Loss of ADAM9 expression impairs β1 integrin endocytosis, focal adhesion formation and cancer cell migration

Mygind, Kasper Johansen; Schwarz, Jeanette; Sahgal, Pranshu; Ivaska, Johanna; Kveiborg, Marie

doi:10.1242/jcs.205393

Cited by 27 publications

(28 citation statements)

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“…Various reports point toward an impact of ADAM9 on cell adhesion (Cominetti et al ., ; Mygind et al ., ). Generally, (tumor) cell–matrix interaction is a crucial aspect of tumor biology, including in PDAC.…”

Section: Resultsmentioning

confidence: 97%

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ADAM9 contributes to vascular invasion in pancreatic ductal adenocarcinoma

et al. 2019

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A disintegrin and a metalloprotease (ADAM)‐9 is a metzincin cell‐surface protease with strongly elevated expression in solid tumors, including pancreatic ductal adenocarcinoma (PDAC). In this study, we performed immunohistochemistry (IHC) of a tissue microarray (TMA) to examine the expression of ADAM9 in a cohort of >100 clinically annotated PDAC cases. We report that ADAM9 is prominently expressed by PDAC tumor cells, and increased ADAM9 expression levels correlate with poor tumor grading (P = 0.027) and the presence of vasculature invasion (P = 0.017). We employed gene expression silencing to generate a loss‐of‐function system for ADAM9 in two established PDAC cell lines. In vitro analysis showed that loss of ADAM9 does not impede cellular proliferation and invasiveness in basement membrane. However, ADAM9 plays a crucial role in mediating cell migration and adhesion to extracellular matrix substrates such as fibronectin, tenascin, and vitronectin. This effect appears to depend on its catalytic activity. In addition, ADAM9 facilitates anchorage‐independent growth. In AsPC1 cells, but not in MiaPaCa‐2 cells, we noted a pronounced yet heterogeneous impact of ADAM9 on the abundance of various integrins, a process that we characterized as post‐translational regulation. Sprout formation of human umbilical vein endothelial cells (HUVECs) is promoted by ADAM9, as examined by transfer of cancer cell conditioned medium; this finding further supports a pro‐angiogenic role of ADAM9 expressed by PDAC cancer cells. Immunoblotting analysis of cancer cell conditioned medium highlighted that ADAM9 regulates the levels of angiogenic factors, including shed heparin‐binding EGF‐like growth factor (HB‐EGF). Finally, we carried out orthotopic seeding of either wild‐type AsPC‐1 cells or AsPC‐1 cells with silenced ADAM9 expression into murine pancreas. In this in vivo setting, ADAM9 was also found to foster angiogenesis without an impact on tumor cell proliferation. In summary, our results characterize ADAM9 as an important regulator in PDAC tumor biology with a strong pro‐angiogenic impact.

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Section: Resultsmentioning

confidence: 97%

“…At the same time, non‐catalytic functions of ADAM9 are equally important. For example, interaction of ADAM9 with β1 integrin may prevent integrin endocytosis, thereby increasing integrin surface expression (Mygind et al ., ).…”

Section: Introductionmentioning

confidence: 97%

ADAM9 contributes to vascular invasion in pancreatic ductal adenocarcinoma

et al. 2019

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“…ADAMs have been shown to regulate receptor signaling through a variety of mechanisms many of which involve the catalytically active MP domain (27, 28, 30, 31, 33, 35, 42, 63-65). This raises the possibility that ADAM9 may function as a cofactor for EMCV infection.…”

Section: Discussionmentioning

confidence: 99%

Identification of A Disintegrin and Metalloproteinase 9 domain (ADAM9) required in the early stages of encephalomyocarditis virus infection

