Abstract 1465: Analysis of tumor-infiltrating lymphocytes (TILs) reveals biologically different subgroups of breast ductal carcinoma in situ
Authors: Beguinot-Cornillon Marie , Dauplat Marie-Melanie , Kwiatkowski Fabrice , Lebouedec Guillaume , Tixier Lucie , Pomel Christophe , Penault-Llorca Frederique , Radosevic-Robin Nina 
Published: July 2016

Abstract
Background: Treatment of breast ductal carcinoma in situ (DCIS) aims to prevent invasive recurrence. Recent studies have shown an important impact of TILs on the outcome of invasive breast cancer, however their influence on breast DCIS prognosis has not been fully explored. In this study we investigated whether the amount and/or phenotype of TILs can help recognizing DCIS subgroups of different biology and recurrence risk.
Methods: The study included 134 patients, diagnosed and treated for a DCIS from 2001 to 2005 in our institution. Formalin-fixed and paraffin-embedded (FFPE) cancer tissue samples, taken before any treatment, were retrospectively collected. H&E whole tissue sections served for assessment of the cancer size, grade, histotype, architecture, mitotic index and amount of stromal lymphocytic (Ly) infiltrate. The latter was semi-quantitatively graded into 4 grades (0 - absent, 1 - mild, 2 - moderate, 3 - intense). Ly-phenotype was assessed by immunohistochemistry (IHC) on tissue microarrays (TMAs), constructed by sampling each case at the area of the densest Ly-infiltration (3 cores of 0.6 mm diameter per case). The number/mm2 of the CD8+, CD4+, FOXP3+, CD20+ and CD38+ mononuclear cells was determined. TMAs served also for estrogen receptor (ER), progesterone receptor (PR), HER2 and Ki67 IHC staining, as well as for HER2 amplification status.
Results: There were 97 DCIS and 37 microinvasive DCIS (micDCIS). The micDCIS displayed significantly more diffuse architecture, frequent HER2 amplification (HER2amp), higher grade, lower ER expression (0.029&lt;p&lt;0.044 for each) and more peritumoral Ly-infiltrate (grades 2+3, micDCIS vs DCIS, 51.5% vs. 39.0%, p = 0.036). All but CD20+ cells were more numerous in micDCIS than in DCIS (0.0016&lt;p&lt;0.05). Within the entire cohort, the cases having the (CD8+/CD4+):(CD20+/CD38+) ratio higher than 1 had a significantly greater risk of containing a microinvasive component (OR 3.47 (1.26-9.57), p = 0.029). Interestingly, that ratio was significantly higher (p = 0.012) in micDCIS than in the DCIS with grade 2 or 3 Ly-infiltrate (Ly-DCIS, n = 38). On the other side, there was no difference between micDCIS and Ly-DCIS in architecture, histograde, HER2amp rate and ER expression. Cluster analysis further confirmed significant similarities between micDCIS and Ly-DCIS, putting them both apart from non-Ly-DCIS (p = 0.0034).
The overall 10-year recurrence rate was 13% (18/134 pts). No parameter significantly correlated with recurrence risk, however the micDCIS have received significantly more treatment than DCIS (axillary lymphadenectomy (p&lt;10-7), chemotherapy (p = 0.031) or hormonal therapy (p = 5.4×10-6)).
Conclusion: These results indicate that Ly-DCIS might be biologically and immunologically similar to micDCIS. TILs in DCIS are worth investigating in larger studies, as they could be a marker of microinvasion and help tailoring the initial treatment of the disease.
Citation Format: Marie Beguinot-Cornillon, Marie-Melanie Dauplat, Fabrice Kwiatkowski, Guillaume Lebouedec, Lucie Tixier, Christophe Pomel, Frederique Penault-Llorca, Nina Radosevic-Robin. Analysis of tumor-infiltrating lymphocytes (TILs) reveals biologically different subgroups of breast ductal carcinoma in situ. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1465.