Triple-negative Ductal Carcinoma Research Articles

Germline BRCA 1 Positive Breast Ovarian Cancer Syndrome

Background: BRCA1 (Beast Cancer gene 1) and BRCA2 (Breast Cancer gene 2) are genes that produce proteins that help repair damaged DNA. Mutations in these genes predispose the person to a substantiable risk of developing breast and ovarian cancers among others. Approximately 10 to 15% of all cases of epithelial ovarian cancer and les than 10 % of Breast cancers are caused by a mutation in the BRCA1 or BRCA2 genes, which when occurs together results in hereditary breast ovarian cancer syndrome. Case Presentation: This case report presents the clinical course of a 57-year-old female patient who presented with a mass in her right breast in June 2022. The patient’s past medical history is significant with a diagnosis of triple-negative non-metastatic left breast cancer that was treated with surgery followed by adjuvant chemotherapy containing Doxorubicin and Taxanes with loco-regional radiotherapy in May 2009. On June 2022 (later after 13 years) patient noticed a Rt breast mass and was evaluated further. A mammography showed an ill-defined micro lobulated lesion 4 cm from the nipple, measuring 43 x 27 mm with a few microcalcifications. Later, an ultrasound right breast confirmed the presence of an ill-defined lesion in her right breast with no significant axillary nodes, True-cut biopsy performed on October 26th, 2022 revealed invasive triple negative ductal carcinoma. Staging work up with Positron Emission tomography and computerized tomography scan showed in addition to the right breast mass, an ovarian mass with peritoneal deposits. This prompted a peritoneal biopsy which confirmed a high grade primary serous carcinoma of ovary. Additionally, BRCA1/2 germline testing showed a positive BRCA1 germline mutation. Following this, the patient was started on neoadjuvant keynote 522 protocol specifically with Epirubicin, Cyclophosphamide, Paclitaxel carboplatin with pembrolizumab. As of the date of reporting she has completed CARBOPLATIN-TAXOL and is now on the third ........

P165 - Is metaplastic breast cancer more aggressive than “classic” triple-negative ductal carcinoma? Recent prognostic and therapeutic consideration

P165 - Is metaplastic breast cancer more aggressive than “classic” triple-negative ductal carcinoma? Recent prognostic and therapeutic consideration

EZH2 and MMSET Were Identified as Potentially Useful Therapeutic Targets in Metaplastic Breast Carcinoma.

Metaplastic breast carcinoma (MBC) is a rare malignancy, which is often triple-negative for the hormone receptors and human epidermal growth factor receptor 2, and thus, does not benefit from targeted therapy. In this study, we examined the expression of methylation and demethylation enzymes by immunostaining MBC and the adjacent normal tissues or triple-negative ductal carcinoma (TNDC), and identified alterations that may be used as therapeutic targets. We retrospectively studied surgical specimens from 15 patients who underwent surgery for MBC at Kanagawa Cancer Center between 2005 and 2016, and similarly from 14 patients with TNDC. The frequencies of high methylation/demethylation enzyme expression were compared among them. The frequencies of high enhancer of zeste homolog 2 (EZH2) and multiple myeloma SET domain (MMSET) expression were significantly higher in both MBC and TNDC than in normal tissue. EZH2 and MMSET may be useful therapeutic targets in MBC.

DNA methylation landscape of triple-negative ductal carcinoma in situ (DCIS) progressing to the invasive stage in canine breast cancer

Triple-negative breast cancer (TNBC) is a subtype of breast cancer unresponsive to traditional receptor-targeted treatments, leading to a disproportionate number of deaths. Invasive breast cancer is believed to evolve from non-invasive ductal carcinoma in situ (DCIS). Detection of triple-negative DCIS (TN-DCIS) is challenging, therefore strategies to study molecular events governing progression of pre-invasive TN-DCIS to invasive TNBC are needed. Here, we study a canine TN-DCIS progression and investigate the DNA methylation landscape of normal breast tissue, atypical ductal hyperplasia (ADH), DCIS and invasive breast cancer. We report hypo- and hypermethylation of genes within functional categories related to cancer such as transcriptional regulation, apoptosis, signal transduction, and cell migration. DNA methylation changes associated with cancer-related genes become more pronounced at invasive breast cancer stage. Importantly, we identify invasive-only and DCIS-specific DNA methylation alterations that could potentially determine which lesions progress to invasive cancer and which could remain as pre-invasive DCIS. Changes in DNA methylation during TN-DCIS progression in this canine model correspond with gene expression patterns in human breast tissues. This study provides evidence for utilizing methylation status of gene candidates to define late-stage (DCIS and invasive), invasive stage only or DCIS stage only of TN-DCIS progression.

