Oral Presentation ESA-SRB-APEG-NZSE 2022

PUMA blockade protects oocytes from chemotherapy-induced damage (#218)

Lauren R Alesi 1 , Roseanne Rosario 2 , Amy L Winship 1 , Jessica M Stringer 1 , Richard A Anderson 2 , Karla J Hutt 1
  1. Ovarian Biology Laboratory, Monash Biomedicine Discovery Institute, Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia
  2. MRC Centre for Reproductive Health, Queen’s Medical Research Institute, University of Edinburgh, Edinburgh, United Kingdom

Irreversible ovarian damage and depletion of oocytes are devastating side effects of many cancer treatments; often leaving female cancer survivors infertile and at risk of premature menopause. Unfortunately, current fertility preservation options have significant drawbacks, with no strategies available to protect both fertility and long-term endocrine function in young girls and women receiving cancer treatment. PUMA, an apoptotic protein, triggers oocyte death following exposure to DNA-damaging insults, like chemotherapy. In fact, genetic loss of PUMA preserves fertility post-chemotherapy without compromising offspring health. Excitingly, a small molecule PUMA inhibitor (PUMAi) has recently become available, making PUMA blockade for fertility preservation a real therapeutic possibility for the first time.

To assess whether PUMA blockade can prevent oocyte apoptosis post-chemotherapy, postnatal day 5 C57BL6/J ovaries were cultured ex vivo for 5 days in media containing the cyclophosphamide metabolite 4-HC (2µM) ± PUMAi (200µM). Whilst 4-HC alone significantly depleted primordial follicles by 85% (p<0.01), primordial follicle numbers in ovaries from the 4-HC+PUMAi group were not significantly different from controls. Next, adult mice were administered 10mg/kg PUMAi 2 hours before and 20 hours after 150mg/kg cyclophosphamide. This regimen was based on a previous study in which PUMAi prevented intestinal stem cell apoptosis post-chemotherapy. Remarkably, primordial follicle numbers were approximately doubled in the cyclophosphamide + PUMAi group versus cyclophosphamide alone (858±122 vs. 386±106, p<0.05). This is extremely promising, as genetic knockout models of PUMA prove that partial protection of just 12% of follicles sustains female fertility.

Collectively, these data demonstrate that PUMA blockade is a promising avenue for fertility preservation prior to chemotherapy treatment. Further studies are already underway to optimise the PUMAi regimen for maximal ovarian protection; ensure that PUMAi does not impact the anti-tumour efficacy of chemotherapy treatment; and to determine whether PUMAi can prevent primordial follicle apoptosis in vitro in human ovarian tissue.