Positive Subtypes Research Articles

Implications of Persistent HPV52 and HPV58 Positivity for the Management of Cervical Lesions.

ObjectiveThis study aimed to compare the variability of HPV16/18/52/58 subtype infections in patients with different cervical lesions, to explore the guiding significance of persistent positive HPV subtypes 52 and 58 in the stratified management of cervical lesions, and to determine the appropriate management model.MethodThis study was conducted through a retrospective analysis of 244,218 patients who underwent HPV testing at the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University from September 2014 to December 2020 to examine the distribution of different types of HPV infection. From March 2015 to September 2017, 3,014 patients with known HPV underwent colposcopy to analyze high-risk HPV infection for different cervical lesions. Meanwhile, from September 2014 to December 2020, 1,616 patients positive for HPV16/18/52/58 alone with normal TCT who underwent colposcopy in our hospital were retrospectively analyzed for the occurrence of cervical and vulvovaginal lesions, with colposcopic biopsy pathology results serving as the gold standard for statistical analysis.ResultAnalysis of 244,218 patients who had HPV tested revealed that the top 3 high-risk HPV types were HPV52, HPV58, and HPV16. Further analysis of 3,014 patients showed that 78.04% of patients referred for colposcopy had HPV16/18/52/58 alone. Among high-grade squamous intraepithelial lesions (HSIL) and cervical cancer, the most common is HPV16, followed by HPV58 and then HPV52 (p < 0.05). A total of 1,616 patients with normal TCT who were referred for colposcopy due to HPV16/18/52/58 infection were further analyzed. Based on pathological findings in lesions of HSIL and CC, HPV16 is the most common, followed by HPV58 and then HPV18 (p < 0.05). In the 1,616 patients analyzed, high-grade vulvovaginal lesions were detected, with HPV58 being the most common, followed by HPV16 and then HPV52 (p < 0.05).Conclusion1. In patients with positive HPV58 alone and normal TCT, the indications for colposcopy may be relaxed, with particular attention paid to the possibility of vulvar and vaginal lesions.2. Patients with a positive HPV type 52 alone and normal TCT may be considered for a follow-up review and, if necessary, a colposcopy.3. The development of a more suitable HPV vaccine for the Asian population, such as HPV16/18/52/58, may better protect women’s health.

The Role of 18F-FDG PET/CT in Predicting the Neoadjuvant Treatment Response in Patients with Locally Advanced Breast Cancer

Purpose: Patients with locally advanced breast cancer (LABC) should be treated with neoadjuvant chemotherapy (NAC). Pathological complete response (pCR) is related to better disease-free survival (DFS). The best strategy for assessing the efficacy of NAC has not been established yet, but several studies have shown that 18F-FDG PET/CT is a potential imaging tool for assessing pCR. The aim of this study is to investigate the merits of 18F-FDG PET/CT imaging in predicting pCR in both axillary and breast tissue and to establish a threshold maximum standard uptake value (SUVmax) for predicting the response after completion of NAC. Methods: A total of 186 LABC patients, treated with an NAC regimen according to tumor subtype, were retrospectively analyzed in this study. All patients underwent 18F-FDG PET/CT imaging before and after completion of NAC. PET parameters were measured in the most FDG avid breast tissue and axillary lymph nodes. We analyzed the correlation between the tumor SUVmax of the PET/CT response and the pCR after surgery. DFS was also evaluated with respect to pCR. Results: Higher pCR rates were significantly associated with a higher tumor grade, an initial Ki-67 ≥20% (p = 0.03 and p = 0.003, respectively), a triple-negative subtype (32.9%), and a human epidermal growth factor receptor 2 (HER-2)-positive subtype (24.7%) (p < 0.001). There was a significant correlation between the pCR and a complete response in 18F-FDG PET/CT (p < 0.001). The overall sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of 18F-FDG PET/CT to determine the pCR after NAC were 100%, 72.2%, 85%, 75.2%, and 100%, respectively. We demonstrated a 1.1 cutoff SUVmax for breast tumors after NAC (OR: 3.94, 95% CI: 1.14–5.05, p = 0.004), the 18F-FDG PET/CT response to NAC (OR: 0.50, 95% CI: 0.25–0.99, p = 0.003), and the molecular subtype of breast tumors (OR: 0.58, 95% CI: 0.38–0.88, p = 0.011). Conclusion: Our results confirm that 18F-FDG PET/CT is a useful method for predicting the NAC response in LABC.

ADGRF1 promotes tumorigenesis via the activation of STAT3 pathway in HER2+ and triple‐negative breast cancers