Adenoid cystic carcinoma of the breast - Discordant size on imaging and pathology: A case report and review of literature

IntroductionAdenoid cystic carcinoma (ACC) is an uncommon tumour of the breast. It is known for its rare lymph node involvement and distant metastasis. A triple-negative breast cancer that has a favorable prognosis compared to other triple negative ductal carcinomas, it accounts for approximately 0.1–1% of all breast cancers.Presentation of caseWe report a case of a 69-year-old female with a palpable left breast mass who underwent multiple imaging modalities with significant size variance between the studies. Breast conserving therapy (BCT) was performed with axillary sentinel lymph node biopsy (SLNB) followed by radiation therapy (RT). Pathological examination confirmed the tumour as ACC.DiscussionACC, known as an persistent if low-grade malignant tumour of salivary gland, is considered to have low-malignant potential in the breast. It is a very rare subtype and from this scant data, there is minimal mention about size discrepancy between imaging modalities such as ultrasound and MRI.No consistent MRI features have been demonstrated, with the exception of T2 hyperintensity in larger lesions and T2 iso-intensity in smaller lesions. Ultrasound demonstrates primarily a hypoechoic or heterogenous mass with minimum vascularity, consistent with our radiographic findings.ConclusionACC is a rare entity in breast cancer pathology. Its size can be highly variable as measured by various radiographic modalities, and final Pathology from the surgical specimen is, as always, required for an accurate tumoral diameter. With that caveat, careful utilization of pre-operative imaging modalities is critical in pre-surgical planning to choose the appropriate surgery.

The Relationship Between Mismatch Repair Deficiency and PD-L1 Expression in Breast Carcinoma.

Mismatch repair (MMR) deficiency in solid tumors has recently been linked to susceptibility to immunotherapies targeting the programmed cell death-1 (PD-1)/programmed cell death-1 ligand (PD-L1) axis. Loss of MMR proteins has been shown to correlate with tumoral PD-L1 expression in colorectal and endometrial carcinomas, but the association between expression of MMR proteins and PD-L1 has not previously been studied in breast carcinoma, where MMR deficiency is less common. We assessed the relationship between PD-L1 and MMR protein expression by immunohistochemistry in 245 primary and 40 metastatic breast carcinomas. Tumoral staining for PD-L1 was positive in 12% of all cases, including 32% of triple-negative cancers. MMR deficiency was observed in 0.04% of breast cancers; the single MMR-deficient case was a high-grade, triple-negative ductal carcinoma which showed dual loss of MLH1 and PMS2 proteins and expressed PD-L1. Two ER carcinomas initially were scored with MMR protein loss in tissue microarray format but were subsequently shown to be MMR-intact on whole sections. Analysis of MMR gene mutation in The Cancer Genome Atlas corroborates low frequency of MMR deficiency for invasive breast cancer. MMR protein expression is therefore unlikely to show utility as a screen for immunotherapeutic vulnerability in this tumor type, and may provoke unwarranted genetic testing in patients unlikely to have a heritable cancer syndrome. PD-L1 may be a more clinically relevant biomarker for anti-PD-1/PD-L1 therapies in this setting.

