Receptor-negative Breast Cancer Patients Research Articles

Prediction of Successful Ovarian Protection Using Gonadotropin-Releasing Hormone Agonists During Chemotherapy in Young Estrogen Receptor-Negative Breast Cancer Patients.

Background: It is important to identify factors predicting successful ovarian protection using gonadotropin-releasing hormone (GnRH) agonists during chemotherapy. However, only a few studies have prospectively assessed this issue in young breast cancer patients.Objective: This study evaluated the predictive factors for successful ovarian protection with GnRH agonists during chemotherapy in young estrogen receptor-negative breast cancer patients.Materials and Methods: This prospective study analyzed 67 estrogen receptor-negative breast cancer patients ≤40 years of age who were longitudinally assessed after receiving GnRH agonists during cyclophosphamide-based chemotherapy for ovarian protection. Associations between clinical characteristics or pretreatment hormones and successful ovarian protection [resumption of menstruation and anti-Müllerian hormone (AMH) ≥1 ng/ml].Results: The mean age and pretreatment serum level of AMH were 33.2 years and 4.57 ng/ml, respectively. At 12 months after the completion of chemotherapy, most women (97%) experienced the resumption of menstruation. However, the proportion of patients with AMH ≥1 ng/ml at 12 months was 70.1%. In multivariate analyses, only the pretreatment serum AMH level (P < 0.001) was predictive for AMH ≥1 ng/ml at 12 months. Receiver operating characteristic curve analyses of pretreatment AMH exhibited an area under the curve of 0.866 (95% CI = 0.777–0.955) for AMH ≥1 ng/ml at 12 months. The cutoff value for the prediction of AMH concentration ≥1 ng/ml at 12 months was 2.87 ng/ml of pretreatment AMH with a sensitivity of 0.87 and a specificity of 0.75.Conclusions: Pretreatment AMH (2.87 ng/ml) is a useful predictor for AMH ≥1 ng/ml at 12 months after receiving GnRH agonists in young estrogen receptor-negative breast cancer patients. This finding can help improve decision-making regarding fertility preservation.

The EndoPredict score predicts response to neoadjuvant chemotherapy and neoendocrine therapy in hormone receptor-positive, human epidermal growth factor receptor 2-negative breast cancer patients from the ABCSG-34 trial

BackgroundNeoadjuvant chemotherapy (NaCT) and neoadjuvant endocrine therapy (NET) can reduce pre-operative tumour burden in patients with estrogen receptor (ER)-positive, human epidermal growth factor receptor 2 (HER2)-negative early-stage breast cancer. This prospective translational study assessed the ability of a 12-gene molecular score (MS; EndoPredict®) to predict response to NaCT or NET within the ABCSG-34 trial. Patients and methodsHormone receptor (HR)-positive, HER2-negative samples from patients in the ABCSG-34 randomized phase II trial were selected and EndoPredict testing was performed to generate a 12-gene MS. ABCSG-34 patients were assigned to receive either NaCT or NET based on menopausal status, HR expression, grade and Ki67. Response was measured by residual cancer burden (RCB). ResultsPatients selected for NaCT generally had high-risk disease by 12-gene MS (125/134), while slightly more patients treated with NET had low-risk disease (44/83). Low-risk NaCT-treated and high-risk NET-treated tumours responded poorly (NPV 100% [95% CI 66.4%–100%] and NPV 92.3% [95% CI 79.1%–98.4%], respectively]. The 12-gene MS significantly predicted treatment response for NaCT (AUC 0.736 [95% CI 0.63–0.84]) and NET (AUC 0.726 [95% CI 0.60–0.85]). ConclusionsThe 12-gene MS predicted RCB after treatment with neoadjuvant therapies for patients with HR-positive, HER2-negative early-stage breast cancer. Tumours with low MS were unlikely to benefit from NaCT, whereas a high MS predicted resistance to NET. This additional biologic information can aid personalized treatment selection in daily practice and builds a strong rationale to use EndoPredict in biomarker-driven studies in the neoadjuvant setting.

