Research ReportAdenoviral gene transfer of hepatocyte growth factor prevents death of injured adult motoneurons after peripheral nerve avulsion
Introduction
Hepatocyte growth factor (HGF) was initially identified and purified as a potent mitogen of primary cultured hepatocytes (Nakamura et al., 1984, Nakamura et al., 1989). HGF is a heterodimeric protein composed of α and β chains and induces proliferation, migration, differentiation of target cells as well as organogenesis and neovascularization (Funakoshi and Nakamura, 2003). In the nervous system, HGF exhibits strong neurotrophic activities for motoneurons both in vitro and in vivo (Caton et al., 2000, Ebens et al., 1996, Funakoshi and Nakamura, 2003, Honda et al., 1995, Koyama et al., 2003, Maina and Klein, 1999, Naeem et al., 2002, Novak et al., 2000, Okura et al., 1999, Sun et al., 2002, Wong et al., 1997, Yamamoto et al., 1997). There have been no reports, however, concerning the neurotrophic effects of HGF on adult motoneuron death after proximal nerve injury. In animal models of adult motoneuron injury, avulsion of cranial and spinal nerves causes marked motoneuron degeneration in adult rats (Koliatsos et al., 1994, Moran and Graeber, 2004, Ruan et al., 1995, Sakamoto et al., 2000, Sakamoto et al., 2003a, Sakamoto et al., 2003b, Søreide, 1981, Watabe et al., 2000, Watabe et al., 2005, Wu, 1993), so that these animal models can be useful for therapeutic evaluation of neurotrophic factors or neuroprotective molecules against adult motoneuron death (Ikeda et al., 2003, Sakamoto et al., 2000, Sakamoto et al., 2003a, Sakamoto et al., 2003b, Watabe et al., 2000, Watabe et al., 2005). We have recently shown that adenoviral gene transfer of glial-cell-line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), transforming growth factor-β2 (TGFβ2) and growth inhibitory factor (GIF)/metallothionein-III (MT-III) prevented the death of adult rat facial and spinal motoneurons after facial nerve and cervical spinal root avulsion (Sakamoto et al., 2000, Sakamoto et al., 2003a, Sakamoto et al., 2003b, Watabe et al., 2000). In the present study, we investigated whether HGF protects injured motoneurons after facial nerve or spinal root avulsion by using a recombinant adenoviral vector encoding human HGF.
Section snippets
Bioassay of recombinant human HGF
In this study, we constructed a recombinant adenoviral vector encoding human HGF (AxCAhHGF). To test the ability of AxCAhHGF to induce human HGF expression in vitro, COS1 cells were infected with AxCAhHGF and the conditioned media (CMs) were harvested at 3 days postinfection. The levels of human HGF in uninfected and infected CMs analyzed by enzyme-linked immunosorbent assay (ELISA) were 1.9 ± 0.4 ng/ml and 2004.8 ± 160 ng/ml, respectively (n = 3). Western blot analysis of the CM harvested at 3 days
Discussion
HGF binds to tyrosine kinase receptor c-Met and triggers diverse biological responses that include cell motility, proliferation, morphogenesis, neurite extension and anti-apoptotic activities in a variety of cells (Funakoshi and Nakamura, 2003, Maina and Klein, 1999). Although the function of HGF in the nervous system has not been fully elucidated, it has recently been shown that HGF plays a strong neuroprotective role for motoneurons both in vitro and in vivo (Caton et al., 2000, Ebens et al.,
Adenovirus preparation
The human HGF cDNA was excised from pBS-hHGF with deletion of 15 base pairs (Seki et al., 1990) and subsequently cloned into SwaI cloning site of a cassette cosmid pAxCAwt (TaKaRa, Osaka, Japan) carrying an adenovirus type-5 genome lacking the E3, E1A and E1B regions to prevent the virus replication. The cosmid pAxCAwt contains the CAG (cytomegalovirus-enhancer-chicken β-actin hybrid) promoter on the 5′ end and a rabbit globin poly (A) sequence on the 3′ end. The cosmid was then cotransfected
Acknowledgments
We are grateful to Dr. Jing-Song Shen (Jikei University School of Medicine) for adenovirus preparation. This work was supported by Grants-in-Aid for Ministry of Education, Culture, Sports, Science and Technology, Japan, and Research on Psychiatric and Neurological Diseases and Mental Health, H16-kokoro-017, Ministry of Health, Labor and Welfare, Japan.
