scholarly journals Heterogeneity of normal human breast stem and progenitor cells as revealed by transcriptional profiling

2017 ◽  
Author(s):  
Justin A. Colacino ◽  
Ebrahim Azizi ◽  
Michael D. Brooks ◽  
Shamileh Fouladdel ◽  
Sean P. McDermott ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Single Cell ◽  
Progenitor Cells ◽  
Ex Vivo ◽  
Breast Cancers ◽  
Human Breast ◽  
Human Mammary Gland ◽  
Normal Human Breast ◽  
Stem And Progenitor Cells ◽  
Normal Human

AbstractDuring development and pregnancy, the human mammary gland undergoes extensive remodeling in processes driven by populations of stem and progenitor cells. We recently reported that breast cancers are also hierarchically organized and driven by distinct populations of cancer stem cells characterized as CD44+CD24low/−or by expression of Aldehyde dehydrogenase (ALDH). These sets of markers identify largely non-overlapping mesenchymal and epithelial populations, each of which is capable of tumor initiation when transplanted into immunosuppressed mice. Less is known about these two populations, individually or their overlap, in the normal human mammary gland. The goal of this study was to understand the biology of the ALDH+and CD44+CD24−populations in the normal human breast, using flow cytometry based sorting paired with functionalex vivoanalyses, RNA-sequencing, and single cell RNA expression profiling. ALDH+cells and ALDH−CD44+CD24−cells, generally, have epithelial-like and mesenchymal-like characteristics, respectively. Despite this, there are substantial similarities in the biological pathways activated in both populations when compared to differentiated cells. Additionally, we found a substantial proportion of cells that simultaneously express ALDH+and CD44+CD24−whose abundance varies between individuals. At the single cell level, these cells have the greatest mammosphere forming capacity and express high levels of stemness and EMT-associated genes includingID1, SOX2, TWIST1, and ZEB2.Through unbiased analysis of individual ALDH+ cells, we find cells with either epithelial or mesenchymal expression phenotypes. We also identify a subpopulation of cells with a hybrid epithelial/mesenchymal expression phenotype that overexpress genes associated with aggressive triple negative breast cancers. These results highlight the utility of single cell analyses to characterize tissue heterogeneity, even in marker enriched cell populations, and further identifies the genes and pathways that define this heterogeneity.

scholarly journals Acquisition, processing, and single-cell analysis of normal human breast tissues from a biobank

2022 ◽  
Vol 3 (1) ◽  
pp. 101047
Author(s):  
Poornima Bhat-Nakshatri ◽  
Natascia Marino ◽  
Hongyu Gao ◽  
Yunlong Liu ◽  
Anna Maria Storniolo ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Human Breast ◽  
Normal Human Breast ◽  
Normal Human ◽  
Breast Tissues

Progesterone stimulates progenitor cells in normal human breast and breast cancer cells

2014 ◽  
Vol 143 (3) ◽  
pp. 423-433 ◽  
Author(s):  
Heidi N. Hilton ◽  
N. Santucci ◽  
A. Silvestri ◽  
S. Kantimm ◽  
L. I. Huschtscha ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Progenitor Cells ◽  
Breast Cancer Cells ◽  
Human Breast ◽  
Normal Human Breast ◽  
Normal Human

scholarly journals A Novel Method for Growing Human Breast Epithelium in Vivo Using Mouse and Human Mammary Fibroblasts

Endocrinology ◽  
2002 ◽  
Vol 143 (12) ◽  
pp. 4886-4896 ◽  
Author(s):  
Hema Parmar ◽  
Peter Young ◽  
Joanne T. Emerman ◽  
Richard M. Neve ◽  
Shanaz Dairkee ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Nude Mice ◽  
Milk Fat ◽  
Mammary Epithelium ◽  
Collagen Gels ◽  
Human Breast ◽  
Human Mammary Gland ◽  
Human Mammary Epithelium ◽  
Normal Human

Abstract A novel system is described for studying the growth of normal human mammary epithelium in vivo as grafts in athymic nude mice. The key feature of this model is reconstitution of the epithelial-stromal interactions required for normal growth and differentiation of the human mammary epithelium, which produces ducts that are comparable to those in the normal human mammary gland. Human breast epithelial organoids were combined with mammary fibroblasts from mouse or human origin in collagen gels, which were subsequently transplanted under the renal capsule of female nude mice hosts. The resulting grafts showed an increase in the ductal density compared with that observed previously. These ducts expressed appropriate markers for luminal and myoepithelial cells and steroid receptors. Treatment of the host with diethylstilbestrol or estradiol and progesterone significantly increased the number of ducts observed and increased cell proliferation. The grafts also displayed production of β-casein and milk fat globule membrane protein when the hosts were allowed to become pregnant. This model allows for a variety of epithelial and stromal cells to be used in combination, which would aid in understanding key factors that regulate normal human mammary gland development.

