- Title
- Levels of steroids and endometrial proteins as infertility markers in women with reproductive pathologies of immune origin
- Creator
- Al-Juboori, Aminah Ali Abid
- Relation
- University of Newcastle Research Higher Degree Thesis
- Resource Type
- thesis
- Date
- 2021
- Description
- Research Doctorate - Doctor of Philosophy (PhD)
- Description
- Uterine tissue has been an important area of research given its vital role in the propagation of species. In contrast to most other tissues, the inner part of the uterus (endometrium) undergoes many biological events across the menstrual cycle for maintaining the homeostasis and success of the implantation process. These events physiologically regulate by a sequentially timed interplay of female sex hormones and microenvironmental cues. Dysregulation of the fine-tuned signalling of hormones and microenvironment maintaining the endometrial homeostasis ends up in many uterine pathologies such as endometriosis and infertility, recurrent implantation failure (RIF) and endometrial cancer (EC). Till date, the assessments of endometrial receptivity and diagnosis of diseases mostly rely on the morphological methods. These methods require using invasive techniques such as Dilation and curettage (D&C) to sample the endometrial tissue for biopsy and histological confirmation. Moreover, the laparoscopic visualising technique is needed in women with endometriosis for giving the protean appearance of endometrial deposits outside the uterus. The laparoscopic technique has some limitations when used as a diagnostic tool. That is because of its cost which is considered as an economic load burdened the health care system and its surgical risks. Therefore, there is an urgent need to discover less invasive techniques for the diagnosis of endometriosis and infertility, recurrent implantation failure (RIF) and endometrial cancer (EC). Moreover, development of biomarkers for prediction these common endometrial diseases and infertility will open new avenues of investigations to improve fertility and treated the endometrial diseases. The challenging to find an appropriate model met the complexity of endometrial tissue and disease setting is behind the inaccurate diagnostic tests used in clinical trials. To address this issue, this project aimed to establish in vitro systems modelling the physiological events and diseases setting of human endometrium. Furthermore, I used the more sophisticated, high-throughput technology of proteomic approach particularly the analysis of secretome tocharacterise and quantify proteins secreted by cells under the physiological and specific medical condition. Comparison of the cells-based secretome in this study and patient secretome might help in developing many of promising less invasive, accurate diagnostic markers for endometrial diseases. In this study, I have established systems represented the in vivo physiological processes across the menstrual cycle including the decidualisation and polyploidisation as well as the pathological conditions like endometriosis and endometrial cancer (EC). These systems were developed of epithelial and stromal cells including fibroblast and macrophages to reflect the complexity of human endometrial tissue. The system modelled the polyploidisation process exhibited the successful transformation of fibroblastic cells into polyploid decidual cells. Using the proteomic approach, the data showed that most of the proteins expressed and secreted of human polyploid decidual cells (HPDCs) have essential functions in the embryo implantation process. The functions of HPDCs proteins ranged from regulating the embryo attachment and invasion into protecting the embryo from maternal immune rejection and oxidative stress. Next, the system modelled the decidualisation process was utilised to examine the impacts of endometriosis conditions of estrogen dominance and chronic inflammation on decidualisation. The results showed that the inflammatory condition of endometriosis which is represented with conditioned medium (CM) of polarised Mɸs phenotype 1 defects the decidualisation process, whereas the estrogen dominance lacks this direct effect on the decidualisation. The ingenuity pathways and biological functions analysis of the polarised Mɸ phenotype 1 uncovered a set of the pro-inflammatory cytokines implicated in inducing the inflammation and defective decidualisation in human endometrial stromal cells (HESCs). The interactome analysis revealed that the phosphoinositide 3 kinase (PI3K) is the signal operated downstream the inflammatory cues of macrophages (MΦs) phenotype 1, which is the predominant phenotype in endometriosis, and being behind failed the decidualisation. Treated the decidualised cells with a well-known PI3K inhibitor, wortmannin, attenuated the inflammation and restored the decidualisation in inflammatory milieu of endometriosis. Thereafter, the system modelled endometrial cancer (EC) was established of the human endometrial stromal cells (HESCs) with human endometrial epithelial cancerous cells (Ishikawa, IK). The co-culture spheroids reflected the biological architecture of human endometrial tissue including stromal cluster surrounding with epithelial layers. Moreover, the proteomic data revealed the similarity in our co-culture spheroids and human endometrial cancer tissues compared with IK spheroids in expressing the tumorigenesis signals of proliferation, invasion and metastasis as well as the downregulation of tumour repressor protein like phosphatase 1 regulatory subunit 12A (PPP1R12A). In conclusion, the established in vitro models in this study have recapitulated the physiological functions and diseases setting of human endometrium. These models could be considered as valuable preclinical systems for developing promising less invasive diagnostic markers and treatment options for endometrial diseases such as endometriosis and infertility, recurrent implantation failure (RIF), and EC.
- Subject
- 3D modeling; decidualisation; epithelium-stromal crosstalk; endometrium cancer; endometriosis and infertility; polyploidisation; proteomic; recurrent implantation failure; secretome
- Identifier
- http://hdl.handle.net/1959.13/1428337
- Identifier
- uon:38619
- Rights
- Copyright 2021 Aminah Ali Abid Al-Juboori
- Language
- eng
- Full Text
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