Expression of nuclear XIAP associates with cell growth and drug resistance and confers poor prognosis in breast cancer

Evasion from apoptosis is one of the hallmarks of cancer. X-linked inhibitor of apoptosis protein (XIAP) is known to modulate apoptosis by inhibiting caspases and ubiquitinating target proteins. XIAP is mainly found at the cytoplasm, but recent data link nuclear XIAP to poor prognosis in breast cancer. Here, we generated a mutant form of XIAP with a nuclear localization signal (XIAPNLS-C-term) and investigated the oncogenic mechanisms associated with nuclear XIAP in breast cancer. Our results show that cells overexpressing XIAPΔRING (RING deletion) and XIAPNLS-C-term exhibited XIAP nuclear localization more abundantly than XIAPwild-type. Remarkably, overexpression of XIAPNLS-C-term, but not XIAPΔRING, conferred resistance to doxorubicin and increased cellular proliferative capacity. Interestingly, Survivin and c-IAP1 expression were not associated with XIAP oncogenic effects. However, NFκB expression and ubiquitination of K63, but not K48 chains, were increased following XIAPNLS-C-term overexpression, pointing to nuclear signaling transduction. Consistently, multivariate analysis revealed nuclear, but not cytoplasmic XIAP, as an independent prognostic factor in hormone receptor-negative breast cancer patients. Altogether, our findings suggest that nuclear XIAP confers poor outcome and RING-associated breast cancer growth and chemoresistance.

GREM1 is associated with metastasis and predicts poor prognosis in ER-negative breast cancer patients

BackgroundIn breast cancer, activation of bone morphogenetic protein (BMP) signaling and elevated levels of BMP-antagonists have been linked to tumor progression and metastasis. However, the simultaneous upregulation of BMPs and their antagonist, and the fact that both promote tumor aggressiveness seems contradictory and is not fully understood.MethodsWe analyzed the transcriptomes of the metastatic 66cl4 and the non-metastatic 67NR cell lines of the 4T1 mouse mammary tumor model to search for factors that promote metastasis. CRISPR/Cas9 gene editing was used for mechanistic studies in the same cell lines. Furthermore, we analyzed gene expression patterns in human breast cancer biopsies obtained from public datasets to evaluate co-expression and possible relations to clinical outcome.ResultsWe found that mRNA levels of the BMP-antagonist Grem1, encoding gremlin1, and the ligand Bmp4 were both significantly upregulated in cells and primary tumors of 66cl4 compared to 67NR. Depletion of gremlin1 in 66cl4 could impair metastasis to the lungs in this model. Furthermore, we found that expression of Grem1 correlated with upregulation of several stem cell markers in 66cl4 cells compared to 67NR cells. Both in the mouse model and in patients, expression of GREM1 associated with extracellular matrix organization, and formation, biosynthesis and modification of collagen. Importantly, high expression of GREM1 predicted poor prognosis in estrogen receptor negative breast cancer patients. Analyses of large patient cohorts revealed that amplification of genes encoding BMP-antagonists and elevation of the corresponding transcripts is evident in biopsies from more than half of the patients and much more frequent for the secreted BMP-antagonists than the intracellular inhibitors of SMAD signaling.ConclusionIn conclusion, our results show that GREM1 is associated with metastasis and predicts poor prognosis in ER-negative breast cancer patients. Gremlin1 could represent a novel target for therapy.

Advanced glycation end products are elevated in estrogen receptor-positive breast cancer patients, alter response to therapy, and can be targeted by lifestyle intervention

PurposeLifestyle factors associated with personal behavior can alter tumor-associated biological pathways and thereby increase cancer risk, growth, and disease recurrence. Advanced glycation end products (AGEs) are reactive metabolites produced endogenously as a by-product of normal metabolism. A Western lifestyle also promotes AGE accumulation in the body which is associated with disease phenotypes through modification of the genome, protein crosslinking/dysfunction, and aberrant cell signaling. Given the links between lifestyle, AGEs, and disease, we examined the association between dietary-AGEs and breast cancer.MethodsWe evaluated AGE levels in bio-specimens from estrogen receptor-positive (ER+) and estrogen receptor-negative (ER−) breast cancer patients, examined their role in therapy resistance, and assessed the ability of lifestyle intervention to reduce circulating AGE levels in ER+ breast cancer survivors.ResultsAn association between ER status and AGE levels was observed in tumor and serum samples. AGE treatment of ER+ breast cancer cells altered ERα phosphorylation and promoted resistance to tamoxifen therapy. In a proof of concept study, physical activity and dietary intervention was shown to be viable options for reducing circulating AGE levels in breast cancer survivors.ConclusionsThere is a potential prognostic and therapeutic role for lifestyle derived AGEs in breast cancer. Given the potential benefits of lifestyle intervention on incidence and mortality, opportunities exist for the development of community health and nutritional programs aimed at reducing AGE exposure in order to improve breast cancer prevention and treatment outcomes.

