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Methods for diagnosis and prognosis of epithelial cancers

Inactive Publication Date: 2008-03-13
THE GENERAL HOSPITAL CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0006] The present invention is based on the surprising discovery that three proteins, Cystatin B, Chaperonin 10, and Profilin (also referred to as “epithelial cancer markers”), are present in the urine of patients with bladder cancer, a cancer of epithelial origin. Accordingly, the present invention is directed to methods for prognostic evaluation of cancers of epithelial origin and to methods for facilitating diagnosis of cancers of epithelial origin by monitoring the presence of these markers in biological samples. The invention is also directed to markers for therapeutic efficacy. In particular, the amount of Cystatin B detected in urine correlates with disease status such that Cystatin B levels can be used to predict the presence of invasive bladder cancer. Thus, measuring the level of Cystatin B, Chaperonin 10, and / or Profilin proteins in urine provides a quick, easy, and safe screen that can be used to both diagnose and prognose bladder cancer in a patient. Alternatively, the absence of these markers can provide an indication that the patient does not have bladder cancer.
[0010] In one embodiment, a method for facilitating the diagnosis of bladder cancer in a patient is provided. The method comprises obtaining a biological sample, preferably a voided urine specimen, from a patient and detecting the presence or absence of at least one epithelial cancer biomarker (Cystatin B, Chaperonin 10, or Profilin) in urine sample, wherein the presence of at least one epithelial cancer biomarker is indicative of bladder cancer.

Problems solved by technology

For example, to date there are relatively few options available for the diagnosis of breast cancer using easily detectable biomarkers.
These biomarkers offer alternative methods of diagnosis: however, they are not widely used.
Furthermore, despite the use of a number of histochemical, genetic, and immunological markers, clinicians still have a difficult time predicting which tumors will metastasize to other organs.

Method used

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  • Methods for diagnosis and prognosis of epithelial cancers
  • Methods for diagnosis and prognosis of epithelial cancers
  • Methods for diagnosis and prognosis of epithelial cancers

Examples

Experimental program
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Effect test

example i

Proteomic Analysis of Voided Urine, Bladder Cancer Tissue and Cell Lines for Biomarker Discovery in Transitional Cell Carcinoma

Introduction

[0102] There is a need for new biomarkers to aid in the diagnosis and management of cancers of epithelial origin. Urine can serve as an excellent medium for epithelial cancer biomarker discovery and analysis. Proteomic analysis by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) is one effective tool to analyze the proteome of human specimens. 2D PAGE was used to analyze voided urine, human bladder tumor and normal tissue, and human derived bladder cancer cell lines as a method for biomarker discovery.

Methods

Urine

[0103] Under IRB approved protocol, voided urine specimens were collected from sixty-three patients prior to diagnostic cystoscopy with biopsy and twenty-two age-matched control patients with no clinical evidence of bladder cancer and no history of malignancy. Total urinary protein was isolated and quantified. Equival...

example ii

Immunostaining for Cystatin B in Bladder Cancer Tissue

Methods

[0109] Normal bladder and bladder cancer tissue were immunostained using mouse monoclonal anti-cystatin B antibody and counterstained with Haematoxylin. Immunostaining was performed using the bladder cancer tissue microarray BL801 (US Biomax Inc, Rockville, Md.). The tissues were deparaffinized, endogenous peroxide blocked in 3% hydrogen peroxide in methanol, and microwave antigen retrieval performed using Antigen Unmasking Solution. Blocking was performed using 5% normal horse serum and endogenous biotin blocked using Avidin / Biotin kit. Tissue was incubated with mouse monoclonal anti-cystatin B / Stefin B antibody, clone A6 / 2 (GeneTex, Inc, San Antonio, Tex.), followed by anti-mouse biotinylated secondary antibody, amplified using ABC kit, and developed using DAB. Tissue was counterstained using Gill's Hematoxylin #3 (Sigma-Aldrich, St. Louis, Mo.), and blued using Tacha's Bluing Solution (Biocare, Concord, Calif.). All ...

example iii

Cystatin B as a Tissue and Urinary Biomarker of Bladder Cancer Recurrence and Disease Progression

Introduction

[0111] Bladder cancer is the second most common genitourinary malignancy in the United States. In 2006, there were an estimated 61,420 newly diagnosed cases of bladder cancer and an estimated 13,060 deaths due to cancer of the bladder (Jemal, A. et al. CA Cancer J Clin 2006;56: 106-30). Among all newly diagnosed cases, approximately 70% present as superficial tumors (stages Ta, T1, or Tis); however, up to 50-70% of those cases will recur after resection and approximately 10-20% will progress to invasive disease (T2 or greater) (Rubben, H. et al. J Urol 1988;139: 283-5). Pathologic data, including grade, stage, and associated carcinoma in-situ at initial presentation have provided some insight into the risk of disease progression to muscularis propria invasion (Althausen, A. F. et al. J Urol 1976;116: 575-80; Herr, H. W. J Urol 2000;163: 60-1). Nevertheless, the ability to ...

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Abstract

The present invention is based on the discovery that three proteins, Cystatin B, Chaperonin 10, and Profilin are present in the urine of patients with bladder cancer, a cancer of epithelial origin. Accordingly, the present invention is directed to methods for prognostic evaluation of cancers of epithelial origin and to methods for facilitating diagnosis of cancers of epithelial origin by monitoring the presence of these markers in biological samples. The invention is also directed to markers for therapeutic efficacy.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation-in-part of International (PCT) Patent Application No. PCT / US2006 / 003049 filed on Jan. 30, 2006, which claims the benefit under 35 U.S.C. § 19(e) of U.S. Provisional Patent Application No. 60 / 648,110 filed Jan. 28, 2005; the contents of each application are incorporated herein by reference in their entirety.GOVERNMENT SUPPORT [0002] This work was supported by National Institute of Health grant number 2R37 CA37393. The government has certain rights to the invention.BACKGROUND OF THE INVENTION [0003] One of the most important factors in the survival of cancer is detection at an early stage. Clinical assays that detect the early events of cancer offer an opportunity to intervene and prevent cancer progression. With the development of gene profiling and proteomics there has been significant progress in the identification of molecular markers or “biomarkers” that can be used to diagnose and prognose specific...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/50G01N33/535
CPCG01N33/6893G01N33/57407G01N33/57488G01N2333/8139
Inventor ZETTER, BRUCE R.FELDMAN, ADAM S.MCDOUGAL, W. SCOTT
Owner THE GENERAL HOSPITAL CORP
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