NHERF1/EBP50 controls morphogenesis of 3D colonic glands by stabilizing PTEN and ezrin-radixin-moesin proteins at the apical membrane

Maria Magdalena Georgescu, Gilbert Cote, Nitin Kumar Agarwal, Charles L. White

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Na+/H+ exchanger 3 regulating factor 1/ezrin-radixin-moesin (ERM)-binding phosphoprotein 50 (NHERF1/EBP50), an adaptor molecule that interacts with the ERM-neurofibromatosis type 2 family of cytoskeletal proteins through its ERM-binding region and with phosphatase and tensin homolog (PTEN) and β-catenin through its PDZ domains, has been recently implicated in the progression of various human malignancies, including colorectal cancer (CRC). We report here that NHERF1 controls gland morphogenesis, as demonstrated in three-dimensional (3D) human intestinal glands developing from a single nonpolarized cell. Starting from the early two-cell developmental stage, NHERF1 concentrates at the cellular interface in a central membrane disc that marks the apical pole delimiting the forming lumen. NHERF1 depletion leads to severe disruption of the apical-basal polarity,with formation of enlarged and distorted cell spheroids devoid of a central lumen. This characteristic and the increased number of mitoses in NHERF1-depleted spheroids, including multipolar ones, mimic high-grade dysplasia lesions observed in CRC progression. NHERF1 ERM-binding or PDZ-domain mutants fail to localize apically and impair gland formation most likely by outcompeting endogenous ligands, with the latter mutant completely aborting gland development. Examination of NHERF1 ligands showed that even if both ezrin andmoesin colocalized with NHERF1at the apicalmembrane,moesin but not ezrin depletion disrupted morphogenesis similarly to NHERF1. NHERF1 depletion resulted also in membrane displacement of PTEN and nuclear translocation of β-catenin, events contributing to polarity loss and increased proliferation. These findings reveal an essential role of NHERF1 in epithelialmorphogenesis and polarity and validate this 3D system for modeling the molecular changes observed in CRC.

Original languageEnglish (US)
Pages (from-to)365-374
Number of pages10
JournalNeoplasia (United States)
Volume16
Issue number4
DOIs
StatePublished - 2014

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Sodium-Hydrogen Antiporter
Phosphoproteins
Morphogenesis
Phosphoric Monoester Hydrolases
Membranes
PDZ Domains
Catenins
Colorectal Neoplasms
Proteins
Neurofibromatosis 2
Ligands
Cytoskeletal Proteins
Intestinal Mucosa
Mitosis
moesin
radixin
Tensins
ezrin
Neoplasms

ASJC Scopus subject areas

  • Cancer Research

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NHERF1/EBP50 controls morphogenesis of 3D colonic glands by stabilizing PTEN and ezrin-radixin-moesin proteins at the apical membrane. / Georgescu, Maria Magdalena; Cote, Gilbert; Agarwal, Nitin Kumar; White, Charles L.

In: Neoplasia (United States), Vol. 16, No. 4, 2014, p. 365-374.

Research output: Contribution to journalArticle

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abstract = "Na+/H+ exchanger 3 regulating factor 1/ezrin-radixin-moesin (ERM)-binding phosphoprotein 50 (NHERF1/EBP50), an adaptor molecule that interacts with the ERM-neurofibromatosis type 2 family of cytoskeletal proteins through its ERM-binding region and with phosphatase and tensin homolog (PTEN) and β-catenin through its PDZ domains, has been recently implicated in the progression of various human malignancies, including colorectal cancer (CRC). We report here that NHERF1 controls gland morphogenesis, as demonstrated in three-dimensional (3D) human intestinal glands developing from a single nonpolarized cell. Starting from the early two-cell developmental stage, NHERF1 concentrates at the cellular interface in a central membrane disc that marks the apical pole delimiting the forming lumen. NHERF1 depletion leads to severe disruption of the apical-basal polarity,with formation of enlarged and distorted cell spheroids devoid of a central lumen. This characteristic and the increased number of mitoses in NHERF1-depleted spheroids, including multipolar ones, mimic high-grade dysplasia lesions observed in CRC progression. NHERF1 ERM-binding or PDZ-domain mutants fail to localize apically and impair gland formation most likely by outcompeting endogenous ligands, with the latter mutant completely aborting gland development. Examination of NHERF1 ligands showed that even if both ezrin andmoesin colocalized with NHERF1at the apicalmembrane,moesin but not ezrin depletion disrupted morphogenesis similarly to NHERF1. NHERF1 depletion resulted also in membrane displacement of PTEN and nuclear translocation of β-catenin, events contributing to polarity loss and increased proliferation. These findings reveal an essential role of NHERF1 in epithelialmorphogenesis and polarity and validate this 3D system for modeling the molecular changes observed in CRC.",
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