Multi-omics Analysis to Elucidate Mechanisms of Chemotherapy-Induced Primary Ovarian Insufficiency

Abstract ID 94248 Poster Board 413 Background and Purpose: The advances in cancer survival rates greatly enhance the awareness of the side effects of cancer therapy and long-term life quality after cancer. Primary ovarian insufficiency (POI) and infertility caused by gonadotoxic chemotherapeutic drugs are significant side effects in young female cancer patients. This condition is characterized by a reduction in ovarian follicles, especially primordial follicles, before the age of 40. Our research teams have demonstrated that a commonly used chemotherapeutic drug, doxorubicin (DOX), primarily induces DNA damage and apoptosis of the oocytes in primordial follicles, resulting in the entire primordial follicle death, POI, infertility, and endocrine disorders. However, the underlying mechanism remains largely unknown. Therefore, our study aims to use a multi-omics approach to investigate the mechanisms responsible for chemotherapy-induced POI. Methods: Five-day-old CD-1 female mice were intraperitoneally (IP) administrated with a one-time clinically relevant dose of 10 mg/kg DOX, a commonly used chemotherapeutic drug for multiple cancers, especially leukemic patients. The ovaries were collected 6 hours post-injection for multi-omics analysis, including transcriptomics, proteomics, phospho-proteomics, and metabolomics. Additionally, the ovaries treated with the same condition were dissociated into single cells for single ovarian somatic cells and oocytes SMART-Seq2 analysis. Results: The ovary transcriptomic and proteomic analyses revealed the activation of the p53 signaling pathway following DOX treatment. The phosphor-proteomic analysis confirmed the activation of DNA damage response (DDR) signaling indicated by the hyperphosphorylation of ataxia telangiectasia mutated (ATM), checkpoint kinase 1 and 2 (CHEK1/2), and TAp63α, a p53 family member protein abundantly expressed in oocytes. Moreover, multi-omics analysis of ovaries identified several molecules and signaling pathways that have not been reported to regulate DOX-induced ovarian toxicities, including MRN complex, protein degradation, SWI/SNF chromatin remodeling complex, histone modification, and oxidative phosphorylation. The single-cell SMART-seq 2 analysis revealed distinctive transcriptomic changes between oocytes and pre-granulosa cells in response to DOX treatment. Several signaling pathways were selectively altered in oocytes or pre-granulosa cells. For instance, the NFκB signaling pathway was uniquely activated in oocytes, while the cytoskeleton-related pathways were exclusively changed in pre-granulosa cells. Conclusions: The multi-omics analysis provides information on classic and novel signaling pathways crucial for elucidating the mechanisms of chemotherapy-induced POI. This is critically important for the development of therapeutic strategies to preserve young female cancer patients' fertility and ovarian endocrine functions.