Adhesion G protein‐coupled receptors (aGPCR) regulate many cellular processes involved in cancer. ADGRF1, a class VI aGPCR, is overexpressed and/or predicts poor survival in many cancer types. We previously reported that in breast cancer (BC), ADGRF1 is highly expressed in human epidermal growth factor receptor‐2 positive (HER2+) and basal‐like subtype, 85% of which are triple‐negative. Also, higher ADGRF1 expression predicts worse BC‐specific survival in both HER2+ and triple‐negative BCs patients. We previously showed that inducible ADGRF1 overexpression promoted and knockdown inhibited tumorigenesis in HER2+ BC cells. Consistently, ADGRF1 overexpression promoted tumor growth in vivo. We also reported that ADGRF1 coupling to Gαs was responsible for its pro‐tumorigenic effects. However, the downstream mediators of tumorigenesis promoted by ADGRF1 signaling remain largely unknown in cancer. Since STAT3 is a well‐known mediator of tumorigenesis in many cancers, and previous studies have shown that ADGRF1 modulates STAT3 activation in some cancers, we aimed to evaluate the effects of genetic and pharmacologic modulation of ADGRF1 on STAT3 phosphorylation (Y705) in HER2+ and triple‐negative breast cancers models. In a HER2+ BC cell line model (BT474), we found that ADGRF1 overexpression increased STAT3 phosphorylation (Y705) in vitro and in vivo. ADGRF1 knockdown also decreased STAT3 phosphorylation in BT474 cells and MDA‐MB‐231 cells, a triple‐negative BC cell line with high endogenous ADGRF1 gene expression and amplification. ADGRF1 knockdown inhibited cell growth as well as mammosphere and colony formation (a measure of tumorigenesis) in triple‐negative BC cell lines. Synaptamide increased secondary mammosphere formation, STAT3 phosphorylation (Y705), and cAMP but not IP1 in MDA‐MB‐231 cells while it had no effect on BT474 cells with inducible ADGRF1 overexpression. Synaptamide‐induced STAT3 phosphorylation was reversed by SQ22536, an adenylyl cyclase inhibitor. Our data suggest that ADGRF1 promotes tumorigenesis by coupling to Gas, which in turn activates STAT3 in HER2+ and triple‐negative BCs.

Bird’s eye view analysis of in situ cholesterol metabolic pathways in breast cancer patients and its clinicopathological significance in their subtypes

Obesity has been known to increase the risks of breast cancer (BC) development and also to be associated with adverse clinical outcome of the patients. Abnormalities of cholesterol metabolism are not only related to obesity but also to biological or clinical behavior of BC patients. However, which metabolites or pathways of cholesterol metabolism could represent the characteristics of BC patients have remained virtually unknown. Therefore, in this study, we attempted to perform bird’s eye view or comprehensive analysis of in situ or intra-tumoral cholesterol metabolic pathways using the multimodal approaches in order to elucidate the possible significance of cholesterol metabolites and its metabolic enzymes including CYP27A1, CYP7A1, and CYP46A1. GC-MS study using BC specimens was first performed in 60 BCE patients to evaluate cholesterol metabolism from cholesterol through oxysterols in both BC and normal tissues. Results of those analyses above lead to evaluating immunoreactivity and mRNA expression of CYP27A1, CYP7A1 and CYP46A1 in 213 and 153 BCE cases, respectively. Results of comprehensive GC-MS analysis did reveal that three oxysterols, 27-HC, 7α-HC and 24-HC were all related to malignant phenotypes in BC. 27-HC abundance was significantly associated with higher tumor stage (P = 0.0475) of BC patients. Luminal B type BC patients harboring high CYP27A1, the enzyme responsible for production of 27-HC were significantly associated with worse disease-free survival than those with low CYP27A1 (P = 0.0463). 7α-HC tended to be more abundant in HER2 positive and TNBC subtypes and higher levels of 7α-HC were also significantly associated with higher Ki-67 labeling index (P = 0.0022) and histological grade (P = 0.0286). CYP7A1, the enzyme involved in production of 7α-HC, was significantly more abundant in TNBC than other subtypes (vs Luminal A; P = 0.0321, vs Luminal B; P = 0.0048, vs HER2; P = 0.0103). The levels of 24-HC in BC were lower than normal breast tissues regardless of its subtypes. CYP46A1, the enzyme involved in the production of 24-HC, was detected only in 33 (15.5%) out of 213 BCE cases examined in this study. Results of our bird’s eye view analysis of in situ or intra-tumoral cholesterol metabolism in BC patients did firstly reveal BC subtype dependent involvement of its different pathways. Results also indicated the therapeutic possibility of subtype dependent modification of cholesterol metabolizing pathways in BC patients.

Relation between tumor SUVmax, TLR and TSR and breast carcinoma molecular subtypes in PET CT

Breast cancer is known to be one of the most cancer affecting women around the globe and the second most common cancer in general. In third worlds countries, breast cancer is the most cause of cancer death. Early diagnosis and accurate follow-up of these patients affect the management. There are multiple prognostic factors most important one is the immunohistochemical molecular markers in the specimens including human epidermal growth factor, progesterone, and estrogen receptors (HER 2, PR, ER). In breast cancer, the HER2 positive molecular subtype is associated with bad prognosis and aggressive histological features, yet while following neoadjuvant chemotherapy, it achieves an increased pathological complete response rate. 18F-fluoro-2-deoxy-d-glucose positron emission tomography (FDG PET) has proved to be an effective and accurate imaging technique for lymph node and distant metastasis assessment, tumor staging, restaging of recurrence, treatment response, and follow-up. In breast cancer, tumor molecular subtype, tumor size, proliferation index, and histological grade correlated with 18F-fluoro-2-deoxy-d-glucose (FDG) uptake. This study evaluates the possible correlation between tumor SUV max ,tumor to liver and tumor to spleen (standardized uptake value) SUV max ratio and the four different molecular subtypes in patients with pathologically proven breast cancer.

