Identification of a Competitive HGF Antagonist Encoded by an Alternative Transcript

Chan, Andrew M.; Rubin, Jeffrey S.; Bottaro, Donald P.; Hirschfield, D; Chedid, Marcio; Aaronson, Stuart A.

doi:10.1126/science.1720571

Cited by 206 publications

(145 citation statements)

References 32 publications

(15 reference statements)

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“…We found no change in proliferation of liver cells treated with HGF-based fusion proteins, as assessed by a cell viability assay (data not shown). This is consistent with previous studies conducted on the binding properties of HGF, which revealed that ␣HGF and other smaller fragments of this factor can enter the cell without causing other downstream responses related to HGF (17,18). This enables the use of the HGF-␣ subunit as a targeting moiety, while avoiding undesired biological responses that are induced by the growth factor itself.…”

Section: Discussionsupporting

confidence: 92%

A Novel Approach for Enzyme Replacement Therapy

Eavri

Lorberboum-Galski

2007

Journal of Biological Chemistry

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Metabolic diseases arise from mutations in key enzymes of major metabolic pathways. One promising approach for the treatment of such diseases is based on the administration of a wild-type enzyme to substitute the activity of the impaired enzyme by the use of enzyme replacement therapy, yet it is important to deliver this enzyme to the specific deficient tissue. We suggest a new concept for the treatment of metabolic diseases using fusion proteins. We examined the feasibility of this concept in the well characterized metabolic disease, phenylketonuria (PKU), which results from a mutation in the liver enzyme phenylalanine hydroxylase (PAH). PAH is a key enzyme in the metabolic pathway of phenylalanine. Deficiency in PAH leads to high and persistent levels of this amino acid in the plasma of PKU patients, causing permanent neurological damage. Currently a low protein diet is still considered the only effective treatment for most PKU patients. To restore PAH activity in the liver of PKU patients, we constructed PAH-based fusion proteins with delivery moieties based on the HIV-transactivator of transcription peptide, and fragments of human hepatocyte growth factor aiming to specifically target PAH to the liver. We show that these new fusion proteins can be delivered into a variety of human liver cell lines and retain PAH activity after being internalized. We also show that plasma phenylalanine levels were dramatically lowered in mice treated with PAHbased fusion proteins after intravenous administration. We therefore suggest an alternative concept for the treatment of PKU using targeted fusion proteins, which may also be applied to the treatment of other metabolic diseases.

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Section: Discussionsupporting

confidence: 92%

A Novel Approach for Enzyme Replacement Therapy

Eavri

Lorberboum-Galski

2007

Journal of Biological Chemistry

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“…In addition to mitogenesis, HGF and dHGF possess a diverse array of biologic activities including motogenesis and morphogenesis (Brinkmann et al, 1995;Chan et al, 1993;Stoker et al, 1987;Tsarfaty et al, 1992). HGF and dHGF exert their biological e ects on target cells through binding and stimulating a speci®c transmembrane tyrosine kinase cell surface receptor (HGFR) known as Met, the product of the proto-oncogene c-met Naldini et al, 1991;Rubin et al, 1993;Weidner et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

The five amino acid-deleted isoform of hepatocyte growth factor promotes carcinogenesis in transgenic mice

et al. 1999

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Hepatocyte growth factor (HGF) is a polypeptide with mitogenic, motogenic, and morphogenic e ects on di erent cell types including hepatocytes. HGF is expressed as two biologically active isotypes resulting from alternative RNA splicing. The roles of each HGF isoform in development, liver regeneration and tumorigenesis have not yet been well characterized. We report the generation and analysis of transgenic mice overexpressing the ®ve amino acid-deleted variant of HGF (dHGF) in the liver by virtue of an albumin expression vector. These ALB-dHGF transgenic mice develop normally, have an enhanced rate of liver regeneration after partial hepatectomy, and exhibit a threefold higher incidence of hepatocellular carcinoma (HCC) beyond 17 months of age. Moreover, overexpression of dHGF dramatically accelerates diethyl-nitrosamine induced HCC tumorigenesis. These tumors arise faster, are signi®cantly larger, more numerous and more invasive than those appearing in non-transgenic littermates. Approximately 90% of female dHGF-transgenic mice had multiple macroscopic HCCs 40 weeks after injection of DEN; whereas the non-transgenic counterparts had only microscopic nodules. Liver tumors and cultured tumor cell lines from dHGF transgenics showed high levels of HGF and c-Met mRNA and protein. Together, these results reveal that in vivo dHGF plays an active role in liver regeneration and HCC tumorigenesis.

