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Fibroblasts Direct Differentiation Of Human Breast Epithelial Progenitors

Mikkel Morsing, J. Kim, R. Villadsen, Nadine Goldhammer, A. Jafari, M. Kassem, O. W. Petersen, L. Rønnov-Jessen
Published 2020 · Biology, Medicine

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Background Breast cancer arises within specific regions in the human breast referred to as the terminal duct lobular units (TDLUs). These are relatively dynamic structures characterized by sex hormone driven cyclic epithelial turnover. TDLUs consist of unique parenchymal entities embedded within a fibroblast-rich lobular stroma. Here, we established and characterized a new human breast lobular fibroblast cell line against its interlobular counterpart with a view to assessing the role of region-specific stromal cues in the control of TDLU dynamics. Methods Primary lobular and interlobular fibroblasts were transduced to express human telomerase reverse transcriptase (hTERT). Differentiation of the established cell lines along lobular and interlobular pathways was determined by immunocytochemical staining and genome-wide RNA sequencing. Their functional properties were further characterized by analysis of mesenchymal stem cell (MSC) differentiation repertoire in culture and in vivo. The cells’ physiological relevance for parenchymal differentiation was examined in heterotypic co-culture with fluorescence-activated cell sorting (FACS)-purified normal breast primary luminal or myoepithelial progenitors. The co-cultures were immunostained for quantitative assessment of epithelial branching morphogenesis, polarization, growth, and luminal epithelial maturation. In extension, myoepithelial progenitors were tested for luminal differentiation capacity in culture and in mouse xenografts. To unravel the significance of transforming growth factor-beta (TGF-β)-mediated crosstalk in TDLU-like morphogenesis and differentiation, fibroblasts were incubated with the TGF-β signaling inhibitor, SB431542, prior to heterotypic co-culture with luminal cells. Results hTERT immortalized fibroblast cell lines retained critical phenotypic traits in culture and linked to primary fibroblasts. Cell culture assays and transplantation to mice showed that the origin of fibroblasts determines TDLU-like and ductal-like differentiation of epithelial progenitors. Whereas lobular fibroblasts supported a high level of branching morphogenesis by luminal cells, interlobular fibroblasts supported ductal-like myoepithelial characteristics. TDLU-like morphogenesis, at least in part, relied on intact TGF-β signaling. Conclusions The significance of the most prominent cell type in normal breast stroma, the fibroblast, in directing epithelial differentiation is largely unknown. Through establishment of lobular and interlobular fibroblast cell lines, we here demonstrate that epithelial progenitors are submitted to stromal cues for site-specific differentiation. Our findings lend credence to considering stromal subtleties of crucial importance in the development of normal breast and, in turn, breast cancer.
This paper references
Defining Skin Fibroblastic Cell Types Beyond CD90
Dongsheng Jiang (2018)
RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome
Bo Li (2011)
Legumain Regulates Differentiation Fate of Human Bone Marrow Stromal Cells and Is Altered in Postmenopausal Osteoporosis
A. Jafari (2017)
Aldehyde dehydrogenase and estrogen receptor define a hierarchy of cellular differentiation in the normal human mammary epithelium
G. Honeth (2013)
Targeting the cancer-associated fibroblasts as a treatment in triple-negative breast cancer
Ken Takai (2016)
Preservation of defined phenotypic traits in short-term cultured human breast carcinoma derived epithelial cells.
O. W. Petersen (1987)
Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers
E. Lim (2009)
Diversity, topographic differentiation, and positional memory in human fibroblasts
H. Chang (2002)
The origin of the myofibroblasts in breast cancer. Recapitulation of tumor environment in culture unravels diversity and implicates converted fibroblasts and recruited smooth muscle cells.
L. Rønnov-Jessen (1995)
A method for quantifying normal human mammary epithelial stem cells with in vivo regenerative ability
P. Eirew (2008)
Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2
M. Love (2014)
CAF Subpopulations: A New Reservoir of Stromal Targets in Pancreatic Cancer.