Abstract P1-21-04: The real word study on clinical features and prognostic factors of Chinese breast cancer patients with brain metastasis

Abstract Purpose: we used the real-world data of Chinese patients to describe clinicopathological characteristics of patients with breast cancer brain metastases (BCBM) and to investigate survival after diagnosis of brain metastases (BM). Methods: The clinicopathological characteristics of 700 patients with BCBM at the fifth Medical Center of Chinese PLA General Hospital between 2003 and 2021 were retrospectively reviewed. Information was collected from the electronic medical records, including patient demographics, tumor characteristics, and dates of diagnosis of original breast carcinoma and subsequent metastases. Sites and number of metastatic lesions were also recorded, along with data on treatments and outcomes. The prognostic and predictive effects of these clinicopathological variables in BCBM were analyzed. Results: The median age at diagnosis of primary breast cancer was 44 years (range, 22-80 years) and all patients were female. Among the 700 patients with BCBM, 30.0%, 49.7%, 18.9%, 1.4% had Luminal, HER2 positive, triple-negative and unknown subtypes, respectively. Based on available clinical information, the proportions of extracranial metastasis at diagnosis included bone (63.7%), lymph nodes (61.6%), lung (55.1%), liver (50.1%), the soft tissues of the thoracic wall (26.1%) and bone marrow (4.9%).The median time from the diagnosis of breast cancer to the development of BM was 52.0 months [95% confidence interval (CI), 44.3-59.7], 36.0 months (95%CI, 31.9-40.1), and 29.0 months (95%CI, 22.4-35.6) for patients with Luminal, HER2 positive and triple-negative, respectively (p&amp;lt;0.0001) (Figure 1). The median time from the first recurrence to the diagnosis of BM was 18 months for Luminal subtype (95% CI, 15.3-20.7), 14 months for HER2 positive subtype (95% CI, 12.0-16.0), and 12 months for triple-negative subtype (95% CI, 9.0-15.0), respectively (p&amp;lt;0.0001) (Fig 2). The median survival after the diagnosis of BCBM in Luminal, HER2 positive and triple-negative patients was 11 months (95% CI, 8.7-13.2), 13 months (95% CI, 11.4-14.6) and 7 months (95% CI, 5.7-8.3), respectively(p&amp;lt;0.05) (Fig 3). Patients presenting with a solitary brain metastasis had a longer median survival after the diagnosis of BM than patients who presented with multiple brain metastases (14.0 months versus 9 months, p&amp;lt;0.0001). The median survival time of patients with and without anti-HER2 treatment was 14 months (95% CI, 12.2-15.8) and 9 months (95% CI, 6.1-11.9), respectively (p=0.01) (Fig 4). Patients presenting with leptomeningeal metastasis had shorter time after the diagnosis of BM than patients without leptomeningeal metastasis, which were 5 months (95% CI, 1.8-8.2) and 11 months (95% CI, 9.8-12.2), respectively (p=0.002). Conclusion: Despite the improvement noted with more recent year of diagnosis and treatment, survival following a diagnosis of BCBM remains poor and still needs attention. It is necessary to further explore the comprehensive treatment mode of patients with different types and degrees of brain metastasis in order to provide reference for clinical treatment. Table 1.patient characteristics (n=700)Parametern (%)Mean age, y, at diagnosis (range)44(22-80)Stage at initial diagnosis of cancerI89(12.7)II342(48.9)III167(23.9)IV69(9.9)unknown33(4.7)Molecular subtypesLuminal A/B211(30.0)HER2-positive347(49.7)TNBC132(18.9)Unknown10(1.4)Site of first metastasisBrain128(18.3)Others572(81.7)Number of BM at time of BM diagnosis1189(27.0)248(6.9)≥3454(64.9)Unknown9(1.3)Visceral metastases at diagnosis of BMliver351(50.1)lung386(55.1)bone446(63.7)lymph nodes431(61.6)the soft tissues of the thoracic wall183(26.1)bone marrow34(4.9) Citation Format: Tao Wang, Huiqiang Zhang, Feng Li, Xia Wu, Jinmei Zhou, Shao hua Zhang, Li Bian, Ze fei Jiang. The real word study on clinical features and prognostic factors of Chinese breast cancer patients with brain metastasis [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-21-04.

ULTRASONOGRAPHY AND MAMMOGRAPHIC FEATURES ACTING AS PROXY TO MOLECULAR SUBTYPING OF BREAST CARCINOMA: A CROSS SECTIONAL STUDY

1. AIMS AND OBJECTIVES : Ÿ To determine the correlation between and predict the molecular subtypes of breast carcinoma based on mammography and ultrasound ndings. Ÿ To correlate the different epidemiological features of breast cancer patients with the molecular subtypes of breast carcinoma. 2. Materials And Methods: In our cross-sectional study, 84 biopsy proven cases of breast carcinoma were referred to radiology department. After taking informed consent and in presence of a female attendant, ultrasonography and mammography of the patients were done. All these patients went for surgical excision of the neoplastic mass; and the mass were sent for IHC analysis. Epidemiological data, clinical features, mammographic features and sonographic features were correlated with IHC subtype of the mass and P-Values were found out. P-Value&lt;0.05 was considered to be statistically signicant. 3. Results: Ÿ Among 84 patients, 22 were of Triple Basal Negative subtype, 17 were of Luminal-A subtype, 16 were of Luminal-B (Her-2 negative) subtype, 15 were of Her-2 enriched subtype and 14 out of 84 patients were of Luminal-B (Her-2 positive) subtype. Ÿ IHC correlation between sex distribution, age distribution, age of menarche distribution, achievement of menopause and age of menopause distribution, parity distribution, history of breast feeding, family history of breast carcinoma, history of breast pain, breast ulceration, nipple discharge, quadrant location of breast mass and size of mass showed P-Value &gt;0.05; which was statistically insignicant. Ÿ IHC corelation with mammographic microcalcications, shape of breast mass, margins of breast mass, posterior acoustic pattern of breast masses and vascularity of the breast mass showed P-Value &lt;0.05; which was statistically signicant. 4. Conclusion: Mammographic microcalcication and other features on sonography like shape of the mass, margins of the mass, posterior acoustic features and vascularity of the mass are strongly correlated in predicting the molecular subtype of breast carcinoma.

