination revealed firm, non-tender, and immobile skin lesions. Magnetic resonance imaging scan of the head showed soft tissue nodules in different sequences. A punch biopsy was taken from the largest scalp lesion and showed metastatic invasive ductal carcinoma. A panel of immunohistochemistry stains was applied, because a single specific marker for differentiating primary cutaneous adnexal tumors or other malignant neoplasms from breast cancer has not yet been identified. The panel showed positive estrogen receptor 95%, progesterone receptor 5%, negative human epidermal growth factor receptor 2, positive GATA binding protein 3, positive cytokeratin-7, negative P63, and negative KIT (CD117).ConclusionBreast cancer metastases to the scalp are extremely uncommon. When a scalp metastasis is present, it might be the only symptomatic sign of disease progression or widespread metastatic lesions. However, such lesions warrant a comprehensive radiologic and pathologic workup to rule out other possibilities of skin pathologies, such as sebaceous skin adenocarcinoma as it effects the management plan.

Abstract P6-01-24: Long-term outcome data using Endopredict® as risk stratification and chemotherapy decision biomarker in hormone receptor positive, HER2-negative early breast cancer

Abstract Background: The Endopredict test is used for estimating risk of distant recurrence for women presenting with early-stage breast cancer with a positive estrogen receptor (ER) and negative human epidermal growth factor receptor 2 (HER2) status. The current ASCO Guideline Update on biomarkers confirms the value of the Endopredict test to guide decisions of adjuvant endocrine and chemotherapy. This study shows prospective long-term outcome data of early breast cancer patients whose chemotherapy decision was guided by the Endopredict test result (EPclin). Methods: ER-positive and HER2-negative early breast cancer patients with 0-3 positive lymph nodes treated between March 2012 and March 2015 were included in this single institution study. The Endopredict® test was carried out on all tumour samples. Demographic, clinical and pathological data were assessed for each patient at baseline. Treatment compliance, local recurrence, distant metastases and survival was evaluated. Risk estimates were obtained by the Kaplan-Meier method and cumulative risk functions in case of competing risks. Group comparisons were performed by Cox proportional hazards regression models and quantified through hazard ratios. Median Follow-Up was estimated by the inverse Kaplan-Meier method. Exploratory hypothesis testing was conducted at two-sided 5% significance levels. Results: In a cohort of 368 consecutive cases the median follow-up time was 8.2 years. Endopredict allocated 238 pts (64%) in the EPclin low risk and 130 pts (34%) in the EPclin high risk group. The 5-year distant metastasis free survival (DMFS) in the EPclin low risk group was 96.6% (95% CI 0.943-0.989) and 85.5% (95% CI 0.796-0.920) in the EPclin high risk group. With a hazard ratio (HR) of 2.21 (95% CI: 1.27-3.88; p=0.005) the risk for distant metastasis in EPclin high risk patients was more than two-fold higher in comparison with EPclin low risk patients. 87 pts. (66.9%) of the EPclin high risk group underwent chemotherapy (compliant), whereas 43 pts (33.1 %) opposed the recommended chemotherapy (non-compliant). Kaplan-Meier plots in the EPclin high risk subgroups compliant vs non-compliant showed a significant disease-free survival (DFS) benefit towards the patients following the chemotherapy recommendation (HR 0.46; 95%CI 0.23-0.95; p=0.036). The 5-year DFS for the high risk compliant subgroup was 89.1% (95% CI: 0.827-0.961) vs. the high risk non-compliant subgroup with 68.9% (95% CI: 0.562-0.845). Regarding the subgroups pre- and postmenopausal, patients with a EPclin high risk test result were at significant higher risk of experiencing distant metastases than patients with a EPclin low risk test result in both subgroups (premenopausal: HR 3.55; 95%CI 1.17-12.32; p=0.025; postmenopausal: HR 1.19; 95%CI 0.99-3.7; p=0.054). We analyzed the EPclin categorization in context of the ki67 subtypes luminal A (low; 0-10%) and luminal B (high; 25-100%). The EPclin-based risk stratification was significantly associated with improved DFS in both ki67 subtypes (ki67 low: HR 4; 95%CI 1.25-12.04; p=0.021 and ki67 high: HR 3.77; 95%CI 1.19-18.93; p=0.022). 33.3% (21 pts) of all tumor samples classified as luminal B (63 pts), were reclassified towards the low risk group via Endopredict, sparing chemotherapy recommendation. Contrary 19.2% (14 pts) of all luminal A (73 pts) were categorized to high risk EPclin. Conclusion: These first long term prospective outcome data confirm, that Endopredict can guide decisions on adjuvant chemotherapy in early ER positive, HER2 negative breast cancer. Pts categorized as EPclin high risk benefitted from an adjuvant chemotherapy. Our results show that Endopredict risk stratification is also applicable in premenopausal women. Furthermore the Endopredict test showed a better classification accuracy in comparison to ki67 subtypes, resulting in a more precise estimation of prognosis. Citation Format: Evelyn Klein, Adriana Josipovic, Aurelia Noske, Sophie Anders, Carolin Mogler, Wilko Weichert, Alexander Hapfelmeier, Marion Kiechle, Johannes Ettl. Long-term outcome data using Endopredict® as risk stratification and chemotherapy decision biomarker in hormone receptor positive, HER2-negative early breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-01-24.

