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3) Development of Human Embryo Viability Biomarker Diagnostics For

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3) Development of Human Embryo Viability Biomarker Diagnostics For 1) Understanding the impact of sperm genetic and epigenetic abnormalities on the ART derived human embryos: metabolomic and morphokinetic approach Assisted Reproductive Technology (ART), as a treatment for infertile couple, has been successfully helping in delivery of a healthy child. However, recent studies have shown that ART born children are at increased risk of having imprinting disorders. It has been earlier shown that poor quality sperm carry aberrant epigenetic signature hence can transmit the same to the resulting embryos and fetus. Limitations over conventional gamete and embryo assessment methods in determining sperm derived genetic and epigenetic abnormalities made us to look into the impact of sperm mediated (epi) genetic errors on embryonic behaviour. Further it is proposed to develop a non-invasive tool to identify such errors in sperm and embryos through proteomic/metabolomic approach. Funded by: Indian Council of Medical Research, Govt. of India (ICMR) 2) Non-invasive approach to assess embryo quality using oxygen consumption Assisted Reproductive Technology (ART) is now being widely used in many countries to help the infertile couples to achieve pregnancy. Currently, embryo selection for ART is based on the morphology of embryo and the rate of embryo development in culture, which are highly subjective and not always very accurate. Hence there is a need for an objective method of embryo selection, which is highly reliable. As oxygen consumption is a key indicator of embryo metabolism and it is directly proportional to the growth of embryo, this study is focused on using the oxygen consumption of embryo as non-invasive approach to select the best quality embryos. The oxygen measurement methods that are currently used in embryo metabolism research are difficult to commercialize due to complexity, cost, maintenance and expert handling issues. So, for the first time, this study is going to use simple, effective and optical sensor based non-invasive oxygen depletion method for the selection of good quality embryo, which may help to build a better life. Funded by: Science and Engineering Research Board, Govt. of India (SERB) 3) Development of human embryo viability biomarker diagnostics for elective embryo transfer to improve pregnancy outcome: potential application in art clinic (IMPRINT project in collaboration with IISc, Bangalore) To treat human infertility, in-vitro fertilization (IVF) and embryo transfer (ET) technologies (ART) are practiced. But, the live-birth rate is <15%. Among other causes, poor biological viability of cultured-embryos substantially contributes to embryonic loss. Hence, there is an urgent-unmet need to assess viability of embryos by analyzing embryo-derived biomarkers in the spent medium. Using immuno-diagnostic or/and embryomics technology platforms, a point-of-care (POC) molecular diagnostics would be developed which would be potential implemented as an adjunct to embryo-scores, practiced in ART-clinics. Goals expected to be achieved are (1) embryo viability-biomarker immunodiagnostic development, (2) retrospective correlative analysis between biomarkers’ prediction potential and pregnancy outcomes and (3) embryomics-based biomarkers discovery with potential translational application in clinical (ART) setting. The outcome will benefit infertile couples in the country in terms of (a) clinicians-advised strategic medical choice, improved live-birth rates and substantial cost-savings. Funded by: Ministry of Human Resource Development, Govt. of India (MHRD) 4) Embryo spent medium analysis and machine learning in IVF: Understanding the functional competence of gametes and evaluating implantation potential of embryos (SERB project in collaboration with NMR centre, IISc, Bangalore) There has been growing interest in understanding energy metabolism in human embryos generated using assisted reproductive techniques (ART) for improving the overall success rate of the method. Using NMR spectroscopy as a noninvasive tool, this study is aimed to understand the human embryo metabolism in identifying specific biomarkers to assess the quality of embryos for their implantation potential in conjunction with application of infertility based machine learning. Validation of these techniques is expected to help ART to professionals in predicting successful reproductive outcome. Funded by: Science and Engineering Research Board, Govt. of India (SERB) 5) Fertility preservation in cancer affected prepubertal boys: An experimental approach using testicular tissue cryopreservation. Gonadal damage is a relatively common consequence of the treatments used to cure paediatric cancer as the testes have a low