CAREER: Dynamic transcription factor activity profiling for investigating mechanisms guiding early stage ovarian folliculogenesis

Regents Of The University Of Michigan - Ann Arbor, Ann Arbor MI

Investigators

Abstract

PI: Shikanov, Ariella Proposal Number: 1552580 Young women and prepubescent girls undergoing toxic treatments for cancer or autoimmune diseases have limited options to preserve their fertility. Fertility preservation is especially challenging in female patients, because females are born with an ovarian reserve comprised of a finite number of follicles that carry immature eggs. Cryopreservation of ovarian tissue prior to toxic treatments and subsequent culture of immature follicles is a promising and novel way to preserve fertility. The challenge is that follicles have low survival and maturation rates in culture, which are attributed to the complex and poorly understood signaling and interactions between the cells in a follicle. The central objective of this CAREER proposal is to discover the key signaling factors involved in the early stage development of ovarian follicles by using systems biology approach. This system will lead to the development of culture conditions for successful follicle culture to be translated to human follicles for fertility preservation, serve as a platform to screen therapeutic compounds, identify pathways with aberrant activity that could be targeted for therapy, or design environments that promote specific cellular processes in regenerative medicine and toxicology. The proposed research will identify numerous transcription factors and establish their role in the growth of early-stage ovarian follicles in vitro. These fundamental studies might provide the foundation for the development of a culture system that can support the development of early stage follicles. In the bench-to-bed translation, this knowledge may be used for fertility preservation in young female cancer survivors suffering from chemotherapy-induced infertility. Our overall research objective is to: (a) establish the dynamic activity of transcription factors during early-stage follicle development in vitro; (b) identify signaling pathways that are responsible for promoting follicle survival and growth through follicle-follicle and follicle-support cells interactions; (c) determine the secretome of cultured early-stage follicles in conditions that allow follicle-follicle and follicle-stroma interactions and correlate the functional paracrine signaling factors with the activity of transcription factors (TFs) to identify the key factors and processes driving successful folliculogenesis. To achieve this objective, we will employ a systems level approach utilizing quantitative experimental characterization with multivariate analysis and modeling. Previous investigator's work has already demonstrated how optimizing biomaterial design can enhance follicle survival rates. We will build upon this system and use the new TRACER technology (TRanscription Activity CEll aRray) to discover the identity, timing and activity pattern of TFs and secreted growth factors involved in folliculogenesis. The key transcription and secreted factors during the development of primary follicles will be investigated in vitro and compared to what is already known for in vivo development. The generated network of transcription factors predicted to be located downstream of the identified secreted factors will describe currently unknown signaling pathway active during early folliculogenesis. The broader research impact of this work will result in culture systems that can maximize ovarian follicle development and provide high quality oocytes for fertility preservation and toxicology screening. This approach has a strong potential to shed light on the mechanism of biological processes of ovarian follicle maturation, which are largely unknown and might benefit basic biologists, bioengineers, clinicians and women facing infertility altogether. The educational outreach program of this proposal integrates middle and high school students with undergraduate and graduate students to the benefit of both groups. The program will incorporate reproductive tissue engineering and biomaterials-related content and projects and will inspire K-12 students to pursue science and engineering careers through conducting K-12 outreach. The investigator is also committed to support women and minority retention in science and engineering through organizational activities and one-on-one mentoring.