Ductal Carcinoma In Situ Recurrence Research Articles

The clinical significance of oestrogen receptor expression in breast ductal carcinoma in situ

BackgroundOestrogen receptor (ER) in invasive breast cancer (BC) predicts response to endocrine therapy (ET) and provides prognostic value. In this study, we investigated the value of ER expression in ductal carcinoma in situ (DCIS) in terms of outcome and the impact on ET decision.MethodsIn total, 643 pure DCIS, diagnosed at Nottingham University Hospitals, were assessed for ER. Clinicopathological data were correlated against ER status, together with assessment of recurrence rate.ResultsER positivity was observed in 74% (475/643) of cases. ER positivity was associated with clinicopathological variables of good prognosis; however, outcome analysis revealed that ER status was not associated with local recurrence. In the intermediate- and high-grade ER-positive DCIS, 58% (11/19) and 63% (15/24) of the recurrences were invasive, respectively, comprising 7% and 6% of all ER-positive DCIS, respectively. Invasive recurrence in low-grade DCIS was infrequent (2%), and none of these patients died of BC. The ER status of the recurrent invasive tumours matched the primary DCIS ER status (94% in ipsilateral and 90% of contralateral recurrence).ConclusionThe strong correlation between DCIS and invasive recurrence ER status and the clinical impact of ET justify discussion of the use of ET in ER-positive DCIS treated by breast-conserving surgery. The excellent outcome of low-grade DCIS, which was almost always ER-positive, does not, in the opinion of authors, justify the use of risk-reducing ET. Therefore, the decision on ET for DCIS should be personalised and consider grade, ER status and other characteristics.

The prognostic significance of immune microenvironment in breast ductal carcinoma in situ

BackgroundThe role of different subtypes of tumour infiltrating lymphocytes (TILs) in breast ductal carcinoma in situ (DCIS) is still poorly defined. This study aimed to assess the prognostic significance of B and T lymphocytes and immune checkpoint proteins expression in DCIS.MethodsA well characterised DCIS cohort (n = 700) with long-term follow-up comprising pure DCIS (n = 508) and DCIS mixed with invasive carcinoma (IBC; n = 192) were stained immunohistochemically for CD20, CD3, CD4, CD8, FOXP3, PD1 and PDL1. Copy number variation and TP53 mutation status were assessed in a subset of cases (n = 58).ResultsCD3+ lymphocytes were the predominant cell subtype in the pure DCIS cohort, while FOXP3 showed the lowest levels. PDL1 expression was mainly seen in the stromal TILs. Higher abundance of TILs subtypes was associated with higher tumour grade, hormone receptor negativity and HER2 positivity. Mutant TP53 variants were associated with higher levels of stromal CD3+, CD4+ and FOXP3+ cells. DCIS coexisting with invasive carcinoma harboured denser stromal infiltrates of all immune cells and checkpoint proteins apart from CD4+ cells. Stromal PD1 was the most differentially expressed protein between DCIS and invasive carcinoma (Z = 5.8, p < 0.0001). Dense TILs, stromal FOXP3 and PDL1 were poor prognostic factors for DCIS recurrence, while dense TILs were independently associated with poor outcome for all recurrences (HR = 7.0; p = 0.024), and invasive recurrence (HR = 2.1; p = 0.029).ConclusionsImmunosuppressive proteins are potential markers for high risk DCIS and disease progression. Different stromal and intratumoural lymphocyte composition between pure DCIS, DCIS associated with IBC and invasive carcinoma play a potential role in their prognostic significance and related to the underlying genomic instability. Assessment of overall TILs provides a promising tool for evaluation of the DCIS immune microenvironment.

Abstract P6-10-21: Evaluation of genomic changes in ductal carcinoma in situ as potential biomarkers of recurrence risk

Abstract Routine breast cancer screening has increased detection of pre-invasive lesions such as ductal carcinoma in situ (DCIS). Currently, DCIS cases are mostly treated with wide local excision combined with adjuvant radiotherapy and/or hormonal therapy, even though only 25% of DCIS treated with surgery alone recur with 50% of recurrences being invasive. At present, prediction of recurrence risk is based on histopathological features and patient characteristics but reliable identification of individual recurrence risk is still challenging. Certain genetic aberrations (i.e. copy number variation, somatic mutations) are associated with breast cancer progression. While most recurrences are assumed to be genetically related to the primary tumour (“clonal recurrence”), the prevalence of ipsilateral independent primary events (“non-clonal recurrence”) after treatment for DCIS is uncertain. If this is a substantial proportion of cases, their presence would compromise attempts to find tumour intrinsic biomarkers of recurrence. To enable the discovery of better recurrence biomarkers, this project initially aims to identify cases with primary DCIS and later clonal ipsilateral recurrence. Sections from formalin-fixed paraffin-embedded blocks from primary DCIS cases and their subsequent recurrences were obtained from Nottingham City Hospital (UK). Two pathologists reviewed representative sections to identify areas for microdissection. Tumour epithelial cells were needle-microdissected and DNA was isolated. Sequencing libraries were prepared using a) a custom capture panel of 110 breast cancer-related genes or b) NEBNext Ultra II for low-coverage whole-genome sequencing (LC-WGS) depending on DNA quality and quantity. Sequencing was then performed on Illumina NextSeq 500 system with an average coverage of a) 400x and b) 1.3x, respectively. Data was processed by an in-house bioinformatics pipeline to detect and filter for high confidence somatic variants. Copy number profiles were obtained using CopywriteR (panel off-target reads), PureCN (panel on-target reads) and Control-FREEC (LC-WGS). Of 33 patients initially ascertained on the basis of a recurrent breast event, 30 had an ipsilateral recurrent tumour after 4.7 years on average, and 24 primary/recurrence pairs had sufficient data available to determine clonality. Eighteen pairs (75%) were classified as clonal by the following criteria: occurring in similar locations, showing a similar or higher tumour grade, and sharing ≥ 3 copy number breakpoints or likely somatic variants in the genes included in the panel. Six pairs (25%) did not meet this criteria and were classified as non-clonal. Preliminary data analysis shows that invasive recurrences (n = 12) were detected on average 3 years later than DCIS recurrences (n = 6), despite constant surveillance. The most commonly observed somatic mutations occurred in the TP53 and PIK3CA genes. Genetic variants in TP53 and PIK3CA were equally common in primary tumours with clonal and non-clonal recurrence. Frequent copy number alterations involved losses on 16q as well as gains on 1q, 8q and 17q, whereat the 8q gain was less profound. The presence of 25% non-clonal recurrences shows that differentiating between clonal and non-clonal recurrences might be crucial for further recurrence risk biomarker discovery. Thus, this project will proceed with comparing genetic variation in primary DCIS with clonal recurrence to DCIS cases without later recurrence. These samples are currently being obtained and analysed. Citation Format: Dorothea Lesche, Hugo Saunders, Sakshi Mahale, Magnus Zenthoven, Siobhan Huges, Tanjina Kader, Niko Thio, Islam M Miligy, Jia-Min Pang, Andrew R Green, Emad A Rakha, Ian G Campbell, Stephen B Fox, Kylie Gorringe. Evaluation of genomic changes in ductal carcinoma in situ as potential biomarkers of recurrence risk [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 P6-10-21.

