Endometrium On-a-Chip Reveals Insulin-and Glucose-induced Alterations in the Transcriptome and Proteomic Secretome

Author(s)

De Bem, Tiago H. C. ^{[1, 2]} ; Tinning, Haidee ^[2] ; Vasconcelos, Elton J. R. ^[3] ; Wang, Dapeng ^[3] ; Forde, Niamh ^{[2, 3]}

Affiliation

^[1] Univ Sao Paulo, Fac Zootecnia & Engn Alimentos, Dept Med Vet, Pirassununga, SP - Brazil

^[2] Univ Leeds, Sch Med, Leeds Inst Cardiovasc & Metab Med, Discovery & Translat Sci Dept, Leeds, W Yorkshire - England

^[3] Univ Leeds, LeedsOm, Leeds, W Yorkshire - England

• Document type: Journal article
• Source: Endocrinology; v. 162, n. 6 JUN 2021.
• Web of Science Citations: 0

Abstract

The molecular interactions between the maternal environment and the developing embryo are key for early pregnancy success and are influenced by factors such as maternal metabolic status. Our understanding of the mechanism(s) through which these individual nutritional stressors alter endometrial function and the in utero environment for early pregnancy success is, however, limited. Here we report, for the first time, the use of an endometrium-on-a-chip microfluidics approach to produce a multicellular endometrium in vitro. Isolated endometrial cells (epithelial and stromal) from the uteri of nonpregnant cows in the early luteal phase (Days 4-7) were seeded in the upper chamber of the device (epithelial cells; 4-6 x 10(4) cells/mL) and stromal cells seeded in the lower chamber (1.5-2 x 10(4) cells/mL). Exposure of cells to different concentrations of glucose (0.5, 5.0, or 50 mM) or insulin (Vehicle, 1 or 10 ng/mL) was performed at a flow rate of 1 mu L/minute for 72 hours. Quantitative differences in the cellular transcriptome and the secreted proteome of in vitro-derived uterine luminal fluid were determined by RNA-sequencing and tandem mass tagging mass spectrometry, respectively. High glucose concentrations altered 21 and 191 protein-coding genes in epithelial and stromal cells, respectively (P < .05), with a dose-dependent quantitative change in the protein secretome (1 and 23 proteins). Altering insulin concentrations resulted in limited transcriptional changes including transcripts for insulin-like binding proteins that were cell specific but altered the quantitative secretion of 196 proteins. These findings highlight 1 potential mechanism by which changes to maternal glucose and insulin alter uterine function. (AU)

• FAPESP's process: 18/14137-1 - A new culture system of endometrial cells to study maternal/fetal communication in cattle
• Grantee: Tiago Henrique Camara De Bem
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 16/22790-1 - Extracellular vesicles in inter-embryonic and maternal-embryonic communication during in vitro development of bovine embryos
• Grantee: Tiago Henrique Camara De Bem
• Support Opportunities: Scholarships in Brazil - Post-Doctoral