SP 1: Endometrium, Implantation and Placentation
Head of the Lab
Research questions, aims
The CEPRE Medical Scientist project will be integrated in studies on the cellular and molecular base of the endometrium and its role in embryo implantation and placentation including the interaction of endometrial immune cells, glands, vessels, stroma and invading trophoblast. Also the effects of ovarian factors, including hormones, follicular fluid and cells will be investigated. The aim is an improved understanding of the optimal conditions for embryo implantation and its translation to clinical diagnostics and treatments leading to successful pregnancies.
Scientific Background
The cellular composition of the endometrium is influenced by individual variations, age, vaginal and endometrial microbiome, maternal diseases, drugs, toxicants, sperm and others (2). The periimplantation period is controlled by soluble factors, cell-to-cell contacts and extracellular vesicles from the endometrium and the embryo, but the optimal composition is not known (3).
Own preceding work
The Placenta Lab has an expanding endometrium biobank of >20,000 well documented biopsies in paraffin from patients treated in German and European fertility centers. Mostly, natural killer and plasma cells have been studied as markers of immunological disorders or chronic endometritis (4, 5). Follicular fluid and granulosa cells have been analyzed by proteomic approaches (6). The impact of cancer and its treatment before and in pregnancy on ovary and placenta has been investigated (7, 8). 3D models from different cell types, including trophoblastic cells, have been used for confrontation or implantation-like experiments (9, 10). Extracellular vesicles from manifold cellular sources have been analyzed with emphasis on their non-coding RNA content and their potential target cell interactions (1, 11).
Method spectrum, involvement of the Medical Scientist
The MS will use the large pool of endometrial samples by imaging techniques (multiplex staining, immunohistochemistry, in-situ-hybridization) and high-end technologies available through the “medical optics and photonics” focus at the JUH. Immune and aging markers and micro- and nanomorphological characteristics will be investigated. Day-of-cycle dependent factors will be identified to define the optimal time range for implantation. Machine learning and artificial intelligence will be used for reliable image analyses. Functional and implantation assays will be performed by using complex matrices composed of different endometrial cell types, soluble factors and 3D trophoblastic spheroids. Primary endometrial cells can be obtained from donors using menstrual cups. Extracellular vesicles will be enriched from endometrial, granulosa and trophoblast cells by ultracentrifugation and analyzed by (glyco)proteomic technologies, RNA sequencing, single cell sequencing and respective confirmative methods (e.g., Western blotting, qPCR). The MS will transfect the listed cells to modify their EVs, their membrane structure, protein content and RNA load for testing the impact on potential target cells.
Relevance for CEPRE and for Reproductive Health
This subprogram will cover the earliest aspects of reproductive health in CEPRE. The MS will intensify the knowledge of peri-implantation processes in the uterus and apply methods and technologies which allow their experimental analyses.
References
1. Zarkovic M, Hufsky F, Markert UR, Marz M. The Role of Non-Coding RNAs in the Human Placenta. Cells. 2022;11(9).
2. Blois SM, Ilarregui JM, Tometten M, Garcia M, Orsal AS, Cordo-Russo R, Toscano MA, Bianco GA, Kobelt P, Handjiski B, Tirado I, Markert UR, Klapp BF, Poirier F, Szekeres-Bartho J, Rabinovich GA, Arck PC. A pivotal role for galectin-1 in fetomaternal tolerance. Nat Med. 2007;13(12):1450-7.
3. Fitzgerald JS, Poehlmann TG, Schleussner E, Markert UR. Trophoblast invasion: the role of intracellular cytokine signalling via signal transducer and activator of transcription 3 (STAT3). Hum Reprod Update. 2008;14(4):335-44.
4. Chiokadze M, Bar C, Pastuschek J, Dons'koi BV, Khazhylenko KG, Schleussner E, Markert UR, Favaro RR. Beyond Uterine Natural Killer Cell Numbers in Unexplained Recurrent Pregnancy Loss: Combined Analysis of CD45, CD56, CD16, CD57, and CD138. Diagnostics. 2020;10(9).
5. Li YY, Yu SY, Huang CY, Lian RC, Chen C, Liu S, Li LF, Diao LH, Markert UR, Zeng Y. Evaluation of peripheral and uterine immune status of chronic endometritis in patients with recurrent reproductive failure. Fertility and Sterility. 2020;113(1):187-+.
6. Lehmann R, Schmidt A, Pastuschek J, Muller MM, Fritzsche A, Dieterle S, Greb RR, Markert UR, Slevogt H. Comparison of sample preparation techniques and data analysis for the LC-MS/MS-based identification of proteins in human follicular fluid. Am J Reprod Immunol. 2018;80(2):e12994.
7. Froehlich K, Schmidt A, Heger JI, Al-Kawlani B, Aberl CA, Jeschke U, Loibl S, Markert UR. Breast cancer, placenta and pregnancy. Eur J Cancer. 2019;115:68-78.
8. Loibl S, Schmidt A, Gentilini O, Kaufman B, Kuhl C, Denkert C, von Minckwitz G, Parokonnaya A, Stensheim H, Thomssen C, van Calsteren K, Poortmans P, Berveiller P, Markert UR, Amant F. Breast Cancer Diagnosed During Pregnancy: Adapting Recent Advances in Breast Cancer Care for Pregnant Patients. JAMA Oncol. 2015;1(8):1145-53.
9. Froehlich K, Haeger JD, Heger J, Pastuschek J, Photini SM, Yan Y, Lupp A, Pfarrer C, Mrowka R, Schleussner E, Markert UR, Schmidt A. Generation of Multicellular Breast Cancer Tumor Spheroids: Comparison of Different Protocols. J Mammary Gland Biol Neoplasia. 2016;21(3-4):89-98.
10. Heger JI, Froehlich K, Pastuschek J, Schmidt A, Baer C, Mrowka R, Backsch C, Schleussner E, Markert UR, Schmidt A. Human serum alters cell culture behavior and improves spheroid formation in comparison to fetal bovine serum. Exp Cell Res. 2018;365(1):57-65.
11. Zabel RR, Morales-Prieto DM, Schultz R, Hammer M, Schleussner E, Markert UR, Favaro RR. Enrichment and Characterization of Trophoblast-Derived Extracellular Vesicles Isolated from Ex Vivo One-Sided Placenta Perfusion. Placenta. 2019;83:E63-E.