Leukemia disease genes and uses thereof

A leukemia and gene technology, applied in the field of leukemia disease genes and the use of said leukemia disease genes to diagnose and treat leukemia, can solve laborious and time-consuming problems

Inactive Publication Date: 2008-05-14
WYETH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these studies involve positive selection of specific cell subtypes, which is laborious and time-consuming

Method used

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  • Leukemia disease genes and uses thereof

Examples

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Effect test

example 1

[0180] Example 1. Purification of PBMC and RNA

[0181] All blood was collected from diseases from disease volunteers, MDS patients and AML patients. Receive the awareness of the drug dynamics part of these clinical studies and enable the scheme to be approved by the local human test committee (INSTITIONAL REVIEW BOARD) at the clinical site. MDS patients are mainly a caucasian and an average age of 66 years (within 52-84 years). AML patients are only Caucasian and have an average age of 45 (within 19-65 years). Disease-free volunteers are only a caucasian and have an average age of 23 years (within 18-32 years).

[0182]The inclusion standards of AML patients include 20% of the bone marrow, and the morphological diagnosis of AML performed by the FAB classification system; and instructions CD33 + State flow cytometry. The inclusion criteria of MDS patients include MDS morphological diagnosis, and refractory anemia, refractory anemia accompanied by cyclic ferrochromic cells, materna...

example 2

[0184] Example 2. RNA amplification and GeneChip Generation of hybridization probes.

[0185] Use the modified Lockhart et al. (1996) Nature Biotechnology The procedures described in 14: 1675-1680 are used to prepare a labeled target for oligonucleotide arrays. 2 micrograms total RNA were converted to cDNA using oligo-(DT) 24 primers containing T7DNA polymerase promoters. The cDNA was used as a template using a T7DNA polymerase kit (Ambion, Woodlands, TX, USA) and biotinylated CTP and UTP (Enzo, Farmingdale, NY, USA). At 94 ° C, the labeled cRNA was broken for 35 minutes in 40 μl of the final volume of 40 mM Tris-acetate (pH 8.0), 100 mM KOAC, 30 mM mgOAc.

example 3

[0186] Example 3. Hybridization and fluorescence detection with AFFYMETRIX microarray

[0187] Individual illness and disease-free samples with HG-U133a GeneChips (Affymetrix) hybridization. No samples are collected. The tagged target was diluted in 1 × MES buffer having 100 μg / ml of herring DNA and 50 μg / mL acetylation BSA. Such like Hill et al. (2000) Science The arrays are standardized to each other and to evaluate the sensitivity of the oligonucleotide array, such that each hybridization reaction includes 11 bacterial genes in vivo synthesis transcripts. The abundance of these transcripts is within the range of 1: 300,000 (3 ppm) to 1: 1000 (1000 ppm) on the number of control transcripts on the overall transcript. The detection sensitivity of the array can be within the range of from about 1: 300,000 and 1: 100,000 copies of the signal by the signal reactivity of these control transcripts.

[0188] The labeled probe was denatured at 99 ° C for 5 minutes, and then denatur...

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Abstract

The invention provides the use of whole blood samples or samples comprising peripheral blood mononuclear cells (PBMCs) for diagnosing or evaluating the progression or treatment of leukemia. Genes that are differentially expressed in un-fractionated PBMCs of leukemia patients, as compared to in disease-free humans or in patients who have a differential type of leukemia, can be identified according to the invention. These genes are leukemia disease genes and can be used to detect the presence or progression of leukemia in a subj ect of interest. Leukemias that are amenable to the present invention include, but are not limited to, acute myelogenous leukemia (AML) and myelodysplastic syndromes (MDS). Non-limiting examples of AML or MDS disease genes are provided in Tables 2, 4 and 6.

Description

Technical field [0001] The present invention relates to a leukemia disease gene and a method of diagnosing and treating leukemia using the leukemia pathogenesis. Background technique [0002] Myelosaptogenic abnormal syndrome (MDS) is a variable clinical process and results characterized by a group of different types of bone marrow cell precursor clonal disorders. About 30% of patients with MDS ultimately develop acute myeloid leukemia (AML), and particularly applicable to early identification of this group of patients will focus on these individuals. [0003] Recent expression map studies have revealed AC133 in patients with MDS and AML + Differences in hematopoietic stem cells (Miyazato et al. (2001) BLOOD 98: 422-427). CD34 purified from the bone marrow from MDS patients + Similar results have also been observed in the transcription spectrum of the cells, the transcription spectrum from the MDS patient has been related to the CD34 from the health individual. + Cellular transc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q2600/112G01N33/57426C12Q1/6886C12Q2600/106C12Q1/6837A61P35/02C12Q2539/101
Inventor 迈克尔·E·布尔奇斯基珍妮弗·安·斯托弗安德鲁·J·多尔纳纳塔莉·C·特温
Owner WYETH LLC
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