Abstract P3-02-12: CAV1, MCT1, and MCT4 expression in ductal carcinoma in situ of the breast and its association with clinicopathologic features

Abstract Background: Multicompartment tumor metabolism and metabolic coupling are markers of aggressive tumor behavior, clinically manifested as disease recurrence and progression. Loss of caveolin-1(CAV1) in stromal cells and upregulation of monocarboxylate transporters (MCTs), especially MCT1 and MCT4, in both stromal and cancer cells have been identified as playing an important role in the metabolic coupling necessary for release and uptake of metabolites. However, this phenomenon has only scarcely been described in ductal carcinoma in situ (DCIS) of the breast. The purpose of this study was to investigate the role of CAV1, MCT1, and MCT4 in DCIS carcinogenesis and its association with clinicopathologic factors. Materials and methods: CAV1, MCT1, and MCT4 expression was examined in 9 pairs of DCIS tissues and matched normal adjacent tissues at mRNA and protein levels by qRT-PCR, RNAscope in situ hybridization (ISH) and immunohistochemistry. Immunohistochemical staining of CAV1, MCT1, and MCT4 was done in 79 DCIS samples with known hormone receptor (HR) and human epidermal growth factor 2 (HER2) expression using tissue microarray. Results: CAV1 mRNA expression by qRT-PCR analysis was significantly decreased in DCIS tissues compared to their corresponding normal tissues. On the other hand, MCT1 and MCT4 mRNA expression in DCIS tissues were increased compared with corresponding normal tissues and the difference was statistically significant in MCT1. Compared with their corresponding normal tissues, DCIS tissues showed decreased stromal CAV1 mRNA and protein expression and increased epithelial MCT1 and MCT4 mRNA and protein expression by ISH and immunohistochemistry. Low stromal CAV1, high epithelial MCT1, and high epithelial MCT4 expressions were observed in 29 (36.7%), 51 (64.6%) and 15 (18.9%) cases with DCIS, respectively. Low stromal CAV1 expression was significantly associated with high nuclear grade (P &amp;lt; 0.05). High epithelial MCT4 expression was associated with larger tumor size and HER2 positivity (P &amp;lt; 0.05 and P &amp;lt; 0.05, respectively). Although not statistically significant, low stromal CAV1 and high epithelial MCT4 expression tended to be associated with recurrence of DCIS. No associations were observed between stromal CAV1 expression and epithelial MCT 1 or MCT4 expression. Conclusions: These results suggested that changes in CAV1, MCT1, and MCT4 are associated with carcinogenesis of DCIS, and further studies are needed for their clinical usefulness. Keywords: Ductal carcinoma in situ; CAV1; MCT1; MCT4; Metabolism Citation Format: Ji Shin Lee, Nah Ihm Kim, Min Ho Park. CAV1, MCT1, and MCT4 expression in ductal carcinoma in situ of the breast and its association with clinicopathologic features [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 P3-02-12.

Abstract P6-10-20: Ductal carcinoma in situ (DCIS) of the breast: miRNA deregulation correlates with the Oncotype DX risk score