Randomized Multicenter Phase II Trial of Neoadjuvant Therapy Comparing Weekly Nab-paclitaxel Followed by FEC With Docetaxel Followed by FEC in HER2− Early-stage Breast Cancer

BackgroundWeekly nanoparticle albumin-bound paclitaxel (nab-paclitaxel) demonstrated greater efficacy with less toxicity than docetaxel in metastatic breast cancer. We conducted a randomized phase II to compare these regimens as neoadjuvant chemotherapy for HER2− early-stage breast cancer. Patients and MethodsStage I-III human epidermal growth factor receptor-negative (HER2−) breast cancer patients were included in the present trial and received either docetaxel every 3 weeks or nab-paclitaxel on days 1, 8, and 15 every 28 days for 4 cycles, followed by FEC (5-fluorouracil, epirubicin, cyclophosphamide) every 3 weeks for 4 cycles. The primary endpoint was the pathologic complete response (pCR) rate, defined as ypT0 and ypN0. The secondary endpoints were pCR (ypT0/ypTis and ypN0), the clinical response rate (using the Response Evaluation Criteria In Solid Tumors criteria), histologic effect of treatment (using the Japanese Breast Cancer Society classification), breast conservation rate, and adverse events. ResultsA total of 152 eligible patients were enrolled at 6 centers. The baseline characteristics were well balanced. In comparing the 2 regimens (docetaxel/nab-paclitaxel), the pCR rate was 12% and 17% (P = .323). In the Ki67 > 20% group, the pCR rate was greater (24%) for the nab-paclitaxel arm than for the docetaxel arm (16%; P = .432). The most common grade 3/4 adverse event was neutropenia, observed in 40% and 36% of cases in the nab-paclitaxel and docetaxel arms, respectively. The nonhematologic adverse events of any grade were myalgia (34% and 32%), arthralgia (42% and 35%), and peripheral sensory neuropathy (55% and 65%) for the 2 treatment arms. No grade 3/4 peripheral sensory neuropathy was observed in the nab-paclitaxel arm. ConclusionWeekly nab-paclitaxel administered at a dose of 100 mg/m2 showed equivalent efficacy and was well tolerated compared with docetaxel as neoadjuvant therapy. Nab-paclitaxel might be more effective in patients with highly proliferative cancer.

Breast cancer: External communication.

In a new study, researchers show that basal-like breast cancer can be converted into the luminal subtype by inhibiting platelet-derived growth factor (PDGF)-CC signalling in the tumour microenvironment, thereby potentially broadening treatment options for oestrogen receptor-negative breast cancer patients.

SF3B1 mutation is a poor prognostic indicator in luminal B and progesterone receptor-negative breast cancer patients.

The purpose of this study was to explore the relationship between SF3B1 mutations and the prognoses of patients with breast cancer. Clinical and SF3B1 mutation data from The Cancer Genome Atlas were analyzed. SF3B1 mutations were evaluated as prognostic factors in all breast cancer patients and specific subgroups through Cox regression and Kaplan-Meier analyses. We also investigated the relationship between traditional parameters and SF3B1 mutations. Receiver operating characteristics curves were used to analyze common risk factors for their sensitivity and specificity in predicting SF3B1 mutations. SF3B1 mutations were a poor prognostic factor in luminal B and progesterone receptor (PR)-negative breast cancer (P < 0.01). Age at diagnosis and estrogen receptor (ER) status were associated with SF3B1 mutations in all breast cancers (P < 0.01) and in luminal B and PR-negative subgroups (P < 0.01). The age at diagnosis and ER status combined had a higher sensitivity and specificity for predicting SF3B1 mutations than each factor alone. SF3B1 mutations are a poor prognostic factor in luminal B and PR-negative breast cancer patients. These mutations are significantly associated with age at diagnosis and ER status. SF3B1 mutations may therefore be a novel therapeutic target for breast cancer patients.

Outcome after neoadjuvant chemotherapy in estrogen receptor-positive and progesterone receptor-negative breast cancer patients: a pooled analysis of individual patient data from ten prospectively randomized controlled neoadjuvant trials.

The estrogen receptor (ER) is involved in control of progesterone receptor (PgR) expression and lack of PgR may be also a surrogate of altered growth factor signaling. The aim of this study was therefore to investigate PgR expression as predictive factor for response to neoadjuvant therapy and long-term outcome. Five thousand and six hundred and thirteen patients with primary breast cancer and positive ER expression from ten German neoadjuvant trials of anthracycline and taxane-based chemotherapy were included. Pathologic complete response (pCR), disease-free survival (DFS), distant disease-free survival (DDFS), overall survival (OS), and local recurrence-free survival (LRFS) were compared according to PgR expression. The lack of PgR expression (1172 patients) was associated with grade 3 (38.4 vs. 26.3%; p<0.001), nodal involvement (>cN2) (6.8% vs. 4.7%; p=0.004), and HER2 positivity (36.2 vs. 22.3%; p<0.001). pCR rates of PgR-negative tumors were higher in the entire cohort (13.8 vs. 7.5%; p<0.001) and in the HER2-negative subgroup (11.2 vs. 5.8%; p<0.001). In multivariable logistic regression, PgR negativity was an independent predictive factor for pCR overall (OR 1.76; p<0.001) and in the HER2-negative patients (OR 1.99; p<0.001). Patients with PgR-negative disease had significantly worse outcome (p<0.001, respectively). Multivariable Cox regression analysis revealed that PgR was an independent prognostic factor for DFS, OS, DDFS, and LRFS. ER-positive/PgR-negative breast carcinomas are associated with higher response but also worse long-term outcome after neoadjuvant therapy. PgR negativity is an independent predictive factor for pCR after neoadjuvant chemotherapy in ER-positive HER2-negative breast cancer.