Sub-Classification of Triple-Negative Breast Cancer using Androgen Receptor and Cytokeratin 5/6

Background: Triple negative breast cancer (TNBC) is a unique heterogenous subtypes of breast cancer which is characterized by negative estrogen, progesterone, and human epidermal growth factor receptor (HER-2) status. TNBC displays different molecular phenotype with which basal-like tumour can be identified using high molecular weight basal cytokeratin 5/6 (CK5/6).&#x0D; Methods: Ninety-five (95) formalin fixed cases from Korle Bu Teaching Hospital in Ghana’s (KBTH) archives were sampled in a retrospective study from 2012-2016. Blocks of these triple-negative breast cancer was subclassified using CK5/6 and Androgen Receptor (AR) antibodies. Subclasses were also identified.&#x0D; Results and Conclusion: In all ninety-five (95) TNBC cases, hormonal subtyping was sub-classified using CK 5/6 and AR. The mean ±SD of these cases was recorded as 53.96 (±13.56) years and the age range of these cases was 22-104 years. The average size (±SD) of the tumour was recorded to be 14.43(±7.62) and it had a range of 2.4-45cm. lymph nodes retrieved also had a mean ± SD of 10.35(±6.05) with an average tumour lymph nodes involvement of 2.6(± 3.697).&#x0D; Invasive Ductal carcinoma was identified as the commonest histologic type of TNBC with approximately 95% of the cases. This was followed by invasive lobular (2.1%), medullary carcinoma (2.1%) and metaplastic carcinoma (1.1%).&#x0D; Approximately 30% of TNBC stained positive for CK5/6.&#x0D; It can however be concluded that, most TNBC are not basal-like when the basal marker CK5/6 is used.

Using American Type Culture Collection Cell Lines to Evaluate Interlaboratory Variables for Estrogen Receptor and Human Epidermal Growth Factor Receptor 2 Immunostaining.

Most laboratories currently use patient tissues for validating immunohistochemical stains. To explore advantages of using cell lines with known antigenicity as a validation method. Five American Type Culture Collection (ATCC) cell lines with known negative, low positive, and moderate to strong estrogen receptor (ER) expression as well as negative, equivocal, and positive human epidermal growth factor receptor 2 (HER2) expression were cultured and made into cell blocks. One block from each cell line was fixed in formalin and another in ethanol before cell block preparation. Two sets of paired unstained slides from each block were sent to 10 different laboratories for HER2 and ER staining to be stained on runs from different days according to each laboratory's defined protocol. The 10 study participants evaluated 40 slides in a blinded fashion. For ER expression, all 80 interpretations for the ER strong and moderate positive cell lines had the target ER-positive result, and 74 of 80 ER-negative cell lines (92.5%) had agreement with the intended negative result. The ER low positive cell line showed varied but positive expression among all observers. The HER2 (3+)-positive cell lines yielded a target interpretation of 3+ in 65 of 80 interpretations (81.2%). For the HER2-negative cell line 69 of 78 interpretations (88.5%) were consistent with the target response (0 or 1+). No significant variation was observed between the ethanol- and non-ethanol-exposed cell lines, or between runs by the same laboratory. Variation from target results clustered within laboratories. This study indicates that variability between laboratories can be identified by using cell lines for quantitative or semiquantitative immunohistochemistry when using cultured cell lines of known antigenicity. These cell lines could potentially play a role in aiding anatomic pathology laboratories in validating immunohistochemistry tests for formalin- and ethanol-fixed tissues.

Prediction of Breast Cancer Histological Outcome by Radiomics and Artificial Intelligence Analysis in Contrast-Enhanced Mammography

