MT2-MMP-dependent release of collagen IV NC1 domains regulates submandibular gland branching morphogenesis

TY - JOUR

T1 - MT2-MMP-dependent release of collagen IV NC1 domains regulates submandibular gland branching morphogenesis

AU - Rebustini, Ivan T

AU - Myers, Christopher

AU - Lassiter, Keyonica S

AU - Surmak, Andrew

AU - Szabova, Ludmila

AU - Holmbeck, Kenn

AU - Pedchenko, Vadim

AU - Hudson, Billy G

AU - Hoffman, Matthew P

PY - 2009/10

Y1 - 2009/10

N2 - Proteolysis is essential during branching morphogenesis, but the roles of MT-MMPs and their proteolytic products are not clearly understood. Here, we discover that decreasing MT-MMP activity during submandibular gland branching morphogenesis decreases proliferation and increases collagen IV and MT-MMP expression. Specifically, reducing epithelial MT2-MMP profoundly decreases proliferation and morphogenesis, increases Col4a2 and intracellular accumulation of collagen IV, and decreases the proteolytic release of collagen IV NC1 domains. Importantly, we demonstrate the presence of collagen IV NC1 domains in developing tissue. Furthermore, recombinant collagen IV NC1 domains rescue branching morphogenesis after MT2-siRNA treatment, increasing MT-MMP and proproliferative gene expression via beta1 integrin and PI3K-AKT signaling. Additionally, HBEGF also rescues MT2-siRNA treatment, increasing NC1 domain release, proliferation, and MT2-MMP and Hbegf expression. Our studies provide mechanistic insight into how MT2-MMP-dependent release of bioactive NC1 domains from collagen IV is critical for integrating collagen IV synthesis and proteolysis with epithelial proliferation during branching morphogenesis.

AB - Proteolysis is essential during branching morphogenesis, but the roles of MT-MMPs and their proteolytic products are not clearly understood. Here, we discover that decreasing MT-MMP activity during submandibular gland branching morphogenesis decreases proliferation and increases collagen IV and MT-MMP expression. Specifically, reducing epithelial MT2-MMP profoundly decreases proliferation and morphogenesis, increases Col4a2 and intracellular accumulation of collagen IV, and decreases the proteolytic release of collagen IV NC1 domains. Importantly, we demonstrate the presence of collagen IV NC1 domains in developing tissue. Furthermore, recombinant collagen IV NC1 domains rescue branching morphogenesis after MT2-siRNA treatment, increasing MT-MMP and proproliferative gene expression via beta1 integrin and PI3K-AKT signaling. Additionally, HBEGF also rescues MT2-siRNA treatment, increasing NC1 domain release, proliferation, and MT2-MMP and Hbegf expression. Our studies provide mechanistic insight into how MT2-MMP-dependent release of bioactive NC1 domains from collagen IV is critical for integrating collagen IV synthesis and proteolysis with epithelial proliferation during branching morphogenesis.

KW - Animals

KW - Blotting, Western

KW - Cell Proliferation

KW - Collagen Type IV/metabolism

KW - Epithelial Cells/metabolism

KW - Fluorescent Antibody Technique

KW - Heparin/metabolism

KW - Heparin-binding EGF-like Growth Factor

KW - Immunoprecipitation

KW - Integrin beta1/genetics

KW - Intercellular Signaling Peptides and Proteins/genetics

KW - Matrix Metalloproteinase 14/physiology

KW - Matrix Metalloproteinase 15/genetics

KW - Matrix Metalloproteinase Inhibitors

KW - Mice

KW - Mice, Knockout

KW - Morphogenesis

KW - Phosphatidylinositol 3-Kinases/genetics

KW - Proto-Oncogene Proteins c-akt/genetics

KW - RNA, Messenger/genetics

KW - RNA, Small Interfering/pharmacology

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Submandibular Gland/growth & development

U2 - 10.1016/j.devcel.2009.07.016

DO - 10.1016/j.devcel.2009.07.016

M3 - Journal article

C2 - 19853562

SN - 1534-5807

VL - 17

SP - 482

EP - 493

JO - Developmental Cell

JF - Developmental Cell

IS - 4

ER -