Background: The NRG Oncology/NSABP B-52 neoadjuvant clinical trial was conducted to test if the addition of estrogen deprivation (ED) would improve the pCR rate in HER2+/ER+ breast cancer patients (pts) treated with docetaxel, carboplatin, trastuzumab, and pertuzumab (TCHP). A numerical increase in pCR rate was observed with ED (46.1% v 40.9%), but the difference was not statistically significant. We have previously quantitated T cells (CD8, FOXP3), macrophages (CD68), and immune checkpoint proteins (PD-1, PD-L1) with multiplex immunofluorescence in B-52 and shown that CD68 and FOXP3 cells were associated with pCR but not CD8 cells. Our purpose was to determine the associations of FCGR genotypes and immune cells with pCR. Methods: A single baseline, pre-treatment FFFPE tissue section per case (N=181) was used to perform a 7-plex multiplex immunofluorescence procedure using opal fluorophores for staining. The Vectra Pathology System and inForm analysis software (Akoya Biosciences) was used for imaging and quantitation of CD8, CD68, FOXP3, PD-1, and PD-L1 cells in both the tumoral and stromal regions. Stromal data is reported here. Favorable- and unfavorable- FcGγR genotypes for FCGR2A-131H/R and FCGR3A-158V/F alleles were determined via the Sequenom MassARRAY iPLEX platform. Rates of pCR with pts with 1 or 2 favorable alleles was compared to pts who were homozygous for the unfavorable allele. Within each genotype, Wilcoxon rank sum test was used to test the association of markers with pCR and within each treatment. Results: No significant association of FCGR2A and 3A alleles with pCR was detected in the entire B-52 cohort, however, among pts with favorable FCGR genotypes (FCGR2A-131-HH, or H/R, FCGR3A-158- VV, or VF HR) the median value of the % CD68 cells was significantly higher in tumors that achieved pCR v those that did not (p=0.0004, p=0.0006), respectively. In pts who were homozygous for the FCGR2A or FCGR3A unfavorable alleles, there was no significant difference in the median values of the % of CD68 cells between pCR and no-pCR tumors. Further stratification of tumors by treatment showed that pts with an FCGR2A or FCGR3A favorable genotype and whose tumors achieved pCR had a higher median value of CD68 only in the TCHP + ED arm (p=0.0007, p=0.0003), respectively and not in the TCHP arm (p=0.059; p=0.21). Higher levels of PD-L1 were associated with pCR in pts with FCGR3A- favorable genotypes, but higher levels of FOXP3 were associated with pCR regardless of genotype. In contrast to the other cell types, higher PD-1 or CD8 cells showed no association with genotypes. Conclusions: This is an exploratory study examining the potential role of ADCP in HER2+/ER+ breast cancer and supports the notion that ADCP may be one mechanism that promotes the elimination of tumor cells in a subset of pts in the neoadjuvant setting. Tumors that achieve pCR have higher % of CD68 cells, in pts with favorable FCGR2A and 3A genotypes than pts who do not. However, in pts with unfavorable FCGR3A or FCGR2A genotypes there was no difference in the median CD68 levels in pCR v no-pCR tumors. When tumors were further stratified by CD68 levels, FCGR3A genotypes, and treatment, the association of pCR in tumors with high CD68 and FCGR3A favorable genotypes was seen only in the TCHP+ED arm. This may indicate that ED may improve pCR rates in some tumors with more macrophages and favorable genotypes. Macrophages are known to have estrogen receptors, and estrogen has been shown to promote the alternative activation of macrophages, potentially dampening down the immune response. Thus, one could speculate that ED may block the estrogen-induced alternative activation of macrophages, allowing the classically activated macrophages to phagocytize tumor cells. Support: BCRF, U10CA180868 & Admin Sup, U24CA196067, Genentech, NSABP Foun.

Citation Format: Katherine L Pogue-Geile, Ying Wang, Huichen Feng, Corey Lipchick, Patrick Gavin, Rim S Kim, Reena S Cecchini, Samuel A Jacobs, Ashok Srinivasan, Sandra M Swain, Eleftherios Mamounas, Charles E Geyer, Jr, Priya Rastogi, Peter C Lucas, C. Kent Osborne, Soonmyung Paik, Norman Wolmark, Mothaffar F Rimawi. Potential role of the antibody-dependent cellular phagocytosis (ADCP) in tumors achieving pCR in NRG Oncology/NSABP B-52 [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-07-04.