Simple SummaryThe assessment of breast lesions through mammographic images is currently challenging, especially in dense breasts. Contrast-enhanced mammography has been shown to overcome the limitations of standard mammography but it greatly depends on the interpretative skills of the physician. The aim of this study was to evaluate the potentialities of statistical and artificial intelligence algorithms as a tool for helping the radiologists in the interpretation of images. The most remarkable results were achieved in discriminating benign from malignant lesions and in the identification of the presence of the hormone receptor. A tool to support the physician’s decision-making process may be designed starting from simple logistic regression and tree-based algorithms. This type of tool may help the radiologist in assessing the investigated breast and in choosing the appropriate follow-up without resorting to histology.Purpose: To evaluate radiomics features in order to: differentiate malignant versus benign lesions; predict low versus moderate and high grading; identify positive or negative hormone receptors; and discriminate positive versus negative human epidermal growth factor receptor 2 related to breast cancer. Methods: A total of 182 patients with known breast lesions and that underwent Contrast-Enhanced Mammography were enrolled in this retrospective study. The reference standard was pathology (118 malignant lesions and 64 benign lesions). A total of 837 textural metrics were extracted by manually segmenting the region of interest from both craniocaudally (CC) and mediolateral oblique (MLO) views. Non-parametric Wilcoxon–Mann–Whitney test, receiver operating characteristic, logistic regression and tree-based machine learning algorithms were used. The Adaptive Synthetic Sampling balancing approach was used and a feature selection process was implemented. Results: In univariate analysis, the classification of malignant versus benign lesions achieved the best performance when considering the original_gldm_DependenceNonUniformity feature extracted on CC view (accuracy of 88.98%). An accuracy of 83.65% was reached in the classification of grading, whereas a slightly lower value of accuracy (81.65%) was found in the classification of the presence of the hormone receptor; the features extracted were the original_glrlm_RunEntropy and the original_gldm_DependenceNonUniformity, respectively. The results of multivariate analysis achieved the best performances when using two or more features as predictors for classifying malignant versus benign lesions from CC view images (max test accuracy of 95.83% with a non-regularized logistic regression). Considering the features extracted from MLO view images, the best test accuracy (91.67%) was obtained when predicting the grading using a classification-tree algorithm. Combinations of only two features, extracted from both CC and MLO views, always showed test accuracy values greater than or equal to 90.00%, with the only exception being the prediction of the human epidermal growth factor receptor 2, where the best performance (test accuracy of 89.29%) was obtained with the random forest algorithm. Conclusions: The results confirm that the identification of malignant breast lesions and the differentiation of histological outcomes and some molecular subtypes of tumors (mainly positive hormone receptor tumors) can be obtained with satisfactory accuracy through both univariate and multivariate analysis of textural features extracted from Contrast-Enhanced Mammography images.

Brain metastasis in de novo breast cancer: An updated population-level study from SEER database

PurposeAlthough there are current studies on breast cancer brain metastasis, population-level analysis is still lacking. As treatment for metastatic breast cancer has improved, an updated population-level analysis is necessary. Our aim was to use the SEER database to characterize the incidence and survival of patients with brain metastases at the initial diagnosis of breast cancer. Patients and methodsPatients with breast cancer from 2010 to 2018 were identified using the SEER database. The stratified incidence and median survival of patients with BM at diagnosis were described. Multivariate logistic and Cox regression were performed to determine the covariates associated with brain metastasis and survival outcomes, respectively. Multiple comparisons based on Cox proportional hazards model were performed for the analysis of interactive effects on overall survival. ResultsA total of 2,248 patients with brain metastases at the initial diagnosis of breast cancer were identified, accounting for 0.40% of all patients with breast cancer, and 7.26% of patients with metastatic disease. Incidence proportions were highest, and survival outcomes were worst among patients with hormone receptor (HR)-negative human epidermal growth factor receptor 2 (HER2)-positive and triple-negative subtypes. For patients with brain metastases, the prognostic differences among different molecular subtypes have been gradually narrowing, and the survival benefits from various treatment methods have been all increased over time. ConclusionOur study provides an updated population-level estimate of the incidence and survival for patients with brain metastases at the diagnosis of breast cancer, thus may help early identification, prognostic stratification and treatment planning for such patients.

Abstract PS10-52: Adjuvant treatment with paclitaxel plus trastuzumab for node negative human epidermal growth factor receptor 2-positive breast cancer: Real life experience

