Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of...
Reexamination Certificate
2001-07-02
2004-06-29
Eyler, Yvonne (Department: 1646)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
C435S252300, C435S254110, C435S254200, C435S320100, C536S023500, C530S350000
Reexamination Certificate
active
06756228
ABSTRACT:
BACKGROUND OF THE INVENTION
Throughout this application, various publications are referenced by author and year. Citations for these references may be found at the end of the specification immediately preceding the claims. The disclosures of these publications in their entireties are hereby incorporated by reference into this application to describe more fully the art to which this invention pertains.
The genetics of atherosclerosis has been the focus of intense investigation. A subset of cases is caused by uncommon Mendelian mutations that predispose individuals to atherosclerosis (Breslow 2000; Keating and Sanguinetti 1996; Lifton 1996). The mutated genes include low-density lipoprotein receptor (LDLR) (Hobbs et al. 1992), cystathionine beta-synthase (CBS) (Kraus 1999), and, in some cases, ATP-binding cassette-A1 (Bodzioch et al. 1999; Brooks-Wilson et al. 1999; Rust et al. 1999) among others. Identification of these genes has shed light on biochemical pathways involved in atherogenesis and provided the basis for current therapeutic interventions. However, the common forms of atherosclerosis are multifactorial in origin. Attempts to map the common susceptibility loci have been hampered by genetic heterogeneity, polygenic inheritance, incomplete pedigrees, and environmental influences. The fact that few of the genome-wide linkage studies have reported loci with large effects points to the existence of multiple loci each having small to moderate effects (Aouizerat et al. 1999; Hixson and Blangero 2000; Rice et al. 2000; Shearman 2000). The modest nature of susceptibility gene effects will likely require extremely large sample sizes or very densely-spaced genetic markers for successful linkage mapping (Risch and Merikangas 1996).
Mouse models offer significant advantages for genetic dissection of complex diseases. The ability to perform selective breeding, produce many offspring, determine inheritance of alleles without ambiguity, and control the environment is a critical factor. Early studies of murine atherosclerosis indicated that there was a clear genetic component. Inbred strains of mice exhibited a spectrum of aortic fatty streak lesion areas following the feeding of atherogenic diets high in cholesterol, fat, and cholic acid (Paigen et al. 1985; Qiao et al. 1994; Roberts and Thompson 1977). A number of susceptibility loci (Ath1-8) were reported based on phenotypic analyses of recombinant inbred strains derived from “resistant” and “susceptible” parents (Paigen 1995; Paigen et al. 1987, 1989; Stewart-Phillips et al. 1989). Although these studies were instrumental in pointing out strain-specific variations, none of the loci have been confirmed by more rigorous analyses of large genetic crosses.
A shortcoming of the diet-fed, inbred mouse model (in terms of carrying out quantitative genetic studies) is that aortic lesion development is minimal even in susceptible strains. Recently, Dansky et al. (1999) showed that the strain-related differences in susceptibility could be accentuated when a gene-targeted disease model was employed. Thus, C57BL/6J mice homozygous for the apolipoprotein E knockout allele exhibited 7-9 fold greater aortic root lesion area relative to FVB/NJ mice homozygous for the allele without any overlap of the phenotypic values. To provide candidate susceptibility loci for human atherosclerosis, we have performed a genome scan of an interspecific cross using the low-density lipoprotein receptor knockout model (Ishibashi et al. 1993). In this model, feeding of a Western-style diet results in elevated plasma LDL levels (similar to levels in humans) and development of human-like complicated fibrous plaques (Masucci-Magoulas et al. 1997). Two significant susceptibility loci were localized to chromosome (Chr) 4 and 6. The effects of these loci were independent of common risk factors for human disease including plasma lipoprotein levels, plasma insulin levels, and body weight.
SUMMARY OF THE INVENTION
This invention provides an isolated nucleic acid encoding a mammalian LOX-1 receptor protein, wherein the receptor protein comprises consecutive amino acids having the following sequence: —S, X, X, E, L, K, X, X, I, X, T, X, X, X, K, L, X, E, K, S, K, E, Q, X, E, L, X, X, X, X, X, N, L, Q, E, X, L, X, R, X, A, N, X, S— (SEQ ID NO: 39), wherein X is any amino acid.
