MUCPLUG
Abstract
Single-Cell Characterization of Gestational Cervical Epithelial Reprogramming
The cervical mucus plug serves as a critical physical and immunological barrier during pregnancy, yet the cellular mechanisms governing its formation and maintenance remain poorly defined. Current gaps in reproductive biology highlight an immediate need for a reliable cellular reference map of the pregnant cervix before mechanistic perturbations or translational interventions can be effectively developed. This project aims to define how pregnancy reprograms cervical epithelial cells into specialized mucus-producing states and to identify the molecular signatures that distinguish these populations across gestation.
The central hypothesis of this study is that pregnancy induces a discrete set of specialized cervical goblet-cell states characterized by distinct transcriptional programs. These programs are hypothesized to regulate mucin biosynthesis, secretion, cellular stress adaptation, innate defense, and epithelial maintenance. We propose that the relative abundance and functional identity of these states shift dynamically throughout gestation, resulting in a responsive, evolving barrier rather than a static anatomical structure.
To test this hypothesis and establish a foundational framework for cervical biology, the project will pursue three specific objectives:
Generation of a Single-Cell Atlas: We will utilize single-cell RNA sequencing (scRNA-seq) to profile cervical epithelial cells harvested from non-pregnant mice and pregnant mice at precisely defined gestational stages. This atlas will serve as the baseline for identifying pregnancy-induced cellular shifts.
Molecular Definition of Secretory Subtypes: We will identify and molecularly characterize specific goblet-cell subtypes and transitional secretory states that emerge exclusively during pregnancy. This includes the identification of unique marker genes and the pathways driving their differentiation.
Reconstruction of Gestational Trajectories: Using computational modeling, we will reconstruct the developmental trajectories of these epithelial states. This analysis will nominate candidate transcriptional regulators and identify sortable cell populations, providing the necessary tools for future functional and mechanistic validation.
The successful completion of this research will provide the first mechanistically useful framework for understanding cervical mucus-plug biology. By transitioning from a bulk-tissue perspective to a high-resolution single-cell analysis, this project reduces conceptual uncertainty regarding the cellular composition of the pregnant cervix. Furthermore, it will establish a validated set of molecular targets and markers to investigate barrier regulation and the pathogenesis of infection-associated preterm birth. Ultimately, this work ensures that subsequent therapeutic and diagnostic studies are constructed upon an accurate cellular model of the gestational cervical environment.
