Mutations in long-lived epithelial stem cells and their clonal progeny in pre-malignant lesions and in oral squamous cell carcinoma

Abstract Oral squamous cell carcinomas (OSCCs) are the most common cancers of the oral cavity, but the molecular mechanisms driving OSCC carcinogenesis remain unclear. Our group previously established a murine OSCC model based on a 10-week carcinogen [4-nitroquinoline 1-oxide (4-NQO)] treatment. Here we used K14CreERTAM;Rosa26LacZ mice to perform lineage tracing to delineate the mutational profiles in clonal cell populations resulting from single, long-lived epithelial stem cells, here called LacZ+ stem cell clones (LSCCs). Using laser-capture microdissection, we examined mutational changes in LSCCs immediately after the 10-week 4-NQO treatment and >17 weeks after 4-NQO treatment. We found a 1.8-fold ±0.4 (P = 0.009) increase in single-nucleotide variants and insertions/deletions (indels) in tumor compared with pre-neoplastic LSCCs. The percentages of indels and of loss of heterozygosity events were 1.3-fold±0.3 (P = 0.02) and 2.2-fold±0.7 (P = 0.08) higher in pre-neoplastic compared with tumor LSCCs. Mutations in cell adhesion- and development-associated genes occurred in 83% of the tumor LSCCs. Frequently mutated genes in tumor LSCCs were involved in planar cell polarity (Celsr1, Fat4) or development (Notch1). Chromosomal amplifications in 50% of the tumor LSCCs occurred in epidermal growth factor receptor, phosphoinositide 3-kinase and cell adhesion pathways. All pre-neoplastic and tumor LSCCs were characterized by key smoking-associated changes also observed in human OSCC, C>A and G>T. DeconstructSigs analysis identified smoking and head and neck cancer as the most frequent mutational signatures in pre-neoplastic and tumor LSCCs. Thus, this model recapitulates a smoking-associated mutational profile also observed in humans and illustrates the role of LSCCs in early carcinogenesis and OSCCs.

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Perivascular-Derived Mesenchymal Stem Cells

Journal of Dental Research ◽

10.1177/0022034519862258 ◽

2019 ◽

Vol 98 (10) ◽

pp. 1066-1072 ◽

Cited By ~ 4

Author(s):

V. Yianni ◽

P.T. Sharpe

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Dental Pulp ◽

Molecular Mechanisms ◽

Lineage Tracing ◽

Specific Gene ◽

Genetic Lineage ◽

Differentiation Potential

Cells have been identified in postnatal tissues that, when isolated from multiple mesenchymal compartments, can be stimulated in vitro to give rise to cells that resemble mature mesenchymal phenotypes, such as odontoblasts, osteoblasts, adipocytes, and myoblasts. This has made these adult cells, collectively called mesenchymal stem cells (MSCs), strong candidates for fields such as tissue engineering and regenerative medicine. Based on evidence from in vivo genetic lineage–tracing studies, pericytes have been identified as a source of MSC precursors in vivo in multiple organs, in response to injury or during homeostasis. Questions of intense debate and interest in the field of tissue engineering and regenerative studies include the following: 1) Are all pericytes, irrespective of tissue of isolation, equal in their differentiation potential? 2) What are the mechanisms that regulate the differentiation of MSCs? To gain a better understanding of the latter, recent work has utilized ChIP-seq (chromatin immunoprecipitation followed by sequencing) to reconstruct histone landscapes. This indicated that for dental pulp pericytes, the odontoblast-specific gene Dspp was found in a transcriptionally permissive state, while in bone marrow pericytes, the osteoblast-specific gene Runx2 was primed for expression. RNA sequencing has also been utilized to further characterize the 2 pericyte populations, and results highlighted that dental pulp pericytes are already precommitted to an odontoblast fate based on enrichment analysis indicating overrepresentation of key odontogenic genes. Furthermore, ChIP-seq analysis of the polycomb repressive complex 1 component RING1B indicated that this complex is likely to be involved in inhibiting inappropriate differentiation, as it localized to a number of loci of key transcription factors that are needed for the induction of adipogenesis, chondrogenesis, or myogenesis. In this review, we highlight recent data elucidating molecular mechanisms that indicate that pericytes can be tissue-specific precommitted MSC precursors in vivo and that this precommitment is a major driving force behind MSC differentiation.