Abstract Background: Ductal carcinoma in situ (DCIS) is considered a non-obligate precursor of invasive breast cancer (IBC). It is estimated that 5-14% of women diagnosed with DCIS experience an ipsilateral IBC. Current clinical criteria do not discriminate well between women with DCIS who will and those who will not develop ipsilateral IBC or a DCIS recurrence. The Oncotype DX DCIS assay (12 genes) was derived from a multigene expression array (Oncotype DX, 21 genes), and includes 5 proliferation genes, PGR, GSTM1, and 5 reference genes. This assay has been shown to predict risk of subsequent ipsilateral IBC and DCIS, and is used by clinicians to evaluate both the risk of local recurrence and need for radiation therapy. Since abnormal expression of microRNAs (miRNAs) is thought to contribute to the development of IBC, and that very few studies have evaluated miRNA expression deregulation in DCIS lesions, we sought to determine if specific miRNA expression changes may help identify DCIS lesions with different Oncotype DX DCIS scores. Methods: For this study, we selected archived specimens from 52 women diagnosed with DCIS between 2012 and 2018, whose DCIS lesions were tested with the Oncotype DX DCIS assay (low (n=35), intermediate (n=11), high (n=6) risk scores). Total RNA was extracted by macrodissection of the DCIS lesions. MiRNA expression differences between DCIS lesions were evaluated using small-RNA next-generation sequencing. Quantitative PCR validations were performed using Taqman miRNA assays on the top 15 differentially expressed miRNAs Results: Small-RNA expression data obtained by next-generation sequencing of 43 DCIS RNA specimens identified miRNAs differentially expressed between the risk score groups. Fifteen differentially expressed miRNAs were selected for their association based on Oncotype DX DCIS score differences (miR-142-3p, miR-142-5p, miR-744, miR-155, miR-30c, miR-150, miR-146a, miR-190b, miR-135b, miR-193b) or on age of the patients (miR-19b, miR-19a, miR-135a, miR-132). QPCR validation of these 15 miRNAs, using RNA from 30 DCIS lesions with known Oncotypes DX DCIS scores, of which 21 were samples that had been sequenced (low (n=8), intermediate (n=9), and high (n=4)), and 9 were not sequenced (low (n=7), intermediate (n=1), high (n=1)) and identified 5 miRNAs (miR-135a (p=&amp;lt;0.05), miR-190b (p=0.043), miR-205 (p=&amp;lt;0.001), miR-30c (p=&amp;lt;0.01), miR-744 (p=&amp;lt;0.038)) that emerged for their association with the score groups. These 5 miRNAs provided a good discrimination tool between low and intermediate-high score samples. A composite score including these 5 miRNAs showed a highly significant association with the Oncotype DX DCIS Score (p=&amp;lt;0.001). Conclusions: Our analyses identified a subset of 5 miRNAs that is significantly associated with the Oncotype DX DCIS score of DCIS lesions. Interestingly, three of these miRNAs (miR-205, miR-744 and miR-135b) have been reported to regulate proliferation pathways, while the other two (miR-30c, and miR-190b) have been associated with estrogen expression pathways in breast cancer cells. Importantly, increased expression of miR-30c in IBC has been associated with benefit from endocrine therapy while increased expression of miR-205 has been associated with benefit from tyrosine kinase inhibitors in IBC, as well. Mir-205 has also been reported as a radiosensitizer in breast cancer cells. Altogether, our data suggest that evaluation of miRNA expression levels may add predictive and prognostic value to the diagnosis of DCIS. Citation Format: Olivier Loudig, Iddo Ben-Dov, Christina Liu, Miglena Komforti, Susan Fineberg. Ductal carcinoma in situ (DCIS) of the breast: miRNA deregulation correlates with the Oncotype DX risk score [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 P6-10-20.

Abstract P3-08-22: The mutational landscape of cancer driver genes in matched primary ductal carcinoma in situ and recurrent ductal carcinoma in situ or recurrent invasive cancers

Abstract Due to breast screening, ductal carcinoma in situ (DCIS) accounts for approximately 25% of all newly diagnosed breast neoplasms. Believed to be a precursor to invasive carcinoma, a significant number of patients diagnosed with DCIS are effectively managed by surgery alone or in conjunction with radiotherapy and endocrine therapy. In general, there is an 8-11% relative risk for a subsequent invasive carcinoma over a period of 10 year, with 98% breast cancer-specific survival after 10 years of follow up. Although mastectomy, breast conserving surgery, and radiotherapy can reduce the risk of recurrence, there are ongoing lifetime consequences of treatment. Thus, there is a clinical need to identify those patients who are at risk of an invasive recurrence from those who might not recur or experience a subsequent DCIS recurrence. In this study, 60 patients with pure primary DCIS, treated with only with breast conserving therapy, across three different clinical outcome groups were examined: patients who did not experience a recurrence within 5 years (n=20); patients who experienced a recurrent DCIS within 5 years (n=20); and patients that recurred with an invasive cancer within 5 years (n=20). Pure primary DCIS lesions, as well as the matched DCIS recurrence or invasive recurrence, were macrodissected from formalin fixed paraffin embedded tissues and subjected to nucleic acid extraction. All samples were profiled using Thermo Fisher Scientific’s validated targeted sequencing panel, the Oncomine Comprehensive Assay v3.0 (OCAv3.0). This assay is comprised of common cancer driver genes shown to be prognostic and predictive to targeted therapies in use or in late-phase clinical trials, and is currently being used in the NCI-MATCH trial (NCT02465060). While the OCA panel and accompanying Oncomine Knowledgebase Reporter provides information regarding the targeted treatments linked to known actionable mutations, this study utilized all somatic mutations and copy number changes to reveal the genomic landscape of DCIS and their matched recurrences across these pan-cancer driver genes Amongst all primary DCIS samples across the three different clinical outcome groups, PIK3CA, was frequently found to be affected by SNV and Indels (32.8%) in addition to TP53 (26.2%), NF1 (21.3%), CREBBP (16.4%), ATM (14.8%), PALB2 (14.8%). DNA repair genes, including CHEK1, RAD50, RAD51B, MRE11A, BRCA1 and BRCA2, were found to be frequently subject to mutation in these primary DCIS samples ranging from 5%-15%. Similarly, copy number gains were frequently detected in HER2 (26.2%), CDK12 (18%), FGFR1 (4.9%), GNAS (4.9%), MYC, CCNE1, AR, RAD51C and RNF43 (1.6% each), and loses at H3F3A and KNSTRN (each 1.6%). Matched primary DCIS and their recurrent DCIS or invasive lesions exhibited similar changes with invasive cancers suggesting that in some cases, the primary DCIS gives rise to the invasive cancer. We will present the preliminary findings mapping the mutational and copy-number landscape of primary DCIS and matched recurrences to identify putative genomic changes defining these clinical outcome groups and to investigate the genomic progression of DCIS to invasive carcinoma. Citation Format: Jane Bayani, Quang M Trinh, Mary Anne Quintayo, Cheryl Crozier, Ilinca Lungu, Dan Dion, Joema Felipe-Lima, Giancarlo Pruneri, Jonas Bergh, Fredrik Warnberg, Giuseppe Viale, Lincoln D Stein, John MS Bartlett. The mutational landscape of cancer driver genes in matched primary ductal carcinoma in situ and recurrent ductal carcinoma in situ or recurrent invasive cancers [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 P3-08-22.

