Primary Mammary Organoid Model of Lactation and Involution

Sumbal, Jakub; Chiche, Aurélie; Charifou, Elsa; Koledová, Zuzana; Li, Han

doi:10.3389/fcell.2020.00068

Cited by 61 publications

(46 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…S2C , bottom panel). In fact, it was recently demonstrated that pregnancy-induced mammary involution can be recapitulated in vitro [ 6 ], thus supporting an involution-like state to organoid cultures treated for 48 h with pregnancy hormones, an observation that we are now supplementing with molecular signatures. Moreover, we also observed that a fraction of all pregnancy hormone exclusive peaks were shared across all 3 timepoints, suggesting sustainability of epigenetic changes established by pregnancy signals throughout the treatment time-course (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

“…By analyzing markers of active regulatory regions (H3K27ac), our current method allowed for a snapshot of epigenetic changes in a timely fashion, thus illustrating some of the immediate molecular responses to pregnancy hormones. Given that additional organoid methods can mimic phenotypic alterations observed during involution [ 6 ], a deep molecular analysis of such developmental stage would provide further insights into how the epigenome is shaped to assume a post-involution state. Moreover, incorporating additional histone marks of poised enhancers (H3K4me1) or repressed enhancers (H3K27me) will illustrate parts of the epigenome that may became less active after pregnancy.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of Organoid Cultures to Study the Effects of Pregnancy Hormones on the Epigenome and Transcriptional Output of Mammary Epithelial Cells

Ciccone

Trousdell

Santos

2020

J Mammary Gland Biol Neoplasia

View full text Add to dashboard Cite

The use of mouse derived mammary organoids can provide a unique strategy to study mammary gland development across a normal life cycle, as well as offering insights into how malignancies form and progress. Substantial cellular and epigenomic changes are triggered in response to pregnancy hormones, a reaction that engages molecular and cellular changes that transform the mammary epithelial cells into “milk producing machines”. Such epigenomic alterations remain stable in post-involution mammary epithelial cells and control the reactivation of gene transcription in response to re-exposure to pregnancy hormones. Thus, a system that tightly controls exposure to pregnancy hormones, epigenomic alterations, and activation of transcription will allow for a better understanding of such molecular switches. Here, we describe the characterization of ex vivo cultures to mimic the response of mammary organoid cultures to pregnancy hormones and to understand gene regulation and epigenomic reprogramming on consecutive hormone exposure. Our findings suggest that this system yields similar epigenetic modifications to those reported in vivo, thus representing a suitable model to closely track epigenomic rearrangement and define unknown players of pregnancy-induced development.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Characterization of Organoid Cultures to Study the Effects of Pregnancy Hormones on the Epigenome and Transcriptional Output of Mammary Epithelial Cells

Ciccone

Trousdell

Santos

2020

J Mammary Gland Biol Neoplasia

View full text Add to dashboard Cite

show abstract

“…An unexpected finding in our microcontainer system is that organoids exhibit contractility. Although contractility is a known functional behavior of mammary tissue and has been observed in mouse mammary explants ( Mroue et al., 2015 ; Sumbal et al., 2020 ), it has not been observed in reconstituted mammary organoids. Time-lapse microscopy showed that organoids in microcontainers gradually dilate across a span of one or more hours and then rapidly contract ( Figure 3 A).…”

Section: Resultsmentioning

confidence: 99%

Volume-constrained microcontainers enable myoepithelial functional differentiation in highly parallel mammary organoid culture

et al. 2021

View full text Add to dashboard Cite

Summary A long-standing constraint on organoid culture is the need to add exogenous substances to provide hydrogel matrix, which limits the study of fully human or fully native organoids. This paper introduces an approach to culture reconstituted mammary organoids without the impediment of exogenous matrix. We enclose organoids in nanoliter-scale, topologically enclosed, fluid compartments surrounded by agar. Organoids cultured in these “microcontainers” appear to secrete enough extracellular matrix to yield a self-sufficient microenvironment without exogenous supplements. In microcontainers, mammary organoids exhibit contractility and a high-level, physiological, myoepithelial (MEP) behavior that has not been previously reported in reconstituted organoids. The presence of contractility suggests that microcontainers elicit MEP functional differentiation, an important milestone. Microcontainers yield thousands of substantially identical and individually trackable organoids within a single culture vessel, enabling longitudinal studies and statistically powerful experiments, such as the evaluation of small effect sizes. Microcontainers open new doors for researchers who rely on organoid models.

