Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for...
Reexamination Certificate
2000-11-03
2003-05-06
Yucel, Remy (Department: 1633)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
C530S350000
Reexamination Certificate
active
06558932
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to cardiovascular diseases.
Background of the Invention
The vascular system contains vessels that are morphologically and functionally distinct. For example, arteries carry oxygenated blood at high pressure from the heart, while veins serve as capacitance vessels for blood return. While some of the differences within the vascular system occur after the onset of function, a complete vascular loop, formed by the trunk artery and vein, is required to accommodate the output of the first heart beat. The major trunk vessels, the aorta and the axial vein, of vertebrate embryos are formed by the coalescence of scattered mesodermal angioblasts into simple endothelial tubes by a process termed “vasculogenesis,” which is distinguished from “angiogenesis,” the formation of vessels by sprouting and remodeling (Fishman, “Assembly of blood vessels in the embryos,” Lippincott-Raven Publishers, Philadelphia, 1996; Folkman et al.,
Cell
87:1153-1155, 1996; Risau,
Nature
386:671-674, 1997; Yancopolos et al.,
Cell
93:661-664, 1998).
The aorta, the main trunk of the systemic arterial network, is subject to several congenital and acquired disorders, that may lead to severe complications in infancy and adulthood. Coarctation of the aorta is one of the most common human congenital cardiovascular diseases. In coarctation, a discrete, localized vascular malformation partially obstructs the descending aorta, the major artery to the body, and most frequently occurs distal to the origins of vessels supplying the head and arms. Its effects often become more physiologically severe at birth, when closure of the ductus can exacerbate the restriction to aortic blood flow. As a consequence of coarctation, affected individuals suffer from high blood pressure in the upper extremities and head, and from low pressure in the trunk and legs. Survival often depends on the development of collateral blood vessels, which facilitate blood circulation in a manner so as to bypass the lesion.
Another serious cardiovascular disease that affects large numbers of individuals is atherosclerosis. This condition is characterized by the deposition of lipids in the intima of large and medium-sized arteries. Such deposits are associated with fibrosis and calcification.
The gridlock mutation (grl
m145
) is a recessive mutation that was identified in the zebrafish system and that causes a focal vascular malformation resembling coarctation of the aorta in humans. In grl
m145
mutant embryos, fluorescent dextran injected into the heart, to outline patent vessels, is blocked at the origin of the dorsal aorta at the region where the two anterior dorsal aortae merge to form the single aorta of the trunk. The fluorescent beads circulated normally in the head (Weinstein et al.,
Nature Medicine
1:1143-1147, 1995). Thus, the grl mutant embryos lack circulation to the trunk.
SUMMARY OF THE INVENTION
The invention provides a novel gene, gridlock, and its encoded protein. gridlock plays a role in vascular development and modeling, and a mutation in gridlock has been associated with an aortic arch disease, coarctation. Thus, gridlock nucleic acid molecules and polypeptides can be used in methods of diagnosing, treating, and preventing gridlock-related diseases and conditions, such as aortic arch diseases.
In one aspect, the invention provides a substantially pure gridlock polypeptide. In preferred embodiments of this aspect, the gridlock polypeptide includes the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4, or includes an amino acid sequence that is substantially identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4. In other preferred embodiments of this aspect, the polypeptide is derived from a mammal, e.g., a human.
In another aspect, the invention provides a substantially pure nucleic acid molecule, e.g., DNA, including a sequence encoding a gridlock polypeptide. In a preferred embodiment of this aspect, the nucleic acid molecule encodes a human gridlock polypeptide. In other preferred embodiments of this aspect, the nucleic acid molecule encodes a polypeptide including the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4, or encodes a polypeptide including an amino sequence that is substantially identical to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4. In other preferred embodiments of this aspect, the nucleic acid molecule includes a nucleotide sequence that is substantially identical to the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:3.
In a related aspect, the invention provides a nucleic acid molecule having at least 55% nucleic acid sequence identity to a sequence encoding a gridlock polypeptide or a fragment thereof, where the fragment includes at least six amino acids and the nucleic acid molecule hybridizes under high stringency conditions to at least a portion of a gridlock nucleic acid molecule, and does not hybridize under high stringency conditions to a nucleic acid molecule encoding a hairy-related protein family member. In a preferred embodiment of this aspect, the nucleic acid molecule has 100% complementarity to a nucleic acid molecule encoding a gridlock polypeptide or a fragment thereof including at least six amino acids, and the nucleic acid molecule hybridizes under high stringency conditions to at least a portion of a gridlock nucleic acid molecule, and does not hybridize under high stringency conditions to a nucleic acid molecule encoding a hairy-related protein family member.
In a related aspect, the invention provides a nucleic acid molecule including a sequence that is antisense to a gridlock coding strand or a fragment thereof. In a preferred embodiment of this aspect, the antisense sequence is specific for a mutated gridlock coding region.
In another related aspect, the invention provides a vector, e.g., a gene therapy vector including a gridlock nucleic acid molecule, and a cell including this vector.
In other aspects, the invention provides a non-human transgenic animal, e.g., a zebrafish, including a gridlock nucleic acid molecule, and a non-human animal having a knockout mutation in one or both alleles encoding a gridlock polypeptide. The invention also provides a cell from a non-human animal having a knockout mutation in one or both alleles encoding a gridlock polypeptide.
In other aspects, the invention provides a probe for analyzing the gridlock nucleic acid molecules of an animal, the probe having at least 55% nucleic acid sequence identity to a sequence encoding a gridlock polypeptide or a fragment thereof. The fragment includes at least six amino acids and the probe hybridizes under high stringency conditions to at least a portion of a gridlock nucleic acid molecule, and does not hybridize under high stringency conditions to a nucleic acid molecule encoding a hairy-related protein family member. In a preferred embodiment, the probe has 100% complementarity to a nucleic acid molecule encoding a gridlock polypeptide or a fragment thereof including at least six amino acids, and the probe hybridizes under high stringency conditions to at least a portion of a gridlock nucleic acid molecule, and does not hybridize under high stringency conditions to a nucleic acid molecule encoding a hairy-related protein family member.
In another aspect, the invention provides an antibody that specifically binds to a gridlock polypeptide, e.g., a polypeptide including the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4.
In another aspect, the invention provides a method of detecting the presence of a gridlock polypeptide in a sample. The method involves contacting the sample with an antibody that specifically binds to a gridlock polypeptide and assaying for binding of the antibody to the polypeptide.
In another aspect, the invention provides a method of detecting the presence of a mutant gridlock polypeptide in a sample. The method involves contacting the sample with an antibody that specifically binds to a mutant gridlock polypeptide and assaying for binding of the antibody to the mutant polypeptide.
In another aspect, the invention provide
Fishman Mark C.
Zhong Tao P.
Clark & Elbing LLP
Katcheves Konstantina
The General Hospital Corp.
Yucel Remy
LandOfFree
Gridlock nucleic acid molecules, polypeptides, 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 Gridlock nucleic acid molecules, polypeptides, and..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Gridlock nucleic acid molecules, polypeptides, and... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3034876