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“…To date, both the full-length and the deleted isoforms of HGF/SF have been shown to have comparable mitogenic, cytotoxic, and scatter activities (27,28). The remaining splice variants result in much smaller protein products and consist of the N-terminal N domain followed by either the first kringle (HGF/NK1) (29) or the first two kringles (HGF/NK2) (30). These two splice variants have been described as having agonistic as well as antagonistic activities.…”

mentioning

confidence: 99%

“…In addition to sharing several structural motifs, HGF/SF shares approximately a 40% overall homology to plasminogen (22). The common structural motifs are best defined at the gene sequence level and include an N-terminal N domain (Glu 30 (1). The kringle domain, first identified in prothrombin (23), is a stretch of 78 largely hydrophobic amino acids that form a globular structure that is stabilized by three highly conserved disulfide bonds.…”

mentioning

confidence: 99%

Differential Mitogenic Effects of Single Chain Hepatocyte Growth Factor (HGF)/Scatter Factor and HGF/NK1 following Cleavage by Factor Xa

Pediaditakis

Monga

Mars

et al. 2002

Journal of Biological Chemistry

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Hepatocyte growth factor/scatter factor (HGF/SF) is a multifunctional cytokine that is involved in many normal as well as pathological conditions. HGF/NK1, a splice variant of HGF/SF, has been reported to have either antagonistic or agonistic effects with regard to c-Met signaling depending on the cell type. In these experiments, we have determined that HGF/NK1 is a potent mitogen for rat hepatocytes in culture. Furthermore, we have found that coagulation factor Xa (fXa) is capable of cleaving HGF/NK1 and single chain HGF/SF (scHGF/SF). The products resulting from cleavage of HGF/NK1 or scHGF/SF by fXa appear as single bands under non-reducing conditions. The reaction products from the digestion of HGF/NK1 by fXa were separated under reducing conditions, and the cleavage site, as determined by N-terminal sequencing, was located C-terminal to arginine 134. Previous work established that the heparin-binding domain for HGF/SF is located in the N domain of HGF/SF. Additionally, the dimerization of the HGF/SF receptor (c-Met) by the ligand HGF/NK1 is facilitated by heparin and related sulfonated sugars on the cell surface, whereas heparin is not required for HGF/SF-mediated dimerization. Cleavage of single chain HGF/SF or HGF/NK1 by factor Xa does not alter the affinity of the respective molecules for heparin, but it did variably affect the associated mitogenic activity of these factors. The associated mitogenic activity of HGF/ NK1 was reduced by more than 90%, whereas the mitogenic activity of scHGF/SF was unaffected. This suggests mandatory maintenance of a steric interaction of the N domain and the first kringle domain for HGF/NK1 to act as an agonist for rat hepatocyte growth but is not required by full-length HGF/SF.

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Identification of a Competitive HGF Antagonist Encoded by an Alternative Transcript

Cited by 206 publications

References 32 publications

A Novel Approach for Enzyme Replacement Therapy

A Novel Approach for Enzyme Replacement Therapy

The five amino acid-deleted isoform of hepatocyte growth factor promotes carcinogenesis in transgenic mice

Differential Mitogenic Effects of Single Chain Hepatocyte Growth Factor (HGF)/Scatter Factor and HGF/NK1 following Cleavage by Factor Xa

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