Aggressive Type Of Breast Cancer Research Articles

Analysis of the radio-pathological factors of triple negative breast cancer and determination of risk profiles

ObjectiveTriple-negative tumours are the most aggressive type of breast cancer. We aimed to analyse the main radiologic and histopathologic factors of these tumours to create a risk profile. Materials and methodsWe analysed data from 140 patients diagnosed with triple-negative breast cancer between January 2007 and December 2016, with follow-up through April 2018. We analysed the following variables in the breast MRI done for staging: size, necrosis, associated findings, adenopathies, and perfusion and diffusion parameters. We analysed the following variables in histopathologic studies of biopsy specimens: histological type, Scarf-Bloom, Ki67, and p53 in the infiltrating component as well as in the in situ component. We analysed the following variables in histopathologic studies of positive lymph nodes and surgical specimens: size, lymphovascular/perineural invasion, and microglandular adenosis. We analysed the relation between the radiologic and histopathologic factors and recurrence and disease-free survival. ResultsMRI tumour size ≥25mm, non-nodular enhancement, breast oedema, areola-nipple complex retraction, and lymph-node involvement were associated with recurrence and lower disease-free survival. Invasive lobular carcinoma, postsurgical size ≥20mm, and p53<15% were also associated with recurrence and lower disease-free survival. Histologically positive lymph nodes were associated with a greater percentage of recurrence and lymphovascular invasion and with lower disease-free survival. The multivariate analysis found that the variables MRI size ≥25mm, non-nodular enhancement, adenopathies on MRI, and p53 expression <15% were independent predictors of lower disease-free survival. ConclusionsIn triple-negative breast tumours, factors associated with lower disease-free survival are non-nodular enhancement, size ≥25mm, and adenopathies on MRI, and p53 expression <15% on histopathologic study.

Targeting Signaling Pathways in Inflammatory Breast Cancer.

Simple SummaryInflammatory breast cancer (IBC) is the most lethal and aggressive form of breast cancer; it is highly likely to spread to other sites in the body. There is an urgent need to establish novel treatment strategies to reduce IBC recurrence and metastasis. The aim of this work is to provide a comprehensive overview of signaling pathways in IBC, covering understanding of their function in IBC tumor cells and cells surrounding tumor, and clinical efforts to target these pathways for patients with IBC. The findings described in this work will help guide the development of effective therapies through preclinical and clinical research, eventually improving the treatment of patients with IBC.Inflammatory breast cancer (IBC), although rare, is the most aggressive type of breast cancer. Only 2–4% of breast cancer cases are classified as IBC, but—owing to its high rate of metastasis and poor prognosis—8% to 10% of breast cancer-related mortality occur in patients with IBC. Currently, IBC-specific targeted therapies are not available, and there is a critical need for novel therapies derived via understanding novel targets. In this review, we summarize the biological functions of critical signaling pathways in the progression of IBC and the preclinical and clinical studies of targeting these pathways in IBC. We also discuss studies of crosstalk between several signaling pathways and the IBC tumor microenvironment.

EcTI impairs survival and proliferation pathways in triple-negative breast cancer by modulating cell-glycosaminoglycans and inflammatory cytokines.

Breast cancer is the most common malignant tumor among women worldwide, and triple-negative breast cancer is the most aggressive type of breast cancer, which does not respond to hormonal therapies. The protease inhibitor, EcTI, extracted from seeds of Enterolobium contortisiliquum, acts on the main signaling pathways of the MDA-MB-231 triple-negative breast cancer cells. This inhibitor, when bound to collagen I of the extracellular matrix, triggers a series of pathways capable of decreasing the viability, adhesion, migration, and invasion of these cells. This inhibitor can interfere in the cell cycle process through the main signaling pathways such as the adhesion, Integrin/FAK/SRC, Akt, ERK, and the cell death pathway BAX and BCL-2. It also acts by reducing the main inflammatory cytokines such as TGF-α, IL-6, IL-8, and MCP-1, besides NFκB, a transcription factor, responsible for the aggressive and metastatic characteristics of this type of tumor. Thus, the inhibitor was able to reduce the main processes of carcinogenesis of this type of cancer.

Abstract 3704: MicroRNA-3977 acts as a tumor suppressor and inhibits triple-negative breast cancer tumorigenesis by targeting FoxM1