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Fibroblast growth factor receptor-3 regulates Paneth cell lineage allocation and accrual of epithelial stem cells during murine intestinal development

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.90589.2008 ◽

2009 ◽

Vol 297 (1) ◽

pp. G168-G178 ◽

Cited By ~ 29

Author(s):

Alda Vidrich ◽

Jenny M. Buzan ◽

Brooks Brodrick ◽

Chibuzo Ilo ◽

Leigh Bradley ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Fibroblast Growth Factor Receptor ◽

Paneth Cell ◽

Cell Lineage ◽

Growth Factor Receptor ◽

Intestinal Development ◽

Fibroblast Growth ◽

Epithelial Stem Cells

Fibroblast growth factor receptor 3 (FGFR-3) is expressed in the lower crypt epithelium, where stem cells of the intestine reside. The role of FGFR-3 signaling in regulating features of intestinal morphogenesis was examined in FGFR-3-null (FGFR-3−/−) mice. FGFR-3−/− mice had only about half the number of intestinal crypts and a marked decrease in the number of functional clonogenic stem cells, as assessed by an in vivo microcolony-forming assay, compared with wild-type littermates. A marked deficit in allocation of progenitor cells to Paneth cell differentiation was noted, although all the principal epithelial lineages were represented in FGFR-3−/− mice. The total cellular content and nuclear localization of β-catenin protein were reduced in FGFR-3−/− mice, as was expression of cyclin D1 and matrix metalloproteinase-7, major downstream targets of β-catenin/T cell factor-4 (Tcf-4) signaling. Activation of FGFR-3 in Caco-2 cells, an intestinal epithelial cell line, abrogated the fall in β-catenin/Tcf-4 signaling activity that is normally observed in these cells as cultures become progressively more confluent. These findings are consistent with the hypothesis that, during intestinal development, FGFR-3 signaling regulates crypt epithelial stem cell expansion and crypt morphogenesis, as well as Paneth cell lineage specification, through β-catenin/Tcf-4-dependent and -independent pathways.

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The differentiation and transdifferentiation of epithelial cells in vitro – is it a new strategy in regenerative biomedicine?

Medical Journal of Cell Biology ◽

10.2478/acb-2018-0005 ◽

2018 ◽

Vol 6 (1) ◽

pp. 27-32 ◽

Cited By ~ 3

Author(s):

Maurycy Jankowski ◽

Marta Dyszkiewicz-Konwińska ◽

Joanna Budna ◽

Sandra Knap ◽

Artur Bryja ◽

...

Keyword(s):

Stem Cells ◽

Molecular Mechanisms ◽

Proliferative Potential ◽

Clinical Environment ◽

Epithelial Stem Cells ◽

Proliferation And Differentiation ◽

New Strategy ◽

Adult Organism ◽

The Moment

AbstractIn modern medical research, stem cells are one of the main focuses, believed to be able to provide the solution to many currently unsolvable medical cases. However, their extraordinary potential for differentiation creates much obstacles in their potential application in clinical environment, without understanding the whole array of molecular mechanisms that drive the processes associated with their development and maturation. Because of that, there is a large need for studies that concern the most basic levels of those processes. Progenitor stem cells are a favorable target, as they are relatively lineage committed, making the amount of signaling required to reach the final form much lower. Their presence in the adult organism is also an advantage in their potential use, as they can be extracted without the need for storage from the moment of pre-natal development or birth. Epithelial tissues, because of their usual location or function, exhibit extraordinary level of plasticity and proliferative potential. That fact makes them one of the top candidates for use in applications such as tissue engineering, cell based therapies, regenerative and reconstructive medicine. The potential clinical application, however, need to be based on well developed methods, in order to provide an effective treatment without causing major side effects. To achieve that goal, a large amount of research, aiming to analyze the molecular basics of proliferation and differentiation of epithelial stem cells, and stem cells in general, needs to be conducted.