Abstract 2761: Adjuvant chemotherapy outcomes of node negative, T1a,T1b,T1c hormone receptor-negative HER2-positive breast cancer patients

Abstract Background: Patients (pts) with early stage HER2neu-positive breast cancers have a higher risk of recurrence and death compared to hormone receptor positive patients. NCCN guidelines recommends adjuvant chemotherapy for Stage II and above HER2-positive breast cancer patients. It is unclear what benefit patients with Stage I HER2-positive breast cancer receive from adjuvant chemotherapy. The current study evaluates the effects of adjuvant chemotherapy on the survival of early stage ER/PR negative, HER2-positive breast cancer patients registered to the National Cancer Data base (NCDB) from 2010-2012. Patients and Methods: 3416 women from the NCDB were identified with Stage I ER, PR negative, HER2-positive breast cancer diagnosed between 2010-2012. The primary measured outcome was overall survival. Pts were grouped by tumor size (T1a,T1b,T1c) and adjuvant chemotherapy or no adjuvant chemotherapy. Additionally, patients were also grouped into lumpectomy with radiation and mastectomy with or without radiation. Adjusted variables included age, race, Charlson Comorbidity Index (CCI), payer status, income, education, distance travelled, diagnosing/treating facility and treatment delay. Multivariate Cox regression was used to investigate the effect of adjuvant chemotherapy on overall survival while adjusting for other factors. Results: There were a total of 3416 patients, mean age at diagnosis of 57.9 (range 22-90). The mean age was 56.2 years for the chemotherapy group and 61.8 for the no chemotherapy group. Pathological stage distribution was T1a (24.9%), T1b (24.7%) and T1c (50.2%). Adjuvant chemotherapy was received by 49.5% of T1a, 83.5% of T1b and 95% of T1c pts. Mean patient follow up was 2.7 years (range 0.05-5 years) In univariate analysis, the hazard ratio (HR) of death for chemotherapy vs no chemotherapy in each T stage was: T1a; 1.22 (0.46-3.28), T1b; 0.221 (0.095-0.512), T1c; 0.217 (0.094-0.498). In multivariate analysis, adjusting for other factors in the model, HR of death was 0.51 (95%CI: 0.26-1.03) for chemotherapy vs no chemotherapy. The HR of death was 1.92 (95%CI: 1.11-3.32) for pts who received mastectomy without radiation compared to lumpectomy with radiation. In addition, we observed a HR of 5.16 for CCI score of 2 or above as compared to score of 0. There was no difference in outcome based on payer status or race. Conclusion: Patients with early stage T1 a-c HER2-positive breast cancer had a marginal improvement in overall survival with the addition of chemotherapy. Patients who underwent lumpectomy with radiation were also found to have better overall survival compared to mastectomy without radiation. However, the relatively short follow up and the lack of cancer recurrence information preclude making definitive conclusions relative to adjuvant therapy for pts with these tumors. Longer follow-up and prospective controlled trials will be needed to quantify these potential benefits. Citation Format: Anu Paul, Runhua Shi, Prakash Peddi, Gary Burton. Adjuvant chemotherapy outcomes of node negative, T1a,T1b,T1c hormone receptor-negative HER2-positive breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2761. doi:10.1158/1538-7445.AM2017-2761

Impact of adjuvant chemotherapy on the prognosis of hormone receptor negative breast cancer with residual lymph node disease after neoadjuvant chemotherapy

