Endometriotic Organoids: A Novel In Vitro Model of Endometriotic Lesion Development

Yong Song, Gregory W. Burns, Niraj Joshi, Ripla Arora, Julie Kim, Asgerally T. Fazleabas
In: bioRxiv · 2022 · doi:10.1101/2022.02.15.480583 · W4212854385
preprint OA: green CC0 ⤵ 1 in-corpus citation
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Endometriotic organoids derived from epithelial and stromal cells self-organized and recapitulated gene expression changes found in human lesions, modeling invasion with pro-inflammatory factors.

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Abstract

ABSTRACT The development and progression of endometriotic lesions are poorly understood, but immune cell dysfunction and inflammation are closely associated with the pathophysiology of endometriosis. A lack of suitable 3D in vitro models permitting the study of interactions between cell types and the microenvironment is a contributing factor. To address this limitation, we developed endometriotic organoids (EO) to explore the role of epithelial-stromal interactions and model peritoneal cell invasion associated with lesion development. Using a non-adherent microwell culture system, spherical organoids were generated with endometriotic epithelial cells (12Z) combined with immortalized endometriotic stromal cells (iEc-ESC) or immortalized uterine stromal cells (iHUF). Organoids self-organized with stromal cells occupying the center and epithelial cells on the periphery of the organoid. Endometriotic organoids (EO), containing iEc-ESC, resulted in the development of stratified 12Z epithelial cells compared to those with iHUF where the 12Z cells developed as a single layered epithelium. Transcriptomic analysis found 4,522 differentially expressed genes (DEG) between EO and 12Z/iHUF organoids, and the top DEG included increased expression of interleukins and prostaglandin synthase enzymes. An overlap of the EO DEG with baboon endometriotic lesions was highly significant. Finally, to mimic invasion of endometrial tissue into the peritoneum, a model was developed using EO and extracellular matrix containing human peritoneal mesothelial cells (LP9). Invasion of EO into the extracellular matrix-LP9 layer was increased in presence of estrogen or THP1-derived proinflammatory macrophages. Taken together, our results strongly support the concept that EO are an appropriate model for dissecting mechanisms that contribute to endometriotic lesion development. One Sentence Summary Endometriotic organoids are an appropriate model to study epithelial-stromal interactions and model cell invasion associated with lesion development.

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endometriosis

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