The invention provides an isolated nucleic acid encoding a mammalian membrane-bound LOX-1 receptor protein, wherein the nucleic acid encodes a protein selected from the group consisting of:
(a) a LOX-1 receptor protein comprising consecutive amino acids having a sequence identical to that set forth for Isoform 1 in SEQ ID NO: 20,
(b) a LOX-1 receptor protein comprising consecutive amino acids having a sequence identical to that set forth for Isoform 3 in SEQ ID NO: 24, and
(c) a LOX-1 receptor protein comprising consecutive amino acids having a sequence identical to that set forth for Isoform 4 in SEQ ID NO: 26.
The invention provides an isolated nucleic acid encoding a mammalian soluble LOX-1 receptor protein, wherein the nucleic acid encodes a protein selected from the group consisting of:
(a) a LOX-1 receptor protein comprising consecutive amino acids having a sequence identical to that set forth for Isoform 7 in SEQ ID NO: 14,
(b) a LOX-1 receptor protein comprising consecutive amino acids having a sequence identical to that set forth for Isoform 8 in SEQ ID NO: 16, and
(c) a LOX-1 receptor protein comprising consecutive amino acids having a sequence identical to that set forth for Isoform 9 in SEQ ID NO: 18.
The invention provides an isolated nucleic acid encoding a mammalian LOX-1 receptor protein, wherein the nucleic acid comprises:
(a) a nucleic acid sequence given in any one of SEQ ID Nos: 13, 15, 17, 19, 21, 23, 25, 27, or 28; or
(b) a nucleic acid sequence degenerate to a sequence of (a) as a result of the genetic code.
The invention provides a method involving competitive binding for identifying a chemical compound which specifically binds to a mammalian LOX-1 receptor, which comprises contacting cells expressing on their cell surface the mammalian LOX-1 receptor with both the chemical compound and a second chemical compound known to bind to the receptor, and separately with only the second chemical compound, under conditions suitable for binding of both compounds, and measuring specific binding of the second chemical compound to the mammalian LOX-1 receptor, a decrease in the binding of the second chemical compound to the mammalian LOX-1 receptor in the presence of the chemical compound indicating that the chemical compound binds to the mammalian LOX-1 receptor.
The invention provides a method of screening a plurality of chemical compounds not known to bind to a mammalian LOX-1 receptor to identify a compound which specifically binds to the mammalian LOX-1 receptor, which comprises:
(a) contacting cells expressing on their cell surface the mammalian LOX-1 receptor with the plurality of compounds not known to bind specifically to the mammalian LOX-1 receptor, under conditions permitting binding of compounds known to bind to the mammalian LOX-1 receptor;
(b) determining whether the binding of a compound known to bind to the mammalian LOX-1 receptor is reduced in the presence of the compounds within the plurality of compounds, relative to the binding of the compound in the absence of the plurality of compounds; and if so
(c) separately determining the binding to the mammalian LOX-1 receptor of compounds included in the plurality of compounds, so as to thereby identify the compound which specifically binds to the mammalian LOX-1 receptor.
The invention provides a method of identifying a compound which activates a mammalian LOX-1 receptor which comprises contacting cells expressing on their cell surface the mammalian LOX-1 receptor with the compound under conditions permitting activation of the LOX-1 receptor, and detecting activation of the LOX-1 receptor, thereby identifying the compound as a compound which activates a mammalian LOX-1 receptor.
The invention provides a method of identifying a co
Liang Chien-Ping
Tall Alan R.
Welch Carrie L.
Cooper & Dunham LLP
Eyler Yvonne
Li Ruixiang
The Trustees of Columbia University in the City of New York
White John P.
LandOfFree
Atherosclerosis susceptibility gene locus 1 (Athsq1) and... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Atherosclerosis susceptibility gene locus 1 (Athsq1) and..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Atherosclerosis susceptibility gene locus 1 (Athsq1) and... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3352178