Proteolytic Activation of Pro-Macrophage-Stimulating Protein by Hepsin

Ganesan, Rajkumar; Kolumam, Ganesh; Lin, Shuei Liong; Xie, Ming-Hong; Santell, Lydia; Wu, Thomas D.; Lazarus, Robert A.; Chaudhuri, Amitabha; Kirchhofer, Daniel

doi:10.1158/1541-7786.mcr-11-0004

Cited by 64 publications

(54 citation statements)

References 56 publications

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Section: Discussionmentioning

confidence: 99%

“…We next tested the selectivity of KD1-KD2/1-Fc against a panel of naturally soluble or cell anchored serine proteases, including trypsin 3 (51), urokinase (11), kallikrein 4 (30), and hepsin (28,29); each of these proteases have a range of native affinities to wild-type HAI-1. We found that none of the proteins tested could inhibit trypsin 3 or urokinase activity (Fig.…”

Section: Kd1-kd2/1-fc Is a Potent And Selective Inhibitor Of Matriptasementioning

confidence: 99%

“…Like matriptase, the relative fold difference in Ki values for hepsin between KD1-KD2/1-Fc and wild-type KD1 monomer is also 4-fold, which further supports the stoichiometric inhibition hypothesis stated above. Hepsin is reported to share a similar role as matriptase in cancer progression, and thus dual targeting of hepsin and matriptase by KD1-KD2/1-Fc may serve as an attractive therapeutic feature (28,29,35).Cell based activity assays further demonstrate the superior potency of KD1-KD2/1-Fc in inhibiting matriptase activity both soluble and cell associated form. The huPro-HGF activation assay qualitatively confirmed that KD1-KD2/1-Fc inhibits matriptase activation of the pro-domain form of HGF, with reduced MDCK cell scattering at lower inhibitor concentrations than wild-type KD1 monomer (Fig.…”

mentioning

confidence: 92%

“…1A). HAI-1 is primarily expressed on the surface of epithelial cells and naturally blocks the substrate-activating properties of matriptase (24)(25)(26), as well as other structurally related proteases such as the hepatocyte growth factor activator (HGFA) (9,27), hepsin (28,29), and kallakrein-4/5 (9,30). The balance between substrate activation and protease inhibition is critical to the metastatic potential of tumor cells (Fig.…”

Section: Introductionmentioning

confidence: 99%

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Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Mitchell

Kannan

Hunter

et al. 2018

Journal of Biological Chemistry

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Abstract:Dysregulated matriptase activity has been established as a key contributor to cancer progression through its activation of growth factors including the hepatocyte growth factor (HGF). Despite its critical role and prevalence in many human cancers, limitations to developing an effective matriptase inhibitor include weak binding affinity, poor selectivity, and short circulating half-life. We applied rational and combinatorial approaches to engineer a potent inhibitor based on the hepatocyte growth factor activator inhibitor type 1 (HAI-1), a natural matriptase inhibitor. The first Kunitz domain (KD1) of HAI-1 has been well established as a minimal matriptase binding and inhibition domain, while the second Kunitz domain (KD2) is inactive and involved in negative regulation. Here, we replaced the inactive KD2 domain of HAI-1 with an engineered chimeric variant of KD2/KD1 domains, and fused the resulting construct to an antibody Fc domain to increase valency and circulating serum half-life. The final protein variant contains 4 stoichiometric binding sites that we showed were needed to effectively inhibit matriptase with a Ki =70 ± 5 pM, an increase of 120-fold compared to the natural HAI-1 inhibitor, to our knowledge making it one of the most potent matriptase inhibitors identified to date. Furthermore, the engineered inhibitor demonstrates a protease selectivity profile similar to wild-type KD1 but distinct from HAI-1. It also inhibits activation of the natural pro-HGF substrate and matriptase expressed on cancer cells with at least an order of magnitude greater efficacy than KD1. ________________________________________ High expression and dysregulated activity of the type II, membrane-anchored serine protease matriptase in the local tumor environment has been shown to correlate with poor patient prognosis in many human cancers including breast, colorectal, pancreatic, cervical, and prostate cancers (1-10). This dysregulation is partly driven by the high proteolytic processing and turnover of pro-hepatocyte growth factor (Pro-HGF) (9, 11) to a form of HGF that activates its cognate receptor, c-Met (12) (Fig. 1A). Matriptase is also known to activate other proteases and growth factors including urokinase plasminogen activator (uPA) (11), pro-macrophage stimulating protein (Pro-MSP) (13), and platelet derived growth factor-D (PDGF-D) (14), all of which play key roles in cancer growth and metastasis. Furthermore, matriptase has been identified as a critical driver of other diseases, including iron overload disease (15) and osteoarthritis (16), and has been shown to activate the human airway influenza virus (H1N1) (17) and human immunodeficiency virus (HIV) (18 (19)(20)(21)(22)(23). The activity of matriptase is regulated in healthy tissue by the serine protease inhibitor hepatocyte growth factor activator inhibitor type-1 (HAI-1) (Fig. 1A). HAI-1 is primarily expressed on the surface of epithelial cells and naturally blocks the substrate-activating properties of matriptase (24)(25)(26), as well as other struc...

