Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
1999-10-12
2004-02-17
Chan, Christina (Department: 1644)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C530S362000
Reexamination Certificate
active
06693169
ABSTRACT:
BACKGROUND OF THE INVENTION
The invention relates to two novel laminins, i.e., laminin 13 and 14, and methods of making and using these molecules. The invention also relates to the use of laminin 5 to treat neural disorders, e.g., to induce or promote retinal adhesion and the viability of retina photoreceptors.
The laminins are large heterotrimeric glycoproteins of the extracellular matrix. Each laminin heterotrimer is composed of an &agr;, a &bgr;, and a &ggr; chain, chosen from a number of possible homologues of each chain. Currently, eleven laminin chains have been identified: five &agr; chains, three &bgr; chains, and three &ggr; chains (reviewed in [1]).
SUMMARY OF THE INVENTION
The invention is based, in part, on the discovery of two novel members of the laminin family, laminin 13 and laminin 14. Accordingly, the invention features a purified or isolated preparation, a recombinant preparation, or a composition of laminin 13, which includes laminin chains &agr;3, &bgr;2, and &ggr;3. In a preferred embodiment, the laminin 13 is a trimer of an &agr;3, &bgr;2, and &ggr;3 chain.
In a preferred embodiment, the &agr;3 chain has a molecular weight of about 300 kD, 200 kD, or 165 kD, the &bgr;2 chain has a molecular weight of about 190 kD or 170 kD, the &ggr;3 chain has a molecular weight of about 200 kD or 170 kD.
In another preferred embodiment, the &agr;3 chain is reactive or specifically binds to mouse monoclonal antibody BM-2 or any other antibody which can compete for the BM-2 epitope. In another preferred embodiment, the &bgr;2 chain is reactive or specifically binds to guinea pig polyclonal GP1 [47], mouse monoclonal C4 [46], R49, D5, D79, or any other antibody which can compete for the GP1 or C4 epitope.
In another aspect, the invention features, a purified or isolated preparation, a recombinant preparation, or a composition of laminin 14, which includes laminin chains &agr;4, &bgr;2, and &ggr;3. In a preferred embodiment, the laminin 13 is a trimer of an &agr;4, &bgr;2, and &ggr;3 chain.
In a preferred embodiment, the &agr;4 chain has a molecular weight of about 185 kD, the &bgr;2 chain has a molecular weight of about 190 kD or 170 kD the &ggr;3 chain has a molecular weight of about 200 kD or 170 kD.
In another preferred embodiment, the &agr;4 chain is renactive or specifically binds to a &agr;4 rabbit polyclonal antibody disclosed in J. Cell Biol 1997, 137:685-701 or any other antibody which can compete for the epitope of the &agr;4 rabbit polyclonal antibody. In another preferred embodiment, the &bgr;2 chain is reactive or specifically binds to guinea pig polyclonal GP1 [47], mouse monoclonal C4 [46], R49, D5, D79, or any other antibody which can compete for the GP1 or C4 epitope.
The laminin chains of any laminin as disclosed herein can be the initial translation product or a degradation product, e.g., a naturally occurring degradation product of a laminin chain.
In another aspect, the invention features a composition which includes a purified isolated or recombinant laminin 13, 14, or both. The invention includes pharmaceutical preparations, e.g., a pharamaceutical preparation which include a pharmaceutically acceptable carrier.
In another aspect, the invention features an isolated nucleic acid, e.g., DNA, RNA, or cDNA encoding laminin 13. The isolated nucleic acid can be a combination of nucleic acids each encoding one or more laminin chains or a single nucleic acid. The isolated nucleic acid can be expressed in a vector, e.g., an expression vector or expressed directly in a cell. A vector containing a sequence corresponding to the sequence of the isolated nucleic acid can express the isolated nucleic acid in a suitable cell or a suitable in vitro environment.