Abstract BackgroundInvasive breast cancers are characterized by overexpression or amplification of the human epidermal growth factor receptor 2 (HER2) approximately 15 to 25%. Survival outcomes was improved after introduction of Trastuzumab treatment. Trastuzumab treatment was applied mostly with combination chemotherapy regimens at pivotal trials. There were some efforts to avoid the toxicity of combination chemotherapies and reduce the amount of treatment given especially in stage I HER 2 overexpressed breast cancer patients. Adjuvant paclitaxel plus trastuzumab was shown excellent disease free survival (DFS) and overall survival (OS) in node negative, 3 cm and smaller HER 2 overexpressed breast cancer patients. MethodsAll breast cancer patients that treated in Medical Oncology departments of Hacettepe University (Ankara, Turkey), Dr. Abdurrahman Yurtaslan Oncology Training and Research Hospital (Ankara, Turkey), Nicosia Dr. Burhan Nalbantoglu State Hospital (Nicosia, Cyprus) and Near East University (Nicosia, Cyprus) were retrospectively reviewed from patient files, center’s databases and chemotherapy files.ResultsWe retrospectively analyzed 173 patients who treated with adjuvant paclitaxel plus trastuzmab between April 1, 2012 and April 10, 2020. Median age was 52 years (range, 25 to 84 years) and 62.4% had estrogen receptor positive disease. 68.8% of the tumors were high-grade. 14.4% of patients had tumors 1 cm or smaller. 45.0% of patients had tumors larger than 2 cm and 32.3% of patients had tumors larger than 2 cm and up to 3 cm. 12.6% of patients had tumors larger than 3 cm. Mean tumor size was 2.2 cm. 88.4% of patients had N0 disease and 2.9% of patients had microscopic nodal metastasis. 164 of 173 patients completed all 52 weeks of adjuvant treatment. Median follow up of 43 months there were 8 DFS events observed: four distant recurrences (2.3%), three locoregional recurrences (1.7%) and one died without documented recurrence. 3-year DFS rate was 96.6%. There was no recurrences in patients who had tumors 1 cm or smaller. There were 5 DFS events in patients had tumors larger than 2 cm and up to 3 cm subgrup. ConclusionThis real life experience with paclitaxel plus trastuzumab demonstrated few distant recurrences and further supports the APT trial findings. Table 2. Events Observed for the Disease-free Survival.EventsPatients (N=173)Time to Eventno. (%)moAny recurrence or death8 (4.6)Local or regional recurrenceIpsilateral axilla, HER2-positive2 (1.1)25, 64Ipsilateral breast, HER2-positive1 (0.6)48Distant recurrence4 (2.3)7, 35, 36, 42DeathNot breast-cancer–related1 (0.6)52 Citation Format: Omer Diker, Burak Yasin Aktas, Recep Ak, Bahadir Koylu, Onur Bas, Polat Olgun, Hakan Taban, Deniz Can Guven, Neyran Kertmen, Berna Oksuzoglu, Sercan Aksoy. Adjuvant treatment with paclitaxel plus trastuzumab for node negative human epidermal growth factor receptor 2-positive breast cancer: Real life experience [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS10-52.

Diagnostic and Predictive Values of Strain Ratios in the Regions of Interests in Reference Tissue for Breast Tumor.

PurposeTo investigate the diagnostic and predictive value of strain ratios in the regions of interests (ROIs) in reference tissue for breast tumor.Patients and MethodsA total of 707 lesions in 665 consecutive patients were examined with B-mode Breast Imaging-Reporting and Data System (BI-RADS) and Ultrasonic elastography (UE). Elasticity score (ES) and strain ratio (SR) in each lesion were calculated. Receiver operating characteristic (ROC) curves were used to assess the diagnostic value of BI-RADS, ES, SR1, SR2, BI-RADS combined with ES (BI-RADS+ES), BI-RADS combined with SR1 (BI-RADS+SR1), and BI-RADS combined with SR2 (BI-RADS+SR2). The sensitivity, specificity, and areas under the ROC curves (Az) were obtained. Scatter plots were generated to demonstrate the correlation between SR1 and SR2. Kruskal-Walls H-test, Mann–Whitney U-test and one-way ANOVA were performed to evaluate SRs and tumor-related variables. Multiple linear regression analysis was carried out to determine variables independently associated with SRs.ResultsBI-RADS had high sensitivity and low specificity in the diagnosis of breast tumor. The specificity of BI-BADS combined with ES or SR was even higher. The Az value of BI-RADS+ES or BI-RADS+SRs was higher than that of BI-RADS (P < 0.001). The Az value of ES was higher than those of SR1 and SR2 (P < 0.001), and those of SR1 and SR2 were similar. SR1 and SR2 were highly positively correlated. There was no statistical difference between Az values of BI-RADS+ES, BI-RADS+SR1, and BI-RADS+SR2. Indistinct margin, high histologic grade, histological type, and negative human epidermal growth factor receptor (Her-2) were associated with SR1 and SR2. Progesterone receptor (PR) status and molecular subtype were associated with SR2. Histologic grade and tumor margin were significantly associated with SR1, and tumor margin was associated with SR2.ConclusionSRs in different ROIs in the reference tissue at the same depth showed no different diagnostic value for breast tumor. Both SR1 and SR2 could be useful in assessing the biological characteristics of invasive breast carcinoma.