Abstract Triple negative breast cancer (TNBC) is an aggressive type of breast cancer characterized by the absence of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). TNBC is associated with poor prognosis, high rates of relapse and distant metastasis despite surgery and adjuvant chemotherapy. The lack of effective targeted therapies for TNBC requires new therapeutic approaches. The Forkhead box protein M1 (FoxM1) is a member of forkhead superfamily transcription factors that has been associated with normal cell growth and proliferation through cell cycle transition and mitotic spindle control. FoxM1 is also identified as a master regulator of tumor metastasis. Based on analysis of pooled clinical datasets derived from online accessible databases as Breast Cancer Gene-Expression Miner v4.3 and Kaplan-Meier plotter, the expression and prognostic values of FoxM1 increase in TNBC compare with other breast cancer subtypes. Directly binding to the 3'-untranslated region (3' UTR) of FoxM1 mRNAs suppress its high expression in TNBC that leads to inhibition of cell proliferation, motility, and invasion. These results implied that FoxM1 is an essential poor prognostic factor and promising candidate target in the treatment of TNBC. We identified miR-3977 as a potential regulator of FoxM1 by using in silico prediction algorithms, and we investigated its role in TNBC. In this study, we found miR-3977 expression is significantly suppressed in TNBC cell lines and inversely correlated with FoxM1 levels. We demonstrate that miR-3977 directly bound to the 3′ UTR of FoxM1 mRNA and repressed its expression. We also find reduced miR-3977 expression is associated with shorter patient survival in TNBC based on Kaplan-Meier plotter database. Moreover, in vivo delivery of miR-3977 nanoparticles inhibited FoxM1 expression and tumor growth in TNBC xenograft mouse models. In conclusion, we provide the first evidence that miR-3977 functions as a tumor suppressor and gain of miR-3977 expression leads to decrease in FoxM1 expression and inhibits to cell growth, colony formation, invasion, and progression of TNBC. Taken together, our data suggest that miR-3977-based gene therapy is a potential therapeutic strategy in TNBC. Citation Format: Selda Karamil, Nuray Altintas, Bulent Ozpolat. MicroRNA-3977 acts as a tumor suppressor and inhibits triple-negative breast cancer tumorigenesis by targeting FoxM1 [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3704.

Abstract 4212: SJP1602, a selective and dual inhibitor of FAK and Pyk2 for treatment of triple negative breast cancer

Abstract Triple negative breast cancer (TNBC) is the most aggressive type of breast cancer, classified by a lack of ER and PR, and HER2 overexpression. Attributable to its tumor heterogeneity and a high rate of relapse, TNBC carries poor prognosis accounting for 30% mortality of breast cancer patients. Nevertheless, the current therapeutic options for patients with TNBC are limited to chemotherapy, lacking effective targeted therapies. Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase mainly known for its role in tumor progression to a malignant phenotype and its involvement in resistance to conventional therapy. Its over-expression and/or activation is associated with poor survival in various types of cancer including TNBC. With increasing evidence of FAK as a promising therapeutic target in cancer, several small molecule inhibitors targeting FAK are currently on development and among them are defactinib (VS-6063) and GSK2256098, which are currently undergoing clinical investigation in combinatorial approach in solid tumors. In this study, we represent a novel small molecule inhibitor, SJP1602, targeting FAK and its homolog Proline-rich tyrosine kinase 2 (Pyk2). Biochemical kinase profiling of SJP1602 showed its greater selectivity for FAK and Pyk2, compared with VS-6063. Moreover, in vitro evaluation of SJP1602 using 3D spheroid models demonstrated that GI50 values of SJP1602 for spheroid growth of various TNBC cell lines were significantly lower than GI50 values of VS-6063 and GSK2256098. SJP1602 also attenuated spheroid formation along with decreased CD44 expression in MDA-MB-231 cells, implicating its capability to suppress the activity of cancer stem cells. In addition, SJP1602 remarkably reduced the invasive potential of MDA-MB-231 cells in 3D conditions, decreasing levels of the EMT-related protein, snail. More importantly, we confirmed the in vivo antitumor activity of SJP1602 compared with VS-6063 in MDA-MB-231 and BT-549 xenograft mouse models that display mesenchymal tumors with invasive and stem-like phenotype. As the result, SJP1602 exhibited substantial inhibitory effects on tumor growth in both xenograft models. In addition, assessment studies of ADME revealed the excellent plasma stability and liver microsomal metabolic stability of SJP1602 in human. Furthermore, therapeutic index estimated by comparison of AUC at toxic and effective doses was higher than 10, rendering its potential clinical safety and efficacy. Taken together, these findings suggest SJP1602 as a small molecule inhibitor targeting FAK and Pyk2 with its outstanding selectivity and anti-tumor efficacy. Therefore, SJP1602 could be a promising therapeutic agent for TNBC patients that are resistant to conventional therapy as well as for cancer patients with drug resistance involving FAK activation. Citation Format: Kug Hwa Lee, Hyoung Min Cho, Kwangwoo Hwang, Yongbin Park, Sungpyo Hong, Soon Kil Ahn, Min-Hyo Ki. SJP1602, a selective and dual inhibitor of FAK and Pyk2 for treatment of triple negative breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4212.