B. Pereira (2019)
Identification and isolation of a dermal lineage with intrinsic fibrogenic potential
Yuval Rinkevich (2015)
Syndecan-1 expression is induced in the stroma of infiltrating breast carcinoma.
M. Stanley (1999)
Carcinoma-associated fibroblast-like differentiation of human mesenchymal stem cells.
Pravin J Mishra (2008)
CD36 repression activates a multicellular stromal program shared by high mammographic density and tumor tissues.
R. DeFilippis (2012)
RIPK1 prevents aberrant ZBP1-initiated necroptosis
T. Berghe (2017)
Dipeptidyl peptidase iv expression identifies a functional sub‐population of breast fibroblasts
A. Atherton (1992)
CD146/MCAM defines functionality of human bone marrow stromal stem cell populations
L. Harkness (2015)
Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line.
J. Morgenstern (1990)
Rapid Research Autopsy: Piecing the Puzzle of Tumor Heterogeneity.
M. Krook (2019)
Modelling breast cancer requires identification and correction of a critical cell lineage-dependent transduction bias
W. Hines (2015)
Dissociation among in vitro telomerase activity, telomere maintenance, and cellular immortalization.
C. M. Counter (1998)
Single-cell analysis reveals fibroblast clusters linked to immunotherapy resistance in cancer.
Y. Kieffer (2020)
There is more than one kind of myofibroblast: analysis of CD34 expression in benign, in situ, and invasive breast lesions
H. Chauhan (2003)
Proof of region-specific multipotent progenitors in human breast epithelia
Agla J. Fridriksdottir (2017)
Establishment of a normal-derived estrogen receptor-positive cell line comparable to the prevailing human breast cancer subtype
Branden M. Hopkinson (2017)
Targeting the Interplay Between Cancer Fibroblasts, Mesenchymal Stem Cells, and Cancer Stem Cells in Desmoplastic Cancers
T. Chan (2019)
The Role of the TGF-β Coreceptor Endoglin in Cancer
E. Pérez-Gómez (2010)
Tumor Necrosis Factor Receptor Superfamily Member 19 (TNFRSF19) Regulates Differentiation Fate of Human Mesenchymal (Stromal) Stem Cells through Canonical Wnt Signaling and C/EBP*
W. Qiu (2010)
TAFA2 Induces Skeletal (Stromal) Stem Cell Migration Through Activation of Rac1‐p38 Signaling
A. Jafari (2019)
Pro-Fibrotic CD26-Positive Fibroblasts are Present in Greater Abundance in Breast Capsule Tissue of Irradiated Breasts.
M. Borrelli (2019)
Pharmacological Inhibition of Protein Kinase G1 Enhances Bone Formation by Human Skeletal Stem Cells Through Activation of RhoA‐Akt Signaling
A. Jafari (2015)
Development of the human breast.
J. Russo (2004)
Ectoenzyme regulation by phenotypically distinct fibroblast sub-populations isolated from the human mammary gland.
A. Atherton (1994)
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
An atlas of subgross pathology of the human breast with special reference to possible precancerous lesions.
S. Wellings (1975)
Mesenchymal stem cells within tumour stroma promote breast cancer metastasis
A. Karnoub (2007)
Identification, paracrine generation, and possible function of human breast carcinoma myofibroblasts in culture
L. Rønnov-Jessen (2007)
Fast gapped-read alignment with Bowtie 2
B. Langmead (2012)
Evidence for a stem cell hierarchy in the adult human breast
R. Villadsen (2007)
Differential expression of type XIV collagen/undulin by human mammary gland intralobular and interlobular fibroblasts
A. Atherton (1998)
CD36 repression activated a multicellular stromal program shared by high mammagraphic density and tumor tissues
RA DeFilippis (2012)
Evidence of two distinct functionally specialized fibroblast lineages in breast stroma
Mikkel Morsing (2016)
Lineage Identity and Location within the Dermis Determine the Function of Papillary and Reticular Fibroblasts in Human Skin.