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EGFR signaling promotes self-renewal through the establishment of cell polarity in Drosophila follicle stem cells

eLife ◽

10.7554/elife.04437 ◽

2014 ◽

Vol 3 ◽

Cited By ~ 29

Author(s):

Angela Castanieto ◽

Michael J Johnston ◽

Todd G Nystul

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Polarity ◽

Growth Factor Receptor ◽

Egfr Signaling ◽

Epithelial Stem Cells ◽

Cell Fates ◽

Liver Kinase B1 ◽

Follicle Stem Cells ◽

Drosophila Ovary

Epithelial stem cells divide asymmetrically, such that one daughter replenishes the stem cell pool and the other differentiates. We found that, in the epithelial follicle stem cell (FSC) lineage of the Drosophila ovary, epidermal growth factor receptor (EGFR) signaling functions specifically in the FSCs to promote the unique partially polarized state of the FSC, establish apical–basal polarity throughout the lineage, and promote FSC maintenance in the niche. In addition, we identified a novel connection between EGFR signaling and the cell-polarity regulator liver kinase B1 (LKB1), which indicates that EGFR signals through both the Ras–Raf–MEK–Erk pathway and through the LKB1–AMPK pathway to suppress apical identity. The development of apical–basal polarity is the earliest visible difference between FSCs and their daughters, and our findings demonstrate that the EGFR-mediated regulation of apical–basal polarity is essential for the segregation of stem cell and daughter cell fates.

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Tracing epithelial stem cells during development, homeostasis, and repair

The Journal of Cell Biology ◽

10.1083/jcb.201201041 ◽

2012 ◽

Vol 197 (5) ◽

pp. 575-584 ◽

Cited By ~ 50

Author(s):

Alexandra Van Keymeulen ◽

Cédric Blanpain

Keyword(s):

Stem Cells ◽

Tissue Regeneration ◽

External Environment ◽

Lineage Tracing ◽

The Body ◽

Genetic Lineage ◽

Epithelial Stem Cells ◽

Differentiation Potential ◽

Physiological Conditions ◽

Genetic Lineage Tracing

Epithelia ensure many critical functions of the body, including protection against the external environment, nutrition, respiration, and reproduction. Stem cells (SCs) located in the various epithelia ensure the homeostasis and repair of these tissues throughout the lifetime of the animal. Genetic lineage tracing in mice has allowed the labeling of SCs and their progeny. This technique has been instrumental in characterizing the origin and heterogeneity of epithelial SCs, their tissue location, and their differentiation potential under physiological conditions and during tissue regeneration.

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Bipotent stem cells support the cyclical regeneration of endometrial epithelium of the murine uterus

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1814597116 ◽

2019 ◽

Vol 116 (14) ◽

pp. 6848-6857 ◽

Cited By ~ 14

Author(s):

Shiying Jin

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Population ◽

Lineage Tracing ◽

Tissue Loss ◽

Stem Cell Population ◽

Epithelial Stem Cells ◽

Mouse Uterus ◽

Epithelial Regeneration ◽

Endometrial Epithelium

The endometrial epithelium of the uterus regenerates periodically. The cellular source of newly regenerated endometrial epithelia during a mouse estrous cycle or a human menstrual cycle is presently unknown. Here, I have used single-cell lineage tracing in the whole mouse uterus to demonstrate that epithelial stem cells exist in the mouse uterus. These uterine epithelial stem cells provide a resident cellular supply that fuels endometrial epithelial regeneration. They are able to survive cyclical uterine tissue loss and persistently generate all endometrial epithelial lineages, including the functionally distinct luminal and glandular epithelia, to maintain uterine cycling. The uterine epithelial stem cell population also supports the regeneration of uterine endometrial epithelium post parturition. The 5-ethynyl-2′-deoxyuridine pulse-chase experiments further reveal that this stem cell population may reside in the intersection zone between luminal and glandular epithelial compartments. This tissue distribution allows these bipotent uterine epithelial stem cells to bidirectionally differentiate to maintain homeostasis and regeneration of mouse endometrial epithelium under physiological conditions. Thus, uterine function over the reproductive lifespan of a mouse relies on stem cell-maintained rhythmic endometrial regeneration.

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IRF2 maintains the stemness of colonic stem cells by limiting physiological stress from interferon

Scientific Reports ◽

10.1038/s41598-020-71633-3 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 1

Author(s):

Kana Minamide ◽

Taku Sato ◽

Yusuke Nakanishi ◽

Hiroshi Ohno ◽

Tamotsu Kato ◽

...

