Radiotherapy exposure directly damages the uterus and causes pregnancy loss
Meaghan J. Griffiths, … , Amy L. Winship, Karla J. Hutt
Meaghan J. Griffiths, … , Amy L. Winship, Karla J. Hutt
Published March 22, 2023
Citation Information: JCI Insight. 2023;8(6):e163704. https://doi.org/10.1172/jci.insight.163704.
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Research Article Reproductive biology Vascular biology

Radiotherapy exposure directly damages the uterus and causes pregnancy loss

Abstract

Female cancer survivors are significantly more likely to experience infertility than the general population. It is well established that chemotherapy and radiotherapy can damage the ovary and compromise fertility, yet the ability of cancer treatments to induce uterine damage, and the underlying mechanisms, have been understudied. Here, we show that in mice total-body γ-irradiation (TBI) induced extensive DNA damage and apoptosis in uterine cells. We then transferred healthy donor embryos into ovariectomized adolescent female mice that were previously exposed to TBI to study the impacts of radiotherapy on the uterus independent from effects to ovarian endocrine function. Following TBI, embryo attachment and implantation were unaffected, but fetal resorption was evident at midgestation in 100% of dams, suggesting failed placental development. Consistent with this hypothesis, TBI impaired the decidual response in mice and primary human endometrial stromal cells. TBI also caused uterine artery endothelial dysfunction, likely preventing adequate blood vessel remodeling in early pregnancy. Notably, when pro-apoptotic protein Puma-deficient (Puma–/–) mice were exposed to TBI, apoptosis within the uterus was prevented, and decidualization, vascular function, and pregnancy were restored, identifying PUMA-mediated apoptosis as a key mechanism. Collectively, these data show that TBI damages the uterus and compromises pregnancy success, suggesting that optimal fertility preservation during radiotherapy may require protection of both the ovaries and uterus. In this regard, inhibition of PUMA may represent a potential fertility preservation strategy.

Authors

Meaghan J. Griffiths, Sarah A. Marshall, Fiona L. Cousins, Lauren R. Alesi, Jordan Higgins, Saranya Giridharan, Urooza C. Sarma, Ellen Menkhorst, Wei Zhou, Alison S. Care, Jacqueline F. Donoghue, Sarah J. Holdsworth-Carson, Peter A.W. Rogers, Evdokia Dimitriadis, Caroline E. Gargett, Sarah A. Robertson, Amy L. Winship, Karla J. Hutt

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Figure 6

Loss of PUMA protects against radiotherapy-mediated uterine damage.

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Loss of PUMA protects against radiotherapy-mediated uterine damage. Adol...
Adolescent female Puma+/– or Puma–/– mice were exposed to 7 Gy TBI or left as nonirradiated controls (Non-IRR). (A) Uterine Puma mRNA expression was localized by in situ hybridization (bars = 50 μm) and (B) quantified by qPCR 24 hours postirradiation. (C) Representative images of immunofluorescence and TUNEL-stained uterine sections from wild-type or Puma–/– mice at 24 hours postirradiation. (D) Ovariectomized (OVX) adolescent female Puma–/– mice exposed to 7 Gy TBI or nonirradiated controls were hormone-primed with estradiol (E2) and a progesterone pellet (P4) before undergoing artificial decidualization. Representative images of uteri collected 4 days after artificial decidualization are shown. (E) Uterine weight (UW) to body weight (BW) was quantified. (F) Female Puma+/– or Puma–/– mice were exposed to 7 Gy TBI or left as nonirradiated controls, then 4 weeks postirradiation had uterine artery function assessed by wire myography. Vessel relaxation (F and G) and vasoconstriction (H–K) were assessed by area under the curve (AUC). Ovariectomized adolescent female Puma+/– or Puma–/– mice were hormone-primed with estradiol and progesterone and received healthy donor embryo transfers 4 weeks postirradiation. The number of viable (L) and resorbing (M) implantation sites were quantified. Scale bars are 50 μm (A and C) and 5 mm (D); → luminal epithelium; S stroma; * endothelium. Data are mean ± SEM; unpaired t test (2 groups; parametric distribution) or Mann-Whitney test (2 groups; nonparametric distribution), *P < 0.05, **P < 0.01, ***P < 0.001. n = 3–8/group.