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Section: Discussionmentioning

confidence: 99%

Section: Kd1-kd2/1-fc Is a Potent And Selective Inhibitor Of Matriptasementioning

confidence: 99%

mentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Mitchell

Kannan

Hunter

et al. 2018

Journal of Biological Chemistry

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“…Moreover, basement membrane proteins often contain cryptic sites, which are exposed after extracellular matrix digestion and can consequently act as pro-migratory, pro-invasive or angiogenic cues [2,114]. Hepsin can proteolytically activate many factors in the microenvironment of epithelial cells, including hepatocyte growth factor, urokinase-type plasminogen activator, macrophage-stimulating protein and EGF receptor, all of which are well-known players in cancer [110,[115][116][117][118][119][120].…”

Section: (B) Lkb1 and Proteolytic Moulding Of Basement Membrane (I) Lmentioning

confidence: 99%

Breaking the epithelial polarity barrier in cancer: the strange case of LKB1/PAR-4

Partanen

Tervonen

Klefström

2013

Phil. Trans. R. Soc. B

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The PAR clan of polarity regulating genes was initially discovered in a genetic screen searching for genes involved in asymmetric cell divisions in the Caenorhabditis elegans embryo. Today, investigations in worms, flies and mammals have established PAR proteins as conserved and fundamental regulators of animal cell polarization in a broad range of biological phenomena requiring cellular asymmetries. The human homologue of invertebrate PAR-4, a serine–threonine kinase LKB1/STK11, has caught attention as a gene behind Peutz–Jeghers polyposis syndrome and as a bona fide tumour suppressor gene commonly mutated in sporadic cancer. LKB1 functions as a master regulator of AMP-activated protein kinase (AMPK) and 12 other kinases referred to as the AMPK-related kinases, including four human homologues of PAR-1. The role of LKB1 as part of the energy sensing LKB1-AMPK module has been intensively studied, whereas the polarity function of LKB1, in the context of homoeostasis or cancer, has gained less attention. Here, we focus on the PAR-4 identity of LKB1, discussing the weight of evidence indicating a role for LKB1 in regulation of cell polarity and epithelial integrity across species and highlight recent investigations providing new insight into the old question: does the PAR-4 identity of LKB1 matter in cancer?

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Hepsin promotes breast tumor growth signaling via the TGFβ‐EGFR axis

Belitškin,

Munne,

Pant

et al. 2023

Molecular Oncology

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Hepsin, a type II transmembrane serine protease, is commonly overexpressed in prostate and breast cancer. The hepsin protein is stabilized by the Ras‐MAPK pathway, and, downstream, this protease regulates the degradation of extracellular matrix components and activates growth factor pathways, such as the hepatocyte growth factor (HGF) and transforming growth factor beta (TGFβ) pathway. However, how exactly active hepsin promotes cell proliferation machinery to sustain tumor growth is not fully understood. Here, we show that genetic deletion of the gene encoding hepsin (Hpn) in a WAP‐Myc model of aggressive MYC‐driven breast cancer inhibits tumor growth in the primary syngrafted sites and the growth of disseminated tumors in the lungs. The suppression of tumor growth upon loss of hepsin was accompanied by downregulation of TGFβ and EGFR signaling together with a reduction in epidermal growth factor receptor (EGFR) protein levels. We further demonstrate in 3D cultures of patient‐derived breast cancer explants that both basal TGFβ signaling and EGFR protein expression are inhibited by neutralizing antibodies or small‐molecule inhibitors of hepsin. The study demonstrates a role for hepsin as a regulator of cell proliferation and tumor growth through TGFβ and EGFR pathways, warranting consideration of hepsin as a potential indirect upstream target for therapeutic inhibition of TGFβ and EGFR pathways in cancer.

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Proteolytic Activation of Pro-Macrophage-Stimulating Protein by Hepsin

Cited by 64 publications

References 56 publications

Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Breaking the epithelial polarity barrier in cancer: the strange case of LKB1/PAR-4

Hepsin promotes breast tumor growth signaling via the TGFβ‐EGFR axis

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