Abstract 2745: Drug repositioning for triple negative breast and colorectal cancers treatment: Exploring the use of metformin and propranolol

Abstract According to new global cancer data, breast and colorectal cancer (CRC) are part of the 5 types of tumors with the highest incidence and mortality. Triple negative breast cancer (TNBC) is a particularly aggressive type of breast cancer, associated with younger age, recurrence, high metastatic rate, poor prognosis and limited therapeutic options. Regarding CRC, this type of tumor is often diagnosed at advanced stages, when probability of development of distal or local recurrence due to chemotherapy resistance (usually 5-FU) is higher. For both types of tumors, the available treatment modalities are expensive and produce severe side effects. Drug repositioning refers to new uses outside the scope of the original medical indications for existing drugs or compounds; this approach fastens the process of drug development and offers more effective, cheaper and safer drugs with less side effects. We studied the effect of combining Metformin (M; antidiabetic drug) and Propranolol (P; antihypertensive agent) on in vitro and in vivo models of TNBC and CRC. We performed an in vitro screening of drugs under reposition on different TNBC and CCR cell lines and determined that M+P combination was particularly efficient in reducing proliferation in all cell lines tested. Indeed, M+P showed synergistic effect on proliferation, affecting significantly clonogenicity, apoptosis, migration and invasion. In order to explore the mechanisms associated with tumor growth/metastasis development involved in the treatment we evaluated the impact of M+P on cell signaling after proliferative stimuli and epithelial/mesenchymal transition. M+P combination reduced Erk phosphorylation levels after serum induction and promoted the expression of epithelial markers as β-catenin and E-cadherin. In vivo models (4T1, M-406, HCT116 and CT26 cells) demonstrated that M+P [M(2g/l)+P(25mg/l)] was effective reducing tumor growth and metastasis development. To further characterize the cellular events associated to these processes, we evaluated in 4T1 TNBC model the impact of M+P on different steps associated to tumor progression. The treatment affected intravasation (P=0.06), survival (P&amp;lt;0.05) and extravasation (P&amp;lt;0.05). Indeed, we developed an adjuvant model (M+P treatment after tumor excision) in which the drugs combination was effective preventing metastasis development (P&amp;lt;0.05). Interestingly, regarding CCR we found in vitro that M+P reduced proliferation and clonogenicity of 5-FU-resistant CRC cells compared to non-resistant cells (P&amp;lt;0.01). To conclude, M+P treatment decreased tumor and metastasis growth of TNBC and CRC, involving effects in several steps of tumor signaling and tumor dissemination. Our data suggest that M+P could be useful for adjuvant therapy and treatment of resistant tumors. Citation Format: M Virginia Baglioni, Luciano E. Anselmino, Nahuel Cesatti Laluce, Florencia Malizia, Gustavo F. Chapo, Leandro E. Mainetti, Viviana R. Rozados, O Graciela Scharovsky, M. José Rico, Mauricio A. Menacho Márquez. Drug repositioning for triple negative breast and colorectal cancers treatment: Exploring the use of metformin and propranolol [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2745.

#BlackBreastsMatter: Process Evaluation of Recruitment and Engagement of Pregnant African American Women for a Social Media Intervention Study to Increase Breastfeeding.

BackgroundIn the United States, there are lower rates of breastfeeding among African American mothers, particularly those who are younger women. Recent epidemiological studies have shown a strong association of more aggressive types of breast cancer (estrogen receptor negative) among African American women, with a higher risk in African American women who did not breastfeed their children.ObjectiveThis study aims to describe the process evaluation of recruitment and educational strategies to engage pregnant African American participants for a pilot study designed to determine whether social media messaging about breast cancer risk reduction through breastfeeding may positively influence breastfeeding rates.MethodsThis pilot study is conducted in collaboration with a local Women, Infants, and Children (WIC) organization and hospital and prenatal clinics of a local health care network. To engage African American women to enroll in the study, several methods and monitoring processes were explored, including WIC electronic text-based messages sent out to all phones of current WIC recipients (referred to as e-blasts); keyword responses to texts from flyers and posters in local community-based organizations, hospitals, and prenatal clinics; keyword responses using electronic links posted in established Facebook groups; and snowball recruitment of other pregnant women by current participants through Facebook. Once enrolled, participants were randomized to 2 study conditions: (1) an intervention group receiving messages about breast cancer risk reduction and breastfeeding or (2) a control group receiving breastfeeding-only messages. Data were obtained through electronic monitoring, SurveyMonkey, qualitative responses on Facebook, focus groups, and interviews.ResultsMore than 3000 text messages were sent and received through WIC e-blasts and keyword responses from flyers. A total of 472 women were recruited through WIC e-blast, and 161 responded to flyers and contacts through the local health care network, community-based organizations, Facebook, and friend referrals. A total of 633 women were assessed for eligibility to participate in the study. A total of 288 pregnant African American women were enrolled, consented, and completed presurvey assessments (102.8% of the goal), and 22 participants attended focus groups or interviews reporting on their experiences with Facebook and the educational messages.ConclusionsThis process evaluation suggests that using electronic, smartphone apps with social media holds promise for both recruitment and conduct of health education intervention studies for pregnant African American women. Providing messaging and resources through social media to reinforce and educate women about breastfeeding and potentially provide lactation support is intriguing. Convenience (for researchers and participants) is an attribute of social media for this demographic of women and worthy of further research as an educational tool.Trial RegistrationClinicalTrials.gov NCT03680235; https://clinicaltrials.gov/ct2/show/NCT03680235

Construction and Comprehensive Analysis of a ceRNA Network to Reveal Potential Novel Biomarkers for Triple-Negative Breast Cancer.