Objective: To explore the influence of adjuvant chemotherapy on the prognosis of hormone receptor negative breast cancer with residual lymph node disease(RLND)after neoadjuvant chemotherapy. Methods: A total of 110 hormone receptor negative breast cancer patients treated with 4-8 cycles of neoadjuvant chemotherapy were respectively analysed between 2002 and 2012. Residual lymph node disease was comfirmed by subsequent radical mastectomy. Then all these patients were classified into two groups: patients treated with adjuvant chemotherapy(group A) and patients untreated with adjuvant chemotherapy(group B). Results: All patients were female, the median age was 54.5 years old(IQR: 47-59 years). The median follow-up time was 61 months(IQR: 51-88 months). There were 82 patients (74.5%) in group A, and 28 patients (25.5%) in group B. The five-year disease-free survival (DFS) rate was 76.2% in group A and 57.6% in group B. The distant disease-free survival (DDFS) rate was 78.9% in group A and 60.4% in group B. Overall survival (OS) rate was 81.0% in group A and 60.0% in group B. Multivariate analysis showed that there were significant differences for DDFS rate (group A vs group B, P=0.033; hazard ratio [HR], 5.256; 95% confidence interval [95%CI], 1.14-24.17) and OS rates (group A vs group B, P=0.011; HR, 7.478; 95%CI, 1.58-35.30) between two groups. Conclusion: The patients who have hormone receptor negative breast cancer with RLND after neoadjuvant chemotherapy, may benefit from postoperative adjuvant chemotherapy.

AGR2 associates with HER2 expression predicting poor outcome in subset of estrogen receptor negative breast cancer patients

Expression of the AGR2 oncogene was shown to be associated with estrogen receptor positive tumors. This gene contributes to enhanced cellular proliferation, drug resistance, metastasis development and may also serve as a predictor of poor prognosis. However, our analysis of AGR2 expression in a subset of estrogen-receptor negative tumors revealed that AGR2 could also be upregulated in hormone-independent manner. AGR2 expression was shown to be significantly increased in HER2 positive breast tumors on both the mRNA and the protein level. Moreover, in a subset of estrogen- and progesterone-receptor negative and simultaneously HER2-positive cases, increased AGR2 expression significantly correlated with worse patient prognosis. Subsequent analysis of independent data sets either collected in our institute or obtained from Oncomine cancer microarray database confirmed all these findings.

Abstract P1-09-11: Outcome after neoadjuvant chemotherapy in progesterone receptor negative breast cancer patients – A pooled analysis of individual patient data from ten prospectively randomized controlled neoadjuvant trials

Abstract Background The estrogen receptor (ER) as a nuclear transcription factor alters the transcription of estrogen sensitive genes to which the progesterone receptor gene belongs. The ER has also been described to exert non genomic effects by interacting with several cell signalling pathways that do not initially involve increases in gene transcription. These different patterns of action of the ER lead to the assumption that in tumors that utilize the non-genomic ER activity in order to stimulate tumorigenesis and proliferation progesterone receptor (PgR) expression would be decreased or absent. Therefore lack of PgR expression could be a surrogate marker of altered growth factor signalling. The aim of this study was to investigate if PgR expression may act as a predictive factor for response to neoadjuvant chemotherapy and long-term outcome in breast cancer patients. Methods 5613 patients with primary breast cancer, follow-up, positive ER expression; HER2+ and HER2- from overall 10 (n=9785) German neoadjuvant trials receiving an anthracycline and taxane based chemotherapy were included. The pathologic complete response (pCR)(ypT0, ypN0), long term survival data (disease free survival (DFS), distant disease free survival (DDFS), overall survival (OS) and local recurrence free survival (LRFS)) and early relapse, defined as DFS &amp;lt;37 months, were compared according to their PgR expression, overall and in subgroups defined by HER2. Results Tumors lacking PgR expression (1172 patients) were more often of grade 3 (38.4% v 26.3%; p&amp;lt;0.001), tended to have an advanced clinical nodal involvement (6.8% v 4.7%; p=0.004) and were more likely to demonstrate HER2 positivity (36.2% v 22.3%; p&amp;lt;0.001). pCR rates were significantly higher in PgR negative patients in the entire cohort (13.8% v 7.5%; p&amp;lt;0.001) as well as in the HER2 negative subgroup (11.2% v 5.8%; p&amp;lt;0.001) whereas there was no significant difference in the HER2 positive (22.1% v 18%; p=0.117). After adjusting for known predictive factors in the multivariable logistic regression analysis PgR negativity was an independent predictive factor for pCR overall (OR 1.76; p&amp;lt;0.001) and in the HER2 negative patients (OR 1.99; p&amp;lt;0.001). PgR negativity was also significantly associated with an early relapse overall (32.8% v 25.7%; p&amp;lt;0.001) and in the subgroups defined by HER2 (HER2- 32.2% v 24.9%; p&amp;lt;0.001 and HER2+ 39.9%v 30.5%; p=0.002). Patients with PgR negative disease had a significantly worse DFS, OS, DDFS and LRFS (p&amp;lt;0.001, respectively). Multivariable Cox regression analysis revealed that PgR was an independent prognostic factor. This was also observed in the HER2+ and- subgroups. Interestingly, in the PgR negative tumors HER2 status did not influence long-term outcome. Conclusion This analysis demonstrates that ER positive and PgR negative tumors represent a specific subset in primary breast cancer patients associated with higher response but also worse long term outcome after neoadjuvant chemotherapy. Interestingly, PgR negativity served as an independent predictive factor for achieving a pCR after neoadjuvant chemotherapy and therefore its status should be considered when deciding on systemic treatment. Citation Format: van Mackelenberg M, Denkert C, Nekljudova V, Karn T, Schem C, Marme F, Stickeler E, Jackisch C, Hanusch C, Huober J, Fasching P, Blohmer J-U, Kümmel S, Müller V, Schneeweiss A, Untch M, von Minckwitz G, Weber K, Loibl S. Outcome after neoadjuvant chemotherapy in progesterone receptor negative breast cancer patients – A pooled analysis of individual patient data from ten prospectively randomized controlled neoadjuvant trials [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-09-11.