The invention also features an isolated nucleic acid, e.g., DNA, RNA, or cDNA encoding laminin 14. The isolated nucleic acid can be a combination of nucleic acids each encoding one or more laminin chains or a single nucleic acid. The isolated nucleic acid can be expressed in a vector, e.g., an expression vector or expressed directly in a cell. A vector containing a sequence corresponding to the sequence of the isolated nucleic acid can express the isolated nucleic acid in a suitable cell or a suitable in vitro environment.
In another aspect, the invention features a recombinant laminin 13 or laminin 14 which can be produced, e.g., by expressing the laminin chains of laminin 13 or laminin 14 in a suitable cell host and under a condition suitable for the laminin chains to form laminin 13 or laminin 14.
In a preferred embodiment, the laminin 13 differs from a naturally occurring laminin 13 at at least 1, but less than 5, 10, or 15 amino acid residues. In another embodiment, one, two, or each laminin chain of a laminin, differs from its naturally occurring counterpart at at least 1, but less than 5, 10, or 15 amino acid residues.
In a preferred embodiment, the laminin 14 differs from a naturally occurring laminin 14 at at least 1, but less than 5, 10, or 15 amino acid residues. In another embodiment, one, two, or each laminin chain of laminin 14, differs from its naturally occurring counterpart at at least 1, but less than 5, 10, or 15 residues.
In another aspect, the invention features, a method of isolating a laminin 13 or 14. The method includes:
providing retinal tissue, e.g., a tissue selected from the group consisting of retina interphotoreceptor matrix, retina outer plexiform layer, neural retina, Müller cell, and a preparation of retinal neurons, and isolating the laminin 13, 14, or a preparation of both. The laminins can be isolated by the use of immuno affinity columns which use one or more mabs which are specific for the subunits of laminin 13 or 14.
In another aspect, the invention features, a method for producing laminin 13. The method includes:
providing recombinant nucleic acid which encodes a laminin &agr;3 chain, a laminin &bgr;2 chain, and a laminin &ggr;3 chain, and expressing the nucleic acid to provide recombinant laminin 13.
In a preferred embodiment a single cell includes nucleic acid which encodes the laminin &agr;3 chain, a laminin chain &bgr;2, and a laminin &ggr;3 chain.
In another aspect, the invention features, a method for producing laminin 14. The method includes:
providing recombinant nucleic acid which encodes a laminin &agr;4 chain, a laminin &bgr;2 chain, and a laminin &ggr;3 chain, and expressing the nucleic acid to provide recombinant laminin 13.
In a preferred embodiment a single cell includes nucleic acid which encodes the laminin &agr;4 chain, laminin &bgr;2 chain, and laminin &ggr;3 chain.
The invention still provides a method for treating a disorder associated with abnormal functions of synapses, e.g., insufficient stability, viability, formation, or defective organization of synapses. The method comprises administering to a subject an effective amount of laminin 13, laminin 14, laminin 5, separately or in combination with one another.
The invention provides a method for treating a disorder associated with inadequate neural cell growth, healing and regeneration,e.g., axon outgrowth, a disorder associated with abnormal subretinal space or interphotoreceptor matrix (IPM) such as inadequate stability of IPM, a disorder associated with retina contact, continuity, and/or adhesion, a disorder associated with abnormal or insufficient formation of synapses, and a disorder associated with viability of a neural cell, e.g., photoreceptor or an element thereof, e.g., outer segment, inner segment, cell body, and synapses. The method comprises administering to a subject an effective amount of laminin 13, laminin 14, laminin 5, separately or in combination with one another.
Still yet another feature of the present invention provides a method to treat a disorder associated with retinal abnormality, e.g., rod dystrophy, rod-cone dystrophy, macular degeneration, and retinal detachment. The method includes administering to a subject an effective amount of laminin 13, laminin 14, laminin 5, separately or in combination with one another.
Anot
Brunken William J.
Burgeson Robert E.
Hunter Dale D.
Libby Richard R.
Chan Christina
Fish & Richardson
The General Hospital Corporation
VanderVegt F. Pierre
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