Clinicopathological relation of CD10 and cyclooxygenase-2 expression in invasive breast cancer no special type

Background In breast cancer, which is the most frequent female cancer, new histopathologic parameters, such as tumor-infiltrating lymphocytes and stromal factors, are now introduced for prognosis and new strategies of therapy. This study was designed to explore CD10 and cyclooxygenase-2 (COX2) expression levels in invasive duct carcinoma (IDC) no special type (NST) and their relations with clinicopathological data. Methods This work was conducted on 40 cases of IDC of NST that were collected as paraffin-embedded blocks. For each case, sections of paraffin-embedded blocks were submitted to be stained with routine hematoxylin and eosin and other sections were stained using the immunohistochemistry technique with CD10 and COX2 antibodies and then evaluated and scored. Results CD10 was expressed in nearly half of the cases (45%), whereas COX2 was detected in all of the cases (100%). COX2 was strongly expressed in more than two-thirds of the cases. A significant association was detected between CD10 and lymph node status (P=0.017). Although no significant association could be detected, about one-fourth and one-third of cases with marked estrogen receptor and progesterone receptor expressions, respectively, had strong CD10. No significant association could be detected between COX2 and all studied clinical and histological data, including CD10 expression. However, COX2 was expressed more frequently in negative human epidermal growth factor receptor 2/neu and high Ki-67 cases. Cases with associated carcinoma in situ, detected vascular emboli, and lymph node positivity expressed COX2 score 4. Conclusion A significant association was found between CD10 and lymph node staging. COX2 was detected in all studied cases, with tendency to be associated with bad prognostic indicators. These findings suggest that CD10 and COX2 could contribute to tumor aggressiveness and progression in IDC NST.

Water Triggered Synthesis of Highly Stable and Biocompatible 1D Nanowire, 2D Nanoplatelet, and 3D Nanocube CsPbBr3 Perovskites for Multicolor Two-Photon Cell Imaging.

Two-photon imaging in the near-infrared window holds huge promise for real life biological imaging due to the increased penetration depth. All-inorganic CsPbX3 nanocrystals with bright luminescence and broad spectral tunability are excellent smart probes for two-photon bioimaging. But, the poor stability in water is a well-documented issue for limiting their practical use. Herein, we present the development of specific antibody attached water-resistant one-dimensional (1D) CsPbBr3 nanowires, two-dimensional (2D) CsPbBr3 nanoplatelets, and three-dimensional (3D) CsPbBr3 nanocubes which can be used for selective and simultaneous two-photon imaging of heterogeneous breast cancer cells in the near IR biological window. The current manuscript reports the design of excellent photoluminescence quantum yield (PLQY), biocompatible and photostable 1D CsPbBr3 nanowires, 2D CsPbBr3 nanoplatelets, and 3D CsPbBr3 nanocubes through an interfacial conversion from zero-dimensional (0D) Cs4PbBr6 nanocrystals via a water triggered strategy. Reported data show that just by varying the amount of water, one can control the dimension of CsPbBr3 perovskite crystals. Time-dependent transition electron microscopy and emission spectra have been reported to find the possible pathway for the formation of 1D, 2D, and 3D CsPbBr3 nanocrystals from 0D Cs4PbBr6 nanocrystals. Biocompatible 1D, 2D, and 3D CsPbBr3 nanocrystals were developed by coating with amine–poly(ethylene glycol)–propionic acid. Experimental data show the water-driven design of 1D, 2D, and 3D CsPbBr3 nanocrystals exhibits strong single-photon PLQY of ∼66–88% as well as excellent two-photon absorption properties (σ2) of ∼8.3 × 105–7.1 × 104 GM. Furthermore, reported data show more than 86% of PL intensity remains for 1D, 2D, and 3D CsPbBr3 nanocrystals after 35 days under water, and they exhibit excellent photostability of keeping 99% PL intensity after 3 h under UV light. The current report demonstrates for the first time that antibody attached 1D and 2D perovskites have capability for simultaneous two-photon imaging of triple negative breast cancer cells and human epidermal growth factor receptor 2 positive breast cancer cells. CsPbBr3 nanocrystals exhibit very high two-photon absorption cross-section and good photostability in water, which are superior to those of commonly used organic probes (σ2 = 11 GM for fluorescein), and therefore, they have capability to be a better probe for bioimaging applications.

Immune Checkpoint Blockade in Patients with Triple-Negative Breast Cancer.