Derivation of a nuclear heterogeneity image index to grade DCIS

Abnormalities in cell nuclear morphology are a hallmark of cancer. Histological assessment of cell nuclear morphology is frequently used by pathologists to grade ductal carcinoma in situ (DCIS). Objective methods that allow standardization and reproducibility of cell nuclear morphology assessment have potential to improve the criteria needed to predict DCIS progression and recurrence. Aggressive cancers are highly heterogeneous. We asked whether cell nuclear morphology heterogeneity could be incorporated into a metric to classify DCIS. We developed a nuclear heterogeneity image index to objectively, and quantitatively grade DCIS. A whole-tissue cell nuclear morphological analysis, that classified tumors by the worst ten percent in a duct-by-duct manner, identified nuclear size ranges associated with each DCIS grade. Digital image analysis further revealed increasing heterogeneity within ducts or between ducts in tissues of worsening DCIS grade. The findings illustrate how digital image analysis comprises a supplemental tool for pathologists to objectively classify DCIS and in the future, may provide a method to predict patient outcome through analysis of nuclear heterogeneity.

FAP-a and GOLPH3 Are Hallmarks of DCIS Progression to Invasive Breast Cancer.

Biological markers that could predict the progression of ductal carcinoma in-situ (DCIS) to invasive breast cancer (IDC) are required urgently for personalized therapy for patients diagnosed with DCIS. As stroma was invaded by malignant cells, perturbed stromal-epithelial interactions would bring about tissue remodeling. With the specific expression of the fibroblast activation protein-alpha (FAP-a), Carcinoma-associated fibroblasts (CAFs) are the main cell populations in the remodeled tumor stroma. Golgi phosphoprotein 3 (GOLPH3), a documented oncogene possessing potent transforming capacity, is not only up-regulated in many tumors but also an efficient indicator of poor prognosis and more malignant tumors. The present study aimed to retrospectively evaluate the pathological value of FAP-a and GOLPH3 in predicting the recurrence or progression of DCIS to invasive breast cancer. Immunohistochemical techniques were applied to investigate the expression of FAP-a GOLPH3 in 449 cases of DCIS patients received extensive resection and with close follow-up, but not being treated with any form of chemo- or radio-therapy. The combination of FAP-a and GOLPH3 in predicating the recurrence or progression of DCIS into invasive breast cancer was specifically examined. The study demonstrated that the overexpression of FAP-a in stromal fibroblasts and GOLPH3 in carcinoma cells are highly predictive of DCIS recurrence and progression into invasive breast cancer. Both FAP-a and GOLPH3 have high specificity and sensitivity to predict the recurrence of DCIS. Moreover, the combination of FAP-a and GOLPH3 tends to further improve the specificity and sensitivity of DCIS recurrence by 9.72–10.31 and 2.72–3.63%, respectively. FAP-a and GOLPH3 serve as novel markers in predicting the recurrence or progression of DCIS into invasive breast cancer.

Collagen (XI) alpha-1 chain is an independent prognostic factor in breast ductal carcinoma in situ

Collagen11A1 (COL11A1) is a fibrillary type collagen constituting a minor component of the extracellular matrix and plays role in tissue tensile strength. Overexpression of COL11A1 expression is associated with aggressive behavior and poor outcome in several human malignancies. In this study, we evaluated the association between COL11A1 expression and clinicopathological parameters of the breast ductal carcinoma in situ (DCIS) and its prognostic value. COL11A1 protein expression was assessed immunohistochemically in a large well-characterized cohort of DCIS including pure (n = 776) and DCIS associated with invasive carcinoma (DCIS-mixed, n = 239). COL11A1 expression was assessed in tumor cells and surrounding stromal cells, and correlated with clinicopathological parameters, immunoprofile and disease outcome. In pure DCIS, high COL11A1 expression was observed in tumor cells and surrounding stromal cells in 25 and 13% of cases, respectively. Higher COL11A1 expression within the stromal cells was associated with hormone receptor negative, HER2 enriched and triple negative molecular subtypes and showed a positive linear correlation with proliferation index, dense tumor infiltrating lymphocytes and hypoxia-inducible factor 1 alpha. COL11A1 expression in tumor and stromal cells was significantly higher in DCIS associated with invasive carcinoma than in pure DCIS, and within the DCIS-mixed cohort, the invasive component showed higher COL11A1 expression than the DCIS component (all, p < 0.0001). Overexpression of stromal COL11A1 was an independent predictor of shorter local recurrence-free interval for all recurrences (HR = 13.2, 95% CI = 6.9–25.4, p < 0.0001) and for invasive recurrences (HR = 11.2, 95% CI = 4.9–25.8, p < 0.0001). When incorporated with other risk factors, stromal COL11A1 provided better patient risk stratification. DCIS with higher stromal COL11A1 expression showed poor outcome even with adjuvant radiotherapy management. In conclusion, overexpression of stromal COL11A1 is associated with invasive recurrence in DCIS and is a potential marker to predict the response to radiotherapy.

Geometric characteristics of collagen have independent prognostic significance in breast ductal carcinoma in situ: an image analysis study

Collagen plays a key role in normal and malignant tissue homeostasis. While the prognostic significance of collagen fiber remodeling in invasive breast cancer has been studied, its role in ductal carcinoma in situ (DCIS) remains poorly defined. Using image analysis, we aimed to evaluate the prognostic significance of the geometric characteristics of collagen surrounding DCIS. A large well-characterized cohort of DCIS comprising pure DCIS (n = 610) and DCIS coexisting with invasive carcinoma (n = 180) were histochemically stained for collagen using picrosirius red. ImageJ software was used to assess collagen density, degree of collagen fiber dispersion and directionality in relation to DCIS ducts' boundary. We developed a collagen prognostic index and evaluated its prognostic significance. A poor index was observed in 24% of the pure DCIS and was associated with determinants of high-risk DCIS including higher nuclear grade, comedo type necrosis, hormonal receptor negativity, HER2 positivity and high proliferation index. High collagen prognostic index was associated with the collagen remodeling protein prolyl-4-hydroxlase alpha subunit 2 and the hypoxia-related protein hypoxia inducible factor 1α. DCIS coexisting with invasive breast cancer had a higher collagen prognostic index than pure DCIS ( p < 0.0001). High index was an independent poor prognostic factor for DCIS recurrence for all recurrences (HR = 2.3, p = 0.005) and just invasive recurrences (HR = 3.4, p = 0.003). Interaction between collagen prognostic index and radiotherapy showed that the index was associated with poor outcome even with adjuvant radiotherapy ( p = 0.0001). Collagen reorganization around DCIS is associated with poor outcome and provides a potential predictor for disease progression and resistance to radiotherapy. Mechanistic studies are warranted to decipher the underlying mechanisms.