BackgroundTriple-negative breast cancer (TNBC) is the most common and aggressive type of breast cancer with an unfavourable outcome worldwide. Novel therapeutic targets are urgently required to explore this malignancy. This study explored the ceRNA network and the important genes for predicting the therapeutic targets.MethodsIt identified the differentially expressed genes of mRNAs, lncRNAs and miRNAs between TNBC and non-TNBC samples in four cohorts (TCGA, GSE38959, GSE45827 and GSE65194) to explore the novel therapeutic targets for TNBC. Downstream analyses, including functional enrichment analysis, ceRNA network, protein–protein interaction and survival analysis, were then conducted by bioinformatics analysis. Finally, the potential core protein of the ceRNA network in TNBC was validated by immunohistochemistry.ResultsA total of 1,045 lncRNAs and 28 miRNAs were differentially expressed in the TCGA TNBC samples, and the intersections of 282 mRNAs (176 upregulations and 106 downregulations) between the GEO and TCGA databases were identified. A ceRNA network composed of 7 lncRNAs, 62 mRNAs, 12 miRNAs and 244 edges specific to TNBC was established. The functional assay showed dysregulated genes, and GO, DO and KEGG enrichment analysis were performed. Survival analysis showed that mRNA LIFR and lncRNA AC124312.3 were significantly correlated with the overall survival of patients with TNBC in the TCGA databases (P < 0.05). Finally, the LIFR protein was validated, and immunohistochemical results showed the upregulated expression of LIFR in TNBC tissues.ConclusionThus, our study presents an enhanced understanding of the ceRNA network in TNBC, where the key gene LIFR may be a new promising potential therapeutic target for patients with TNBC.

Molecular mechanism of gossypol mediating CCL2 and IL‑8 attenuation in triple‑negative breast cancer cells.

Chronic inflammation associated with cancer is characterized by the production of different types of chemokines and cytokines. In cancer, numerous signaling pathways upregulate the expression levels of several cytokines and evolve cells to the neoplastic state. Therefore, targeting these signaling pathways through the inhibition of distinctive gene expression is a primary target for cancer therapy. The present study investigated the anticancer effects of the natural polyphenol gossypol (GOSS) in triple-negative breast cancer (TNBC) cells, the most aggressive breast cancer type with poor prognosis. GOSS effects were examined in two TNBC cell lines: MDA-MB-231 (MM-231) and MDA-MB-468 (MM-468), representing Caucasian Americans (CA) and African Americans (AA), respectively. The obtained IC50s revealed no significant difference between the two cell lines' response to the compound. However, the use of microarray assays for cytokine determination indicated the ability of GOSS to attenuate the expression levels of cancer-related cytokines in the two cell lines. Although GOSS did not alter CCL2 expression in MM-468 cells, it was able to cause 30% inhibition in TNF-α-stimulated MM-231 cells. Additionally, IL-8 was not altered by GOSS treatment in MM-231 cells, while its expression was inhibited by 60% in TNF-α-activated MM-468 cells. ELISA assays supported the microarray data and indicated that CCL2 expression was inhibited by 40% in MM-231 cells, and IL-8 expression was inhibited by 50% in MM-468 cells. Furthermore, in MM-231 cells, GOSS inhibited CCL2 release via the repression of IKBKE, CCL2 and MAPK1 gene expression. Additionally, in MM-468 cells, the compound downregulated the release of IL-8 through repressing IL-8, MAPK1, MAPK3, CCDC88A, STAT3 and PIK3CD gene expression. In conclusion, the data obtained in the present study indicate that the polyphenol compound GOSS may provide a valuable tool in TNBC therapy.

Inflammatory Breast Cancer in a Very Young Genetically Susceptible Woman: Case Presentation in a Tumour Board Session, Discussion and Decision-Making

Background: Inflammatory breast cancer is a very aggressive type of breast cancer which invades skin lymphatics. The standard treatment includes neoadjuvant chemotherapy followed by modified radical mastectomy and postsurgical radiation. Preserving the skin or nipple are not considered reasonable due to the involvement of the dermal lymphatics and therefore skin sparing or nipplesparing mastectomy are not usually carried out for this type of breast cancer.&#x0D; Case presentation: In this article, we present a case of inflammatory breast cancer in a very young woman who was positive for BRCA2 genes.&#x0D; Question: The possibility to perform nipple sparing or skin sparing mastectomy and immediate breast reconstruction, and the indication for prophylactic contralateral mastectomy as discussed in the tumor board are presented in this article.&#x0D; Conclusion: Neither of the more conservative techniques of mastectomy was appropriate for this case. Also, the prognosis of the patient mostly depends on the course of the ipsilateral tumor and prophylactic mastectomy would be wise only if the subsequent course of the disease is favourable.