Ana Korosec (2019)
Publisher's Note
H. V. D. Togt (2003)
Elevated CD26 Expression by Skin Fibroblasts Distinguishes a Profibrotic Phenotype Involved in Scar Formation Compared to Gingival Fibroblasts.
Wesley Mah (2017)
Identification of fibroblast heterogeneity in the tumor microenvironment
H. Sugimoto (2006)
Endosialin (TEM1, CD248) is a marker of stromal fibroblasts and is not selectively expressed on tumour endothelium
John R MacFadyen (2005)
Change in Sympathetic Nerve Firing Pattern Associated with Dietary Weight Loss in the Metabolic Syndrome
E. Lambert (2011)
21-Gene Recurrence Score Assay and Outcomes of Adjuvant Radiotherapy in Elderly Women With Early-Stage Breast Cancer After Breast-Conserving Surgery
S. Wu (2019)
Love MI, Huber W, Anders S.. Moderated estimation of fold change and dispersion for RNA-Seq data with DESeq2. Genome Biol 15: 550
M. Love (2014)
Derivation of stromal (skeletal and mesenchymal) stem-like cells from human embryonic stem cells.
Amer Mahmood (2012)
Interlobular and intralobular mammary stroma: Genotype may not reflect phenotype
J. Fleming (2008)
The Stromal Niche for Epithelial Stem Cells: A Template for Regeneration and a Brake on Malignancy.
Kelsey J. Roberts (2017)
Mammary stem cells have myoepithelial cell properties
Michael D. Prater (2014)
Turning foes to friends: targeting cancer-associated fibroblasts
Xueman Chen (2018)
Trafficking mechanism of bone marrow-derived mesenchymal stem cells toward hepatocellular carcinoma HepG2 cells by modulating Endoglin, CXCR4 and TGF-β.
A. Mardomi (2016)
A fibroblast-associated antigen: characterization in fibroblasts and immunoreactivity in smooth muscle differentiated stromal cells.
L. Rønnov-Jessen (1992)
CD26 expression is attenuated by TGF‐β and SDF‐1 autocrine signaling on stromal myofibroblasts in human breast cancers
Yoshihiro Mezawa (2019)
Distinct fibroblast lineages determine dermal architecture in skin development and repair
R. R. Driskell (2013)
Transforming Growth Factor-β and Endoglin Signaling Orchestrate Wound Healing
M. Valluru (2011)
Spatial and Single-Cell Transcriptional Profiling Identifies Functionally Distinct Human Dermal Fibroblast Subpopulations
C. Philippeos (2018)
Stromal directives can control cancer
T. Tlsty (2019)
Induction of alpha-smooth muscle actin by transforming growth factor-beta 1 in quiescent human breast gland fibroblasts. Implications for myofibroblast generation in breast neoplasia.
L. Rønnov-Jessen (1993)
The pCL vector system: rapid production of helper-free, high-titer, recombinant retroviruses.
R. Naviaux (1996)
Anatomic Demarcation by Positional Variation in Fibroblast Gene Expression Programs
John L. Rinn (2006)
Human bone marrow-derived MSCs can home to orthotopic breast cancer tumors and promote bone metastasis.
R. Goldstein (2010)
Disequilibrium of BMP2 Levels in the Breast Stem Cell Niche Launches Epithelial Transformation by Overamplifying BMPR1B Cell Response
M. Chapellier (2015)
Reconstruction of functionally normal and malignant human breast tissues in mice.
C. Kuperwasser (2004)
The Comparative Pathology of Human and Mouse Mammary Glands
R. Cardiff (2004)
Stromal cell diversity associated with immune evasion in human triple‐negative breast cancer
Sunny Z. Wu (2020)
Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells
J. L. Simonsen (2002)

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