Differential efatutazone's impact on mammary neoplasia dependent upon Brca1 dose

in Endocrine-Related Cancer
Correspondence should be addressed to P A Furth: paf3@georgetown.edu
Restricted access

 

      Society for Endocrinology

Related Articles

Article Information

Metrics

All Time Past Year Past 30 Days
Abstract Views 316 316 13
Full Text Views 887 887 1
PDF Downloads 32 32 0

Altmetrics

Figures

  • View in gallery

    Efatutazone exposure starting at age 2 months significantly reduced mammary preneoplasia. (A) Bar graphs comparing percentages of Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/mice demonstrating tertiary branching on fourth (inguinal) mammary gland whole mounts (WMs) following exposure to efatutazone or control diet. *P < 0.05, Fisher’s exact test, one-sided. WM number (n) analyzed, age (months (m), mean ± s.e.m.) and weight (grams (g), mean ± s.e.m.) for each cohort: Brca1WT/fl11/Cre/p53+/ Efatutazone (E): n = 12 (9.0 ± 0.7 m, 38.0 ± 1.9 g). Control (C): n = 12 (10.1 ± 0.5 m, 36.9 ± 2.4 g). Brca1fl11/fl11/Cre/p53+/ E: n = 12 (8.4 ± 0.3 m, 40.3 ± 2.2 g). C: n = 11 (8.9 ± 0.3 m, 40.6 ± 2.4 g). WMs prepared at the time of necropsy for experimental endpoints designated in IACUC-approved protocol. All available fourth gland WMs analyzed. Mean ages and weights at necropsy were not statistically significantly different between cohorts. (B) Kaplan–Meier survival plot of all Brca1WT/fl11/Cre/p53+/ mice on efatutazone (green) and control (blue) and Brca1fl11/fl11/Cre/p53+/mice on efatutazone (red) and control (black) diets enrolled in study. *Log-rank test for trend chi-Square P = 0.0005 between both Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/ cohorts. Cohort numbers (n), median age (m) from survival plot: Brca1WT/fl11/Cre/p53+/ E: n = 17 (12.1 m). C: n = 15 (12.1 m). Brca1fl11/fl11/Cre/p53+/ E: n = 17 (9.1 m). C: n = 19 (9.2). Censored points for death due to non-palpable-tumor-related health reasons prior to age 12 months indicated by vertical ticks (skin ulcer, rectal prolapse, leg paralysis, shaking, lymphoma, found dead). Median survival was not statistically significant between efatutazone and control cohorts for either genotype. (C) Representative phase-contrast images of mammospheres (indicated by black arrows) that formed from primary mammary epithelial cells isolated from Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/ on E and C diets. Images taken at 4×. Scale bar = 0.1 mm. (D) Box and whisker plots (mean and range) illustrating a reduction in mammosphere numbers forming from primary mammary epithelial cells isolated from efatutazone-exposed Brca1fl11/fl11/Cre/p53+/ mice as compared to Brca1fl11/fl11/Cre/p53+/ mice on control diet. bP < 0.05 Mann-Whitney U test, two-tailed. Twelve wells plated and analyzed per mouse. Cohort numbers (n), mean age (m) and weight (g): Brca1WT/fl11/Cre/p53+/ E: n = 7 (4.2 ± 0.0 m, 31.5 ± 1.5 g). C: n = 6 (3.9 ± 0.1 m, 34.6 ± 1.8 g). Brca1fl11/fl11/Cre/p53+/ E: n = 4 (3.9 ± 0.1 m, 32.3 ± 1.7 g). C: n = 5 (4.2 ± 0.0 m, 37.3 ± 2.9 g). Mammosphere numbers forming from Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/ mice on control diet were significantly lower than numbers forming from MMTV-Cre (Cre)c and wild-type (WT)a,d mice on standard facility diet. P < 0.05 Kruskal–Wallis Test, Dunn’s multiple comparison. MMTV-Cre: n = 3 (4.2 ± 0.0 m, 31.2 ± 5.1 g). WT: n = 3 (5.2 ± 0.0 m, 25.6 ± 0.6 g). (E) Box and whisker plots (mean and range) illustrating that mammospheres derived from efatutazone-exposed Brca1fl11/fl11/Cre/p53+/ were significantly smaller than those obtained from efatutazone-exposed Brca1WT/fl11/Cre/p53+/ mice. aP < 0.05 Kruskal–-Wallis Test, Dunn’s multiple comparison. Wells and cohorts analyzed were the same as those analyzed for mammosphere numbers (Panel D). (F) Scatter plots (mean ± s.e.m. indicated) comparing numbers of HANs per WM from Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/mice following exposure to efatutazone or control diet. *P < 0.05, Mann–Whitney U test, one-tailed. WMs and cohorts analyzed were the same as those analyzed for tertiary branching (Panel A). (G) Bar graphs comparing percentages of Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/mice demonstrating HANs on WM following exposure to efatutazone or control diet. *P < 0.05, Fisher’s exact test, one-sided. WMs and cohorts analyzed were the same as those analyzed for tertiary branching (Panel A).