Keyword(s):

Stem Cells ◽

Molecular Mechanisms ◽

Physiological Stress ◽

Poly I:C ◽

Type I ◽

Epithelial Stem Cells ◽

Hematopoietic Stem ◽

Microbial Dysbiosis ◽

Similar Phenotype ◽

Physiological Stressor

Abstract The physiological stresses that diminish tissue stem-cell characteristics remain largely unknown. We previously reported that type I interferon (IFN), which is essential for host antiviral responses, is a physiological stressor for hematopoietic stem cells (HSCs) and small intestinal stem cells (ISCs) and that interferon regulatory factor-2 (IRF2), which attenuates IFN signaling, maintains their stemness. Here, using a dextran sodium sulfate (DSS)-induced colitis model, we explore the role of IRF2 in maintaining colonic epithelial stem cells (CoSCs). In mice with a conditional Irf2 deletion in the intestinal epithelium (hereafter Irf2ΔIEC mice), both the number and the organoid-forming potential of CoSCs were markedly reduced. Consistent with this finding, the ability of Irf2ΔIEC mice to regenerate colon epithelium after inducing colitis was severely impaired, independently of microbial dysbiosis. Mechanistically, CoSCs differentiated prematurely into transit-amplifying (TA) cells in Irf2ΔIEC mice, which might explain their low CoSC counts. A similar phenotype was induced in wild-type mice by repeated injections of low doses of poly(I:C), which induces type I IFN. Collectively, we demonstrated that chronic IFN signaling physiologically stresses CoSCs. This study provides new insight into the development of colitis and molecular mechanisms that maintain functional CoSCs throughout life.

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Commentary: Lineage Tracing Supports Maintenance of the Corneal Epithelium by Limbal Epithelial Stem Cells

Stem Cells ◽

10.1002/stem.1892 ◽

2014 ◽

Vol 33 (1) ◽

pp. 310-311

Author(s):

John D. West

Keyword(s):

Stem Cells ◽

Corneal Epithelium ◽

Lineage Tracing ◽

Epithelial Stem Cells ◽

Limbal Epithelial Stem Cells

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Chemosensitivity of Patient-Derived Cancer Stem Cells Identifies Colorectal Cancer Patients with Potential Benefit from FGFR Inhibitor Therapy

Cancers ◽

10.3390/cancers12082010 ◽

2020 ◽

Vol 12 (8) ◽

pp. 2010

Author(s):

Takehito Yamamoto ◽

Hiroyuki Miyoshi ◽

Fumihiko Kakizaki ◽

Hisatsugu Maekawa ◽

Tadayoshi Yamaura ◽

...

Keyword(s):

Colorectal Cancer ◽

Stem Cells ◽

Cancer Patients ◽

Growth Factor Receptor ◽

Genetic Alterations ◽

Inhibitor Therapy ◽

Epithelial Stem Cells ◽

Colorectal Cancer Patients

Some colorectal cancer patients harboring FGFR (fibroblast growth factor receptor) genetic alterations, such as copy number gain, mutation, and/or mRNA overexpression, were selected for enrollment in several recent clinical trials of FGFR inhibitor, because these genetic alterations were preclinically reported to be associated with FGFR inhibitor sensitivity as well as poor prognosis, invasiveness, and/or metastatic potential. However, few enrolled patients were responsive to FGFR inhibitors. Thus, practical strategies are eagerly awaited that can stratify patients for the subset that potentially responds to FGFR inhibitor chemotherapy. In the present study, we evaluated the sensitivity to FGFR inhibitor erdafitinib on 25 patient-derived tumor-initiating cell (TIC) spheroid lines carrying wild-type RAS and RAF genes, both in vitro and in vivo. Then, we assessed possible correlations between the sensitivity and the genetic/genomic data of the spheroid lines tested. Upon their exposure to erdafitinib, seven lines (7/25, 28%) responded significantly. Normal colonic epithelial stem cells were unaffected by the inhibitors. Moreover, the combination of erdafitinib with EGFR inhibitor erlotinib showed stronger growth inhibition than either drug alone, as efficacy was observed in 21 lines (84%) including 14 (56%) that were insensitive to erdafitinib alone. The in vitro erdafitinib response was accurately reflected on mouse xenografts of TIC spheroid lines. However, we found little correlation between their genetic/genomic alterations of TIC spheroids and the sensitivity to the FGFR inhibitor. Accordingly, we propose that direct testing of the patient-derived spheroids in vitro is one of the most reliable personalized methods in FGFR-inhibitor therapy of colorectal cancer patients.

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