King

et al. 2018

Preprint

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word count: 165 words 21 Main text word count: 5,097 words 22 Abstract 23 Encephalomyocarditis virus (EMCV) is a picornavirus that produces lytic 24 infections in murine and human cells. Employing a genome-wide CRISPR-Cas9 25 knockout screen to find host factors required for EMCV infection, we identified a role for 26 ADAM9 in EMCV infection. CRISPR-mediated deletion of ADAM9 in multiple human cell 27 lines rendered the cells highly resistant to EMCV infection and cell death. Primary 28 fibroblasts from ADAM9 KO mice were also strongly resistant to EMCV infection and cell 29 death. In contrast, ADAM9 KO and WT cells were equally susceptible to infection with 30 other viruses, including the picornavirus Coxsackie virus B. ADAM9 KO cells failed to 31 produce viral progeny when incubated with EMCV. However, bypassing EMCV entry into 32 cells through delivery of viral RNA directly to the cytosol yielded infectious EMCV virions 33 from ADAM9 KO cells, suggesting that ADAM9 is not required for EMCV replication 34 post-entry. These findings establish that ADAM9 is required for the early stage of EMCV 35 infection, likely for virus entry or viral genome delivery to the cytosol. 36 Importance 37 Viral myocarditis is a leading cause of death in the U.S., contributing to 38 numerous unexplained deaths in people ≤ 35 years old. Enteroviruses contribute to 39 many cases of human myocarditis. Encephalomyocarditis virus (EMCV) infection causes 40 viral myocarditis in rodent models but its receptor requirements have not been fully 41 identified. CRISPR-Cas9 screens can identify host dependency factors essential for 42 EMCV infection and enhance our understanding of key events that follow viral infection, 43 potentially leading to new strategies for preventing viral myocarditis. Using a CRISPR-44 Cas9 screen, we identified A Disintegrin and Metalloproteinase 9 Domain (ADAM9) as a 45 major factor required for the early stages of EMCV infection in both human and murine 46 infection. 47 Results 90 CRISPR-Cas9 Screening Identifies EMCV Dependency Factors (EDFs) 91EMCV infection is rapidly lytic in human and murine cells (51-54). We took advantage of 92 the high lytic potential of EMCV and the power of CRISPR genetic screening (53, 55) to 93 discover virus-host interaction genes that mediated virus infection and, thus, rendered 94 the cells susceptible to EMCV-induced cell death. HeLa cells stably expressing Cas9 95 were used for screening (53, 55). In initial optimization experiments, we determined that 96 HeLa cells were killed by EMCV within 24 h of infection at a multiplicity of infection (MOI) 97 ≥ 0.1. The rapid lysis of HeLa cells with EMCV infection allowed us to screen for EDFs 98 using pooled single-guide RNAs (sgRNAs) since we could identify such mutant cells by 99 their resistance to EMCV-induced cell death, i.e., these mutants would no longer be 100 susceptible to EMCV infection and would survive EMCV challenge. 101We screened for EDFs using CRISPR-Cas9 pooled human gene screen (Fig. 1). 102 H1-HeLa cells which stably exp...

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“…In contrast, the direct interaction between ADAM9 and integrin β1 has been confirmed by co-immunoprecipitation, and both proteins co-localize on early endosomes in vitro. These findings suggest ADAM9 mediates integrin β1 stability in a non-catalytic manner [ 78 ].…”

Section: Role Of Adam9 In Cancersmentioning

confidence: 99%

An Overview of ADAM9: Structure, Activation, and Regulation in Human Diseases

Chou

Huang

Kuo

et al. 2020

IJMS

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ADAM9 (A disintegrin and a metalloprotease 9) is a membrane-anchored protein that participates in a variety of physiological functions, primarily through the disintegrin domain for adhesion and the metalloprotease domain for ectodomain shedding of a wide variety of cell surface proteins. ADAM9 influences the developmental process, inflammation, and degenerative diseases. Recently, increasing evidence has shown that ADAM9 plays an important role in tumor biology. Overexpression of ADAM9 has been found in several cancer types and is correlated with tumor aggressiveness and poor prognosis. In addition, through either proteolytic or non-proteolytic pathways, ADAM9 promotes tumor progression, therapeutic resistance, and metastasis of cancers. Therefore, comprehensively understanding the mechanism of ADAM9 is crucial for the development of therapeutic anti-cancer strategies. In this review, we summarize the current understanding of ADAM9 in biological function, pathophysiological diseases, and various cancers. Recent advances in therapeutic strategies using ADAM9-related pathways are presented as well.

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Loss of ADAM9 expression impairs β1 integrin endocytosis, focal adhesion formation and cancer cell migration

Cited by 27 publications

References 50 publications

ADAM9 contributes to vascular invasion in pancreatic ductal adenocarcinoma

ADAM9 contributes to vascular invasion in pancreatic ductal adenocarcinoma

Identification of A Disintegrin and Metalloproteinase 9 domain (ADAM9) required in the early stages of encephalomyocarditis virus infection

An Overview of ADAM9: Structure, Activation, and Regulation in Human Diseases

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