Fibroblast growth factor 15

Disclosed is a human Fibroblast growth factor-15 polypeptide and DNA(RNA) encoding such polypeptide. Also provided is a procedure for producing such polypeptide by recombinant techniques. Also disclosed are methods for utilizing such polypeptide for stimulating re-vascularization, for treating wounds and prevent neuronal damage. Antagonists against such polypeptides and their use as a therapeutic to prevent abnormal cellular proliferation, hyper-vascular diseases and epithelial lens cell proliferation are also disclosed. Diagnostic methods for detecting mutations in the coding sequence and alterations in the concentration of the polypeptides in a sample derived from a host are also disclosed.

This invention relates to newly identified polynucleotides, polypeptides 
encoded by such polynucleotides, the use of such polvnucleotides and 
polypeptides, as well as the production of such polynucleotides and 
polypeptides. More particularly, the polypeptide of the present invention 
have been putatively identified as fibroblast growth factor/heparin 
binding growth factor, hereinafter referred to as "FGF-15". The invention 
also relates to inhibiting the action of such polypeptides. 
Fibroblast growth factors are a family of proteins characteristic of 
binding to heparin and are, therefore, also called heparin binding growth 
factors (HBGF). Expression of different members of these proteins are 
found in various tissues and are under particular temporal and spatial 
control. These proteins are potent mitogens for a variety of cells of 
mesodermal, ectodermal, and endodermal origin, including fibroblasts, 
corneal and vascular endothelial cells, granulocytes, adrenal cortical 
cells, chondrocytes, myoblasts, vascular smooth muscle cells, lens 
epithelial cells, melanocytes, keratinocytes, oligodendrocytes, 
astrocytes, osteoblasts, and hematopoietic cells 
Each member has functions overlapping with others and also has its unique 
spectrum of functions. In addition to the ability to stimulate 
proliferation of vascular endothelial cells, both FGF-1 and 2 are 
chemotactic for endothelial cells and FGF-2 has been shown to enable 
endothelial cells to penetrate the basement membrane. Consistent with 
these properties, both FGF-1 and 2 have the capacity to stimulate 
angiogenesis. Another important feature of these growth factors is their 
ability to promote wound healing. Many other members of the FGF family 
share similar activities with FGF-1 and 2 such as promoting angiogenesis 
and wound healing. Several members of the FGF family have been shown to 
induce mesoderm formation and to modulate differentiation of neuronal 
cells, adipocytes and skeletal muscle cells. 
Other than these biological activities in normal tissues, FGF proteins have 
been implicated in promoting tumorigenesis in carcinomas and sarcomas by 
promoting tumor vascularization and as transforming proteins when their 
expression is deregulated. 
The FGF family presently consists of eight structurally-related 
polypeptides: basic FGF, acidic FGF, int 2, hst 1/k-FGF, FGF-5, FGF-6, 
keratinocyte growth factor, AIGF (FGF-8) and recently a glia-activating 
factor has been shown to be a novel heparin-binding growth factor which 
was purified from the culture supernatant of a human glioma cell line 
(Miyamoto, M. et al., Mol. and Cell. Biol., 13(7):4251-4259 (1993). The 
genes for each have been cloned and sequenced. Two of the members, FGF-1 
and FGF-2, have been characterized under many names, but most often as 
acidic and basic fibroblast growth factor, respectively. The normal gene 
products influence the general proliferation capacity of the majority of 
mesoderm and neuroectoderm-derived cells. They are capable of inducing 
angiogenesis in vivo and may play important roles in early development 
(Burgess, W. H. and Maciag, T., Annu. Rev. Biochem., 58:575-606, (1989)). 
Many of the above-identified members of the FGF family also bind to the 
same receptors and elicit a second message through binding to these 
receptors. 
A eukaryotic expression vector encoding a secreted form of FGF-1 has been 
introduced by gene transfer into porcine arteries. This model defines gene 
function in the arterial wall in vivo. FGF-1 expression induced intimal 
thickening in porcine arteries 21 days after gene transfer (Nabel, E. G., 
et al., Nature, 362:844-6 (1993)). It has further been demonstrated that 
basic fibroblast growth factor may regulate glioma growth and progression 
independent of its role in tumor angiogenesis and that basic fibroblast 
growth factor release or secretion may be required for these actions 
(Morrison, R. S., et al., J. Neurosci. Res., 34:502-9 (1993)). 
Fibroblast growth factors, such as basic FGF, have further been implicated 
in the growth of Kaposi's sarcoma cells in vitro (Huang, Y. Q., et al., J. 
Clin. Invest., 91:1191-7 (1993)). Also, the cDNA sequence encoding human 
basic fibroblast growth factor has been cloned downstream of a 
transcription promoter recognized by the bacteriophage T7 RNA polymerase. 
Basic fibroblast growth factors so obtained have been shown to have 
biological activity indistinguishable from human placental fibroblast 
growth factor in mitogenicity, synthesis of plasminogen activator and 
angiogenesis assays (Squires, C. H., et al., J. Biol. Chem., 263:16297-302 
(1988)). 