threshold for damage by radiation and other cytotoxic agents. Although pubertal patients have established options to preserve their fertility before cancer therapy, the methods for deriving mature gamete from testicular germ cells of prepubertal boys remain at experimental stage. These observations made us to propose a multidisciplinary approach with oncologists, paediatric surgeons and basic scientists to establish fertility preservation facility in tertiary care hospital. This translational approach is expected to understand the community effectiveness and accessibility by the target group which helps in expanding the service to the other parts of the country. Funded by: Indian Council of Medical Research, Govt. of India (ICMR) 6) Experimental and community approach towards addressing challenges and opportunities for fertility preservation. The fertility preservation project is a multidisciplinary study to understand the effect of cancer and its’ treatment on the testicular germ cells and their integrity along with understanding the scope of oncofertility in India. The experimental approach of this study is to elucidate the impact of chemotherapeutic agents on mouse prepubertal gonads, to assess their gonadotoxicity. Secondly, it is planned to address the community approach to understand the scope of oncofertility establishment in India, for effective utilization of fertility preservation services in cancer patients prior to treatment. 7) Effect of maternal exposure of metformin on fetal gonadal development and their reproductive potential. Metformin is a dimethylbiguanide drug prescribed for the treatment of diabetes mellitus due to its hypoglycemic activity. It inhibits gluconeogenesis in the liver and reduces gastrointestinal glucose absorption. It also has an important role in reproductive medicine for the treatment of polycystic ovarian syndrome (PCOS) as it reduces androgen levels and restores ovulation. There are reports indicating that metformin can increase sex hormone binding globulin level and decrease testosterone level in maternal and fetal circulation which might alter the intrauterine environment for the growing fetus. However, it is not known whether these endocrine changes induced by metformin has any effect on the development of the fetus, especially the male progeny. The present study is aimed at elucidating the effect of metformin on hormonal levels, ovarian function, embryo development and reproductive potential of the progeny in PCOS condition. Funded by: Science and Engineering Research Board, Govt. of India (SERB) 8) Understanding the role of paternal factors on early cleavage pattern, metabolism and endoplasmic reticulum stress in preimplantation embryos Parthenogenesis is an abnormal form of asexual reproduction in which an oocyte develops into an embryo without being fertilized by spermatozoa. The parthenotes serve as an excellent model to understand the role of paternal and maternal genome in regulating the early embryo development which is still not clearly understood. However, when cultured under in- vitro conditions, the parthenotes are characterized by delayed development, early embryonic arrest, high degree of fragmentation, poor blastocyst quality and differential cleavage dynamics when compared to the normally fertilized embryos. In this study, we intend to understand the reason for difference in the cleavage pattern in parthenotes using time lapse imaging system and fluorescent live-cell imaging technology. In addition, whether the poor developmental potential of parthenotes is associated with elevated endoplasmic reticulum stress or differential metabolic requirement when compared to normally fertilized embryos are studied. Funded by: Department of Science and Technology (DST-INSPIRE) 9) Designing liposome based freezing medium for testicular tissue cryopreservation to aid in male fertility preservation Testicular tissue cryopreservation is known to have potential application in the field reproductive medicine and fertility preservation, especially for pre-pubertal boys. However, an optimum protocol for cryopreservation of testicular tissue is still lacking due to the complexity of tissue and heterogeneity with respect to the types of cells present. During freeze-thaw process extensive changes occur at the membrane level such as loss of cholesterol and phospholipids, reorganization of lipids and lipid peroxidation which subsequently leads to loss of viability and damage to the macromolecules like DNA. This project aims to design an optimum medium for testicular cryopreservation by using liposomes made up of different membrane lipid components to ameliorate the adverse effects of freeze- thaw process. Funded by: Indian Council of Medical Research, Govt. of India (ICMR) 10) Enhancing the cryosurvival of gametes, embryos and gonadal tissues by stabilizing DNA and membrane
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