  • View in gallery

    Impact of efatutazone exposure on mammary cancer development in Brca1WT/fl11/Cre/p53+/and Brca1fl11/fl11/Cre/p53+/ mice. (A) Stacked bar graphs comparing numbers of mice euthanized for tumor burden with lipoma alone, lipoma and mammary cancer, or mammary cancer alone. *P < 0.05, 2 × 3 Fisher’s exact. Number (n) of mice and age (m, mean ± s.e.m.) for each cohort: Brca1WT/fl11/Cre/p53+/ E: n = 8 (9.8 ± 0.7 m). C: n = 7 (10.4 ± 0.7 m). Brca1fl11/fl11/Cre/p53+/ E: n = 13 (8.5 ± 0.4 m). C: n = 10. (9.0 ± 0.4). (B) Stacked bar graphs comparing distribution of mammary cancer histology identified in the palpable mammary cancers. Number (n) of mice and age (m, mean ± s.e.m.) for each cohort: Brca1WT/fl11/Cre/p53+/ E: n = 3 (7.7 ± 0.7 m). C: n = 7 (10.4 ± 0.7 m). Brca1fl11/fl11/Cre/p53+/ E: n = 11 (8.6 ± 0.4 m). C: n = 10. (9.0 ± 0.4). Representative hematoxylin and eosin images of triple negative mammary adenocarcinoma (C), anaplastic carcinoma (D and F), sarcomatoid carcinoma (E and G) histology from palpable mammary cancers that developed in Brca1WT/fl11/Cre/p53+/ and Brca1fl11/fl11/Cre/p53+/ mice on efatutazone or control diet. Insets to right show representative images of ER (top), PR (middle) and HER2 (bottom) IHC. (H and I) Representative images of the histology and ER, PR and HER2 IHC of the two palpable ER+/PR+ mammary adenocarcinomas that developed in Brca1fl11/fl11/Cre/p53+/ mice on efatutazone diet. Arrows indicate representative mammary cancer cells demonstrating nuclear-localized ER and PR staining. Inset IHC images taken at 40× (C, D, E, F and G). Larger images taken at 20× (C, D, E, F, G, H and I). Scale bar = 0.1 mm.

References

  • AlothmanSJ 2018 Studies of Breast Cancer Chemoprevention in Human High-Risk Primary Cells and Genetically Engineered Mouse Models. Washington, DC, USA: Georgetown University. (available at: http://hdl.handle.net/10822/1050904)

    • Search Google Scholar
    • Export Citation
  • AlothmanSJWangWGoerlitzDSIslamMZhongXKishoreAAzharRIKallakuryBVFurthPA 2017 Responsiveness of Brca1 and Trp53 deficiency-induced mammary preneoplasia to selective estrogen modulators versus an aromatase inhibitor in Mus musculus. Cancer Prevention Research 10 244254. (https://doi.org/10.1158/1940-6207.CAPR-16-0268)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • AraiKEguchiTRahmanMMSakamotoRMasudaNNakatsuraTCalderwoodSKKozakiK-IItohM 2016 A novel high-throughput 3D screening system for EMT inhibitors: a pilot screening discovered the EMT inhibitory activity of CDK2 inhibitor SU9516. PLoS ONE 11 e0162394. (https://doi.org/10.1371/journal.pone.0162394)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • DongJT 2013 Anticancer activities of PPARγ in breast cancer are context-dependent. American Journal of Pathology 182 19721975. (https://doi.org/10.1016/j.ajpath.2013.03.005)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • HeubleinSMayrDMeindlAKircherAJeschkeUDitschN 2017 Vitamin D receptor, retinoid X receptor and peroxisome proliferator-activated receptor γ are overexpressed in BRCA1 mutated breast cancer and predict prognosis. Journal of Experimental and Clinical Cancer Research 36 57. (https://doi.org/10.1186/s13046-017-0517-1)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • KoppenAKalkhovenE 2010 Brown vs white adipocytes: the PPARgamma coregulator story. FEBS Letters 584 32503259. (https://doi.org/10.1016/j.febslet.2010.06.035)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • NaklesREKallakuryBVFurthPA 2013 The PPARgamma agonist efatutazone increases the spectrum of well-differentiated mammary cancer subtypes initiated by loss of full-length BRCA1 in association with TP53 haploinsufficiency. American Journal of Pathology 182 19761985. (https://doi.org/10.1016/j.ajpath.2013.02.006)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • OryVKeitzmanWBoekelmanJKallakuryBWellsteinAFurthPARiegelAT 2018 The PPARγ agonist efatutazone delays invasive progression and induces differentiation of ductal carcinoma in situ (DCIS). Breast Cancer Research and Treatment 169 4757. (https://doi.org/10.1007/s10549-017-4649-y)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • RoyRChunJPowellSN 2011 BRCA1 and BRCA2: different roles in a common pathway of genome protection. Nature Reviews Cancer 12 6878. (https://doi.org/10.1038/nrc3181)

    • Search Google Scholar
    • Export Citation
  • SubbaramaiahKHoweLRZhouXKYangPHudisCALevy KopelovichLDannenbergAJ 2012 Pioglitazone, a PPARg agonist, suppresses CYP19 transcription: evidence for involvement of 15-hydroxyprostaglandin dehydrogenase and BRCA1. Cancer Prevention Research 5 11831194. (https://doi.org/10.1158/1940-6207.CAPR-12-0201)

    • Crossref
    • Search Google Scholar
    • Export Citation

PubMed

Google Scholar