Breast Conserving Surgery and Intraoperative Electron Radiotherapy (IOERT) Among Cases of Ductal Carcinoma In Situ

Background: Ductal Carcinoma In Situ (DCIS) which has recently been renamed into Ductal Intraepithelial Neoplasia (DIN), is a malignant cell proliferation without invasion to basement membrane of ducts or lobules of breast. DCIS consists 20-30% of newly diagnosed breast cancers in some Western countries due to higher diagnosis resulting from screening by mammography. Relative Risk (RR) of invasive ductal carcinoma is 8-10 times in DCIS, although high grading lesions and positive or close surgical margins are two important predictive factors in DCIS recurrences. The adjuvant radiotherapy has decreased the rate of ipsilateral local recurrence about 60%. In this article, we evaluated the recurrence rate as DCIS as well as invasive breast cancer in patients with DCIS undergoing breast conserving surgery (BCS) and intraoperative electron radiotherapy (IOERT).Methods: Data were derived from Cancer Research Center database from 38 pure DCIS cases who had received intraoperative radiation therapy between 2012–2017. Intraoperative electron radiotherapy (IOERT) was performed according to Iran's intraoperative radiation therapy consensus.Results: The median age of the patients was 55 years and median histological lesion size was 1.8 centimeters. Number of extracted lymph nodes had a median of 1 and all extracted nodes were negative. Hormonal therapy was performed in 42.1% of patients. IOERT was done as radical full exposure for 86.9% of cases and as boost dose for 13.1% of cases, who needed to complete radiotherapy by external beam. One case in the group received boost dose and 4 cases in the group received full dose had recurrence. The median follow-up of patients was 31 months. Pathology of recurrence was reported as DCIS in 3 cases and invasive breast cancer in 2 of them.Conclusion: There is not a lot of data on the effectiveness of IOERT in DCIS management. Although there are not large number of cases in our study, the local recurrence (13.1%) was only event in our study with 31 months median follow up with no contralateral metastasis, distant metastasis, or death.

Prognostic role of immune infiltrates in breast ductal carcinoma in situ.

Ductal carcinoma in situ (DCIS) of the breast is often regarded as a non-obligate precursor to invasive breast carcinoma but current diagnostic tools are unable to accurately predict the invasive potential of DCIS. Infiltration of immune cells into the tumour and its microenvironment is often an early event at the site of tumourigenesis. These immune infiltrates may be potential predictive and/or prognostic biomarkers for DCIS. This review aims to discuss recent findings pertaining to the potential prognostic significance of immune infiltrates as well as their evaluation in DCIS. A literature search on PubMed was conducted up to 28th January 2019. Search terms used were "DCIS", "ductal carcinoma in situ", "immune", "immunology", "TIL", "TIL assessment", and "tumour-infiltrating lymphocyte". Search filters for "Most Recent" and "English" were applied. Information from published papers related to the research topic were synthesised and summarised for this review. Studies have revealed that immune infiltrates play a role in the biology and microenvironment of DCIS, as well as treatment response. There is currently no consensus on the evaluation of TILs in DCIS for clinical application. This review highlights the recent findings on the potential influence and prognostic value of immunological processes on DCIS progression, as well as the evaluation of TILs in DCIS. Further characterisation of the immune milieu of DCIS is recommended to better understand the immune response in DCIS progression and recurrence.

Abstract P4-08-17: Expression score (Escore) for the prediction of likelihood of recurrence of DCIS

Abstract Background: Ductal carcinoma in situ (DCIS) accounts for at least 20% of breast cancers. Factors associated with recurrence of DCIS or progression to invasive carcinoma are not well delineated. The goals of the current study were to profile the epithelial cells using the GE Cell DIVE™ immuno-fluorescent based analyses. This was coupled with semi-automated algorithms to characterize the inter-relationships between cell populations and likelihood of recurrence. Patients and Methods: A TMA-based (total 8 TMAs) cohort of cases of DCIS with and without recurrence was obtained from Oxford University. Recurrence in this cohort was defined as ipsilateral DCIS, ipsilateral invasive, contralateral invasive and metastatic. Analysis for 31 epithelial markers (HER4, CK56, ABCG2, PTEN, S6, CKAE1, PR, ER, NaKATPase, CK19, ALDH1, CK PCK26, cMET, CD44v6, HER2, CDCP1, p53, CK15, COX2, VEGFR2, ABCb1, HTF9C, CD10, MRP4, CEACAM5, EGFR, p21, MRP5, SLC7A5, Ki67, DAPI) was performed on a single FFPE TMA section containing cases of DCIS. Briefly, FFPE sections from TMAs containing DCIS were sequentially (cyclically) stained for the markers. Each cycle entailed staining with 2-3 markers followed by imaging, dye inactivation, and re-staining. DAPI was used for nuclear demarcation and for registration of the images, while S6, pan-cadherin, Na+K+ATPase and pan-cytokeratin were used for epithelial segmentation. K-means clustering followed regression analysis was performed to identify inter-relationships between markers and association with likelihood of recurrence. Log-rank analysis was performed and the relapse-free survival data depicted using Kaplan Meier plots. Escore was developed by logistic regression model, classification model on recurrence Results: Filtering of the expression analysis by the quality, specificity, compartment localization and fields entirely composed of DCIS, in addition to availability of clinical data resulted final analysis of 31 markers in 67 cases. Correlation analyses were performed on each of the markers to identify markers that were significantly correlated in univariate analysis. K-means cluster analysis was performed using a set of 4 markers (ER, HER2, SLC7A5 and cMET) to identify 6 clusters. High cMET (cluster 1; low HER2 and SLC7A5) and High ER (low cMET, HER2, SLC7A5; Cluster 5) were associated with low risk of recurrence (p values 0.014 and &amp;lt;0.0001). In contrast, Cluster 2 (High HER2, high SLC7A5, low ER) and Cluster 3 (High HER2, low ER, SLC7A5and cMET) were associated with increased risk of recurrence (P values 0.038 and 0.076). A regression analysis based algorithm was developed using these markers to calculate a numerical score which could predict likelihood of recurrence. As depicted in the KM plots, the HR for recurrence increases significantly (P-value 2.4E-05; p=0.02 with LOOCV) with increase in expression score (Escore). Conclusions: We describe the development of an Escore using expression 4 markers to predict likelihood of recurrence. Additional ongoing studies will seek to validate the utility of the Escore in predicting likelihood of recurrence of DCIS and development of invasive carcinomas and comparison with other scoring systems. Citation Format: Badve SS, Cho S, Gokmen-Polar Y, Zavodszky M, Sui Y, Chadwick C, Tan PH, Gerdes M, Harris AL, Ginty F. Expression score (Escore) for the prediction of likelihood of recurrence of DCIS [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-08-17.