A novel small molecule STAT3 inhibitor SLSI-1216 suppresses proliferation and tumor growth of triple-negative breast cancer cells through apoptotic induction

Triple-negative breast cancer (TNBC) is the most aggressive type of breast cancer, characterized by the lack of expression of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Owing to the absence of molecular targets, there are limited treatment options, and TNBC patients exhibit high mortality rates. Signal transducer and activator of transcription 3 (STAT3) is overexpressed and aberrantly activated in TNBC cells. Therefore, inhibition of STAT3-mediated signaling provides a potential strategy for the treatment of TNBC. In this study, A series of synthetic derivatives of SLSI-1 (a STAT3 inhibitor) were designed and evaluated for antitumor activity in TNBC cells. A novel derivative (SLSI-1216) exhibited the most potent anti-proliferative activity. SLSI-1216 effectively inhibited STAT3 activity and activation of STAT3, leading to the downregulation of AXL, a downstream target of STAT3 and epithelial-mesenchymal transition (EMT) progression. The inhibition of EMT by SLSI-1216 was associated with modulation of E-cadherin and N-cadherin. Furthermore, SLSI-1216 induced apoptosis by targeting STAT3 and effectively inhibited tumor growth in vivo. These findings suggest that SLSI-1216, as a potential inhibitor of STAT3, may be a promising therapeutic agent for TNBC.

Inflammatory breast cancer (IBC) in native Hawaiians and Pacific Islanders: Frequency of IBC and impact of socioeconomic status on survival outcomes.

e19083 Background: IBC is one of the most aggressive types of breast cancer, accounting for only 1-4% of all breast cancer cases while causing 8-10% of breast cancer related deaths. Previous studies have demonstrated that IBC has distinct epidemiological characteristics, but the prevalence of IBC particularly in Native Hawaiian (NH) and Pacific Islander (PI) populations, and the socioeconomic disparities related to survival outcomes, are not fully understood. Our primary objective was to determine the frequency at which NH and PI breast cancer patients are diagnosed with IBC, and the association between socioeconomic status and overall survival (OS). Methods: Patients with newly-diagnosed primary invasive breast cancer were identified from January 1, 2000 through December 31, 2018 using Queen’s Medical Center Tumor Registry. Clinical T4d was used to differentiate IBC and non-IBC. OS was defined as the time from diagnosis to death or last follow-up. Patients who were alive at the date of last follow-up were censored. Univariate and multivariate cox proportional hazard models were used to assess the effects of variables of interest on OS. Results: A total of 3,715 patients were included in analysis. There were 98 (2.6%) patients with IBC and 3,617 (97.4%) with non-IBC. Proportion of IBC relative to non-IBC was significantly higher in PI (9.0%) than in White (W) (2.7%) (P &lt; 0.001). Proportion of IBC was higher in NH (4.7%) than W but the difference was not statistically significant (P = 0.08). In multivariate analysis among the non-IBC group, OS was significantly shorter for both PI and NH than W (HR 1.72, [95%CI, 1.13-2.6]; P = 0.01, HR 1.48 [95% CI, 1.14-1.93]; P = 0.003, respectively). There were no significant racial differences seen in OS among IBC patients. Being under or uninsured was also significantly associated with short OS outcomes among patients with non-IBC but not those with IBC. Conclusions: PI have a significantly high proportion of IBC relative to non-IBC. Race and insurance status were not associated with OS in IBC although they were significantly associated with short OS in non-IBC. This result suggests that in IBC, the potential effect of racial disparity on OS is negated by the aggressive and insidious nature of the cancer. The effect of a relatively small sample size cannot be excluded, and further study in a larger population is warranted.

Epigenetics in Inflammatory Breast Cancer: Biological Features and Therapeutic Perspectives.

Evidence has emerged implicating epigenetic alterations in inflammatory breast cancer (IBC) origin and progression. IBC is a rare and rapidly progressing disease, considered the most aggressive type of breast cancer (BC). At clinical presentation, IBC is characterized by diffuse erythema, skin ridging, dermal lymphatic invasion, and peau d’orange aspect. The widespread distribution of the tumor as emboli throughout the breast and intra- and intertumor heterogeneity is associated with its poor prognosis. In this review, we highlighted studies documenting the essential roles of epigenetic mechanisms in remodeling chromatin and modulating gene expression during mammary gland differentiation and the development of IBC. Compiling evidence has emerged implicating epigenetic changes as a common denominator linking the main risk factors (socioeconomic status, environmental exposure to endocrine disruptors, racial disparities, and obesity) with IBC development. DNA methylation changes and their impact on the diagnosis, prognosis, and treatment of IBC are also described. Recent studies are focusing on the use of histone deacetylase inhibitors as promising epigenetic drugs for treating IBC. All efforts must be undertaken to unravel the epigenetic marks that drive this disease and how this knowledge could impact strategies to reduce the risk of IBC development and progression.

SNHG3 silencing suppresses the malignant development of triple-negative breast cancer cells by regulating miRNA-326/integrin α5 axis and inactivating Vav2/Rac1 signaling pathway.