Utility of Sox10 labeling in metastatic breast carcinomas

Sox10 labeling by immunohistochemistry has been primarily reported in tumors of neural crest origin, such as nerve sheath tumors and melanoma. However, Sox10 also labels primary breast carcinomas, particularly those with the basal-like, triple-negative phenotype. However, the utility of Sox10 labeling in metastatic breast carcinomas has not been reported. Here, we prospectively evaluated Sox10 labeling in surgically resected metastatic breast carcinomas from 26 patients sampled on tissue microarrays. In this cohort, Sox10 labeling was seen in 3 metastatic breast carcinomas (12%), all of which were grade III, triple-negative ductal carcinomas metastatic to the brain (n=2) or lung (n=1). Overall, 38% of triple-negative metastases were Sox10 positive, compared to 0% of estrogen receptor (ER)+ or human epidermal growth factor 2 (HER-2) + metastases (P=.045). In addition, we retrospectively reviewed the use of Sox10 immunohistochemistry in metastatic carcinomas in our clinical practice. We identified 21 cases from January 2012-July 2017 in which Sox10 immunohistochemistry was ordered on clinical sign-out in the work-up of a metastatic carcinoma as being of possible breast origin. Overall, Sox10 labeled 57% (n=12) of all evaluated metastatic carcinomas. All of the Sox10+ tumors were ER-, such that 71% of ER- carcinomas were Sox10+ in comparison to 0% of ER+ carcinomas (P=0.049). In conclusion, the differential diagnosis of a Sox10+ malignancy of unknown origin should not be limited to metastatic melanoma. Sox10 labeling is seen in a subset of metastatic triple-negative breast carcinomas, supporting its use as a marker of breast origin in this setting.

Mutational Profiling Can Establish Clonal or Independent Origin in Synchronous Bilateral Breast and Other Tumors.

BackgroundSynchronous tumors can be independent primary tumors or a primary-metastatic (clonal) pair, which may have clinical implications. Mutational profiling of tumor DNA is increasingly common in the clinic. We investigated whether mutational profiling can distinguish independent from clonal tumors in breast and other cancers, using a carefully defined test based on the Clonal Likelihood Score (CLS = 100 x # shared high confidence (HC) mutations/ # total HC mutations).MethodsStatistical properties of a formal test using the CLS were investigated. A high CLS is evidence in favor of clonality; the test is implemented as a one-sided binomial test of proportions. Test parameters were empirically determined using 16,422 independent breast tumor pairs and 15 primary-metastatic tumor pairs from 10 cancer types using The Cancer Genome Atlas.ResultsWe validated performance of the test with its established parameters, using five published data sets comprising 15,758 known independent tumor pairs (maximum CLS = 4.1%, minimum p-value = 0.48) and 283 known tumor clonal pairs (minimum CLS 13%, maximum p-value <0.01), across renal cell, testicular, and colorectal cancer. The CLS test correctly classified all validation samples but one, which it appears may have been incorrectly classified in the published data. As proof-of-concept we then applied the CLS test to two new cases of invasive synchronous bilateral breast cancer at our institution, each with one hormone receptor positive (ER+/PR+/HER2-) lobular and one triple negative ductal carcinoma. High confidence mutations were identified by exome sequencing and results were validated using deep targeted sequencing. The first tumor pair had CLS of 81% (p-value < 10–15), supporting clonality. In the second pair, no common mutations of 184 variants were validated (p-value >0.99), supporting independence. A plausible molecular mechanism for the shift from hormone receptor positive to triple negative was identified in the clonal pair.ConclusionWe have developed the statistical properties of a carefully defined Clonal Likelihood Score test from mutational profiling of tumor DNA. Under identified conditions, the test appears to reliably distinguish between synchronous tumors of clonal and of independent origin in several cancer types. This approach may have scientific and clinical utility.

Abstract IA22: Investigating breast cancer evolution and diversity with single-cell sequencing

Abstract Sequencing breast tumor cohorts has identified many prevalent mutations, but provides limited insight into the genomic diversity within tumors. Here, we developed a whole-genome and exome single-cell sequencing approach called Nuc-Seq that can achieve 83-97% coverage breadth with low error rates. We applied this method to sequence single normal and tumor cells from an estrogen-receptor positive breast cancer and a triple-negative ductal carcinoma, and performed copy number profiling in parallel. Our data shows that aneuploid rearrangements occurred early in tumor evolution and remained highly stable as the tumor masses expanded. In contrast, point mutations evolved gradually, generating extensive clonal diversity. Many of the diverse mutations were shown to occur at low frequencies (0.03 -10%) in the tumor mass by targeted single-molecule sequencing. Using mathematical modeling we found that the triple-negative tumor cells had an increased mutation rate (13.3X) while the ER+ tumor cells did not. These findings have important implications for the diagnosis, therapeutic treatment and evolution of chemoresistance in breast cancer. Citation Format: Nicholas E. Navin. Investigating breast cancer evolution and diversity with single-cell sequencing. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr IA22.