Role of DNA methylation as a diagnostic biomarker of sporadic breast cancer

The initiation and progression of breast cancer have been recognized for many years to be secondary to the accumulation of genetic mutations which lead to aberrant cellular function. Genetic mutations, either inherited or sporadic, may result in the activation of oncogenes and the inactivation of tumor suppressor genes. The more recent discovery that reversible alterations in histone proteins and deoxyribonucleic acid (DNA) can also lead to tumorigenesis has introduced a novel term to the field of cancer research: epigenetics . Epigenetics refers to the study of heritable changes in gene regulation that do not involve a change in the DNA sequence. The most often studied in epigenetics of breast cancer is DNA methylation. That a promoter methylation result in transcription blockade supports the notion that cellular inhibition takes place. Compared to normal tissues, hypermethylation occurs from double to triple in cancerous ones. DNA methylation plays a crucial role in oncogenesis and is one of the hallmarks of cancer. Detection of aberrantly methylated CpG islands in promoter region of several genes in DNA sample derived from nipple aspirates, serum, or cancer tissue associated with down regulation of expression or loss of function of these genes has been associated with early stages of breast cancer, where hypermethylation of CpG island points to poorer prognosis in breast cancer. DNA methylation has been identified as signature for TNBC. Methylation of BRCA1 gene is frequently demonstrated in young, estrogen receptor-negative breast cancer patients. Methylation of specific genes is known to differ across race and socioeconomic status. BRCA1 methylation in premenopausal women with sporadic breast cancer in West Sumatra region has been higher than in Western women. DNA methylation may be used to enhance current breast cancer classification. There is such a distinction between methylation and gene expression profiles of breast cancer that not all methylation profiles fit within the same molecular subtype. Specific gene methylation profiles are identified for basal-like, luminal A and HER2-overexpressing breast cancers. A number of studies have analyzed the methylation status of BRCA1 , a key player in TNBC. One study demonstrated that BRCA1 promoter was methylated in TNBC. It was discovered that the sensitivity of TNBC cell lines to PARP inhibitors was increased when BRCA1 was methylated. Concurrently, BRCA1 methylation quantity was higher in patients with complete response than in those who are non-responders of neoadjuvant chemotherapy. Epigenetics is now the cutting edge of cancer research. Advances in this field will have major implications in diagnosis, prevention, treatment of cancer, and formulation of new epigenetically targeted cancer drugs.

The Circadian Rhythm Gene Arntl2 Is a Metastasis Susceptibility Gene for Estrogen Receptor-Negative Breast Cancer.

Breast cancer mortality is primarily due to metastasis rather than primary tumors, yet relatively little is understood regarding the etiology of metastatic breast cancer. Previously, using a mouse genetics approach, we demonstrated that inherited germline polymorphisms contribute to metastatic disease, and that these single nucleotide polymorphisms (SNPs) could be used to predict outcome in breast cancer patients. In this study, a backcross between a highly metastatic (FVB/NJ) and low metastatic (MOLF/EiJ) mouse strain identified Arntl2, a gene encoding a circadian rhythm transcription factor, as a metastasis susceptibility gene associated with progression, specifically in estrogen receptor-negative breast cancer patients. Integrated whole genome sequence analysis with DNase hypersensitivity sites reveals SNPs in the predicted promoter of Arntl2. Using CRISPR/Cas9-mediated substitution of the MOLF promoter, we demonstrate that the SNPs regulate Arntl2 transcription and affect metastatic burden. Finally, analysis of SNPs associated with ARNTL2 expression in human breast cancer patients revealed reproducible associations of ARNTL2 expression quantitative trait loci (eQTL) SNPs with disease-free survival, consistent with the mouse studies.

