Year-end Sale % OFF 
Abstract
Inadequate or inappropriate implantation and placentation during the establishment of human pregnancy is thought to lead to first trimester miscarriage, placental insufficiency and other obstetric complications. To create the placental blood supply, specialized cells, the ‘extravillous trophoblast’ (EVT) invade through the differentiated uterine endometrium (the decidua) to engraft and remodel uterine spiral arteries. We hypothesized that decidual factors would regulate EVT function by altering the production of EVT membrane and secreted factors. We used a proteomics approach to identify EVT membrane and secreted proteins regulated by decidual cell factors. Human endometrial stromal cells were decidualized in vitro by treatment with estradiol (10−8 M), medroxyprogesterone acetate (10−7 M) and cAMP (0.5 mM) for 14 days. Conditioned media (CM) was collected on day 2 (non-decidualized CM) and 14 (decidualized CM) of treatment. Isolated primary EVT cultured on Matrigel™ were treated with media control, non-decidualized or decidualized CM for 16 h. EVT CM was fractionated for proteins <30 kDa using size-exclusion affinity nanoparticles (SEAN) before trypsin digestion and HPLC-MS/MS. 43 proteins produced by EVT were identified; 14 not previously known to be expressed in the placenta and 12 which had previously been associated with diseases of pregnancy including preeclampsia. Profilin 1, lysosome associated membrane glycoprotein 1 (LAMP1), dipeptidyl peptidase 1 (DPP1/cathepsin C) and annexin A2 expression by interstitial EVT in vivo was validated by immunhistochemistry. Decidual CM regulation in vitro was validated by western blotting: decidualized CM upregulated profilin 1 in EVT CM and non-decidualized CM upregulated annexin A2 in EVT CM and pro-DPP1 in EVT cell lysate. Here, non-decidualized factors induced protease expression by EVT suggesting that non-decidualized factors may induce a pro-inflammatory cascade. Preeclampsia is a pro-inflammatory condition. Overall, we have demonstrated the potential of a proteomics approach to identify novel proteins expressed by EVT and to uncover the mechanisms leading to disease states.
Highlights
During the establishment of pregnancy, a human blastocyst must implant into the uterine endometrium to facilitate the formation of a functional placenta
13 unique proteins were found expressed by extravillous trophoblast’ (EVT) in response to treatment with non-decidualized Conditioned media (CM), 6 were previously unknown to be expressed by EVT (Table S1) and of the 7 known proteins, 4 are dysregulated in preeclampsia and 1 is associated with endometrial cancer (Table S1 [30,31,32,33,34])
EVT treated with decidualized CM expressed 12 unique proteins; 6 previously unknown to be expressed by EVT and 5 of the 6 known proteins are dysregulated in either preeclampsia (3), IUGR (1) or in IVF and ICSI pregnancies (1; Table S1 [35,36,37,38,39])
Summary
During the establishment of pregnancy, a human blastocyst must implant into the uterine endometrium to facilitate the formation of a functional placenta. To form a functional placenta, specialized cells, the ‘extravillous trophoblast’ (EVT) engraft and remodel uterine spiral arteries, creating the placental blood supply at the end of the first trimester [3]. Prior to implantation and in preparation for pregnancy, stromal cells of the human uterine endometrium undergo ‘decidualization’. Decidualization involves the categorical reprogramming of endometrial stromal cells such that different genes are expressed at different stages of the differentiation process [5]. Decidualization begins spontaneously in stromal cells adjacent to spiral arterioles during the mid-secretory phase of the menstrual cycle (5th–10th day after luteinizing hormone surge) in response to progesterone and regardless of the presence of a functional blastocyst. Decidualization intensifies and continues to form the decidua of pregnancy [6]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
