Phosphodiesterase 4d7 as prostate cancer marker

A phosphodiesterase, prostate cancer technology, applied in the determination/inspection of microorganisms, enzymes, hydrolases, etc., can solve problems such as false positives and elevations

Active Publication Date: 2012-07-04
KONINKLIJKE PHILIPS ELECTRONICS NV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PSA is not prostate cancer-specific and can be elevated in other situations, resulting in a large number of false positives (in about 70% of men with elevated PSA levels who undergo biopsy, no cancer is found)

Method used

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  • Phosphodiesterase 4d7 as prostate cancer marker
  • Phosphodiesterase 4d7 as prostate cancer marker
  • Phosphodiesterase 4d7 as prostate cancer marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0344] Example 1 - Quantitative RT-PCR assay of human cell lines and human tissue xenografts

[0345] From figure 1 From the indicated cell lines and human tissue xenografts (see also Marques et al., 2006, Eur. Urol., 49(2):245-57 for additional details), RNA was isolated and transcribed into cDNA by standard procedures . Prepared cDNAs of samples "PC346P xenografts" to "346Flu2" (which are cell lines) and samples "PC295" to "PC374" (which are xenografts) were tested for the expression level of PDE4D7.

[0346] qRT-PCR.: Materials and methods

[0347] Treat RNA samples with DNase to ensure no DNA contamination. RNA samples were treated with DNase I (In Vitrogen) at 37°C for 30 min before cDNA synthesis. 1 μg of RNA samples were then treated with Superscript Vilo (In Vitrogen) following the manufacturer's instructions to synthesize first-strand DNA for qPCR analysis. DNA samples were then treated with RNase H1 at 37°C for 30 min.

[0348] The resulting DNA was diluted t...

Embodiment 2

[0378] Example 2 - Quantitative RT-PCR assay using human tissue samples

[0379] The relative gene expression of human PDE4D7 was evaluated in prostate cancer tissues derived from hormone-sensitive / responsive patients compared with hormone-resistant / castration-resistant patients.

[0380] Materials and methods

[0381] In Table 1 below, details of the samples used in the qPCR measurements of the PDE gene expression experiments are given:

[0382] Table 1: Patient information used in the experiment

[0383]

[0384] All samples were derived from male patients (52-82 years old). The "Tissue" column identifies the tissue that has been removed during the procedure, be it prostate tissue or lymph nodes for staging. The "Type of Tissue" column describes the tissue resection method. If not stated otherwise, this tissue was excised during prostate surgery (prostatectomy). "TURP" is defined as transurethral resection of the prostate. The cDNA pool included 4 samples from pat...

Embodiment 3

[0406] Effect of embodiment 3-PDE4D7 expression on PC3 proliferation

[0407] To test the effect of heterologous expression of human PDE4D7 on the proliferation of androgen-independent cell lines, several gene constructs were transfected into PC3 cells, and the growth of the tested cells was compared to control-treated cell lines. PC3 cells transfected with a dominant-negative form of PDE4D7 in which the catalytic activity of PDE4D7 has been knocked out by genetic mutation of the coding sequence of PDE4D7, as well as PC3 cells transfected with a different isoform of the PDE4D family (i.e. PDE4D 1) PC3 cells, as control untreated PC3 cells.

[0408] In RPMI1640 supplemented with L-glutamine (2mM final concentration) and PenStrep (1%), in 5% CO 2 , 37°C, culture PC3 cells in 5% fetal bovine serum.

[0409] PC3 cells were seeded at a density of 5000 cells per well of a 96-well plate (MTP). Transfection complexes for the genes of interest (PDE4D7, dominant-negative PDE4D7, an...

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Abstract

The present invention relates to phosphodiesterase 4D7 (PDE4D7) for use as a marker for prostate cancer, and the use of PDE4D7 as a marker for diagnosing, detecting, monitoring or prognosticating prostate cancer or the progression of prostate cancer. The present invention also relates to a composition for diagnosing, detecting, monitoring or prognosticating prostate cancer or the progression of prostate cancer, a corresponding method and immunoassay, a method for diagnosing, monitoring or prognosticating hormone-resistant prostate cancer vs. hormone-sensitive prostate cancer, a corresponding immunoassay, a method of data acquisition, an immunoassay for diagnosing, detecting, monitoring or prognosticating prostate cancer or the progression of prostate cancer, a method of identifying an individual for eligibility for prostate cancer therapy, an immunoassay for stratifying an individual or cohort of individuals with a prostate cancer disease, an immunoassay for stratifying an individual with prostate cancer, as well as a pharmaceutical composition comprising a compound directly stimulating or modulating the activity of PDE4D7, a compound indirectly stimulating or modulating the activity of PDE4D7, the PDE4D7 protein or a biologically active equivalent thereof, a nucleic acid encoding and expressing PDE4D7, a miRNA inhibitor specific for PDE4D7 miRNAs, a demethylation agent and/or a phosphodiesterase displacement factor.

Description

technical field [0001] The present invention relates to phosphodiesterase 4D7 (PDE4D7) as a marker for prostate cancer, and the use of PDE4D7 as a marker for diagnosing, detecting, monitoring or predicting prostate cancer or the progression of prostate cancer. The present invention also relates to compositions for diagnosing, detecting, monitoring or predicting prostate cancer or the progression of prostate cancer, corresponding methods and immunoassays for diagnosing, monitoring or predicting hormone-sensitive prostate cancer relative to hormone-sensitive prostate cancer Method of resistant prostate cancer, corresponding immunoassay, method of data acquisition, immunoassay for diagnosis, detection, monitoring or prediction of prostate cancer or progression of prostate cancer, identification of individual's eligibility for prostate cancer treatment Methods, immunoassays for stratifying an individual or a group of individuals with prostate cancer disease, immunoassays for strat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574C12N9/16C12Q1/44C12N15/11
CPCC12Q2600/112C12N15/115C12N2310/16C12Q1/6886C12N9/16C12N2310/113C12Y301/04017C12N2310/141C12N15/1137C12N2310/14C12Q1/44C12Q2600/106G01N2800/52G01N33/57434G01N2333/916Y10S436/813A61P13/08A61P35/00A61P43/00G01N33/6893G01N2800/54C12Q2600/118G01N33/68
Inventor R·霍夫曼M·D·豪斯利D·J·P·亨德森
Owner KONINKLIJKE PHILIPS ELECTRONICS NV
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