Abstract P5-05-03: Adipocyte associated methionine domain containing (AAMDC): A nuclear envelope protein with predictive and prognostic potential in luminal breast cancer

Abstract Background Estrogen receptor (ER) positive breast cancers (BCs) are less likely to relapse than other types, due to a lower propensity to disseminate and to sensitivity to anti-estrogen drugs in the adjuvant setting. However, combined analysis of DNA mutation and mRNA expression profiles in BC identifies a number of Integrative Clusters (IntClus), one of which, IntClus2, is characterized by ER positivity but high relpase and mortality. A hallmark of IntClus2 tumours is an amplification of chromosome 11, at the center of which lies the gene for the protein AAMDC. Pre-clinical research shows this protein to drive proliferation, migration, colony formation, switching between glucose and lipid metabolism, folate metabolism and anti-estrogen resistance. We postulated that resultant clinical behaviours could include; high relapse and mortality, anti-estrogen resistance, an interaction with the protective effect on relapse from statins and sensitivity to fluorouracil (FU)-based therapies. Methods The retrospective study cohort comprised 1766 ER positive early BC patients treated 1994-2014. Patient and tumour demographic, statin prescription and treatment data were collated. Primary tumour tissue was available with AAMDC expression data on 419 patients. On noting nuclear (N), cytoplasmic (C) and nuclear envelope (NE) expression, separate intensity scores were attributed to each localization. Intensity score cut-points were based on providing statistically useable sub-group sizes for comparison. Results No significant correlations between BC biology and patterns of AAMDC expression were seen. Cytoplasmic AAMDC weakly linked to some aggressive features, including high grade, luminal B sub-type and progesterone receptor negativity, although less lymph node spread. In contrast, nuclear AAMDC associated with some less aggressive features, being less in high grade and luminal B cancers. Nuclear envelope AAMDC was rarer in the HER2 positive sub-type without other correlates. A trend to higher distant relapse was seen for NE +ive BCs (20 v 14%, p=0.09). Overall mortality was statistically higher for NE expression (54 v 44% mortality, p=0.04) and cytoplasmic expression (56 v 47% mortality, p=0.04) and a trend to lower mortality for nuclear expression (46 v 54%, p=0.06). Looking at combinatorial expression, BC deaths were significantly lower in the nuclear +ive/NE -ive phenotype (4.7%) than the nuclear -ive/NE -ive (19.5%, p=0.013), nuclear -ive/NE +ive (17.6%, p=0.017) or nuclear +ive/NE +ive phenotypes (19.3%, p=0.011).Considering the effects of statins on early BC outcome, overall our patients on statins had notably lower BC return rates (HR 0.31, p=0.004). Considering interactions with AAMDC, in high-risk NE +ive cancers statin protection was high (HR for relapse 0.33, p=0.032), compared to no protection in already low risk NE -ive tumours (HR 0.90, p=0.41). The hypothesis that AAMDC confers sensitivity to FU-based agents was confirmed with a correlation observed for longer responses to FU-based chemotherapy for increasing NE AAMDC expression (R=0.334), whereas no link was seen for other agents or other AAMDC expression sub-locations. The mean times on FU-based therapy for low, moderate and high NE expression were 2.0, 5.7 and 12.2 months respectively (p=0.02). This work indicates that AAMDC expression can BC affect outcome, identifying nuclear envelope located AAMDC as both associating with higher mortality and correlating with superior protection from statins as well as response to FU-based chemotherapies. This could guide metastatic chemotherapy selection for affected patients, implies NE positive BCs might preferentially benefit from adjuvant statins and suggests that AAMDC and NE-associated AAMDC partners may be future therapeutic targets. Citation Format: Indunil Weerasena, Lisa Spalding, Benjamin F Dessauvagie, Emily Golden, Eleanor Woodward, Pilar Blancafort, Andrew Redfern. Adipocyte associated methionine domain containing (AAMDC): A nuclear envelope protein with predictive and prognostic potential in luminal breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-05-03.

Abstract P5-16-02: Updated efficacy, safety and translational data from MARIO-3, a phase II open-label study evaluating a novel triplet combination of eganelisib (IPI-549), atezolizumab (atezo), and nab-paclitaxel (nab-pac) as first-line (1L) therapy for locally advanced or metastatic triple-negative breast cancer (TNBC)

