Hyaluronan modulates growth factor induced mammary gland branching in a size dependent manner

Cornelia Tolg; Han Yuan; Sarah M. Flynn; Kaustuv Basu; Jenny Ma; Kenneth Chor Kin Tse; Beatrice Kowalska; Diana Vulkanesku; Mary K. Cowman; James B. McCarthy; Eva A. Turley

doi:10.1016/j.matbio.2017.02.003

Hyaluronan modulates growth factor induced mammary gland branching in a size dependent manner

Cornelia Tolg, Han Yuan, Sarah M. Flynn, Kaustuv Basu, Jenny Ma, Kenneth Chor Kin Tse, Beatrice Kowalska, Diana Vulkanesku, Mary K. Cowman, James B. McCarthy, Eva A. Turley

Laboratory Medicine and Pathology

Research output: Contribution to journal › Article › peer-review

47 Scopus citations

Abstract

Mammary gland morphogenesis begins during fetal development but expansion of the mammary tree occurs postnatally in response to hormones, growth factors and extracellular matrix. Hyaluronan (HA) is an extracellular matrix polysaccharide that has been shown to modulate growth factor-induced branching in culture. Neither the physiological relevance of HA to mammary gland morphogenesis nor the role that HA receptors play in these responses are currently well understood. We show that HA synthase (HAS2) is expressed in both ductal epithelia and stromal cells but HA primarily accumulates in the stroma. HA accumulation and expression of the HA receptors CD44 and RHAMM are highest during gestation when gland remodeling, lateral branch infilling and lobulo-alveoli formation is active. Molecular weight analyses show that approximately 98% of HA at all stages of morphogenesis is > 300 kDa. Low levels of 7–114 kDa HA fragments are also detected and in particular the accumulation of 7–21 kDa HA fragments are significantly higher during gestation than other morphogenetic stages (p < 0.05). Using these in vivo results as a guide, in culture analyses of mammary epithelial cell lines (EpH4 and NMuMG) were performed to determine the roles of high molecular weight, 7–21 kDa (10 kDa MWavg) and HA receptors in EGF-induced branching morphogenesis. Results of these assays show that while HA synthesis is required for branching and 10 kDa HA fragments strongly stimulate branching, the activity of HA decreases with increasing molecular weight and 500 kDa HA strongly inhibits this morphogenetic process. The response to 10 kDa HA requires RHAMM function and genetic deletion of RHAMM transiently blunts lateral branching in vivo. Collectively, these results reveal distinct roles for HA polymer size in modulating growth factor induced mammary gland branching and implicates these polymers in both the expansion and sculpting of the mammary tree during gestation.

Original language	English (US)
Pages (from-to)	117-132
Number of pages	16
Journal	Matrix Biology
Volume	63
DOIs	https://doi.org/10.1016/j.matbio.2017.02.003
State	Published - Nov 2017

Bibliographical note

Funding Information:
This work was supported by a CBCF grant to ET, The Endre A. Balazs Foundation to MC and the Chairman's Fund in Cancer Research and the W. Allen Elsa Pardee Foundation to JBM. We are grateful to Mina J. Bissell for much advice on this project.

Publisher Copyright:
© 2017 Elsevier B.V.

Keywords

Branching morphogenesis
CD44
Hyaluronan
Hyaluronan fragments
Mammary gland
RHAMM

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title = "Hyaluronan modulates growth factor induced mammary gland branching in a size dependent manner",