Antiandrogenic actions of medroxyprogesterone acetate on epithelial cells within normal human breast tissues cultured ex vivo

2014 ◽  
Vol 21 (1) ◽  
pp. 79-88 ◽  
Author(s):  
Aleksandra M. Ochnik ◽  
Nicole L. Moore ◽  
Tanja Jankovic-Karasoulos ◽  
Tina Bianco-Miotto ◽  
Natalie K. Ryan ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Medroxyprogesterone Acetate ◽  
Ex Vivo ◽  
Human Breast ◽  
Normal Human Breast ◽  
Normal Human ◽  
Breast Tissues

scholarly journals A single cell atlas of human cornea that defines its development, limbal stem and progenitor cells and the interactions with the limbal niche

2020 ◽  
Author(s):  
Joseph Collin ◽  
Rachel Queen ◽  
Darin Zerti ◽  
Sanja Bojic ◽  
Nicky Moyse ◽  
...  
Keyword(s):  
Single Cell ◽  
Progenitor Cells ◽  
Immune Cells ◽  
Ocular Surface ◽  
Ex Vivo ◽  
Corneal Stroma ◽  
Human Cornea ◽  
Rna Seq ◽  
Limbal Epithelium ◽  
Stem And Progenitor Cells

SummaryTo study the development and composition of human ocular surface, we performed single cell (sc) RNA-Seq at key embryonic, fetal and adult stages and generated the first atlas of the corneal cell types from development to adulthood. Our data indicate that during development, the conjunctival epithelium is the first to be specified from the ocular surface epithelium, followed by the corneal epithelium and the establishment of proliferative epithelial progenitors, which predate the formation of limbal niche by a few weeks. Bioinformatic comparison of adult cell clusters identified GPHA2, a novel cell-surface marker for quiescent limbal stem cells (qLSCs), whose function is to maintain qLSCs self-renewal. Combining scRNA- and ATAC-Seq analysis, we identified multiple upstream regulators for qLSCs and transit amplifying (TA) cells and demonstrated a close interaction between the immune cells and epithelial stem and progenitor cells in the cornea. RNA-Seq analysis indicated loss of qLSCs and acquisition of proliferative limbal basal epithelial progenitor markers during ex vivo limbal epithelial cell expansion, independently of the culture method used. Extending the single cell analyses to keratoconus, we were able to reveal activation of collagenase in the corneal stroma and a reduced pool of TA cells in the limbal epithelium as two key changes underlying the disease phenotype. Our scRNA- and ATAC-Seq data of developing and adult cornea in steady state and disease conditions provide a unique resource for defining pathways/genes that can lead to improvement in ex vivo expansion and differentiation methods for cell based replacement therapies and better understanding and treatment of ocular surface disorders.Key findingsscRNA-Seq of adult human cornea and conjunctiva reveals the signature of various ocular surface cell populationsscRNA-Seq of human developing cornea identifies stage-specific definitions of corneal epithelial, stromal and endothelial layersscRNA-Seq analysis results in identification of novel markers for qLSCs and TA cellsCombined scRNA- and ATAC-Seq analysis reveals key transcriptional networks in qLSCs and TA cells and close interactions with immune cellsExpansion of limbal epithelium results in downregulation of qLSCs and acquisition of proliferative limbal epithelial progenitor markersscRNA-Seq of keratoconus corneas reveals activation of collagenase in the corneal stroma and a reduced pool of TA cells in the limbal epitheliumGraphical abstractSchematic presentation of main techniques and findings presented in this manuscript.

Effects of oestrogen on gene expression in epithelium and stroma of normal human breast tissue

2006 ◽  
Vol 13 (2) ◽  
pp. 617-628 ◽  
Author(s):  
C L Wilson ◽  
A H Sims ◽  
A Howell ◽  
C J Miller ◽  
R B Clarke
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cell Proliferation ◽  
Breast Tissue ◽  
Epithelial Cell Proliferation ◽  
Human Breast ◽  
Human Breast Tissue ◽  
Tissue Microenvironment ◽  
Normal Human Breast ◽  
Normal Human

Oestrogen (E) is essential for normal and cancer development in the breast, while anti-oestrogens have been shown to reduce the risk of the disease. However, little is known about the effect of E on gene expression in the normal human breast, particularly when the epithelium and stroma are intact. Previous expression profiles of the response to E have been performed on tumour cell lines, in the absence of stroma. We investigated gene expression in normal human breast tissue transplanted into 9–10-week-old female athymic nude (Balb/c nu/nu) mice. After 2 weeks, when epithelial proliferation is minimal, one-third of the mice were treated with 17β-oestradiol (E2) to give human luteal-phase levels in the mouse, which we have previously shown to induce maximal epithelial cell proliferation. RNA was isolated from treated and untreated mice, labelled and hybridized to Affymetrix HG-U133A (human) GeneChips. Gene expression levels were generated using BioConductor implementations of the RMA and MAS5 algorithms. E2 treatment was found to represent the largest source of variation in gene expression and cross-species hybridization of mouse RNA from xenograft samples was demonstrated to be negligible. Known E2-responsive genes (such as TFF1 and AREG), and genes thought to be involved in breast cancer metastasis (including mammoglobin, KRT19 and AGR2), were upregulated in response to E treatment. Genes known to be co-expressed with E receptor α in breast cancer cell lines and tumours were both upregulated (XBP-1 and GREB1) and downregulated (RARRES1 and GATA3). In addition, genes that are normally expressed in the myoepithelium and extracellular matrix that maintain the tissue microenvironment were also differentially expressed. This suggests that the response to oestrogen in normal breast is highly dependent upon epithelial–stromal/myoepithelial interactions which maintain the tissue microenvironment during epithelial cell proliferation.

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