Qualitative Radiogenomics: Association Between BI-RADS Calcification Descriptors and Recurrence Risk as Assessed by the Oncotype DX Ductal Carcinoma In Situ Score.

Treatment of ductal carcinoma in situ (DCIS) is controversial given the variable recurrence and progression to invasive carcinoma. Identifying women who would benefit from adjuvant radiation therapy on the basis of their recurrence risk may allow more individualized management strategies. The Oncotype DX Breast DCIS Score-which we refer to here as the "DCIS score"-is a validated surrogate marker of local recurrence. This study evaluated the association between BI-RADS mammographic calcification descriptors and the DCIS score. Fifty-eight women diagnosed with DCIS presenting with calcifications who had Oncotype DX Breast DCIS assay results were identified. Pretreatment BI-RADS mammographic calcification features were collected including morphology, distribution, and maximum span. The association between calcification descriptors and DCIS score was assessed with logistic regression modeling. Mean DCIS scores were calculated for calcification features significantly associated with DCIS score. All analyses were adjusted for patient age, DCIS grade, and progesterone receptor status. Of the suspicious calcifications that proved to represent DCIS, 19.0% were amorphous; 25.9%, coarse heterogeneous; 39.7%, fine pleomorphic; and 15.5%, fine linear or fine linear branching in morphology. The mean DCIS scores by calcification morphology were 22.3, 35.5, 36.7, and 44.1, respectively. Amorphous calcification morphology had a significantly lower adjusted mean DCIS score compared with fine pleomorphic morphology (p = 0.01) and fine linear or fine linear branching morphology (p = 0.02). The adjusted odds ratio (OR) of intermediate or high risk of recurrence (defined as a DCIS score ≥ 39) was significantly higher for women with fine pleomorphic calcifications (OR = 53.1, p = 0.01) and for those with fine linear or fine linear branching calcifications (OR = 24.0, p = 0.04) than for women with amorphous calcifications. Women with amorphous calcification morphology had the lowest DCIS scores compared with women with fine pleomorphic and fine linear or fine linear branching morphologies. Both fine pleomorphic and fine linear or fine linear branching morphologies were associated with higher odds of intermediate or high risk of recurrence. These findings suggest mammographic features are potential biomarkers of DCIS recurrence and could help individualize treatment decisions.

Endocrine Therapy for Ductal Carcinoma In Situ (DCIS) of the Breast with Breast Conserving Surgery (BCS) and Radiotherapy (RT): a Meta-Analysis.

The management of ductal carcinoma in situ (DCIS) with endocrine therapy remains controversial. A meta-analysis was conducted to evaluate the role of endocrine therapy for DCIS with breast conserving surgery (BCS) and radiotherapy (RT). A total of 7 articles with randomized controlled trials were included. Five articles compared the effects of BCS and RT followed by tamoxifen (TAM) or not (BCS + RT + TAM vs BCS + RT) and 2 compared the effects of TAM and anastrozole (ANA). TAM obviously reduced the rates of recurrence of ipsilateral breast cancer (IBCR), recurrence of contralateral breast cancer (CBCR), recurrence of ipsilateral invasive breast cancer (IBCR-INV) and recurrence of contralateral DCIS (CBCR-DCIS), and increased the rate of event-free survival (EFS). While ANA reduced the rates of CBCR and recurrence of contralateral invasive breast cancer (CBCR-INV). Patients with ANA had higher incidence of arthralgia, osteoporosis, hypercholesteremia, headache and vaginal dryness, but lower incidence of deep-vein thrombosis, pulmonary embolism, vasomotor or gynaecological, hot flushes, vaginal haemorrhage, vaginal discharge and vaginal candidiasis than TAM. In conclusion, DCIS patients with positive hormone receptors should be recommended to receive endocrine therapy. Selection of TAM or ANA is based on clinical characteristics and underlying disease of patients, as well as the side-effects of drugs.

Adjuvant radiation therapy in small ductal carcinoma in situ

BackgroundThe objective of this study was to evaluate ipsilateral breast tumor recurrence (IBTR) rates in patients with small (≤1 cm) ductal carcinoma in situ (DCIS) who were followed up for more than 15 years. MethodsWe identified 209 patients with primary small (≤1 cm) DCIS without invasion who received curative excision with and without adjuvant radiation therapy (RT) from 1996 to 2009. IBTR rates and prognostic factors in all patients were estimated by univariate and multivariate analyses. ResultsWith a median follow-up of 104 months, eight (53.3%) had DCIS recurrence and seven (46.7%) had recurrence of invasive ductal carcinoma. IBTR rate of all patients was 7.5% at 10 years and 12.1% at 15 years. In univariate analysis, age and subtypes were significant factors for IBTR. In multivariate analysis, resection margin, adjuvant RT, and endocrine therapy were significant factors for IBTR. ConclusionsIBTR rate of small (≤1 cm) DCIS following excision with or without adjuvant RT was 12.1% at 15 years. Adjuvant RT and endocrine therapy were associated with lower IBTR rate in small DCIS.