Lower bone mineral density at the spine in oestrogen/progesterone receptor negative breast cancer patients

Lower bone mineral density at the spine in oestrogen/progesterone receptor negative breast cancer patients

Activated human mesenchymal stem/stromal cells suppress metastatic features of MDA-MB-231 cells by secreting IFN-β.

Our recent study showed that human mesenchymal stem/stromal cells (hMSCs) are activated to express tumor necrosis factor (TNF)-α-related apoptosis-inducing ligand (TRAIL) by exposure to TNF-α and these activated hMSCs effectively induce apoptosis in triple-negative breast cancer MDA-MB-231 (MDA) cells in vitro and in vivo. Here, we further demonstrated that activated hMSCs not only induced apoptosis of MDA cells but also reduced metastatic features in MDA cells. These activated hMSC-exposed MDA cells showed reduced tumorigenicity and suppressed formation of lung metastasis when implanted in the mammary fat pad. Surprisingly, the activated hMSC-exposed MDA cells increased TRAIL expression, resulting in apoptosis in MDA cells. Interestingly, upregulation of TRAIL in MDA cells was mediated by interferon-beta (IFN-β) secreted from activated hMSCs. Furthermore, IFN-β in activated hMSCs was induced by RNA and DNA released from apoptotic MDA cells in absent in melanoma 2 (AIM2) and IFN induced with helicase C domain 1 (IFIH1)-dependent manners. These observations were only seen in the TRAIL-sensitive breast cancer cell lines but not in the TRAIL-resistant breast cancer cell lines. Consistent with these results, Kaplan–Meier survival analysis also showed that lack of innate sensors detecting DNA or RNA is strongly associated with poor survival in estrogen receptor-negative breast cancer patients. In addition, cancer-associated fibroblasts (CAF) isolated from a breast cancer patient were also able to express TRAIL and IFN-β upon DNA and RNA stimulation. Therefore, our results suggest that the crosstalk between TRAIL-sensitive cancer cells and stromal cells creates a tumor-suppressive microenvironment and further provide a novel therapeutic approach to target stromal cells within cancer microenvironment for TRAIL sensitive cancer treatment.

Pathological complete response rate in hormone receptor-negative breast cancer treated with neoadjuvant FEC, followed by weekly paclitaxel administration: A retrospective study and review of the literature.

While tumor size, the presence of inflammatory carcinoma and lymph node involvement are the main prognostic factors of women with locally advanced breast cancer, the prognostic value of the estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 (HER2) status has not been fully clarified. The present study examined the therapeutic efficacy of a neoadjuvant fluorouracil, epirubicin and cyclophosphamide regimen (FEC), followed by weekly paclitaxel and/or trastuzumab administration, in the treatment of hormone receptor-negative breast cancer patients. Between April 2012 and February 2014, 14 patients with hormone receptor-negative local breast cancer (triple-negative type, 9 patients; HER2 type, 5 patients) were included in the study. In all cases, the histological type of the primary cancer was invasive ductal carcinoma. Among the 14 women who received the regimen, 5 presented with stage I cancer (35.7%), 3 with stage IIA (21.4%), 3 with stage IIB (21.4%), 1 with stage IIIB (7.1%) and 2 with stage IIIC (14.3%), according to the American Joint Committee on Cancer staging system. With regard to the tumor-node-metastasis classification, 5 patients were T1N0M0 (35.7%), 3 were T2N0M0 (21.4%), 3 were T2N1M0 (21.4%), 2 were T3N3M0 (14.3%) and 1 was T4N1M0 (7.1%). The pathological response was evaluated using resected tissue following neoadjuvant chemotherapy, according to the criteria established by the Japanese Breast Cancer Society. Patients were classified into pathological responders (grades 2 and 3, 71.4% of all patients) and non-responders (grade 1, 28.6% of all patients). A pathological complete response (pCR) was achieved in 50.0% of all cases (7/14); 44.4% of triple-negative-type cases (4/9) and 60.0% of HER2-type cases (3/5). Hematological and non-hematological toxicity was reversible and manageable. No patients withdrew from treatment, and favorable compliance was achieved. The present study demonstrated that neoadjuvant FEC followed by weekly administration of paclitaxel and/or trastuzumab induces a high pathological response and a high pCR rate in patients with hormone receptor-negative breast cancer. Due to the high clinical benefit rate and acceptable safety profile, this regimen should be considered an acceptable neoadjuvant treatment option for hormone receptor-negative breast cancer.