Long non-coding RNA small nucleolar RNA host gene 3 (SNHG3) has been shown to participate in several tumorigenesis. Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer, which is the first leading cause of new cancer diagnoses in women globally. However, the role of SNHG3 remains little known in breast cancers, especially in TNBC. Expression of SNHG3, miRNA-326-5p (miR-326) and integrin α5 (ITGA5) was detected using Real Time-PCR and Western blotting. Cell viability, apoptosis, migration, and invasion were measured by methyl thiazolyl tetrazolium assay, flow cytometry, and transwell assays, respectively. Vav2/Rac1 signaling pathway was evaluated by Western blotting by analyzing Vav2 and Rac1 levels. The interaction among miR-326, SNHG3 and ITGA5 was confirmed by Dual-Luciferase reporter assay. We found that the expression of SNHG3 and ITGA5 was upregulated and miR-326 was downregulated in TNBC tumors and cell lines (MDA-MB-231, BT-549, MDA-MB-468 and SUM159). Functionally, both SNHG3 silencing and miR-326 overexpression enhanced cell apoptosis, but depressed cell viability, migration and invasion in MDA-MB-231 and BT-549 cells, as well as inhibited Vav2 and Rac1 expression. Notably, miR-326 deletion could abolish the tumor-suppressive role of SNHG3 silencing; meanwhile, the similar anti-tumor effect of miR-326 overexpression was abrogated by ITGA5 restoration. Mechanically, SNHG3 silencing downregulated ITGA5 expression by functioning as a molecular "sponge" for miR-326. Silencing of SNHG3 suppressed the malignant development of TNBC cells, at least partially, through miR-326/ITGA5 axis and inhibiting Vav2/Rac1 signaling pathway.

Análisis de los factores radiopatológicos del cáncer de mama triple negativo y determinación de perfiles de riesgo

ObjectiveTriple-negative tumors are the most aggressive type of breast cancer. We aimed to analyze the main radiologic and histopathologic factors of these tumors to create a risk profile. Materials and methodsWe analyzed data from 140 patients diagnosed with triple-negative breast cancer between January 2007 and December 2016, with follow-up through April 2018. We analyzed the following variables in the breast MRI done for staging: size, necrosis, associated findings, adenopathies, and perfusion and diffusion parameters. We analyzed the following variables in histopathologic studies of biopsy specimens: histological type, Scarf-Bloom, Ki67, and p53 in the infiltrating component as well as in the in situ component. We analyzed the following variables in histopathologic studies of positive lymph nodes and surgical specimens: size, lymphovascular/perineural invasion, and microglandular adenosis. We analyzed the relation between the radiologic and histopathologic factors and recurrence and disease-free survival. ResultsMRI tumor size>25mm, non-nodular enhancement, breast edema, areola-nipple complex retraction, and lymph-node involvement were associated with recurrence and lower disease-free survival. Invasive lobular carcinoma, postsurgical size>20mm, and p53<15% were also associated with recurrence and lower disease-free survival. Histologically positive lymph nodes were associated with a greater percentage of recurrence and lymphovascular invasion and with lower disease-free survival. The multivariate analysis found that the variables MRI size>25mm, non-nodular enhancement, adenopathies on MRI, and p53 expression <15% were independent predictors of lower disease-free survival. ConclusionsIn triple-negative breast tumors, factors associated with lower disease-free survival are non-nodular enhancement, size>25mm, and adenopathies on MRI, and p53 expression <15% on histopathologic study.

Abstract P6-10-28: Therapeutic stratification of triple negative breast cancer by integrating chemosensitivity &amp; phospho-proteome profiles

Abstract Triple negative breast cancer (TNBC) is a highly aggressive type of breast cancer with poor prognosis that accounts approximately for 15% of breast cancer cases. The lack of targetable hormonal receptors or HER2/ErbB2 makes TNBC therapeutically challenging and existing drug treatments are often ineffective. TNBC is also a highly heterogeneous class of breast cancer, and different genetic approaches have not been so far successful in identifying clinically useful patient stratification strategies to date. In previous studies, we have discovered a correlation between targeted phospho-proteomics and mitotic inhibitor sensitivities in TNBC cells, suggesting the potential of using phospho-proteome profiles to identifying markers predictive of actionable therapeutic strategies. In the current study, we further explored the potential of phospho-proteomics for TNBC stratification and drug chemo-sensitivity prediction. We screened 25 TNBC cell lines against 517 oncology compounds, and in parallel ran mass spectrometry-based global phospho-proteome profiling of the same cell lines, resulting in 4283 confident phospho-peptide quantifications. Using multitask multiple kernel learning, we integrated the phospho-peptide data with other publicly available omics data for the 25 TNBC cell lines (genomics, transcriptomics, metabolomics, and methylomics), and developed a multi-omics machine learning model to quantify the predictive contribution of each omics profile to drug response prediction. When considering all the 517 compounds, we found that the phospho-proteomic and gene expression data were most informative for the prediction of compound’s chemo-sensitivity. This integrated approach has the potential to both identify panels of multi-omics markers for individual drug classes, as well as to stratify the heterogeneous TNBC cells based on their genomic, molecular and functional profiles. Citation Format: Prson Gautam, Xiangju Qin, Julia Vainonen, Srikar Nagelli, Johanna Lampe, Juha Klefström, Jukka Westermarck, Krister Wennerberg, Tero Aittokallio. Therapeutic stratification of triple negative breast cancer by integrating chemosensitivity &amp; phospho-proteome profiles [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P6-10-28.