Abstract Background: Eganelisib is a first-in-class, oral, immuno-oncology macrophage reprogramming agent. The IMpassion130 randomized trial in 1L advanced TNBC has demonstrated improved efficacy with the addition of atezo to nab-pac in patients with PD-L1+ tumors. Adding eganelisib to the approved doublet of atezo and nab-pac is expected to improve patient benefit regardless of PD-L1 status. Methods: Eligible patients (pts) had measurable unresectable locally advanced or metastatic TNBC and no prior systemic therapy for advanced disease. A safety run-in assessed the safety of the triplet of oral eganelisib 30 mg daily in combination with IV nab-pac 100 mg/m2 given on days 1, 8, &amp; 15, and IV atezo 840 mg given on days 1 &amp; 15 of a 28 day cycle. After establishing tolerability and confirmation of the 30 mg dose in the safety run-in (n=6), the expansion phase of the phase II study was initiated to enroll approximately 60 pts (30 PD-L1+ and 30 PD-L1-). Cycles are repeated until loss of clinical benefit, unacceptable toxicity or consent withdrawal. The primary efficacy endpoint is confirmed complete response (CR) rate per RECIST v1.1. Secondary endpoints include the overall response rate (ORR), progression free survival and safety assessment. Results: As of 6/26/2021, data from the safety run-in and the expansion cohorts are reported for 43 pts evaluable for safety and 38 evaluable for efficacy defined as having at least one post-baseline tumor assessment. 2 CRs (2/38), 19 PRs (19/38), and 11 SDs (11/38) were observed with an ORR of 55.3% (21/38) and a disease control rate (DCR) of 84.2% (32/38). Responses were seen irrespective of PD-L1 status; in PD-L1+ pts, ORR is 66.7% (8/12) and DCR is 91.7% (11/12), and in PD-L1- pts, ORR is 47.8% (11/23) and DCR is 78.3% (18/23). The most common all-grade adverse events were nausea (51.2%), fatigue (48.8%), alopecia (32.6%), diarrhea (32.6%), rash maculo-papular (30.2%), ALT increased (27.9%), and AST increased (25.6%). The most common grade ≥3 adverse events were ALT increased (18.6%), AST increased (14.0%), neutrophil count decreased (9.3%) and rash maculo-papular (9.3%). The safety profile is acceptable and in line with expectations of component drugs with no additive or new safety signals. Translational data from paired biopsies show a shift in tumor associated macrophages towards an immune activated state as well as an increase in cytotoxic T cells. In addition, a subset of tumors with baseline PD-L1 negative status show conversion to PD-L1 positive status post treatment. Consistent with immune activation, increased interferon gamma signature, increased proinflammatory cytokines and T cell invigoration were observed in peripheral blood cells. Conclusions: The novel triplet regimen of eganelisib, atezo, and nab-pac shows promising anti-tumor activity (ORR 55.3% and DCR 84.2%) irrespective of PD-L1 status and has manageable toxicity. . Translational data from paired biopsies and peripheral blood show evidence of immune activation regardless of PD-L1 status as well as conversion from PD-L1 negative to PD-L1 positive subtype within a subset of patients treated with the triplet regimen. The expansion phase of the phase II study is currently enrolling. At the time of presentation, we will have updated efficacy, safety and translational data. Citation Format: Soliman Hatem, Jeffrey Hargis, Anthony Elias, Arielle Lee, Rachel Swart, Shaker Dahkil, Alexandra Drakaki, Vu Phan, Frederic Kass, Melody Cobleigh, Sanil Babu, Katherine Tkaczuk, Brenda O'Connell, Jennifer Roberts, Nora Zizlsperger, Erika Hamilton. Updated efficacy, safety and translational data from MARIO-3, a phase II open-label study evaluating a novel triplet combination of eganelisib (IPI-549), atezolizumab (atezo), and nab-paclitaxel (nab-pac) as first-line (1L) therapy for locally advanced or metastatic triple-negative breast cancer (TNBC) [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-16-02.

Quantitative CK19 biomarker detection in breast cancer cell lines.

Cytokeratin19 (CK19) was detected as the most related marker for circulating tumor cells, which was assessed in specific cell lines. MCF7, SKBR3, T47D, and MDA-MB-231, and HeLa cell line as negative control were used. CK19 expression was confirmed by using mouse monoclonal anti-human CK19 antibody. CK19 detection in MDA-MB-231 was not observed. CK19 marker expression was compared in T47D, MCF7, and SKBR3 cell lines. T47D and MCF7 belonged to the luminal subtype of breast cancer (BC) that CK19 expression regulated with an ER marker. SKBR3 belonged to the HER2 positive subtype of BC. However, MDA-MB-231 belonged to the claudin-low subtype of BC that lack of CK19 expression strongly is related to negative ER, PR, and HER2. Therefore, there are not only quantitative differences in CK19 expression, but its expression could also link to the other markers of BC that should be considered in the molecular classification of breast carcinoma. Different expression levels related to cell classification could be useful in the prognosis and treatment of cancers with epithelial origins.

Human papillomavirus E6E7 mRNA and TERC lncRNA in situ detection in cervical scraped cells and cervical disease progression assessment

BackgroundHuman papillomavirus screen in female cervical cells has demonstrated values in clinical diagnosis of precancerous lesions and cervical cancers. Human papillomavirus tests of cervical cells by utilizing Polymerase Chain Reaction (PCR) method provides human papillomavirus infection status however no further virus in situ information. Although it is well known that the tests of human papillomavirus E6/E7 RNA location in infected cervical cells and cell internal malignancy molecular will provide clues for gynecologists to evaluate disease progression, there are technique difficulties to preserve RNAs in cervical scraped cells for in situ hybridization.MethodsIn current study, after developing a cervical cell collection and preparation method for RNA in situ hybridization, we captured the chance to screen 98 patient cervical cell samples and detected human papillomavirus E6/E7 mRNAs of high-risk subtypes, low-risk subtypes and long non-coding RNA (lncRNA) TERC in the cells.ResultsThere were 69 samples exhibited consistence between human papillomavirus PCR and human papillomavirus RNA in situ hybridization results in cervical collected cells. Among them, 23 were both positive and 46 were both negative. In the rest 29 samples, 8 were HPV RNAscope positive, either high risk or low risk subtypes, however HPV PCR negative. Another 9 samples were HPV PCR results positive whereas RNAscope negative. The last 12 samples were HPV positive detected by both RNAscope and PCR methods, however inconsistent between high-risk and low-risk subtypes. In RNAscope positive samples, viral E6/E7 mRNAs were observed to distribute in cervical scraped cell nucleus and cytoplasm. Moreover, HPV viral RNA gathered clusters were observed outside of cells through human papillomavirus RNA in situ hybridization detection. Varied numbers of human papillomavirus infective cells were detected by RNAscope assay in different patients even though they were all human papillomavirus high-risk subtype positive discovered by human papillomavirus PCR results. A cell malignancy related long non-coding RNA, TERC, has been detected in seven patient samples. The patient follow-up information was further analyzed with RNAscope results which indicated a combination of RNAscope positive signals of TERC and human papillomavirus high risk signals in more than 10 cells (cytoplasm or nucleus) may connect with cervical lesion fast progression which deserves further studies in the future.CConclusionsTaken together, current study has provided an observable clue for gynecologists to evaluate human papillomavirus infection stage and cell malignancy status which may contribute for assessment of cervical disease progression.

