Clonal Analysis via Barcoding Reveals Diverse Growth and Differentiation of Transplanted Mouse and Human Mammary Stem Cells

Nguyen, Long V.; Makarem, Maisam; Carles, Annaïck; Moksa, Michelle; Kannan, Nagarajan; Pandoh, Pawan; Eirew, Peter; Osako, Tomo; Kardel, Melanie; Cheung, Alice; Kennedy, William; Tse, Kane; Zeng, Thomas; Zhao, Yongjun; Humphries, R. Keith; Aparicio, Samuel; Eaves, Connie J.; Hirst, Martin

doi:10.1016/j.stem.2013.12.011

Cited by 60 publications

(76 citation statements)

References 39 publications

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“…The recent development of cellular barcoding technology offers an alternative approach to investigate the diversity of differentiation potential, although this requires an intervening culture period. The transplantation of barcoded basal cells into cleared fat pads resulted in many bilineage cells within the primary outgrowths, compatible with the existence of a common stem cell (Nguyen et al 2014). …”

Section: Matters Arising In Lineage Tracing and Transplantation Studiesmentioning

confidence: 82%

Mammary stem cells and the differentiation hierarchy: current status and perspectives

2014

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The mammary epithelium is highly responsive to local and systemic signals, which orchestrate morphogenesis of the ductal tree during puberty and pregnancy. Based on transplantation and lineage tracing studies, a hierarchy of stem and progenitor cells has been shown to exist among the mammary epithelium. Lineage tracing has highlighted the existence of bipotent mammary stem cells (MaSCs) in situ as well as long-lived unipotent cells that drive morphogenesis and homeostasis of the ductal tree. Moreover, there is accumulating evidence for a heterogeneous MaSC compartment comprising fetal MaSCs, slow-cycling cells, and both long-term and short-term repopulating cells. In parallel, diverse luminal progenitor subtypes have been identified in mouse and human mammary tissue. Elucidation of the normal cellular hierarchy is an important step toward understanding the “cells of origin” and molecular perturbations that drive breast cancer.

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Section: Matters Arising In Lineage Tracing and Transplantation Studiesmentioning

confidence: 82%

Mammary stem cells and the differentiation hierarchy: current status and perspectives

2014

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“…These epigenetic changes impact the biological activity of cells through their modification of transcriptional states and regulatory machinery, and the proportion of cells carrying these mutations is known to vary at the genetic level [12,13]. But, a method for determining the proportion of cells that carry these changes has never been demonstrated at the epigenetic level, even though it has been reported that a subset of the CpG sites vary their methylation level between 20% and 80% [8].…”

Section: Resultsmentioning

confidence: 99%

Dynamic evolution of clonal epialleles revealed by methclone

Li¹,

Garrett-Bakelman²,

Perl³

et al. 2014

Genome Biol

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We describe methclone, a novel method to identify epigenetic loci that harbor large changes in the clonality of their epialleles (epigenetic alleles). Methclone efficiently analyzes genome-wide DNA methylation sequencing data. We quantify the changes using a composition entropy difference calculation and also introduce a new measure of global clonality shift, loci with epiallele shift per million loci covered, which enables comparisons between different samples to gauge overall epiallelic dynamics. Finally, we demonstrate the utility of methclone in capturing functional epiallele shifts in leukemia patients from diagnosis to relapse. Methclone is open-source and freely available at

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“…Our findings establish the normal human luminal epithelium as a distinct site of oxidative stress resistance in the presence of high intracellular ROS levels mediated by a unique enzymatic repertoire. These features distinguish LPs from BCs, which include very primitive bipotent cells with both luminal and myoepithelial differentiation abilities (24). The unique properties of the LPs include up-regulated levels and activity of SODs and their superoxide substrates, with an accompanying switch in mechanisms used to control ROS levels and promote cell survival in the presence of increased ROS levels.…”

Section: Discussionmentioning

confidence: 99%

Glutathione-dependent and -independent oxidative stress-control mechanisms distinguish normal human mammary epithelial cell subsets

Kannan

Nguyen

Makarem

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Mechanisms that control the levels and activities of reactive oxygen species (ROS) in normal human mammary cells are poorly understood. We show that purified normal human basal mammary epithelial cells maintain low levels of ROS primarily by a glutathione-dependent but inefficient antioxidant mechanism that uses mitochondrial glutathione peroxidase 2. In contrast, the matching purified luminal progenitor cells contain higher levels of ROS, multiple glutathione-independent antioxidants and oxidative nucleotide damage-controlling proteins and consume O 2 at a higher rate. The luminal progenitor cells are more resistant to glutathione depletion than the basal cells, including those with in vivo and in vitro proliferation and differentiation activity. The luminal progenitors also are more resistant to H 2 O 2 or ionizing radiation. Importantly, even freshly isolated "steady-state" normal luminal progenitors show elevated levels of unrepaired oxidative DNA damage. Distinct ROS control mechanisms operating in different subsets of normal human mammary cells could have differentiation state-specific functions and long-term consequences.human epithelial stem and progenitor cells | mammary differentiation | 3D clonogenic assay | superoxide dismutase | peroxiredoxin C ellular synthesis of different reactive oxygen species (ROS) results primarily from the incomplete reduction of molecular oxygen in mitochondria to generate free radical superoxide anions. ROS also are produced when cells are exposed to different environmental sources of oxidative stressors, including ionizing radiation. Together, these regulate many normal cellular processes and also contribute to DNA damage, tumorigenesis, and cell death (1-3).In mice, the inner layer of "luminal" epithelial cells of the normal adult mammary gland have been found to contain higher levels of ROS than the outer "basal" layer of epithelial cells (4). The basis for these differences in ROS levels in the two cell types has been attributed to differences in their content of mitochondria; however, their stress response mechanisms have not been defined. Even less is known about ROS levels and their control in the normal adult human mammary gland, which consists of a similar continuous bilayered epithelial network of ducts and terminal alveolae. MCF10A cells are an immortal but nontumorigenic human mammary cell line that, when grown in 3D Matrigel cultures, generates multilayered spheres in which a lumen forms owing to the acquisition of lethal levels of ROS by the inner cells (5). These studies have suggested that ROS regulation plays an important role in the structural morphogenesis and homeostasis of normal adult human mammary tissue.Here we report the results of experiments designed to investigate the levels of ROS and their control in highly purified populations of viable luminal progenitors (LPs) and basal cells (BCs) isolated from normal human breast tissue (6-8). Our results confirm the different levels of ROS identified in the corresponding subsets of mouse mammary cells an...

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Clonal Analysis via Barcoding Reveals Diverse Growth and Differentiation of Transplanted Mouse and Human Mammary Stem Cells

Cited by 60 publications

References 39 publications

Mammary stem cells and the differentiation hierarchy: current status and perspectives

Mammary stem cells and the differentiation hierarchy: current status and perspectives

Dynamic evolution of clonal epialleles revealed by methclone

Glutathione-dependent and -independent oxidative stress-control mechanisms distinguish normal human mammary epithelial cell subsets

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