U.S. Pat. No. 5,155,214 discloses substantially pure mammalian basic 
fibroblast growth factors and their production. The amino acid sequences 
of bovine and human basic fibroblast growth factor are disclosed, as well 
as the DNA sequence encoding the polypeptide of the bovine species. 
Newly discovered FGF-9 has around 30% sequence similarity to other members 
of the FGF family. Two cysteine residues and other consensus sequences in 
family members were also well conserved in the FGF-9 sequence. FGF-9 was 
found to have no typical signal sequence in its N terminus like those in 
acidic and basic FGF. However, FGF-9 was found to be secreted from cells 
after synthesis despite its lack of a typical signal sequence FGF 
(Miyamoto, M. et al., Mol. and Cell. Biol., 13(7):4251-4259 (1993). 
Further, FGF-9 was found to stimulate the cell growth of oligodendrocyte 
type 2 astrocyte progenitor cells, BALB/c3T3, and PC-12 cells but not that 
of human umbilical vein endothelial cells (Naruo, K., et al., J. Biol. 
Chem., 268:2857-2864 (1993). 
Basic FGF and acidic FGF are potent modulators of cell proliferation, cell 
motility, differentiation, and survival and act on cell types from 
ectoderm, mesoderm and endoderm. These two FGFs, along with KGF and AIGF, 
were identified by protein purification. However, the other four members 
were isolated as oncogenes, expression of which was restricted to 
embryogenesis and certian types of cancers. FGF-9 was demonstrated to be a 
mitogen against glial cells. Members of the FGF family are reported to 
have oncogenic potency. FGF-9 has shown transforming potency when 
transformed into BALB/c3T3 cells (Miyamoto, M., et al., Mol. Cell. Biol., 
13(7):4251-4259 (1993). 
Androgen induced growth factor (AIGF), also known as FGF-8, was purified 
from a conditioned medium of mouse mammary carcinoma cells (SC-3) 
stimulated with testosterone. AIGF is a distinctive FGF-like growth 
factor, having a putative signal peptide and sharing 30-40% homology with 
known members of the FGF family. Mammalian cells transformed with AIGF 
show a remarkable stimulatory effect on the growth of SC-3 cells in the 
absence of androgen. Therefore, AIGF mediates androgen-induced growth of 
SC-3 cells, and perhaps other cells, since it is secreted by the tumor 
cells themselves. 
The polypeptide of the present invention has been putatively identified as 
a member of the FGF family as a result of amino acid sequence homology 
with other members of the FGF family. 
In accordance with one aspect of the present invention, there are provided 
novel mature polypeptides as well as biologically active and 
diagnostically or therapeutically useful fragments, analogs and 
derivatives thereof. The polypeptides of the present invention are of 
human origin. 
In accordance with another aspect of the present invention, there are 
provided isolated nucleic acid molecules encoding the polypeptides of the 
present invention, including mRNAs, DNAs, cDNAs, genomic DNA, as well as 
antisense analogs thereof and biologically active and diagnostically or 
therapeutically useful fragments thereof. 
In accordance with still another aspect of the present invention, there are 
provided processes for producing such polypeptides by recombinant 
techniques through the use of recombinant vectors, such as cloning and 
expression plasmids useful as reagents in the recombinant production of 
the polypeptides of the present invention, as well as recombinant 
prokaryotic and/or eukaryotic host cells comprising a nucleic acid 
sequence encoding a polypeptide of the present invention. 
In accordance with a further aspect of the present invention, there is 
provided a process for utilizing such polypeptides, or polynucleotides 
encoding such polypeptides, for screening for agonists and antagonists 
thereto and for therapeutic purposes, for example, promoting wound healing 
for example as a result of burns and ulcers, to prevent neuronal damage 
due to neuronal disorders and promote neuronal growth, and to prevent skin 
aging and hair loss, to stimulate angiogenesis, mesodermal induction in 
early embryos and limb regeneration. 
In accordance with yet a further aspect of the present invention, there are 
provided antibodies against such polypeptides. 
In accordance with yet another aspect of the present invention, there are 
provided antagonists against such polypeptides and processes for their use 
to inhibit the action of such polypeptides, for example, in the treatment 
of cellular transformation, for example, tumors, to reduce scarring and 
treat hyper-vascular diseases. 
In accordance with another aspect of the present invention, there are 
provided nucleic acid probes comprising nucleic acid molecules of 
sufficient length to specifically hybridize to a polynucleotide encoding a 
polypeptide of the present invention 
In accordance with yet another aspect of the present invention, there are 
provided diagnostic assays for detecting diseases or susceptibility to 
diseases related to mutations in a nucleic acid sequence of the present 
invention and for detecting over-expression of the polypeptides encoded by 
such sequences. 
In accordance with another aspect of the present invention, there is 
provided a process for utilizing such polypeptides, or polynucleotides 
encoding such polypeptides, for in vitro purposes related to scientific 
research, synthesis of DNA and manufacture of DNA vectors. 
These and other aspects of the present invention should be apparent to 
those skilled in the art from the teachings herein.