Secretory pathway Ca2+-ATPase SPCA2 regulates mitochondrial respiration and DNA damage response through store-independent calcium entry

A complex interplay between the extracellular space, cytoplasm and individual organelles modulates Ca2+ signaling to impact all aspects of cell fate and function. In recent years, the molecular machinery linking endoplasmic reticulum stores to plasma membrane Ca2+ entry has been defined. However, the mechanism and pathophysiological relevance of store-independent modes of Ca2+ entry remain poorly understood. Here, we describe how the secretory pathway Ca2+-ATPase SPCA2 promotes cell cycle progression and survival by activating store-independent Ca2+ entry through plasma membrane Orai1 channels in mammary epithelial cells. Silencing SPCA2 expression or briefly removing extracellular Ca2+ increased mitochondrial ROS production, DNA damage and activation of the ATM/ATR-p53 axis leading to G0/G1 phase cell cycle arrest and apoptosis. Consistent with these findings, SPCA2 knockdown confers redox stress and chemosensitivity to DNA damaging agents. Unexpectedly, SPCA2-mediated Ca2+ entry into mitochondria is required for optimal cellular respiration and the generation of mitochondrial membrane potential. In hormone receptor positive (ER+/PR+) breast cancer subtypes, SPCA2 levels are high and correlate with poor survival prognosis. We suggest that elevated SPCA2 expression could drive pro-survival and chemotherapy resistance in cancer cells, and drugs that target store-independent Ca2+ entry pathways may have therapeutic potential in treating cancer.

Programmed Death-Ligand 1 (PD-L1) as Immunotherapy Biomarker in Breast Cancer.

Simple SummaryBreast cancer (BC) is the most common malignant neoplasm in women and one of the leading causes of cancer death in women worldwide. Programmed death-ligand 1 (PD-L1) is becoming an emerging biomarker in BC in recent years. It has been correlated with worse outcomes in patients with hormone receptor positive, but it has a predictive role to guide response to systemic treatment in the triple-negative breast cancer (TNBC) subtype, especially in the metastatic setting. Immune checkpoint inhibitors are beginning to be a part of the treatment for many TNBC patients. However, more studies are needed in order to identify wherefore immunotherapy benefits TNBC patients regardless of PD-L1 status in the localized disease, but only offer an improvement for PD-L1 positivity expression in the advanced setting. The aim of this review is to analyze PD-L1 in all BC subtypes, including clinical trials with anti-PD-1/L1 and their results.Breast cancer constitutes the most common malignant neoplasm in women around the world. Approximately 12% of patients are diagnosed with metastatic stage, and between 5 and 30% of early or locally advanced BC patients will relapse, making it an incurable disease. PD-L1 ligation is an immune inhibitory molecule of the activation of T cells, playing a relevant role in numerous types of malignant tumors, including BC. The objective of the present review is to analyze the role of PD-L1 as a biomarker in the different BC subtypes, adding clinical trials with immune checkpoint inhibitors and their applicable results. Diverse trials using immunotherapy with anti-PD-1/PD-L1 in BC, as well as prospective or retrospective cohort studies about PD-L1 in BC, were included. Despite divergent results in the reviewed studies, PD-L1 seems to be correlated with worse prognosis in the hormone receptor positive subtype. Immune checkpoints inhibitors targeting the PD-1/PD-L1 axis have achieved great response rates in TNBC patients, especially in combination with chemotherapy, making immunotherapy a new treatment option in this scenario. However, the utility of PD-L1 as a predictive biomarker in the rest of BC subtypes remains unclear. In addition, predictive differences have been found in response to immunotherapy depending on the stage of the tumor disease. Therefore, a better understanding of tumor microenvironment, as well as identifying new potential biomarkers or combined index scores, is necessary in order to make a better selection of the subgroups of BC patients who will derive benefit from immune checkpoint inhibitors.

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.

