Early dietary exposures epigenetically program mammary cancer susceptibility through Igf1-mediated expansion of the mammary stem cell compartment

This article belongs to the Collection Insulin-Like Growth Factors in Development, Cancers and Aging.Diet is a critical environmental factor affecting breast cancer risk, and recent evidence shows that dietary exposures during early development can affect lifetime mammary cancer susceptibility. To elucidate the underlying mechanisms, we used our established crossover feeding mouse model, where exposure to a high-fat and high-sugar (HFHS) diet during defined developmental windows determines mammary tumor incidence and latency in carcinogen-treated mice. Mammary tumor incidence is significantly increased in mice receiving a HFHS post-weaning diet (high-tumor mice, HT) compared to those receiving a HFHS diet during gestation (low-tumor mice, LT). The current study revealed that the mammary stem cell (MaSC) population was significantly increased in mammary glands from HT compared to LT mice. Igf1 expression was increased in mammary stromal cells from HT mice, where it promoted MaSC self-renewal. The increased Igf1 expression was induced by DNA hypomethylation of the Igf1 Pr1 promoter, mediated by a decrease in Dnmt3b levels. Mammary tissues from HT mice also had reduced levels of Igfbp5, leading to increased bioavailability of tissue Igf1. This study provides novel insights into how early dietary exposures program mammary cancer risk, demonstrating that effective dietary intervention can reduce mammary cancer incidenceThe research was supported by institutional funding from Texas A&M University and the Discovery Foundatio

Dynamic role of the codon 72 p53 single-nucleotide polymorphism in mammary tumorigenesis in a humanized mouse model

Female breast cancer (BrCa) is the most common noncutaneous cancer among women in the United States. Human epidemiological studies reveal that a p53 single-nucleotide polymorphism (SNP) at codon 72, encoding proline (P72) or arginine (R72), is associated with differential risk of several cancers, including BrCa. However, the molecular mechanisms by which these variants affect mammary tumorigenesis remain unresolved. To investigate the effects of this polymorphism on susceptibility to mammary cancer, we used a humanized p53 mouse model, homozygous for either P72 or R72. Our studies revealed that R72 mice had a significantly higher mammary tumor incidence and reduced latency in both DMBAinduced and MMTV-Erbb2/Neu mouse mammary tumor models compared to P72 mice. Analyses showed that susceptible mammary glands from E-R72 (R72 x MMTV-Erbb2/Neu) mice developed a senescence-associated secretory phenotype (SASP) with influx of proinflammatory macrophages, ultimately resulting in chronic, protumorigenic inflammation. Mammary tumors arising in E-R72 mice also had an increased influx of tumor-associated macrophages, contributing to angiogenesis and elevated tumor growth rates. These results demonstrate that the p53 R72 variant increased susceptibility to mammary tumorigenesis through chronic inflammation.This work was funded by the National Institutes of Health grant R01MD006228 (to RF-Y)

The Effect of 7,12-dimethylbenz[a]-anthracene (DMBA) on Physical Activity in Female Mice

BACKGROUND: Regular exercise has been shown to reduce the risk of occurrence for certain cancers. In animal models, DMBA is a synthetic carcinogen that has been established as the gold standard for inducing cancerous tumors in rodents. However, it has yet to be established whether DMBA has an effect on voluntary wheel running in mice. If there is an effect, it would confound any experiment which investigates exercise effects on tumor growth. PURPOSE: The overall purpose of this project was to determine if DMBA altered voluntary wheel running in mice. METHODS: All procedures were approved by TAMU IACUC. SENCAR mice breeder pairs (Charles River) and offspring at 3 weeks of age were group housed and randomly assigned to a group receiving the DMBA (n=69) or not receiving the DMBA treatment (n=22). At 4 weeks of age, two running wheels were placed inside the cages and connected to a computer that measured distance and time. The running wheels were mounted to the cage tops of standard rat cages and equipped with a cycling computer (BC8.12, Sigma Sport) to record running distance and duration. The running wheels were plastic and had a 410mm circumference with a solid running surface. From 8 to 14 weeks of age, mice in the DMBA group were gavaged daily with a DMBA dose (20 µg/mouse) dissolved in corn oil. A two way ANOVA was employed to determine the effect of DMBA on activity with factors of time and treatment. RESULTS: DMBA had no effect on the distance (p=0.51) or duration ran (p=0.12), but significantly decreased the speed at which the mice ran (p=0.02). A post-hoc analysis indicated that significant decreases in speed occurred at weeks 12 (35.2 ±9.2 vs. 46.4 ± 14.6; p=0.0002) and 20 (35.4 ±10.3 vs. 46.2 ± 14.1; p\u3c0.0001) of age. CONCLUSION: Our data suggest that DMBA does not affect the distance or time spent running on a wheel, but does affect the speed at which the mice run. While DMBA decreased speed, the significant effects on speed are minor given that neither distance nor duration were different between the groups. Therefore, we can conclude that DMBA does not prevent voluntary wheel running in mice

Paracrine Overexpression of Insulin-Like Growth Factor-1 Enhances Mammary Tumorigenesis in Vivo

Insulin-like growth factor-1 (IGF-1) stimulates proliferation, regulates tissue development, protects against apoptosis, and promotes the malignant phenotype in the breast and other organs. Some epidemiological studies have linked high circulating levels of IGF-1 with an increased risk of breast cancer. To study the role of IGF-1 in mammary tumorigenesis in vivo, we used transgenic mice in which overexpression of IGF-1 is under the control of the bovine keratin 5 (BK5) promoter and is directed to either the myoepithelial or basal cells in a variety of organs, including the mammary gland. This model closely recapitulates the paracrine exposure of breast epithelium to stromal IGF-1 seen in women. Histologically, mammary glands from transgenic mice were hyperplastic and highly vascularized. Mammary glands from prepubertal transgenic mice had significantly increased ductal proliferation compared with wild-type tissues, although this difference was not maintained after puberty. Transgenic mice also had increased susceptibility to mammary carcinogenesis, and 74% of the BK5.IGF-1 mice treated with 7,12-dimethylbenz[a]anthracene (20 μg/day) developed mammary tumors compared with 29% of the wild-type mice. Interestingly, 31% of the vehicle-treated BK5.IGF-1 animals, but none of the wild-type animals, spontaneously developed mammary cancer. The mammary tumors were moderately differentiated adenocarcinomas that expressed functional, nuclear estrogen receptor at both the protein and mRNA levels. These data support the hypothesis that tissue overexpression of IGF-1 stimulates mammary tumorigenesis