Abstract LB-310: Single cell genome sequencing reveals clonal stability and diversity in breast cancer

Abstract Human breast cancers often display intratumor genomic heterogeneity. This clonal diversity confounds the clinical diagnosis and basic research of cancer, because single samples may not represent that tumor as a whole. Sequencing breast tumor cohorts en masse has identified many prevalent mutations, but has limited ability for resolving subclonal diversity. Here, we developed a whole-genome and exome single-cell sequencing approach (Nuc-Seq) using G2/M cells. To validate our method, we applied Nuc-Seq to sequence the whole genomes of two single cells from a genetically monoclonal breast cancer cell line (SK-BR-3) at high coverage depth (61X ± 5 sem, n=2) and breadth (83.70% ± 3.40 sem, n=2) to detect somatic mutations. Our analysis suggests that Nuc-Seq generates low allelic dropout rates (9.73% ± 2.19%) and low false positive error rates for point mutations (FPR = 1.24e-6). We then applied this method to sequence single normal and tumor cells from an estrogen-receptor positive breast cancer and a triple-negative ductal carcinoma at base-pair resolution. In parallel, we performed single cell copy number profiling. In both tumors, we observed a large number of rare variants that were not detected by sequencing the bulk tumor en masse. In contrast, we find that single cell copy number profiles are highly similar. Our data suggest that aneuploid rearrangements occurred early in tumor evolution and remained highly stable as the tumor mass expanded. In contrast we find that point mutations evolved gradually, generating extensive clonal diversity. Many of the diverse mutations were shown to occur at low frequencies (0.03 -10%) in the tumor mass by targeted duplex sequencing. Mathematical modeling suggests that the triple-negative tumor cells have an increased mutation rate (13.3X), while the ER+ tumor cells do not. These findings have important implications for the diagnosis, therapeutic treatment and evolution of chemoresistance in breast cancer. Citation Format: Yong Wang, Nicholas Navin, Jill Waters, Marco Leung, Anna Unruh, Xiuqing Shi, Whijae Roh, Ken Chen, Paul Scheet, Selina Vattathil, Han Liang, Asha Multani, Hong Zhang, Funda Meric-Bernstam, Franziska Michor, Rui Zhao. Single cell genome sequencing reveals clonal stability and diversity in breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-310. doi:10.1158/1538-7445.AM2014-LB-310

Clonal evolution in breast cancer revealed by single nucleus genome sequencing.

SUMMARYSequencing studies of breast tumor cohorts have identified many prevalent mutations, but provide limited insight into the genomic diversity within tumors. Here, we developed a whole-genome and exome single cell sequencing approach called Nuc-Seq that utilizes G2/M nuclei to achieve 91% mean coverage breadth. We applied this method to sequence single normal and tumor nuclei from an estrogen-receptor positive breast cancer and a triple-negative ductal carcinoma. In parallel, we performed single nuclei copy number profiling. Our data show that aneuploid rearrangements occurred early in tumor evolution and remained highly stable as the tumor masses clonally expanded. In contrast, point mutations evolved gradually, generating extensive clonal diversity. Many of the diverse mutations were shown to occur at low frequencies (<10%) in the tumor mass by targeted single-molecule sequencing. Using mathematical modeling we found that the triple-negative tumor cells had an increased mutation rate (13.3X) while the ER+ tumor cells did not. These findings have important implications for the diagnosis, therapeutic treatment and evolution of chemoresistance in breast cancer.

