Dynamic states of cervical epithelia during pregnancy and epithelial barrier disruption

Cooley, Anne; Madhukaran, ShanmugaPriyaa; Stroebele, Elizabeth; Caraballo, Mariano Colon; Wang, Lei; Akgul, Yucel; Hon, Gary C.; Mahendroo, Mala

doi:10.1016/j.isci.2023.105953

Cited by 10 publications

(13 citation statements)

References 90 publications

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“…We also measured relative transcript levels of various genes associated with different layers of vaginal epithelium at different times post-culture 20 . Transcripts associated with stem cells ( Axin2 ) and proliferation ( Birc5, Ki67 ) were more abundant at earlier times (day 5 post-culture), whereas genes associated with luminal keratinocytes ( Sprr1a ) and cornified cells ( Krt1 ) increased at later times (Fig.1C) 20,22 .…”

Section: Resultsmentioning

confidence: 98%

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Epithelial organoid supports resident memory CD8 T cell differentiation

Ulibarri,

Lin,

Ramprashad

et al. 2023

Preprint

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Resident Memory T cells (TRM) play a vital role in regional immune defense in barrier organs. Although laboratory rodents have been extensively used to study fundamental TRM biology, poor isolation efficiency, sampling bias and low cell survival rates have limited our ability to conduct TRM-focused high-throughput assays. Here, we engineered a murine vaginal epithelial organoid (VEO)-CD8 T cell co-culture system that supports CD8 TRM differentiationin vitro. The three-dimensional VEOs established from murine adult stem cells resembled stratified squamous vaginal epithelium and induced gradual differentiation of activated CD8 T cells into epithelial TRM. Thesein vitrogenerated TRM were phenotypically and transcriptionally similar toin vivoTRM, and key tissue residency features were reinforced with a second cognate-antigen exposure during co-culture. TRM differentiation was not affected even when VEOs and CD8 T cells were separated by a semipermeable barrier, indicating soluble factors’ involvement. Pharmacological and genetic approaches showed that TGF-β signaling played a crucial role in their differentiation. We found that the VEOs in our model remained susceptible to viral infections and the CD8 T cells were amenable to genetic manipulation; both of which will allow detailed interrogation of antiviral CD8 T cell biology in a reductionist setting. In summary, we established a robust model which captures bonafide TRM differentiation that is scalable, open to iterative sampling, and can be subjected to high throughput assays that will rapidly add to our understanding of TRM.

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Section: Resultsmentioning

confidence: 98%

“…Transcripts associated with stem cells (Axin2) and proliferation (Birc5, Ki67) were more abundant at earlier times (day 5 post-culture), whereas genes associated with luminal keratinocytes (Sprr1a) and cornified cells (Krt1) increased at later times (Fig. 1C) 20,22 .…”

Section: Establishment Of Vaginal Epithelial Organoid-cd8 T Cell Co-c...mentioning

confidence: 98%

Epithelial organoid supports resident memory CD8 T cell differentiation

Ulibarri,

Lin,

Ramprashad

et al. 2023

Preprint

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“…In mammals, the cervix serves to retain the conceptus during pregnancy and act as a barrier to infection by generation of a mucosal plug (Nott et al, 2016). There is extensive remodeling of the cervix during mammalian pregnancy known to be partially regulated by precise timing in hormonal fluctuations (Cooley et al, 2023; Timmons et al, 2010). It is highly likely that functionally similar processes occur in lizards as ovarian follicles are ovulated every 10–14 days and fertilized eggs are retained for a precise amount of time in the glandular uterus before laying (Crews, 1980).…”

Section: Discussionmentioning

confidence: 99%

“…Future study of the reptilian reproductive tract would benefit from detailed histological examination of the nonglandular uterus as well as expression of keratin 8 or other markers of ciliated, simple epithelium. However, in mice, a subpopulation of cervical epithelia cells expresses the simple epithelial marker keratin 8 along with a marker of squamous epithelium keratin 5, indicating that there are cells that could transdifferentiate between keratin 8 positive and keratin 5 positive identities (Cooley et al, 2023). This implies that keratin 8 expression alone is insufficient as a marker to fully understand the tissue composition in the nonglandular uterus and that a suite of markers would provide more comprehensive information.…”

