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Baalc expression as a diagnostic marker for acute leukemia

a leukemia and diagnostic marker technology, applied in the field of acute leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia leukemia l

Inactive Publication Date: 2009-06-25
THE OHIO STATE UNIV RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many cancers, including leukemias, CNS and prostate cancers suffer from the problem of late detection in patients.
Also, even when such cancers are detected in a patient, it is often difficult to predict lifespan of, or to determine the optimal therapy for, the particular cancer in the particular patient.

Method used

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  • Baalc expression as a diagnostic marker for acute leukemia
  • Baalc expression as a diagnostic marker for acute leukemia
  • Baalc expression as a diagnostic marker for acute leukemia

Examples

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Effect test

example 1

Alternative Splicing of BAALC in Acute Myeloid Leukemia Samples

[0094]Blasts from BM or peripheral blood (PB) were collected from patients with AML. Total RNA was isolated using Trizol (Invitrogen). Total RNA from AML cells from these patients was reverse-transcribed with avian myeloblastosis virus reverse transcriptase (Roche Diagnostics) using oligo(dT) as primer. The reverse transcriptase reaction mixture contained 10 mM Tris (pH 8.3), 40 mM KCl, 1.5 mM MgCl2, 1 mM dithiothreitol, 200 μM each of dATP, dCTP, dTTP and dGTP, 200 ng of the primer, 10 U of AMV reverse transcriptase from Boehringer Mannheim Biochemicals, and 20 units of RNASIN from Promega. The reverse transcriptase reaction was then performed at a temperature of 60° C.

[0095]PCR was then performed using the cDNA from the reverse transcriptase reaction as template. Primer sequences from exons 1 and 8 of human BAALC were used. The primer from exon 1 was ES100, 5′-GTGCGGTACCAAGCTTCCGCGGCGCAGGAGGATG-3′ (SEQ ID NO: 38), The ...

example 2

BAALC Overexpression in Normal Cells and Various Cancer Samples, Including Glioblastoma Samples

[0098]Real-time PCR (TaqMan®, Applied Biosystems) was used to quantify the relative levels of BAALC transcripts in various tissues and cancers using total RNA isolated from the cells using Trizol (Invitrogen). Initially, RNA samples from several organs including BM, PBL, brain and fetal brain, plus different human tumor samples were analyzed. Additionally, RNA samples from 10 normal human tissues, 10 colorectal cancer-normal colonic mucosa-matched sample pairs, 3 esophageal cancer-normal esophagus-matched sample pairs, 3 lung tumor samples, 5 glioblastoma samples, 2 thyroid carcinoma-normal thyroid-matched sample pairs, 1 thyroid carcinoma sample, and 3 testicular and 3 mammary tumor samples were analyzed for expression of BAALC.

[0099]Reverse transcription was performed using the total RNA isolated from the cells. Separate primers for reverse transcription were used to synthesize cDNA for ...

example 3

BAALC Overexpression in Leukemia Samples

[0106]Real-time PCR (TaqMan®, Applied Biosystems) was used to quantify the relative levels of BAALC transcripts in different leukemia samples, using total RNA isolated from the cells in real-time PCR, as described in Example 2. Blasts from BM or peripheral blood from 130 diverse AML patients, 31 ALL patients, 4 Burkitt's lymphoma (BL) patients, 5 chronic myelogenous leukemia (CML) patients, 5 chronic lymphocytic leukemia (CLL) patients, plus the 7 leukemia cell lines HL-60, KG-1, KG-1a, MC-1010, K-562, D1.1, and RS4;11, were analyzed. Pretreatment BM aspirate or peripheral blood samples from patients with the different leukemias were collected after prior consent. CD34-positive progenitor cells were enriched twice from normal BM aspirates by immunomagnetic separation, using MiniMACS columns (Miltenyi Biotec, Auburn, Calif.). Cell lines were obtained from the American Type Culture Collection. Total RNA was isolated using Trizol (Invitrogen).

[01...

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Abstract

Overexpression of the gene, BAALC, in biological samples from a patient is prognostic for tumor aggressiveness and unfavorable patient outcome. The present invention provides polynucleotide primers and probes for assaying for overexpression of BAALC transcripts. Kits containing the primers and probes are also provided. Also provided are antibodies for assaying for overexpression of BAALC proteins as well as peptide immunogens for producing the anti-BAALC antibodies. The present invention also provides methods for characterizing acute myelogenous leukemia, chronic myelogenous leukemia and prostate cancer in a patient, base on detection of BAALC overexpression.

Description

[0001]This application is a divisional of and claims priority to U.S. application Ser. No. 10 / 293,239, filed on Nov. 12, 2002, which claims priority to U.S. Provisional Application Ser. No. 60 / 348,210, filed Nov. 9, 2001, both of which are incorporated by reference herein in their entirety.[0002]This invention was made, at least in part, with government support under National Institutes of Health Grant No. 5P30CA016058. The U.S. government has certain rights in the invention.BACKGROUND[0003]Leukemias comprise approximately 2% of adult cancers and are a heterogeneous group. There are two broad categories of leukemias. Acute leukemias arise when there is a block in the normal differentiation of cells to mature blood cells that results in large accumulations of immature cells or blasts in the blood. Examples of such cancers are acute myelogenous leukemia (AML; other names are acute myeloid leukemia and acute nonlymphocytic leukemia) and acute lymphoblastic leukemia (ALL). In chronic le...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C07H21/04C07K16/18G01N33/574
CPCG01N33/57426C12Q1/6886C12Q2600/112C12Q2600/118C12Q2600/156C12Q2600/158
Inventor TANNER, STEPHAN M.DE LA CHAPELLE, ALBERT
Owner THE OHIO STATE UNIV RES FOUND
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