Abstract B2-55: Critical role of immune spatial heterogeneity and the molecular scaffold in estrogen receptor-negative breast cancer

Abstract Purpose: The abundance of tumor-infiltrating lymphocytes has been associated with a favorable prognosis in estrogen receptor-negative breast cancer. However, a high degree of spatial heterogeneity in lymphocytic infiltration is often observed in histology samples and its clinical implications and underpinning molecular scaffold remain unclear. Materials and methods: To quantify spatial heterogeneity in the distribution of tumor infiltrating lymphocytes, we combined automated histological image processing with methods of spatial statistics used in ecological data analysis. Hematoxylin and eosin-stained sections from two cohorts of estrogen receptor-negative breast cancer patients (discovery: n=120; validation: n=125) were processed with our automated cell classification algorithm to identify the location of lymphocytes and cancer cells. Subsequently, hotspot analysis (Getis-Ord Gi*) was applied to identify statistically significant hotspots of cancer and immune cells, defined as tumor regions with a significantly high number of cancer or immune cells respectively. To identify molecular aberrations that explain tumor spatial heterogeneity, we integrated our image-based hotspots results with microarray gene expression and copy number data profiled for the same set of tumors. Molecular data were generated with tumor materials sandwiched between these sections, thereby maximizing the biological relevance of multiple data types being generated. Results: We found that the amount of colocalized cancer and immune hotspots weighted by tumor area, rather than number of cancer or immune hotspots, significantly correlates with a better prognosis in estrogen receptor-negative breast cancer in uni- and multivariate Cox analysis. Moreover, colocalization of cancer and immune hotspots further stratified patients with immune cell-rich tumors. Subsequently, we developed a bioinformatics tool, iMmune hOTspoT Omics (iMOTTO), to explain the hotspots as a clinically relevant phenotype using molecular profiling data. Our preliminary analysis revealed significant correlations between this phenotype and expression of immune-specific genes such as CD79, CCL19 and SLAMF1, as well as an immunotherapy target, CTLA4. By incorporating the expression of hotspots-associated genes and copy number alteration data into a multivariate regression model, we aim to define a minimal set of genes to explain the observed degree of cancer-immune hotspot colocalization. Conclusion: Taken together, our study demonstrates the importance of quantifying not only the abundance of lymphocytes but also their spatial variation in the tumor specimen for which methods from other disciplines such as spatial statistics can be successfully applied. Systematic integration of histology and omics data revealed key immune regulators as well as novel genes which warrant further investigation to help elucidate the biological processes underlying immune spatial heterogeneity with potentially important clinical implications. Furthermore, our computational approach can be adapted for studying other cancer types for which immunotherapy has been applied, such as melanoma and non-small cell lung cancer, where our hotspot measures can potentially serve as prognostic and predictive biomarkers. Citation Format: Sidra Nawaz, Andreas Heindl, Andrea Agostinelli, Yinyin Yuan. Critical role of immune spatial heterogeneity and the molecular scaffold in estrogen receptor-negative breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Computational and Systems Biology of Cancer; Feb 8-11 2015; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(22 Suppl 2):Abstract nr B2-55.

Prognostic impact of a single-nucleotide polymorphism near the CTSO gene in hormone receptor-positive breast cancer patients.

Tamoxifen can reduce the occurrence of breast cancer by a half in high-risk women. Recently, a genome-wide association study identified two single-nucleotide polymorphisms (SNPs) nearor in the CTSO and ZNF423 genes that were associated with breast cancer risk during tamoxifen therapy. We hypothesized that these two SNPs could be associated with increased recurrence in breast cancer patients who received adjuvant tamoxifen therapy. A total of 586 breast carcinomas were available for SNP genotyping assays. TaqMan pre-designed SNP genotyping assays were used to identify the presence of CTSO rs10030044 and ZNF423 rs8060157. We then investigated the relationship between CTSO rs10030044 genotypes and mRNA expression levels of CTSO and BRCA1 in 290 breast cancer patients. We found a positive correlation between the variant GG genotype of CTSO rs10030044 and shorter disease-free survival, or overall survival in hormone receptor-positive breast cancer patients receiving adjuvant tamoxifen therapy. In contrast, this genotype was not associated with prognosis in hormone receptor-negative breast cancer patients. Multivariate Cox regression analysis revealed that this genotype was an independent factor indicating a poor prognosis in hormone receptor-positive breast cancer patients receiving adjuvant tamoxifen therapy. No association was found between CTSO genotype and mRNA expression of CTSO and BRCA1. ZNF423 rs8060157 genotype was not associated with prognosis in this study. We show that a SNP near the CTSO gene is a poor prognostic factor in breast cancer although further research might help to reveal the factors linking this genotype and prognosis.