References (34)
- et al.
Hepatocyte growth factor/scatter factor is an axonal chemoattractant and a neurotrophic factor for spinal motor neurons
Neuron
(1996) - et al.
Hepatocyte growth factor: from diagnosis to clinical applications
Clin. Chim. Acta
(2003) - et al.
Localization and functional coupling of HGF and c-Met/HGF receptor in rat brain: implication as neurotrophic factor
Brain Res. Mol. Brain Res.
(1995) - et al.
Efficient gene activation system on mammalian cell chromosomes using recombinant adenovirus producing Cre recombinase
Gene
(1996) - et al.
The facial nerve axotomy model
Brain Res. Brain Res. Rev.
(2004) - et al.
Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats
Biochem. Biophys. Res. Commun.
(1984) - et al.
The role of nitric oxide in facial motoneuronal death
Brain Res.
(1995) - et al.
Isolation and expression of cDNA for different forms of hepatocyte growth factor from human leukocyte
Biochem. Biophys. Res. Commun.
(1990) - et al.
Hepatocyte growth factor promotes motor neuron survival and synergizes with ciliary neurotrophic factor
J. Biol. Chem.
(1997) Expression of nitric-oxide synthase (NOS) in injured CNS neurons as shown by NADPH diaphorase histochemistry
Exp. Neurol.
(1993)
The branchial arches and HGF are growth-promoting and chemoattractant for cranial motor axons
Development
Neurotrophic effect of hepatocyte growth factor on central nervous system neurons in vitro
J. Neurosci. Res.
Neuroprotection by scatter factor/hepatocyte growth factor and FGF-1 in cerebellar granule neurons is phosphatidylinositol 3-kinase/akt-dependent and MAPK/CREB-independent
J. Neurochem.
Oral administration of a neuroprotective compound T-588 prevents motoneuron degeneration after facial nerve avulsion in adult rats
Amyotroph. Lateral. Scler. Other Mot. Neuron Disord.
Gene expression profile of spinal motor neurons in sporadic amyotrophic lateral sclerosis
Ann. Neurol.
Expression of hepatocyte growth factor and c-Met in the anterior horn cells of the spinal cord in the patients with amyotrophic lateral sclerosis (ALS): immunohistochemical studies on sporadic ALS and familial ALS with superoxide dismutase 1 gene mutation
Acta Neuropathol. (Berl.)
Ventral root avulsion: an experimental model of death of adult motor neurons
J. Comp. Neurol.
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2018, Biomedicine and PharmacotherapyCitation Excerpt :After optic nerve injury HGF promotes long-term survival and axonal regeneration of retinal ganglion cells [19]. Adenoviral transfer of HGF gene prevents death of injured adult motoneurons in a rat model of peripheral nerve avulsion [20]. Furthermore, non-viral HGF gene therapy by intramuscular injections in patients with painful diabetic neuropathy provided symptomatic relief with improvement in quality of life [21].
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2007, Brain ResearchCitation Excerpt :Hepatocyte growth factor (HGF) was initially identified and cloned as a mitogen for primary hepatocytes (Nakamura et al., 1984, 1989) and was later found to be a novel neurotrophic factor for various types of neurons in both the CNS and PNS (Funakoshi and Nakamura, 2003; Maina et al., 1998), such as hippocampus (Honda et al., 1995), midbrain dopaminergic neurons (Hamanoue et al., 1996), cerebral cortical neurons (Sun et al., 2002a), sensory neurons (Funakoshi and Nakamura, 2001; Maina et al., 1997), motor neurons (Ebens et al., 1996; Yamamoto et al., 1997), cerebellar granular cells (Zhang et al., 2000), and cortical interneurons (Powell et al., 2001) in vitro. HGF was also found to be a neurotrophic factor in vivo in rodent models of brain ischemia (Ishihara et al., 2005; Miyazawa et al., 1998) and motor nerve injuries (Hayashi et al., 2006; Okura et al., 1999). Anxiolytic effects of HGF are also evident (Isogawa et al., 2005).