Triple-negative breast cancer constitutes ~ 15% of all breast cancer subtypes. Because of the negative hormone receptor and human epidermal growth factor receptor 2 status, therapy is mainly based on chemotherapy with a poor median overall survival in the metastatic setting of ~ 18months. Compared to other breast cancer subtypes, triple-negative breast cancer is characterized by a higher mutational load, which renders the tumor immunogenic and amenable to immunotherapeutic intervention. Based on the promising results of immunotherapy in other cancer entities, including melanoma or non-small cell lung cancer, a vast number of studies are currently assessing immunotherapeutic approaches in patients with triple-negative breast cancer. While monotherapies with antibodies against programmed death-1 and programmed death ligand-1 have shown little efficacy in patients with heavily pretreated metastatic triple-negative breast cancer, treatment efficacy likely depends on the therapeutic setting, the treatment line, and the combination of immunotherapies with other anticancer drugs. Several studies are currently evaluating the safety and efficacy of immune checkpoint inhibition in combination with chemotherapy, angiogenesis inhibitors, poly(ADP-ribose) polymerase inhibitors, as well as radiotherapy in the metastatic and (neo-)adjuvant settings. The US Food and Drug Administration approval of nab-paclitaxel in combination with atezolizumab in 2019 presented a landmark therapeutic development for patients with triple-negative breast cancer, given the limited treatment options available for this highly aggressive disease. In this review, we provide an overview on important ongoing and completed immunotherapeutic studies in triple-negative breast cancer and their possible implications for clinical practice.

Influence of Androgen Receptor on the Prognosis of Breast Cancer.

We investigated the prognostic influence of androgen receptor (AR) on breast cancer. AR status was assessed using immunohistochemistry with tissue microarrays from 395 operable primary breast cancer patients who received curative surgery. The Kaplan–Meier estimator was used to analyze the survival rates and a log-rank test was used to determine the significance of the differences in survival. The Cox proportional hazards model was used to calculate the hazard ratio (HR) and the 95% confidence interval (CI) of survival. There were 203 (51.4%) subjects with a low expression of AR, and 192 patients (48.6%) with a high expression rate. The high AR expression group showed superior overall survival (p = 0.047) and disease-free survival (p = 0.004) when compared with the low AR expression group. The high AR expression group showed superior systemic recurrence-free survival when compared with the low AR expression group (p = 0.027). AR was an independent prognostic factor for both overall survival (HR, 0.586; 95% CI, 0.381–0.901; p = 0.015) and disease-free survival (HR, 0.430; 95% CI, 0.274–0.674; p < 0.001). A high AR expression was a significant favorable prognostic factor only in the subgroups with positive hormone receptors (HRc) and negative human epidermal growth factor receptor 2 (HER2) when considering disease-free survival (p = 0.026). The high AR expression group was significantly associated with superior overall survival and disease-free survival when compared with the low AR expression group with breast cancer patients. AR was a significant independent prognostic factor for both overall survival and disease-free survival. The prognostic impact of AR was valid in the HRc(+)/HER2(−) subtype when considering disease-free survival. These findings suggest the clinical usefulness of AR as a prognostic marker of breast cancer in clinical settings.

Curcumin‐Induced Apoptosis Pathways in Positive and Negative Human Epidermal Growth Factor Receptor Breast Cancer Cells

AimCurcumin, have several advantages and potential health benefits, such as low cytotoxicity and minimal development of resistance, as compared to chemically‐based therapies, and have anti‐cancerous efficacies, including induction of apoptosis, in cancer cells of various tissue origins. The aim of this study was toelucidate the mechanisms of curcumin‐induced apoptosis in human epidermal growth factor receptor positive (BT474) and negative (MCF‐7) breast cancer cell lines.MethodsCytotoxic activity of curcumin was measured by the sulphorhodamine B assay. We studied the induction of apoptosis by flow cytometry and detection of an apoptosis‐associated PARP cleavage product via Western blotting. Westernblotting was also used to measure Cytochrome C translocation, which determines mitochondrial involvement and caspase‐7 activation. The Caspase cascade was measured by activity levels of caspase‐3, caspase‐8, and caspase‐9 using chromogenic substrates.ResultsCurcumin CC50 (concentration that kills 50% of the cells) was 25.92 ±5.13μM for MCF‐7, and 49.96 ± 5.98 μM for BT474 cells. Flow cytometry results indicated a 13.36 ±5.96% expression increase and a 6.87 ± 1.42% expression increase in early stage apoptosis (Annexin V positively stained cells) in MCF‐7 and BT474 cells, respectively. Late stage apoptosis (dual stained Annexin V/Propidium Iodide (PI)cells) in curcumin exposed MCF‐7 cells was increased by 3.45 ± 0.94% expression, but no increase was seen in BT474 cells. PARP cleavage product (89kD) was detected at similar levels in both curcumin exposed MCF‐7 and BT474 cells and in a dose dependent manner. Curcumin‐exposedMCF‐7 cells, but not BT474 cells, showed substantial release of Cytochrome C from the mitochondria into the cytoplasm. Curcumin significantly upregulatedcaspase‐8 activity without modulating the activity of caspase‐9 in both BT474 and MCF‐7 cells. However, Caspase‐3 activity was upregulated only in BT474 cells. MCF‐7 cells reportedly have a gene deletion for Caspase‐3 and therefore do not express it. Caspase‐7 was then tested in MCF‐7 cells, but no upregulation was measured.ConclusionCytotoxicity of Curcumin is low for both MCF‐7 and BT474 cells and concentrations are easily achievable. Apoptosis induction by Curcumin exposure was detected by both PARP cleavage and flow cytometry. Flow cytometry showed that curcumin induced predominantly early stage apoptosis (Annexin V stained) in both BT474 and MCF‐7 cells, with a late‐stage apoptosis increase seen only in MCF‐7. Caspase studies determined an extrinsic pathway (Caspase‐8 and Caspase‐3 positive, Caspase‐9 negative) in BT474 cells. MCF‐7 cells were also Caspase‐8 positive, Caspase‐9 negative, but Caspase‐3 was not detected. In the absence of Caspase‐3, Caspase‐7 might serve as an effector caspase. However, Curcumin exposure did not upregulate its expression in MCF‐7 cells. While curcumin activated the mitochondria‐independent extrinsic pathway in BT474 cells (no translocation of Cytochrome C into the cytoplasm), a mitochondria‐dependent (translocation of Cytochrome C into the cytoplasm), extrinsic with possible intrinsic pathway involvement was activated in MCF‐7 cells. Further studies are needed to elucidate the altered apoptotic pathway utilized in MCF‐7 cells.