show abstract

“…The accumulation of milk at the end of the lactation stage in mammary glands triggers the death of excessive epithelial cells. These cells are promptly removed by neighboring epithelial cells and by infiltrating phagocytes with the help of MFG-E8 protein to ensure that the gland returns to its pre-pregnant state ( Ensslin and Shur, 2007 ; Monks et al, 2008 ; Watson and Kreuzaler, 2011 ; Sumbal et al, 2020 ). Therefore, we focused on determining the pathways associated with MFG-E8 protein in MEC proliferation and oncogenesis processes.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Repression of MFG-E8 Causes Disturbance in the Homeostasis of Cell Cycle Through DOCK/ZP4/STAT Signaling in Buffalo Mammary Epithelial Cells

Verma

Ali

Singh

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

The mammary gland is a unique apocrine gland made up of a branching network of ducts that end in alveoli. It is an ideal system to study the molecular mechanisms associated with cell proliferation, differentiation, and oncogenesis. MFG-E8, also known as Lactadherin, is a vital glycoprotein related to the milk fat globule membrane and initially identified to get secreted in bovine milk. Our previous report suggests that a high level of MFG-E8 is indicative of high milk yield in dairy animals. Here, we showed that MFG-E8 controls the cell growth and morphology of epithelial cells through a network of regulatory transcription factors. To understand the comprehensive action, we downregulated its expression in MECs by MFG-E8 specific shRNA. We generated a knockdown proteome profile of differentially expressed proteins through a quantitative iTRAQ experiment on a high-resolution mass spectrometer (Q-TOF). The downregulation of MFG-E8 resulted in reduced phagocytosis and cell migration ability, whereas it also leads to more lifespan to knockdown vis-a-vis healthy cells, which is confirmed through BrdU, MTT, and Caspase 3/7. The bioinformatics analysis revealed that MFG-E8 knockdown perturbs a large number of intracellular signaling, eventually leading to cessation in cell growth. Based on the directed network analysis, we found that MFG-E8 is activated by CX3CL1, TP63, and CSF2 and leads to the activation of SOCS3 and CCL2 for the regulation of cell proliferation. We further proved that the depletion of MFG-E8 resulted in activated cytoskeletal remodeling by MFG-E8 knockdown, which results in the activation of three independent pathways ZP4/JAK-STAT5, DOCK1/STAT3, and PIP3/AKT/mTOR. Overall, this study suggests that MFG-E8 expression in mammary epithelial cells is an indication of intracellular deterioration in cell health. To date, to the best of our knowledge, this is the first study that explores the downstream targets of MFG-E8 involved in the regulation of mammary epithelial cell health.

show abstract

Primary Mammary Organoid Model of Lactation and Involution

Cited by 61 publications

References 58 publications

Characterization of Organoid Cultures to Study the Effects of Pregnancy Hormones on the Epigenome and Transcriptional Output of Mammary Epithelial Cells

Characterization of Organoid Cultures to Study the Effects of Pregnancy Hormones on the Epigenome and Transcriptional Output of Mammary Epithelial Cells

Volume-constrained microcontainers enable myoepithelial functional differentiation in highly parallel mammary organoid culture

Transcriptional Repression of MFG-E8 Causes Disturbance in the Homeostasis of Cell Cycle Through DOCK/ZP4/STAT Signaling in Buffalo Mammary Epithelial Cells

Contact Info

Product

Resources

About