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Targeted inhibition of cell-surface serine protease Hepsin blocks prostate cancer bone metastasis.

Tang X, Mahajan SS, Nguyen LT, Béliveau F, Leduc R, Simon JA, Vasioukhin V - Oncotarget (2014)

Bottom Line: The development of effective therapies inhibiting prostate cancer progression and metastasis may substantially impact prostate cancer mortality and potentially reduce the rates of invasive treatments by enhancing the safety of active surveillance strategies.Hepsin (HPN) is a cell surface serine protease amplified in a subset of human sarcomas (7.2%), as well as in ovarian (10%), lung adeno (5.4%), lung squamous cell (4.5%), adenoid cystic (5%), breast (2.6%), uterine (1.7%) and colon (1.4%) carcinomas.These findings indicate that inhibition of Hepsin with small-molecule compounds could provide an effective tool for attenuation of prostate cancer progression and metastasis.

View Article: PubMed Central - PubMed

Affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.

ABSTRACT
The development of effective therapies inhibiting prostate cancer progression and metastasis may substantially impact prostate cancer mortality and potentially reduce the rates of invasive treatments by enhancing the safety of active surveillance strategies. Hepsin (HPN) is a cell surface serine protease amplified in a subset of human sarcomas (7.2%), as well as in ovarian (10%), lung adeno (5.4%), lung squamous cell (4.5%), adenoid cystic (5%), breast (2.6%), uterine (1.7%) and colon (1.4%) carcinomas. While HPN is not amplified in prostate cancer, it is one of the most prominently overexpressed genes in the majority of human prostate tumors and genetic experiments in mice indicate that Hepsin promotes prostate cancer metastasis, particularly metastasis to the bone marrow. We report here the development, analysis and animal trial of the small-molecule Hepsin inhibitor HepIn-13. Long-term exposure to HepIn-13 inhibited bone, liver and lung metastasis in a murine model of metastatic prostate cancer. These findings indicate that inhibition of Hepsin with small-molecule compounds could provide an effective tool for attenuation of prostate cancer progression and metastasis.

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Hepsin inhibitors attenuate the Hepsin-mediated cleavage of pro-HGF in cell based activity assays(A) Western blot (WB) analysis of vector control- (Ctrl) or Hepsin-expressing HEK 293 cells with anti-Hepsin and anti-β-actin antibodies. Note that Hepsin is present as both full-length (precursor) and cleaved (activated) enzymes. (B) Western blot (WB) analysis of vector control (Ctrl) or pro-HGF-HA expressing HEK 293 cells and their conditioned media with anti-HA and anti-β-actin antibodies. (C) Cell-based Hepsin activity assay. Control or Hepsin-expressing 293 cells in the presence of DMSO or 10μM of indicated compounds were cultured for 2 hours in media containing pro-HGF-HA, and the remaining uncleaved pro-HGF-HA was detected by Western blotting with anti-HA antibodies. (C') Quantitation of data shown in C. Data are the means of three independent experiments ±SD
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Figure 2: Hepsin inhibitors attenuate the Hepsin-mediated cleavage of pro-HGF in cell based activity assays(A) Western blot (WB) analysis of vector control- (Ctrl) or Hepsin-expressing HEK 293 cells with anti-Hepsin and anti-β-actin antibodies. Note that Hepsin is present as both full-length (precursor) and cleaved (activated) enzymes. (B) Western blot (WB) analysis of vector control (Ctrl) or pro-HGF-HA expressing HEK 293 cells and their conditioned media with anti-HA and anti-β-actin antibodies. (C) Cell-based Hepsin activity assay. Control or Hepsin-expressing 293 cells in the presence of DMSO or 10μM of indicated compounds were cultured for 2 hours in media containing pro-HGF-HA, and the remaining uncleaved pro-HGF-HA was detected by Western blotting with anti-HA antibodies. (C') Quantitation of data shown in C. Data are the means of three independent experiments ±SD

Mentions: To determine whether the identified compounds can suppress the activity of full-length Hepsin, when it is expressed on the surface of live cells, we developed a cell-based Hepsin activity assay. For this purpose, we generated HEK293 cells overexpressing full-length Hepsin (Figure 2, A). HA-tagged human pro-HGF secreted into serum-free conditioned media from stably transduced HEK293 cells was used as a protein substrate in these experiments (Figure 2, B). Hepsin overexpressing, but not the control vector-transduced cells, efficiently cleaved the HA-tagged pro-HGF (Figure 2, C). This cleavage was inhibited in the presence of Hepsin inhibitors (Figure 2, C-C').


