Abstract P5-07-11: Mammary gland specific Stat3 deletion during involution results in distinct histological changes and higher collagen deposition

Abstract

Abstract Rationale: African American women (AAW) have a higher incidence of triple negative breast cancer (TNBC), and higher mortality from breast cancer when compared to Caucasian women (CW). Epidemiological studies have shown that lack, or short duration, of breast-feeding, a more common practice among AAW than CW, may be one of the factors that increase the risk of TNBC. The pregnancy-lactation-involution cycle is a dynamic process where upon pregnancy, breast undergoes extensive proliferation and differentiation for milk production followed by apoptosis as it involutes to near pre-pregnant state. Prolonged breastfeeding results in gradual involution (GI) whereas no or short duration of breastfeeding leads to forced and abrupt involution (AI) of breast. We modeled GI and AI in wild-type FVB/N mice and reported that mammary glands of abruptly involuted mice had expansion of luminal progenitor (LP) cells over time and hyperplastic precancerous changes within 120 days postpartum1. In addition, there was increased Stat3 (Signal Transducer and Activator of Transcription 3) activation (pStat3Y705) in AI glands. Persistent Stat3 activation is linked to tumor cell proliferation, survival, invasion, and tumor-promoting inflammation2. In mice, Stat3 is required for the initiation and acute phase response during mammary gland involution and is crucial for the proliferation of LP cells3-5. We hypothesized that Stat3 activation plays a key role in the development of hyperplasia and fibrosis observed following AI.Methods: We bred Stat3Fl/Flmice with MMTV-Cre mice to induce mammary epithelial specific deletion of Stat3. Experimental (MMTV-Cre+;Stat3Fl/Fl) and control females (Stat3Fl/Fl) were bred once at 8 weeks of age and litter sizes were normalized to 6pups/dam within 24 hours of partum. With day of partum as day0, all 6 pups were removed on day7 and AI was initiated. Mammary glands were harvested on postpartum day28, day56, and day120. Florescence activated cell sorting (FACS) was performed to determine distribution of mammary epithelial subpopulations. Histological effects of Stat3 deletion on mammary gland involution was assessed via staining using haematoxylin and eosin (H&E), Masson’s Trichrome and immunostaining using anti-Ki67 and anti-Stat3 antibodies. Results: Stat3 deletion failed to abrogate the hyperplasia and collagen deposition we initially observed in our AI mice with intact Stat3 as determined by H&E and Trichrome stains. In addition, collagen deposition was further increased following Stat3 deletion, when compared to control cohort. FACS analysis demonstrated that Stat3 deletion resulted in reduction of LP cell population only at early timepoint (day28) but this population rebounded in glands by later timepoints. Stat3 deletion temporarily increased cell proliferation by Ki-67 staining at day28 and 56, with no difference observed between controls at day120. Conclusion: This is the first study showing results of mammary epithelial specific Stat3 deletion on long-term effect of AI. Contrary to our hypothesis, Stat3 deletion did not reduce the hyperplastic changes we observed following abrupt involution of mammary glands. Our findings indicate that Stat3 signaling may limit collagen formation within the mammary gland during abrupt involution. Stat3 is important for the maintenance of LP cells at early timepoints. Further studies are underway to determine the critical downstream events following Stat3 activation that limits long-term effects of abrupt involution. 1. Basree et. al. 2019, in press, Breast Cancer Research 2. Yu et. al. 2014 (PMID: 25342631) 3. Chapman et. al. 1999 (PMID: 10521404) 4. Hughes et. al. 2012 (PMID: 22081431)5. Staniszewska et. al. 2012 (PMID: 23285109) Citation Format: Allen Zhang, Neelam Shinde, Christopher S Koivisto, Mustafa M Basree, Resham S Mawalkar, Hee K Kim, Gustavo Leone, Sarmila Majumder, Bhuvaneswari Ramaswamy. Mammary gland specific Stat3 deletion during involution results in distinct histological changes and higher collagen deposition [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 P5-07-11.