Prognostic factors and molecular subtypes in young women with breast cancer

Introduction: Because of its high incidence, breast cancer is the subject of numerous studies today. Despite being an uncommon disease in young women, when it affects this population, it tends to be more aggressive and has high mortality rates. Objective: The objective of this study was to evaluate the prognostic factors present in the immunohistochemical profile of young women with breast cancer, comparing the age groups of very young women (&lt;35 years old — Group I) and young women (between 35 and 40 years old — Group II), to see if the data obtained match what is reported in the literature. Methods: A cross-sectional study was carried out, analyzing the immunohistochemical tests of 90 female patients with invasive breast carcinoma. The groups were classified on the basis of molecular subtype: luminal A, luminal B, hybrid luminal, human epidermal growth factor receptor 2 positive and triple-negative. Results: The histological type with the highest incidence was invasive breast carcinoma of no special type. The most frequent molecular subtypes were luminal B and triple-negative. With regard to estrogen and progesterone receptors, there was a slight predominance of positive receptors. Ki-67 levels showed that in the triple-negative and human epidermal growth factor receptor 2 positive subtypes, there was a predominance of high cell proliferation index. Conclusion: In the population of young women in this cohort of patients, there was agreement with literature data regarding the predominance of the invasive carcinoma of no special type histological type and the luminal B and triple-negative molecular subtypes, and the presence of high cell proliferation rates, attesting to the higher prevalence of more aggressive tumors in the younger population. There was also no statistically significant difference in all aspects analyzed when comparing Groups I and II. However, a higher frequency of negative hormone receptors or overexpressed human epidermal growth factor receptor 2 molecular subtypes was not detected, characteristics that are common to young women with breast cancer, which has been pointed out in several studies worldwide.

Central Nervous System Metastases from Triple-Negative Breast Cancer: Current Treatments and Future Prospective.

It is estimated that approximately one-third of patients with triple-negative breast cancer (TNBC) will develop brain metastases. The prognosis for patients with breast cancer brain metastasis has improved in the recent past, especially for hormone receptor and human epidermal growth factor receptor 2 (HER) positive subtypes. However, the overall survival rate for patients with triple-negative subtype remains poor. The development of newer treatment options, including antibody-drug conjugates such as Sacituzumab govitecan, is particularly encouraging. This article reviews the clinical outcomes, challenges, and current approach to the treatment of brain metastasis in TNBC. We have also briefly discussed newer treatment options and ongoing clinical trials. The development of brain metastasis significantly decreases the quality of life of patients with TNBC, and newer treatment strategies and therapeutics are the need of the hour for this disease subgroup.

Early on-treatment transcriptional profiling as a tool for improving pathological response prediction in HER2-positive inflammatory breast cancer.

Background:Inflammatory breast cancer (IBC) is a rare and understudied disease, with 40% of cases presenting with human epidermal growth factor receptor 2 (HER2)-positive subtype. The goals of this study were to (i) assess the pathologic complete response (pCR) rate of short-term neoadjuvant dual-HER2-blockade and paclitaxel, (ii) contrast baseline and on-treatment transcriptional profiles of IBC tumor biopsies associated with pCR, and (iii) identify biological pathways that may explain the effect of neoadjuvant therapy on tumor response.Patients and Methods:A single-arm phase II trial of neoadjuvant trastuzumab (H), pertuzumab (P), and paclitaxel for 16 weeks was completed among patients with newly diagnosed HER2-positive IBC. Fresh-frozen tumor biopsies were obtained pretreatment (D1) and 8 days later (D8), following a single dose of HP, prior to adding paclitaxel. We performed RNA-sequencing on D1 and D8 tumor biopsies, identified genes associated with pCR using differential gene expression analysis, identified pathways associated with pCR using gene set enrichment and gene expression deconvolution methods, and compared the pCR predictive value of principal components derived from gene expression profiles by calculating and area under the curve for D1 and D8 subsets.Results:Twenty-three participants were enrolled, of whom 21 completed surgery following neoadjuvant therapy. Paired longitudinal fresh-frozen tumor samples (D1 and D8) were obtained from all patients. Among the 21 patients who underwent surgery, the pCR and the 4-year disease-free survival were 48% (90% CI 0.29–0.67) and 90% (95% CI 66–97%), respectively. The transcriptional profile of D8 biopsies was found to be more predictive of pCR (AUC = 0.91, 95% CI: 0.7993–1) than the D1 biopsies (AUC = 0.79, 95% CI: 0.5905–0.9822).Conclusions:In patients with HER2-positive IBC treated with neoadjuvant HP and paclitaxel for 16 weeks, gene expression patterns of tumor biopsies measured 1 week after treatment initiation not only offered different biological information but importantly served as a better predictor of pCR than baseline transcriptional analysis.Trial Registration:ClinicalTrials.gov identifier: NCT01796197 (https://clinicaltrials.gov/ct2/show/NCT01796197); registered on February 21, 2013.

Neutrophil-lymphocyte Ratio as Independent Prognostic Factor among Breast Cancer Patients in a Tertiary Care Hospital, Kolkata, India