Integrated Bioinformatics Data Analysis Reveals Prognostic Significance Of SIDT1 In Triple-Negative Breast Cancer.

BackgroundTriple-negative breast cancer (TNBC) is a heterogeneous disease with a worse prognosis. However, current therapies have rarely improved the outcome of patients with TNBC. Here we sought to identify novel biomarkers or targets for TNBC.Materials and methodsPatients GSE76275 clinic traits and their corresponding mRNA profiles for 198 TNBC and 67 non-TNBC were obtained from the GEO database. Weighted gene co-expression network analysis (WGCNA) of the GSE76275 keyed out hub genes, and the differentially expressed genes (DEGs) were identified with the cut-off of adjusted P (adj. P) <0.01 and |log2 fold-change (FC)| > 1.5. The hub - DEGs overlapping genes, as key genes, were considered for further study using Kaplan-Meier plotter online analysis. Subsequently, Breast Cancer Gene-Expression Miner v4.0 and tissue microarray analysis were applied to determine the transcriptional and translational levels of every key gene. Following plasmid transfection for overexpression, the proliferation of TNBC cells was determined by CCK8 and colony formation assay. Moreover, xenograft tumor models were canvassed to investigate their effect upon in vivo tumor growth.ResultsFour genes (SIDT1, ANKRD30A, GPR160, and CA12) were found to be associated with relapse-free survival (RFS) in TNBC through WGCNA and DEGs integrated analysis. Patients with a higher level of SIDT1 had significantly better RFS compared to those with lower levels. The transcriptional and translational levels of SIDT1 were validated as downregulated in patients with triple-negative status, negative estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Furthermore, SIDT1 inhibited proliferation of breast cancer cells (MDA-MB-231 and MDA-MB-468) and xenograft studies demonstrated that SIDT1 can suppress tumor growth in vivo.ConclusionThis study suggests that SIDT1 may play a crucial role in TNBC progression and has the potential as a prognostic biomarker of TNBC.

长非编码RNA ZEB1-AS1在乳腺癌中的表达及其临床意义

Objective To detect the expression and clinic significance of long non-coding RNA ZEB1-AS1 in breast cancer. Methods A total of 130 patients with breast cancer in Baoji Central Hospital of Shaanxi Province from June 2007 to April 2015 were selected. Quantitative real-time PCR (qRT-PCR) was used to detect the expression level of ZEB1-AS1 in breast cancer tissues and corresponding normal tissues, and the relationships between the expression level of ZEB1-AS1 and the clinic characteristics of the patients and their overall survival time were analyzed. siRNA was used to disturb the expression of ZEB1-AS1. CCK-8 assay, clone formation assay and Transwell assay were used to detect the proliferation, cloning ability and migration of breast cancer MCF-7 cells in control group, siRNA-1 group and siRNA-2 group. Results The expression level of ZEB1-AS1 in breast cancer tissues was higher than that in corresponding normal tissues [M(QR): 0.001 6 (0.005 1) vs. 0.000 9 (0.001 5); Z=-4.426, P<0.001]. The higher expression of ZEB1-AS1 was correlated with lymphatic metastasis (χ2=9.148, P=0.027), negative human epidermal growth factor receptor 2 (χ2=5.039, P=0.025), triple negative breast cancer (χ2=4.597, P=0.032). The patients with the higher expression of ZEB1-AS1 had a shorter overall survival time compared with the patients with the lower expression of ZEB1-AS1 (χ2=14.340, P<0.001). CCK-8 assay showed that knock down of ZEB1-AS1 after 72 h, the absorbance values of the control group, siRNA-1 group and siRNA-2 group were 0.605±0.049, 0.488±0.054, 0.417±0.038 respectively, with a statistically significant different (F=15.936, P<0.001), and the two siRNA groups were significantly inhibited in cell proliferation compared with the control group (both P<0.05). The colonies of the control group, siRNA-1 group and siRNA-2 group were 297.5±11.4, 192.0±12.1, 204.8±12.8 respectively, with a statistically significant different (F=112.526, P<0.001), and the two siRNA groups were significantly inhibited in the cell clone compared with the control group (both P<0.001). The migratory cells numbers of the control group, siRNA-1 group and siRNA-2 group were 184.5±8.6, 147.5±18.6, 57.6±7.3 respectively, with a statistically significant different (F=12.409, P=0.001), and the two siRNA groups were significantly inhibited in the cell migration (both P<0.001). Conclusion ZEB1-AS1 is overexpressed in breast cancer, overexpression of ZEB1-AS1 induces a shorter overall survival in breast cancer patients, and knock down of ZEB1-AS1 can inhibit the proliferation, migration and colony formation ability of the breast cancer cell line. Key words: Breast neoplasms; Prognosis; ZEB1-AS1

