Interrogating the Hippo/YAP signaling pathway to preserve fertility in cancer survivors
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Citation
Abstract
The advancement in cancer treatments has led to significantly improved survival rates, but fertility, an essential component of the life quality of cancer survivors, deteriorated or was even damaged. There are over 1.27 million cancer survivors in the U.S., and over 51% of women are concerned about becoming infertile upon cancer treatment. Fertility preservation for cancer survivors becomes an evident priority for researchers and clinicians globally. The objective of this study was to understand the relationship between Hippo/YAP signaling and fertility in the context of cancer. This was accomplished through cell culture studies, immunohistochemical staining (IHC), immunofluorescence (IF), and DNA fragmentation assays (TUNEL) to assess the impact of chemotherapy on follicle health and to investigate the role of YAP in this process. Mice were injected with chemotherapy drugs (Cisplatin and Paclitaxel), TRULI (LATS 1/2 inhibitor), and a combination of the two drugs at different dosages. The ovaries were isolated, fixed, sectioned, and stained for IHC and IF. Cell culture studies revealed the timely expression of YAP1 as a critical regulator for granulosa cell development and possible treatment regimens using TRULI. Staining of γH2AX, Ser-139 phosphorylated form of the minor histone H2A variant, showed a positive signal for primordial and primary follicles with granulosa cell involvement in the chemotherapy groups, indicating possible DNA damage for early-stage ovarian follicles. Additionally, the TUNEL assay was conducted on a granulosa cell culture to confirm DNA fragmentation across various experimental groups, and these values were taken as ratios of TUNEL-positive cells divided by the total number of nuclei. The results showed an 11.6% TUNEL-positive value in the group treated with chemotherapy. In contrast, the control group with growth medium only had a 3.2% TUNEL positive value. The TRULI-only group had a 6.2% TUNEL-positive value, and the TRULI + Chemotherapy group had an 8.8% TUNEL-positive assay value. This confirmed that chemotherapy groups induce more damage to granulosa cells surrounding the oocyte compared to other groups. KGN morphology changes revealed that low-dose TRULI (2.5 uM) was optimal for cell growth. Protective and recovery treatment regimens for TRULI administration illustrated normal morphology and active growth. Finally, corpus luteum numbers were counted across various treatment conditions, and we concluded that there were fewer corpus luteum in the chemotherapy group when compared to the control, indicating some ovarian dysfunction due to treatment. We were able to conclude that chemotherapy induces damage to follicles and TRULI, a LATS 1/2 inhibitor, is a good potential therapeutic for this issue that shows promising results.
Description
2025
License
Attribution-NonCommercial-NoDerivatives 4.0 International