Targeted inhibition of cell-surface serine protease Hepsin blocks prostate cancer bone metastasis.

Tang X, Mahajan SS, Nguyen LT, Béliveau F, Leduc R, Simon JA, Vasioukhin V - Oncotarget (2014)

Hepsin inhibitors attenuate the Hepsin-mediated cleavage of pro-HGF in cell based activity assays(A) Western blot (WB) analysis of vector control- (Ctrl) or Hepsin-expressing HEK 293 cells with anti-Hepsin and anti-β-actin antibodies. Note that Hepsin is present as both full-length (precursor) and cleaved (activated) enzymes. (B) Western blot (WB) analysis of vector control (Ctrl) or pro-HGF-HA expressing HEK 293 cells and their conditioned media with anti-HA and anti-β-actin antibodies. (C) Cell-based Hepsin activity assay. Control or Hepsin-expressing 293 cells in the presence of DMSO or 10μM of indicated compounds were cultured for 2 hours in media containing pro-HGF-HA, and the remaining uncleaved pro-HGF-HA was detected by Western blotting with anti-HA antibodies. (C') Quantitation of data shown in C. Data are the means of three independent experiments ±SD
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4012739&req=5

Figure 2: Hepsin inhibitors attenuate the Hepsin-mediated cleavage of pro-HGF in cell based activity assays(A) Western blot (WB) analysis of vector control- (Ctrl) or Hepsin-expressing HEK 293 cells with anti-Hepsin and anti-β-actin antibodies. Note that Hepsin is present as both full-length (precursor) and cleaved (activated) enzymes. (B) Western blot (WB) analysis of vector control (Ctrl) or pro-HGF-HA expressing HEK 293 cells and their conditioned media with anti-HA and anti-β-actin antibodies. (C) Cell-based Hepsin activity assay. Control or Hepsin-expressing 293 cells in the presence of DMSO or 10μM of indicated compounds were cultured for 2 hours in media containing pro-HGF-HA, and the remaining uncleaved pro-HGF-HA was detected by Western blotting with anti-HA antibodies. (C') Quantitation of data shown in C. Data are the means of three independent experiments ±SD
Mentions: To determine whether the identified compounds can suppress the activity of full-length Hepsin, when it is expressed on the surface of live cells, we developed a cell-based Hepsin activity assay. For this purpose, we generated HEK293 cells overexpressing full-length Hepsin (Figure 2, A). HA-tagged human pro-HGF secreted into serum-free conditioned media from stably transduced HEK293 cells was used as a protein substrate in these experiments (Figure 2, B). Hepsin overexpressing, but not the control vector-transduced cells, efficiently cleaved the HA-tagged pro-HGF (Figure 2, C). This cleavage was inhibited in the presence of Hepsin inhibitors (Figure 2, C-C').

Bottom Line: The development of effective therapies inhibiting prostate cancer progression and metastasis may substantially impact prostate cancer mortality and potentially reduce the rates of invasive treatments by enhancing the safety of active surveillance strategies.Hepsin (HPN) is a cell surface serine protease amplified in a subset of human sarcomas (7.2%), as well as in ovarian (10%), lung adeno (5.4%), lung squamous cell (4.5%), adenoid cystic (5%), breast (2.6%), uterine (1.7%) and colon (1.4%) carcinomas.These findings indicate that inhibition of Hepsin with small-molecule compounds could provide an effective tool for attenuation of prostate cancer progression and metastasis.

View Article: PubMed Central - PubMed

Affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.

ABSTRACT
The development of effective therapies inhibiting prostate cancer progression and metastasis may substantially impact prostate cancer mortality and potentially reduce the rates of invasive treatments by enhancing the safety of active surveillance strategies. Hepsin (HPN) is a cell surface serine protease amplified in a subset of human sarcomas (7.2%), as well as in ovarian (10%), lung adeno (5.4%), lung squamous cell (4.5%), adenoid cystic (5%), breast (2.6%), uterine (1.7%) and colon (1.4%) carcinomas. While HPN is not amplified in prostate cancer, it is one of the most prominently overexpressed genes in the majority of human prostate tumors and genetic experiments in mice indicate that Hepsin promotes prostate cancer metastasis, particularly metastasis to the bone marrow. We report here the development, analysis and animal trial of the small-molecule Hepsin inhibitor HepIn-13. Long-term exposure to HepIn-13 inhibited bone, liver and lung metastasis in a murine model of metastatic prostate cancer. These findings indicate that inhibition of Hepsin with small-molecule compounds could provide an effective tool for attenuation of prostate cancer progression and metastasis.

Show MeSH
Related in: MedlinePlus