Abstract PD3-07: A phase II pre-surgical trial of lapatinib for the treatment of women with HER2 positive or EGFR positive ductal carcinoma in situ

Abstract Background: Estrogen receptor (ER)-negative tumors and human epidermal growth factor 2-Neu (HER2) positive breast cancers are known to be more clinically aggressive subtypes of breast cancer and account for 30% of all breast cancers. Women with HER2 + breast cancers, whether ER+ or ER -, require cytotoxic chemotherapy with a HER2-targeting agent, and often have adverse outcomes. Thus, preventive agents are needed to reduce the incidence of these subtypes of aggressive breast cancer. Lapatinib, a dual tyrosine kinase inhibitor, inhibits epidermal growth factor receptors (EGFR) and HER2 kinases and has shown to decrease breast cell proliferation in invasive breast cancer and adjacent premalignant lesions. Therefore, we conducted a multi-institutional randomized Phase II clinical trial to study the effects of the signal transduction inhibitor lapatinib in women with HER2-positive or EGFR-positive ductal carcinoma in situ (DCIS). Methods: Randomized participants received either lapatinib (750mg, 1000mg, or 1500mg) or placebo daily for 2-6 weeks prior to their surgery. After minimal accrual, the trial was later amended to lapatinib 1000mg or placebo. Pre-treatment breast tissue was obtained from initial diagnostic core biopsy and post-treatment breast tissue was obtained from surgical excision specimen. Blood was obtained prior to surgery to assess serum lapatinib level. Participants kept a daily symptom assessment log and had a cardiac assessment at baseline and prior to surgery. Patients were instructed to take drug up to and including the day before surgery. The dual primary endpoint for this study was change in proliferation in pre- versus post-treatment biopsies between the two treatment arms, as measured by Ki67 as well as toxicity assessment. Secondary endpoints included incidence of DCIS at surgery and modulation of tissue biomarker expression in growth factor receptors (EGFR, ErbB2); phosphorylated growth factor receptor (phospho-ErbB2); signal transduction markers (MAPK, phospho-MAPK); hormone receptors (ER, PR); and p27. Results:Twenty-two women (mean age: 51; range: 32-66) with HER2+ or EGFR+ DCIS were treated with lapatinib (1,000 or 1,500 mg) or placebo for 2–6 weeks prior to surgical excision. Ki67 expression was significantly decreased in the lapatinib treatment arms compared to placebo (p=0.0122). Diarrhea, fatigue, and skin reactions were notable adverse events that occurred predominately in the lapatinib arm compared to placebo. No grade 3 or 4 events related to the study drug were noted during the study. No changes were noted in cardiac function. DCIS was present in all surgical specimens in both arms. Invasive breast cancer was noted in 1 patient on lapatinib 1000mg and 3 patients on placebo. No statistically significant changes were noted in signal transduction biomarkers Conclusion:These results demonstrate the effectiveness of lapatinib in reducing proliferation in women with EGFR+ or HER2+ DCIS. Even low-grade toxicities can deter use of an agent in the prevention setting. This and the lack of a risk model for HER2+ and triple negative breast cancer make the development of larger scale clinical prevention trials of lapatinib for the prevention a challenge. Citation Format: Thomas PS, Contreras A, Pruthi S, Krontiras H, Rimawi M, Garber J, Wang T, Hilsenbeck SG, Vornik LA, Gilmer T, Friedman R, Heckman-Stoddard BM, Dunn B, Kuerer H, Brown PH. A phase II pre-surgical trial of lapatinib for the treatment of women with HER2 positive or EGFR positive ductal carcinoma in situ [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD3-07.