Ductal carcinoma in situ associated with triple negative invasive breast cancer: evidence for a precursor–product relationship

AimWe evaluated pathological features of the ductal carcinoma in situ component of 241 triple negative invasive breast cancers.ResultsWe found that 151 (62.6%) in situ lesions were of high nuclear grade,...

Neurotensin Receptor 1 (NTSR1) Overexpression in Breast Carcinomas Is Common and Independent of ER/PR/Her2 Expression

Neurotensin (NT) is a 13-amino acid peptide with trophic effects on some neoplasms. Its bioactivities are mainly mediated by neurotensin receptor 1 (NTSR1). Both NT and NTSR1 were found to be upregulated in breast cancer. NT/NTSR1 thus becomes a potential therapeutic target. We studied whether any correlation exists between the expression of NTSR1 in breast carcinomas and the expression of ER, PR, and Her2. A total 85 cases of invasive ductal (62) and lobular (23) breast carcinomas were studied. Based on their ER/PR profiles, the ductal carcinomas (DCs) were subcategorized into ER+/PR+ (21), ER+/PR﹣ (20), and ER﹣/PR﹣ (21). All of the lobular carcinomas (LCs) were ER+/PR+. 21.57% of all DCs and 5.56% of LCs were Her2 positive. 77.78% of ER﹣/PR﹣ DCs were also Her2 negative (triple negative). The expression of NTSR1 was detected by immunohistochemistry and was semiquantitated (as negative, 1+, 2+, 3+). Both 2+ and 3+ were collectively defined as overexpression. The expression of NTSR1 was weak and focal in non-neoplastic mammary epithelial cells. It is increased in 74.19% of DCs (80.95% in ER+/PR+, 75% in ER+/PR﹣, and 66.67% in ER﹣/PR﹣ group), and in 95.65% of LCs. The overexpression of NTSR1 is similar between ER+ DCs and ER﹣ DCs (75% vs 66.67%, p > 0.05) as well as between PR+ DCs and PR﹣ DCs (80.95% in ER+/PR+ DCs vs 75% in ER+/PR﹣ DCs, p > 0.05). And it was seen in 77.78% of Her2+ DCs, 78.38% of Her2﹣ DCs, 94.12% of Her2﹣ LCs, and 78.57% of triple negative DCs. Overall, NTSR1 is commonly overexpressed in both ductal and lobular breast carcinomas and is independent of the ER/PR/Her2 profiles of the tumors. The present data supports the potential benefit of developing NTSR1 blockers in the adjuvant therapy of breast carcinomas, particularly for those “triple negative” tumors.

Radiographic features for triple negative ductal carcinoma in situ of the breast

Triple negative (TN) breast cancer is characterized as having a high malignancy potential and a poor prognosis. An understanding of the radiological features of TN DCIS will enable the early detection of intractable TN invasive breast cancer. Our cohort of 494 DCIS patients, including 18 TN DCIS cases (3.6%), was diagnosed and treated between January 2006 and November 2009. We reviewed the TN DCIS cases in order to assess mammogram (MMG), ultrasound (US), magnetic resonance imaging (MRI), and pathology findings. Routine diagnostic MMG, US, and MRI were performed before surgery in our institution. The average age of TN DCIS patients was 54.2years (40-73). MMG findings were as follows: no abnormal findings (n=4), masses (n=3), focal asymmetric density (n=2), architectural distortion (n=5), and calcifications (n=4). US findings included low echoic masses (n=17) and architectural distortion (n=4). MRI findings (two patients did not undergo this examination) included mass (n=6) and non-mass-like enhancements (n=10). The average lesion measured 3.3cm in diameter (0.5-8.5cm). The histological findings were non-comedo type (n=9) and mixed type including some comedo components (n=9). Notably, our study revealed that only 22% of TN DCIS cases were detected via mammographic abnormal calcifications. This percentage was less than in non-TN DCIS cases. TN DCIS cases were detected mainly because of masses or asymmetry. US and MRI findings of TN DCIS revealed the same features considered to be common DCIS findings. We diagnosed TN DCIS in 3.6% of our cohort including 494 DCIS patients. Incidences of TN DCIS with mammographic abnormal calcifications were fewer than those of non-TN DCIS cases.