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Identification of genes involved in angiogenesis, and development of an angiogenesis diagnostic chip to identify patients with impaired angiogenesis

A technology of genes and subjects, applied in the field of composition of genetic factors

Inactive Publication Date: 2006-03-15
MEDSTAR RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention overcomes the problems and shortcomings associated with current strategies and designs and provides a kit, combination for determining the "vascular type" of an individual patient to predict whether a given individual will naturally develop good versus poor collaterals Objects and methods

Method used

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  • Identification of genes involved in angiogenesis, and development of an angiogenesis diagnostic chip to identify patients with impaired angiogenesis
  • Identification of genes involved in angiogenesis, and development of an angiogenesis diagnostic chip to identify patients with impaired angiogenesis
  • Identification of genes involved in angiogenesis, and development of an angiogenesis diagnostic chip to identify patients with impaired angiogenesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0074] Microarray Analysis of Mouse Hind Limbs

[0075] isolate RNA

[0076] Mice underwent femoral artery ligation and resection. The control group was treated with sham surgery. Adductor muscles of mice after surgery and sham surgery were collected and snap-frozen. Pooled muscles (30-50 mg) were ground to a powder (collected in liquid nitrogen) with a mortar and pestle and homogenized in 2.5 ml guanidine isothiocyanate. Total RNA was extracted by ultracentrifugation at 4°C for 24 hours on a cesium chloride buffer gradient. See Sambrook et al., supra.

[0077] Target preparation and DNA microarray hybridization

[0078] For first-strand cDNA synthesis reactions, 5.0–8.0 μg of total RNA was incubated with T7-(dT)24 primer for 10 min at 70°C, then placed on ice. For the temperature adjustment step, 5× first-strand cDNA buffer, 0.1 M DTT, and 10 mM dNTP mix was added, and the reaction was incubated at 42°C for 1 hr. SSII reverse transcriptase was added and the reactio...

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Abstract

The invention is directed to methods for angiotyping individual patients to predict the likelihood of whether a given individual will develop good vs. poor collaterals naturally. Accordingly, this can involve obtaining and providing a list of genes involved in collateral development. In particular, angiotyping individual patients can be used to predict the likelihood of whether a given individual will develop good vs. poor collaterals in response to specific angiogenesis therapy. From an array of genes that have been determined through experimental studies as being differentially expressed in tissues in which collaterals are developing in response to arterial occlusion, single nucleotide polymorphisms (SNPs), or other epigenetic changes, such as DNA methylation patterns, can be identified. SNPs and DNA methylation patterns are detected using microchips or similar technology assaying for all, or most, of the genes determined to play a role in collateral development. In addition, abnormally low or abnormally high differential expression of any combination of the candidate genes can be detected in such tissue as peripheral blood cells. The presence of a predisposition to develop poor vs. good collaterals is indicated by the presence of SNPs, and / or alterations in DNA methylation patterns, and / or difference in expression levels involving one or more of the genes.

Description

[0001] This application claims priority to US Provisional Application Serial No. 60 / 432,005, filed December 10, 2002, the contents of which are incorporated herein by reference in their entirety. field of invention [0002] The present invention provides compositions and methods for identifying and isolating genetic factors associated with angiogenesis, and also relates to the production and use of arrays containing the isolated genetic factors. Background of the invention [0003] Coronary artery disease and peripheral vascular disease are common morbidities in Western societies. In these diseases, the arteries that supply blood to the heart muscle or legs become narrowed due to deposits of fatty, fibrous, or calcified material inside the arteries. The buildup of these deposits is called atherosclerosis. Atherosclerosis reduces blood flow to the heart or leg muscles, which causes the muscles to starve of oxygen, which can lead to angina (chest pain), myocardial infarction ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q2600/156C12Q2600/172C12Q2600/154C12Q1/6837C12Q1/6886C12Q2600/106C12Q1/6883C12Q2600/118
Inventor S·E·埃普斯坦M·S·伯内特
Owner MEDSTAR RES INST
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