Abstract P6-15-03: Inflammatory breast cancer (IBC) defined: Proposed common diagnostic criteria and scoring - Moving beyond the subjective ‘clinical diagnosis’ of IBC to advance research

Abstract Spurred by the lack of progress in improving outcomes in patients with inflammatory breast cancer (IBC), a deadly and aggressive type of breast cancer, Susan G. Komen, the Inflammatory Breast Cancer Research Foundation, and the Milburn Foundation convened patient advocates, breast cancer researchers, and clinicians to review the state of IBC care and research globally, and propose specific initiatives to move the field forward. The outcome of the discussions was abundantly clear: the field needs a formal definition of IBC. Without a clear definition, both patient care and research suffer. Diagnosis remains subjective and treatment variable. Clinical trials become inherently underpowered by inclusion of subjects with locally advanced, but not inflammatory, breast cancer. Similarly, the search for a molecular sine qua non or unifying pathway aberration is hindered by inclusion of bio-specimens from subjects with a different disease. The goal is to move beyond the subjective ‘clinical diagnosis’ to a set of specific diagnostic criteria and scoring system that will advance IBC research and facilitate the discoveries that will improve care of IBC patients. Here we propose a definition of IBC, based on a review of clinical, and pathologic features. Table 1. Proposed Scoring System for inflammatory breast cancer diagnosisPriority FactorCharacteristicScore3213Timing of signs/symptoms≤ 3 months3-6 months&amp;gt; 6 months3Skin changesPeau d’orangeSkin edema/thickening (≥ 1/3 of the breast)Focal skin edema/thickening (&amp;lt; 1/3 of the breast)3Swelling/engorgement of the breastClinically apparent enlargement of the breast or new asymmetry in breast sizeBreast edema identified on imaging but not clinically detectable2Erythema or other skin discoloration: pink, red, darkened, bruising/purplish or serpiginous in characterNearly complete involvement of the breastPartial involvement of the breastMinimal involvement or ambiguous color change2Nipple abnormalitiesNew nipple inversionNew nipple flattening or other asymmetryCrusting of the nipple/areola without other nipple changes2Lymphatic emboliDermal lymphatic emboli present without evidence of direct involvement of the dermis or epidermisNon-dermal lymphatic embolic present in breast parenchyma or stroma1Breast imagingDiffuse involvement of breast parenchyma, with or without dominant massEnlargement of non-axillary nodes (internal mammary, supraclavicular, subpectoral etc.) With increased awareness of IBC, patients are coming to medical attention earlier when clinical characteristics may not be as profound as historically depicted. Not all criterion are required for a diagnosis. To determine the total score, multiply the priority factor by the score for each criteria, then add the subtotals for each criteria. A priority factor is used to weigh criterion more heavily that are key to a consistent diagnosis of IBC to better define breast cancers that are ambiguous. While we cannot currently recommend a molecular-genetic profile that unambiguously identifies IBC, we will also provide a panel of prioritized biomarkers that are strongly associated with IBC and deserve further study in cohorts meeting our diagnostic criteria. We acknowledge that our proposed scoring system requires validation and refinement. If accurate, we expect patients with IBC based on these criteria to have a different clinical course and outcome compared to patients with “non-inflammatory” locally advanced breast cancer (LABC). Ultimately, we envision using the final disease classification and scoring system to develop a staging system specific to IBC. The discovery and validation of a distinct molecular or genetic profile that identifies IBC is of utmost importance to advancing research in this disease and cannot be accomplished without a clear and validated definition of IBC. Citation Format: Ginny Mason, Beth A Overmoyer, Wendy A. Woodward, Sunil Badve, Robert J. Schneider, Reshma Jagsi, Julie E. Lang, Mary Alpaugh, Krissa Smith, Kathy Miller. Inflammatory breast cancer (IBC) defined: Proposed common diagnostic criteria and scoring - Moving beyond the subjective ‘clinical diagnosis’ of IBC to advance research [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P6-15-03.

SSi6 promotes cell death by apoptosis through cell cycle arrest and inhibits migration and invasion of MDA-MB-231 human breast cancer cells.