Management and 5-year outcomes in 9938 women with screen-detected ductal carcinoma in situ: the UK Sloane Project

BackgroundManagement of screen-detected ductal carcinoma in situ (DCIS) remains controversial. MethodsA prospective cohort of patients with DCIS diagnosed through the UK National Health Service Breast Screening Programme (1st April 2003 to 31st March 2012) was linked to national databases and case note review to analyse patterns of care, recurrence and mortality. ResultsScreen-detected DCIS in 9938 women, with mean age of 60 years (range 46–87), was treated by mastectomy (2931) or breast conserving surgery (BCS) (7007; 70%). At 64 months median follow-up, 697 (6.8%) had further DCIS or invasive breast cancer after BCS (7.8%) or mastectomy (4.5%) (p < 0.001). Breast radiotherapy (RT) after BCS (4363/7007; 62.3%) was associated with a 3.1% absolute reduction in ipsilateral recurrent DCIS or invasive breast cancer (no RT: 7.2% versus RT: 4.1% [p < 0.001]) and a 1.9% absolute reduction for ipsilateral invasive breast recurrence (no RT: 3.8% versus RT: 1.9% [p < 0.001]), independent of the excision margin width or size of DCIS. Women without RT after BCS had more ipsilateral breast recurrences (p < 0.001) when the radial excision margin was <2 mm. Adjuvant endocrine therapy (1208/9938; 12%) was associated with a reduction in any ipsilateral recurrence, whether RT was received (hazard ratio [HR] 0.57; 95% confidence interval [CI] 0.41–0.80) or not (HR 0.68; 95% CI 0.51–0.91) after BCS. Women who developed invasive breast recurrence had a worse survival than those with recurrent DCIS (p < 0.001). Among 321 (3.2%) who died, only 46 deaths were attributed to invasive breast cancer. ConclusionRecurrent DCIS or invasive cancer is uncommon after screen-detected DCIS. Both RT and endocrine therapy were associated with a reduction in further events but not with breast cancer mortality within 5 years of diagnosis. Further research to identify biomarkers of recurrence risk, particularly as invasive disease, is indicated.

Abstract 634: SOX11 is a potential clinical marker for hormone receptor negative ductal carcinoma in situ

Abstract Background: Ductal carcinoma in situ (DCIS) comprises 20-25% of screen-detected breast cancer and, like invasive ductal carcinoma (IDC), is heterogeneous in terms of underlying biology, presentation and outcome. There are limited potential biomarkers of outcome for DCIS, although estrogen receptor (ER)-positive (pos), progesterone receptor (PgR)-pos, and HER2-negative (neg) DCIS appears to have a better outlook than DCIS neg for ER and PgR but pos for HER2. The aim of this study was to identify markers for DCIS to develop a panel to stratify DCIS into DCIS at low or high risk of further DCIS. Methods and results: To identify genes driving DCIS evolution followed by the progression to IDC, we first used transcriptional data sets (GSE788, GSE16873) which had data from both normal mammary glands (NMG) and DCIS and performed class comparison (NMG vs DCIS). For both GSE7882 and GSE16883, the number of over- and under-expressed genes was 297 and 187. Over-expressed genes in DCIS represented a mitotic/proliferative feature annotated as mitotic spindle and condensed chromosome. Under-expressed genes represented loss of epithelial features annotated as epithelial cell differentiation and development. A total of 484 differentially expressed genes in DCIS were further correlated with recurrence events of IDC using Kessler's breast cancer data set to identify genes contributing aggressive feature across DCIS and IDC. Genes correlating to recurrence events were selected. From 484 genes, 99 were significantly associated with recurrence events of IDC (with P&amp;lt;0.003). Among these 99 genes, proliferation-related genes were depleted but component genes of the Oncotype DCIS score and genes reported as relevant to DCIS biology, such as SOX11, were included for Nanostring transcriptional analysis. The final number of genes-of-interest was 58 including 5 housekeeping genes. 40 DCIS and 8 NMG lesions were macro-dissected from formalin-fix paraffin-embedded blocks (FFPE) and extracted transcripts were subjected for Nanostring analysis. Gene expression data was clustered in an unsupervised manner using R software. Two sample clusters were identified: an ER/PgR-neg cluster and an ER/PgR-pos cluster. Over-expression of SOX11 and HER2 were exclusively seen in the ER/PgR-neg cluster within which samples were further subcategorized into HER2low/SOX11+ and HER2high/SOX11+. These RNA expression findings are undergoing confirmation by immunohistochemistry of FFPE sections. A further independent series of DCIS which has recurred as DCIS or IDC is currently under investigation for validation of an ER/PgR/HER2/SOX11 signature of poor prognosis DCIS. Conclusion: SOX11 is exclusively overexpressed in ER/PgR-neg DCIS and is a candidate clinical marker for recurrence of DCIS or progression of ER/PgR-neg DCIS to IDC. Citation Format: Kazuharu Kai, Wei Lu, Fei Yang, Ximing Tang, Ignacio I. Wistuba, Subrata Sen, Alastair Thompson. SOX11 is a potential clinical marker for hormone receptor negative ductal carcinoma in situ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 634.

