Microrna signatures associated with cytogenetics and prognosis in acute myeloid leukemia (aml) and uses thereof

A technology for acute myeloid and leukemia, applied in the field of molecular biology, can solve the problems of poor understanding, difficult diagnosis and treatment

Inactive Publication Date: 2011-04-06
THE OHIO STATE UNIV RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, little is known about miRNA expression in AML
[0011] Although considerable research has been done on therapies to treat such diseases, they remain difficult to diagnose and treat effectively, and the observed mortality rates in patients suggest a need for improved diagnosis, treatment and prevention of the diseases

Method used

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  • Microrna signatures associated with cytogenetics and prognosis in acute myeloid leukemia (aml) and uses thereof
  • Microrna signatures associated with cytogenetics and prognosis in acute myeloid leukemia (aml) and uses thereof
  • Microrna signatures associated with cytogenetics and prognosis in acute myeloid leukemia (aml) and uses thereof

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Experimental program
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Embodiment I

[0042] Patient and Cell Samples

[0043] Pretreatment bone marrow and blood samples from 182 newly diagnosed AML patients were obtained from the Cell and Tissue Bank of M.D.Anderson Cancer Center (MDACC; (n=172) and Thomas Jefferson University (n=10). Using the microarray platform, the A total of 122 AML samples were analyzed for miRNA expression while using quantitative real-time polymerase chain reaction (RT-PCR; Figure 4 - Table 1 ), 60 untreated AML samples were used to validate the outcome signature.

[0044] A second cohort of 54 AML patients with relapsed (n=34) or refractory (n=20) disease obtained from MDACC was used to determine the difference between newly diagnosed AML patients and relapsed / primary refractory Differences in miRNA expression among AML patients ( Figure 13 - Table S7 ). According to the Declaration of Helsinki, informed consent was obtained from patients to obtain and store cells for future research according to institutional guidelines. Patien...

Embodiment II

[0102] microRNA (miRNA) microarray

[0103] Five micrograms of total RNA were used in quadruplicate on miRNA microarray chips with probes corresponding to 250 human mature and precursor miRNAs (as described in miRBase (microrna.sanger.ac.uk) November 2005). described) hybridization. Total RNA was added separately to a final volume of 12 μl of reaction mixture containing 1 μg of 3′-(N)8-(A)12-biotin-(A)12-biotin-5′ random oligonucleotide primers). The mixture was incubated at 70 °C for 10 min and then cooled on ice. Keep the mixture on ice, add 4 μl 5× first-strand buffer, 2 μl 0.1 M DTT, 1 μl 10 mM dNTP mix, and 1 μl SuperScript II RNaseH-reverse transcriptase (200 units / μl) to a final volume of 20 μl, and dissolve the mixture Incubate for 90 minutes in a 37°C water bath. After incubation for first-strand cDNA synthesis, 3.5 μl of 0.5M NaOH / 50 mM EDTA was added to 20 μl of the first-strand reaction mixture and incubated at 65°C for 15 minutes to denature and synthesize RNA / ...

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Abstract

Methods and compositions for the diagnosis, prognosis and / or treatment of leukemia associated diseases are disclosed.

Description

[0001] Inventors: Carlo M. Croce, Ramiro Garzon [0002] Cross Reference Related Applications [0003] This application claims the benefit of US Provisional Application 61 / 067,419, filed February 28, 2008, the entire disclosure of which is expressly incorporated herein by reference. [0004] Statement Regarding Federally Sponsored Research [0005] This invention was made with government support under NCI grants CA76259 and CA8134. The Government has certain rights in this invention. [0006] Technical Field and Industrial Applicability of the Invention [0007] The present invention relates generally to the field of molecular biology. Certain aspects of the invention include uses in the diagnosis, treatment and prognosis of leukemia-related disorders. Background of the invention [0008] There is no admission that the background art disclosed in this section legally constitutes prior art. [0009] Acute myeloid leukemia (AML) is a cytogenetically and molecularly heterog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48C12Q1/68
CPCC12Q2600/158C12Q2600/136C12Q2600/106C12Q2600/118C12Q2600/178C12Q1/6886A61P35/00A61P35/02A61P35/04
Inventor R·伽赞C·M·克罗斯
Owner THE OHIO STATE UNIV RES FOUND
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