Triple-negative breast cancer subtype is the most aggressive type of breast cancer due to the lack of specific therapeutic targets, having limited treatment options, low survival prognosis and high recurrence rates. In this work, we describe the effects of a semisynthetic derivative of [6]-gingerol (6G) called SSi6, produced by the addition of a 2,4-dinitrophenylhydrazine reagent on several aspects of triple-negative breast cancer biology. Human breast cancer cell lines MDA-MB-231 and MCF-10A were used in the experiments. MTT assays were used to detect cell viability. Cell cycle and apoptosis assay were analyzed using flow cytometer Accuri C6 and analysis of proteins as retinoblastoma Rb and kinases Cdk4/6 were analyzed by western blotting. SSi6 induced cytotoxic effects on triple-negative breast cancer cells, with higher selectivity when compared to the non-tumor MCF-10A cells. In addition, SSi6 inhibited migration and invasion of triple-negative breast cancer cells and was able to arrest cell cycle at the G1-phase, mainly by decreasing Cdk4/6-Rb axis levels. Therefore, SSi6 provoked the induction of apoptosis in triple-negative breast cancer cells. SSi6 was more efficient in producing these effects, compared to the original 6G natural product. This study may contribute to a better understanding of the effects of natural and semisynthetic products on the in-vitro metastatic processes in the MDA-MB-231 triple-negative breast cancer cell line. Additional, it can be useful to understand the effects of chemical modifications on already effective natural compounds aiming at the improvement of their bioactive properties, such as in the increase of the cytotoxic selectivity against tumor cells, compared to non-tumor ones.

Evaluation of Gene Expression Data From Cybrids and Tumours Highlights Elevated NDRG1-Driven Proliferation in Triple-Negative Breast Cancer.

Background:Triple-negative breast cancer is an aggressive type of breast cancer with high risk of recurrence. It is still poorly understood and lacks any targeted therapy, which makes it difficult to treat. Thus, it is important to understand the underlying mechanisms and pathways that are dysregulated in triple-negative breast cancer.Methods:To investigate the role of mitochondria in triple-negative breast cancer progression, we analysed previously reported gene expression data from triple-negative breast cancer cybrids with SUM-159 as the nuclear donor cell and SUM-159 or A1N4 (c-SUM-159, c-A1N4) as the mitochondrial donor cells and with 143B as the nuclear donor cell and MCF-10A or MDA-MB-231 (c-MCF-10A, c-MDA-MB-231) as the mitochondrial donor cells. The role of potential biomarkers in cell proliferation and migration was examined in SUM-159 and MDA-MB-231 cells using sulforhodamine B and wound healing assays.Results:Rank product analysis of cybrid gene expression data identified 149 genes which were significantly up-regulated in the cybrids with mitochondria from the cancer cell line. Analysis of previously reported breast tumour gene expression datasets confirmed 9 of the 149 genes were amplified, up-regulated, or down-regulated in more than 10% of the patients. The genes included NDRG1, PVT1, and EXT1, which are co-located in cytoband 8q24, which is frequently amplified in breast cancer. NDRG1 showed the largest down-regulation in the cybrids with benign mitochondria and was associated with poor prognosis in a breast cancer clinical dataset. Knockdown of NDRG1 expression significantly decreased proliferation of SUM-159 triple-negative breast cancer cells.Conclusions:These results indicate that mitochondria-regulated nuclear gene expression helps breast cancer cells survive and proliferate, consistent with previous work focusing on an Src gene signature which is mitochondria regulated and drives malignancy in breast cancer cybrids. This is the first study to show that mitochondria in triple-negative breast cancer mediate significant up-regulation of a number of genes, and silencing of NDRG1 leads to significant reduction in proliferation.

The ethyl acetate fraction of sponge Stylissa carteri decreases viability in HER2+ trastuzumab-resistant breast cancer cells

Background: HER2+ breast cancer is a very aggressive type of breast cancer. Although trastuzumab, specifically targeted for HER2, has been used for breast cancer treatments, some patients become resistant to trastuzumab. A marine sponge is one of the potential sources of anticancer agents. One of marine sponges commonly found in Indonesia is Stylissa carteri, but it has not been explored extensively. Objective: This study aimed to identify cytotoxic effects of the ethyl acetate fraction of Stylissa carteri on HCC-1954, HER2+ trastuzumab-resistant breast cancer cells, by assessing cell viability and determining IC50 value. Methods: This study was an experimental in vitro study conducted in The Cell Culture Laboratory, Faculty of Medicine Universitas Padjadjaran on February 2018 to June 2018. The Stylissa carteri was collected from Pramuka Island, Kepulauan Seribu National Park Jakarta. HCC-1954 cells were treated by serial concentration of fractions and were incubated for 72 hours. Cell viability were observed under a microscope and analysed with MTT assay. The IC50 value was also determined by using four parametric logistic regression (4PL) method by Sigmaplot version 12.0. Result: Data of this study showed descent of cell viability significantly when exposing the ethyl acetate fraction of Stylissa carteri. There was a decrease of 49% cell viability in 10 µg/ml of the ethyl acetate fraction of Stylissa carteri. The estimated IC50 value was 9.25 µg/ml. Conclusion: This result indicated that the ethyl acetate fraction of Stylissa carteri has cytotoxic effects on HER2+ trastuzumab-resistant breast cancer cells.