Abstract PD7-03: Cost-effectiveness analysis of intraoperative radiotherapy for ductal carcinoma in situ

Abstract Background Whole breast radiation therapy (WBRT) following lumpectomy for ductal carcinoma in situ (DCIS) is standard of care, however, the risk of local recurrence with and without radiation ranges as low as 0.9% vs. 6.7% over 7 years. Intraoperative radiotherapy (IORT) is a potential alternative with advantages of decreased toxicity to adjacent organs, convenience, and improved quality of life. While prospective trials of IORT for DCIS are ongoing, the objective of this study was to estimate the cost-effectiveness of IORT vs. WBRT vs. no radiation for DCIS. Methods We developed a Markov model using TreeAge Pro 2016 to evaluate the cost-effectiveness of WBRT, IORT, and no radiation in patients with DCIS following lumpectomy. Health states included disease free, local recurrence (ipsilateral DCIS or invasive cancer), distant recurrence or death due to breast cancer, and death due to non-breast cancer causes. A 10-year time horizon and societal perspective were used. Model input parameters were derived from the literature. Costs reflected 2016 Medicare rates. The primary endpoint was incremental cost-effectiveness ratio (ICER), defined as the difference in cost, divided by the difference in quality-adjusted life years (QALYs) of two interventions. We performed analyses of subgroups defined according to DCIS risk (histologic grade, Oncotype Dx® DCIS recurrence score, low risk per RTOG 9804 criteria) and endocrine therapy use (none, tamoxifen, aromatase inhibitor). Sensitivity analyses explored uncertainty in the model. Results IORT was the most cost-effective strategy, with an increase of 0.18 QALYs at an incremental cost of $4,728, corresponding to an ICER of $26,943/QALY when compared with no radiation therapy. WBRT resulted in an increase in 0.18 QALYs at an incremental cost of $6859, corresponding to an ICER of $39,085/QALY. For both strategies, the ICERs did not exceed the willingness to pay (WTP) threshold of $100,000. IORT remained the most cost-effective strategy across DCIS risk groups, but was more cost-effective in higher risk patients, as demonstrated by lower ICERs. In low risk DCIS defined by RTOG 9804 criteria, no radiation was most cost-effective. The ICERs for IORT and WBRT, $152,753 and $208,204/QALY, respectively, exceeded the WTP threshold. IORT remained cost-effective in the setting of endocrine therapy use. Incremental Cost-Effectiveness Ratios (ICER) for each radiation strategy for the base case and scenario analyses ICER ($/QALY) No RTIORTWBRTBase Case Analysis 26,94339,085 Scenario Analysis by DCIS Risk GroupHistologic Grade - Low 36,81152,219- High 25,64337,137 Oncotype Dx DCIS Score - Low 92,892126,398- High 32,00345,690 Low Risk DCIS 152,753208,204 Scenario Analysis by Endocrine TherapyNo Tamoxifen 23,38734,373Tamoxifen 47,81166,616 Tamoxifen 31,96146,272Aromatase Inhibitor 41,31658,674 Conclusion IORT was the most cost-effective radiation strategy for DCIS compared to WBRT and no radiation. This applied to all subgroups with the exception of low-risk DCIS defined by RTOG 9804 criteria for whom no radiation was the most cost-effective strategy. These findings provide support for ongoing studies examining the role of IORT for DCIS with high-risk features, as well as alternative treatment strategies for low-risk DCIS. Citation Format: Onishi M, Connolly EP, Wright JD, Vasan S, Gross T, Tsai W-Y, Chen L, Neugut AI, Accordino MK, Kalinsky K, Crew KD, Hershman DL. Cost-effectiveness analysis of intraoperative radiotherapy for ductal carcinoma in situ [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr PD7-03.

Abstract P5-02-02: Second harmonic generation in combination with nuclear morphometry in the evaluation of DCIS

Abstract Purpose/Objective: Collagen is a major extracellular matrix (ECM) constituent in normal breast and is extensively remodeled in breast carcinoma. Therefore, features of remodeled collagen in the stroma adjacent to ductal carcinoma in situ (DCIS) could indicate cancer progression. The major objective of this study is to identify potential tumor-associated collagen signatures unique to DCIS that will allow us to predict progression based on the collagen texture and nuclear morphology. In this present study, we develop two image analysis pipelines (SHG Texture Extraction and H&amp;E Nuclear Morphology Extractor) to quantify 1) stromal changes, 2) collagen signatures and 3) nuclear morphology from normal breast to DCIS in order to predict local breast cancer recurrence. Method: We used second harmonic generation (SHG) images and H&amp;E to analyze collagen features and to study nuclear morphology using a data set of 336 patients (from which 310 normal and 327 DCIS regions were imaged). The 336 patients were a subset of patients with pure DCIS taken from a case-control study. Clinical-pathologic factors were associated with risk of subsequent ipsilateral cancer (DCIS or invasive). The SHG framework consisted of collagen segmentation using 1) adaptive thresholding and 2) morphological operations. The H&amp;E framework consisted of nuclear segmentation using adaptive thresholding and a maker-controlled watershed algorithm; and nuclear feature extractions including intensity, texture and morphology. Overall, the SHG framework segments collagen regions and computes textural features specifically at collagen regions. Furthermore, the H&amp;E framework segments nuclei and computes nuclei morphology and textural features. These features were used in L1-regularized logistic regression to construct classification models to discriminate normal vs DCIS regions; and to distinguish regions from DCIS patients with vs. without local recurrences. Results: In first experiment, we performed L1-regularized logistic regression to construct a classification model to discriminate normal vs DCIS regions. Our results suggest that using only SHG collagen features, this logistic model selected 19 significant features to build a classification model that achieved area under curve (AUC) 90% and accuracy 83% using 5-Fold cross validation. When H&amp;E nuclei features are used, the logistic model selected 88 significant features and achieved AUC 91% and accuracy 86%. By combined both SHG and H&amp;E features, the model achieved classification AUC 93% and accuracy 88%. By using L1-regularized logistic model with combined significant SHG and H&amp;E features, we achieved AUC 59% with an accuracy of 61% for DCIS and recurrent DCIS regions. Conclusions: Our study suggests that SHG and nuclear morphology features extracted from H&amp;E can improve the classification of normal and DCIS regions. Overall, these results suggest that second harmonic generation and H&amp;E nuclear morphology analysis could aid in the assessment of prognosis and risk of progression to invasive breast cancer. Citation Format: Park CC, Irshad H, Ziaee S, Martin-Tuite P, Habel L, Weaver VM, Schnitt SJ, Beck AH. Second harmonic generation in combination with nuclear morphometry in the evaluation of DCIS [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-02-02.