abstract = "Mammary gland morphogenesis begins during fetal development but expansion of the mammary tree occurs postnatally in response to hormones, growth factors and extracellular matrix. Hyaluronan (HA) is an extracellular matrix polysaccharide that has been shown to modulate growth factor-induced branching in culture. Neither the physiological relevance of HA to mammary gland morphogenesis nor the role that HA receptors play in these responses are currently well understood. We show that HA synthase (HAS2) is expressed in both ductal epithelia and stromal cells but HA primarily accumulates in the stroma. HA accumulation and expression of the HA receptors CD44 and RHAMM are highest during gestation when gland remodeling, lateral branch infilling and lobulo-alveoli formation is active. Molecular weight analyses show that approximately 98% of HA at all stages of morphogenesis is > 300 kDa. Low levels of 7–114 kDa HA fragments are also detected and in particular the accumulation of 7–21 kDa HA fragments are significantly higher during gestation than other morphogenetic stages (p < 0.05). Using these in vivo results as a guide, in culture analyses of mammary epithelial cell lines (EpH4 and NMuMG) were performed to determine the roles of high molecular weight, 7–21 kDa (10 kDa MWavg) and HA receptors in EGF-induced branching morphogenesis. Results of these assays show that while HA synthesis is required for branching and 10 kDa HA fragments strongly stimulate branching, the activity of HA decreases with increasing molecular weight and 500 kDa HA strongly inhibits this morphogenetic process. The response to 10 kDa HA requires RHAMM function and genetic deletion of RHAMM transiently blunts lateral branching in vivo. Collectively, these results reveal distinct roles for HA polymer size in modulating growth factor induced mammary gland branching and implicates these polymers in both the expansion and sculpting of the mammary tree during gestation.",

keywords = "Branching morphogenesis, CD44, Hyaluronan, Hyaluronan fragments, Mammary gland, RHAMM",

author = "Cornelia Tolg and Han Yuan and Flynn, {Sarah M.} and Kaustuv Basu and Jenny Ma and Tse, {Kenneth Chor Kin} and Beatrice Kowalska and Diana Vulkanesku and Cowman, {Mary K.} and McCarthy, {James B.} and Turley, {Eva A.}",

note = "Funding Information: This work was supported by a CBCF grant to ET, The Endre A. Balazs Foundation to MC and the Chairman's Fund in Cancer Research and the W. Allen Elsa Pardee Foundation to JBM. We are grateful to Mina J. Bissell for much advice on this project. Publisher Copyright: {\textcopyright} 2017 Elsevier B.V.",

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AU - Vulkanesku, Diana

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AU - Turley, Eva A.

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N2 - Mammary gland morphogenesis begins during fetal development but expansion of the mammary tree occurs postnatally in response to hormones, growth factors and extracellular matrix. Hyaluronan (HA) is an extracellular matrix polysaccharide that has been shown to modulate growth factor-induced branching in culture. Neither the physiological relevance of HA to mammary gland morphogenesis nor the role that HA receptors play in these responses are currently well understood. We show that HA synthase (HAS2) is expressed in both ductal epithelia and stromal cells but HA primarily accumulates in the stroma. HA accumulation and expression of the HA receptors CD44 and RHAMM are highest during gestation when gland remodeling, lateral branch infilling and lobulo-alveoli formation is active. Molecular weight analyses show that approximately 98% of HA at all stages of morphogenesis is > 300 kDa. Low levels of 7–114 kDa HA fragments are also detected and in particular the accumulation of 7–21 kDa HA fragments are significantly higher during gestation than other morphogenetic stages (p < 0.05). Using these in vivo results as a guide, in culture analyses of mammary epithelial cell lines (EpH4 and NMuMG) were performed to determine the roles of high molecular weight, 7–21 kDa (10 kDa MWavg) and HA receptors in EGF-induced branching morphogenesis. Results of these assays show that while HA synthesis is required for branching and 10 kDa HA fragments strongly stimulate branching, the activity of HA decreases with increasing molecular weight and 500 kDa HA strongly inhibits this morphogenetic process. The response to 10 kDa HA requires RHAMM function and genetic deletion of RHAMM transiently blunts lateral branching in vivo. Collectively, these results reveal distinct roles for HA polymer size in modulating growth factor induced mammary gland branching and implicates these polymers in both the expansion and sculpting of the mammary tree during gestation.

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Hyaluronan modulates growth factor induced mammary gland branching in a size dependent manner

Abstract

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Keywords

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