Introduction: The most important prognostic factor for breast cancer is tumour stage and tumour grade. However, the assessment of the above parameters are time-consuming and require expertise. Thus evaluation of the prognosis of breast cancer is still limited to tertiary care hospitals, with appropriate facilities for histopathological techniques. Recently, inflammatory blood markers have shown a role as a prognostic factor. Out of all the inflammatory blood markers, neutrophil-lymphocyte ratio has emerged as the most useful. Abundant evidence suggests the role of NLR as an adverse prognostic factor in breast cancer. NLR is simple and inexpensive. It can be easily obtained, as the differential count of every patient is done routinely. Thus it can act as an indicator of high-risk patients who are likely to show poor prognosis. Though NLR has been found to play a role in prognosis prediction in breast cancer, much is unknown in this field. Aim: To assess the effectiveness of Neutrophil-Lymphocyte Ratio (NLR) as an independent prognostic parameter among breast cancer patients in a tertiary care hospital in Kolkata, India. Materials and Methods: This observational cross-sectional study was conducted in Department of Pathology, Nil Ratan Sircar Medical College and Hospital, Kolkata, West Bengal, India, where 140 female patients undergoing mastectomy for breast cancer were studied from 1st February 2019 to 31st January 2020. The clinicopathological parameters, histopathological parameters, and molecular subtypes were evaluated. NLR was calculated and related with the other prognostic parameters. Data entry was done in Microsoft Excel and analysis was done using Statistical Package for Social Sciences (SPSS) software version 20.0. One way Analysis of Variance (ANOVA) test and Chi-square test was conducted to assess the relationship of NLR with various other prognostic factors. Results: Out of 140 patients, 78 patients showed NLR values within 1.8 to 3.33. Higher NLR (&gt;3.33) was associated with poor prognostic factors like higher T stage (T4) {17 (53.1%)}, higher stage (stage III) 95 (67.8%), skin involvement 19 (47.5%), Lymphovascular involvement in 23 (39.7%), perineural involvement in 10 (71.4%) and in patients with HER2 positive molecular subtype in 8 (5.7%). Conclusion: This study suggests that a high NLR value was associated with poor prognosis in breast cancer patients. Thus, it can be used as an independent marker of poor prognosis and can help guide the treatment of breast cancer patients. The more we study the role of NLR, the more useful it will be in predicting the course of breast cancer as early as possible.

A Comparative Analysis of Clinical and Imaging Features of Aquaporin 4 (AQP4) Antibody Positive, Myelin Oligodendrocyte Glycoprotein (MOG) Antibody Positive and Double Seronegative Subtypes of Neuro Myelitis Optica Spectrum Disorder (NMOSD).

Objectives:Study was conducted with aim of comparing subtypes types of NMOSD based on serology.Methods:In this retrospective study, patients ≥18 years were included satisfying IPND 2015 criteria. Three groups were created based on seropositivity for AQP4 antibody, MOG antibody or double seronegative. Demographic, clinical and imaging were compared using regression analysis.Results:Forty-six patients, 28 (60.9%) AQP4+, 11 (23.9%) MOG + and remaining 7 (15.2%) double seronegative were included. Thirty-seven patients (80.4%) had presenting symptoms localized to optic nerve and/or cord [AQP4 + 22 (78.5%), MOG + 9 (81.8%) and double seronegative 6 (85.7%)]. Presentation with bilateral optic neuritis was more common in AQP4- patients. Twenty (86.8%) out of the 23 patients who had relapsing disease localized to optic nerve and/or spinal cord [AQP4 + 13/14 (92.8%), MOG + 3/5 (60%) and double seronegative 4/4 (100%)]. Relapses were more common in AQP4+ (77% vs 12% vs10%). In AQP4 negative group disability (EDSS 4.2 vs 3.3) and progression index was relatively less (1.6 vs 1.1). CSF pleocytosis (38.8% vs 17.9%) and raised proteins (66.6% vs 32.1%) were also more common. Optic nerve MRI (>50% optic nerve and chiasma involvement) was more commonly abnormal in AQP4 negative (52.9% vs 31.2%). Regression analysis revealed females to be significantly higher in AQP4 positive NMOSD (89.3%) when compared to MOG positive (36.4%) and double seronegative (42.9%).Conclusion:Gender was the only significant difference between the three groups. There was trend towards greater disability and more relapses in AQP4 + groups.

Influence of progesterone receptor on metastasis and prognosis in breast cancer patients with negative HER-2.

In de novo metastatic breast cancer patients, the site of metastasis and prognosis are related to the molecular subtype of breast cancer. There are few relevant reports to explore the clinicopathological and prognostic characteristics of different single positive hormone receptor subtypes [estrogen receptor (ER)+/progesterone receptor (PR)- and ER-/PR+] of metastatic breast cancer. Using the Surveillance, Epidemiology and End Results (SEER) database between 2010 and 2015.We analyzed the metastatic patterns and prognosis of human epidermal growth factor receptor 2 (HER-2)-negative breast cancer patients. Cox analysis was used to analyze the influence of ER+/PR- and ER-/PR+ on the prognosis of patients in different subgroups and the risk factors affecting the prognosis of patients with single hormone receptor positivity. We included 206,187 breast cancer patients, including 7,726 stage IV patients. The loss of ER was a protective factor against bone metastasis (P<0.001) and a risk factor for visceral metastasis (P<0.001). The ER-/PR+ subtype had a similar proportion of de novo metastatic breast cancer, and similar clinicopathological characteristics, prognosis with triple negative breast cancer (TNBC). Single PR positivity was an independent risk factor for cancer specific survival (CSS) in multi-visceral metastasis subgroup comparing to TNBC. Meanwhile, no significant difference in overall survival (OS) or breast cancer specific survival (BCSS) between ER-/PR+ and ER-/PR- patients in all breast cancer patients or in stage IV breast cancer patients. Age [hazard ratio (HR) =2.16], grade (HR =2.36), T stage (T4: HR =3.24), lymph node metastasis (>10: HR =4.33), distant metastasis (HR =4.99), and no chemotherapy or an unknown (HR =1.65) were high-risk factors but surgery (HR <0.5) were protective factors for CSS in ER-/PR+ patients. ER-/PR+ subtype had a high proportion of stage IV patients. Meanwhile, such subtype breast cancer had similar clinicopathological characteristics, metastatic models (prefers to visceral metastasis), similar even worse prognosis compared with TNBC.