Section: Nonglandular Uterusmentioning

confidence: 99%

Anatomy of the female reproductive tract organs of the brown anole (Anolis sagrei)

Kircher

Stanley

Behringer

2023

The Anatomical Record

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Female reproduction in squamate reptiles (lizards and snakes) is highly diverse and mode of reproduction, clutch size, and reproductive tract morphology all vary widely across this group of ~11,000 species. Recently, CRISPR genome editing techniques that require manipulation of the female reproductive anatomy have been developed in this group, making a more complete understanding of this anatomy essential. We describe the adult female reproductive anatomy of the model reptile the brown anole (Anolis sagrei). We show that the brown anole female reproductive tract has three distinct anterior‐to‐posterior regions, the infundibulum, the glandular uterus, and the nonglandular uterus. The infundibulum has a highly ciliated epithelial lip, a region where the epithelium is inverted so that cilia are present on the inside and outside of the tube. The glandular uterus has epithelial ducts that are patent with a lumen as well as acinar structures with a lumen. The nonglandular uterus has a heterogeneous morphology from anterior to posterior, with a highly folded, ciliated epithelium transitioning to a stratified squamous epithelium. This transition is accompanied by a loss of keratin‐8 expression and together, these changes are similar to the morphological and gene expression changes that occur in the mammalian cervix. We recommend that description of the nonglandular uterus include the regional sub‐specification of a “cervix” and “vagina” as this terminology change more accurately describes the morphology. Our data extend histological studies of reproductive organ morphology in reptiles and expand our understanding of the variation in reproductive system anatomy across squamates and vertebrates.

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“…Although the BV syndrome was first described over a century ago, the mechanisms that underlie this condition and the casual linkages between dysbiosis and adverse health outcomes still remain unknown, which has prevented development of more effective treatment and prevention strategies 3,10 . This is in part because most of our current knowledge and therapeutic development approaches are based on results of studies of microbial cultures 11,12 , metagenomic analyses 1,13,14 , in vitro 10 , or animal 15–17 models that fail to faithfully mimic the complex tissue microenvironment and mucus layer of the human reproductive tract.…”

Section: Introductionmentioning

confidence: 99%

Mucus production, host-microbiome interactions, hormone sensitivity, and innate immune responses modeled in human cervix chips

Izadifar

Cotton

Chen

et al. 2023

Preprint

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Modulation of mucus production by the human ecto- and endo-cervical epithelium by steroid hormones and associated interactions with commensal microbiome play a central role in the physiology and pathophysiology of the female reproductive tract. However, most of our knowledge about these interactions is based on results from animal studies or in vitro models that fail to faithfully mimic the mucosal environment of the human cervix. Here we describe microfluidic organ-on-a-chip (Organ Chip) models of the human cervical mucosa that recreate the cervical epithelial-stromal interface with a functional epithelial barrier and produce abundant mucus that has compositional, biophysical, and hormone-responsive properties similar to the living cervix. Use of continuous fluid flow promoted ecto-cervical differentiation, whereas use of periodic flow including periods of stasis stimulated endo-cervical specialization. Similar results with minor differences were obtained using epithelial cells isolated from three donors each from a different ethnic background (African American, Hispanic, and Caucasian). When the endo-Cervix Chips were co-cultured with living Lactobacillus crispatus and Gardnerella vaginalis bacterial communities to respectively mimic the effects of human host interactions with optimal (healthy) or non-optimal (dysbiotic) microbiome, significant differences in tissue innate immune responses, barrier function, cell viability, and mucus composition were detected reminiscent of those observed in vivo. Thus, human Cervix Chips provide a physiologically relevant experimental in vitro model to study cervical mucus physiology and its role in human host-microbiome interactions as well as a potential preclinical testbed for development of therapeutic interventions to enhance women's health.

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Dynamic states of cervical epithelia during pregnancy and epithelial barrier disruption

Cited by 10 publications

References 90 publications

Epithelial organoid supports resident memory CD8 T cell differentiation

Epithelial organoid supports resident memory CD8 T cell differentiation

Anatomy of the female reproductive tract organs of the brown anole (Anolis sagrei)

Mucus production, host-microbiome interactions, hormone sensitivity, and innate immune responses modeled in human cervix chips

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