Decreased fertility in later reproductive age in women is an increasing trend due to career-building, as ages of first marriage and first birth increase. In addition, acute cancer treatment and chronic disease also lead to loss or decrease in fertility. In order to preserve fertility, advances in our understanding of oocyte and embryo health are required during cryo-preservation and thawing. An understanding of the health of the oocyte or embryo depends on knowledge of the stress during cryopreservation, handling, thawing, and reintroduction into culture. AMPK regulates oocyte maturation during stress and regulates a transcription factor maintaining potency in 2-cell embryos, blastocysts, and blastocyst-derived placental trophoblast stem cells (TSC). AMPK is primarily an energy sensor and suppresses anabolic processes and enhances catabolic processes to regain ATP homeostasis. In an arctic frog species, AMPK is induced during freeze and thaw to levels similar to those our laboratory has observed in stressed embryos and TSC. We hypothesize that AMPK is the key enzyme regulating homeostatic and developmental processes during oocyte freeze-down and thaw. Thus, we also hypothesize that measuring and managing the levels of AMPK activation will facilitate the optimization of oocyte cryopreservation and culture. To test this hypothesis, we will measure AMPK activity in oocytes during pre-freeze handling, after freeze and after thaw and match these levels with those of age matched oocytes in vivo. We will also add doses of AMPK agonist or antagonist during prefreeze handling, freeze down medium, and during culture after thaw, IVF and culture to blastocyst stage. Several landmarks of IVF and embryo development during culture will be measured to determine efficacy of AMPK agonists and antagonists in optimizing cryopreservation. This proposal is innovative in managing AMPK that is emerging as the primary enzyme coregulating homeostatic and developmental responses to stress and known to be regulated during freeze and thaw in arctic frogs.

Public Health Relevance

Every year, nearly 300,000 American women of reproductive age undergo cancer treatment or experience chronic diseases that threaten fertility. These women need a robust technique for preserving fertility, and oocyte cryopreservation will fulfill this need if it can be made more reliable. AMPK activity is important in normal and stressed oocyte maturation. AMPK is important in freezing and thawing of an arctic frog species suggesting it is part of a natural process regulating responses to freeze and thaw. We hypothesize that AMPK is the key enzyme regulating homeostatic and developmental processes during oocyte freeze-down and thaw and will measure AMPK activity during cryopreservation and manage AMPK activity to optimize oocyte cryopreservation.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Small Research Grants (R03)
Project #
1R03HD061431-01
Application #
7706471
Study Section
Pediatrics Subcommittee (CHHD)
Program Officer
Lamar, Charisee A
Project Start
2009-09-30
Project End
2011-08-31
Budget Start
2009-09-30
Budget End
2010-08-31
Support Year
1
Fiscal Year
2009
Total Cost
$76,000
Indirect Cost
Name
Wayne State University
Department
Obstetrics & Gynecology
Type
Schools of Medicine
DUNS #
001962224
City
Detroit
State
MI
Country
United States
Zip Code
48202
Yang, Yu; Bolnick, Alan; Shamir, Alexandra et al. (2017) Blastocyst-Derived Stem Cell Populations under Stress: Impact of Nutrition and Metabolism on Stem Cell Potency Loss and Miscarriage. Stem Cell Rev 13:454-464
Bolnick, Alan; Abdulhasan, Mohammed; Kilburn, Brian et al. (2017) Two-cell embryos are more sensitive than blastocysts to AMPK-dependent suppression of anabolism and stemness by commonly used fertility drugs, a diet supplement, and stress. J Assist Reprod Genet 34:1609-1617
Yang, Yu; Jiang, Zhongliang; Bolnick, Alan et al. (2017) Departure from optimal O2 level for mouse trophoblast stem cell proliferation and potency leads to most rapid AMPK activation. J Reprod Dev 63:87-94
Bolnick, Alan; Awonuga, Awoniyi O; Yang, Yu et al. (2017) Using stem cell oxygen physiology to optimize blastocyst culture while minimizing hypoxic stress. J Assist Reprod Genet 34:1251-1259
Yang, Yu; Arenas-Hernandez, Marcia; Gomez-Lopez, Nardhy et al. (2016) Hypoxic Stress Forces Irreversible Differentiation of a Majority of Mouse Trophoblast Stem Cells Despite FGF4. Biol Reprod 95:110
Yang, Yu; Parker, Graham C; Puscheck, Elizabeth E et al. (2016) Direct reprogramming to multipotent trophoblast stem cells, and is pluripotency needed for regenerative medicine either? Stem Cell Investig 3:24
Li, Quanwen; Gomez-Lopez, Nardhy; Drewlo, Sascha et al. (2016) Development and Validation of a Rex1-RFP Potency Activity Reporter Assay That Quantifies Stress-Forced Potency Loss in Mouse Embryonic Stem Cells. Stem Cells Dev 25:320-8
Xie, Yufen; Zhou, Sichang; Jiang, Zhongliang et al. (2014) Hypoxic stress induces, but cannot sustain trophoblast stem cell differentiation to labyrinthine placenta due to mitochondrial insufficiency. Stem Cell Res 13:478-91
Slater, Jill A; Zhou, Sichang; Puscheck, Elizabeth Ella et al. (2014) Stress-induced enzyme activation primes murine embryonic stem cells to differentiate toward the first extraembryonic lineage. Stem Cells Dev 23:3049-64
Awonuga, Awoniyi O; Yang, Yu; Rappolee, Daniel A (2013) When stresses collide. Biol Reprod 89:74

Showing the most recent 10 out of 14 publications