Method of diagnosing and monitoring bladder cancer and melanoma

Abstract

The present invention provides a method for the diagnosing and monitoring of melanoma

Claims

1. A method for diagnosing, monitoring, and staging of melanoma comprising: the use of an analytical method such as, but not limited to, ELISA, radioimmunoassay, competitive binding assay, Western blot, or dot blot, to measure the quantity of TYRP1 in any human bodily fluid wherein TYRP1 polypeptide level higher than three standard deviations above normal control is associated with the presence of melanoma

Description

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING

[0016] FIG. 1, shows the results of a dot blot showing the plasma level of TYRP 1 in two newly-diagnosed patients with melanoma and five Black and five White healthy blood donors.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

[0017] The present invention relates to diagnostic assays, both quantitative and qualitative for detecting levels of TYRP1 or its polypeptides in bodily fluids including empirical determination of normal and abnormal levels. Thus, for instance, a diagnostic assay in accordance with the invention of detecting over-expression of TYRP1 in bodily fluids of individuals who may have melanoma compared to levels in healthy individuals. Further, the present method of quantifying the TYRP1 levels is particularly useful for discriminating between melanoma and other diseases of the skin since some of the other methods for discriminating melanoma and other ailments of the skin require a biopsy and examination by a pathologist. The current invention could be performed in a physician's office and have a reliable result within three hours. Assay techniques that can be used to determine levels of gene expression of antigens like TYRP1, of the present invention, in a sample derived from a host are well-known to those of skill in the art. Such assay methods include, but are not limited to, radioimmunoassay, competitive binding assays, Western Blot analyses, dot blots, and ELISAs.

[0018] Without limiting the type of assay in the instant invention, ideally for a quantitative diagnostic assay a positive result indicating the disease is one in which blood levels are higher than three standard deviations above the mean level for a normal healthy population of individuals (99.865% of the population). A best case result would yield 100% clinical sensitivity and 100% clinical specificity, which means that the test correctly identifies every individual with the disease as having the disease and also identifying 100% of those who do not as not having the disease, respectively.

[0019] The present invention is further described by the following examples. These examples are provided solely to illustrate the invention by reference to specific embodiments. These examples, while illustrating certain specific aspects of the invention, do not portray the limitations or circumscribe the scope of the disclosed invention.

Example 1

[0020] Demonstrating that the blood level of TYRP1 is elevated in melanoma patients relative to healthy individuals.

[0021] FIG. 1, shows the results of a dot blot showing the plasma level of TYRP 1 in two patients with melanoma and in ten healthy blood donors.

[0022] The two melanoma patients were newly diagnosed White males with melanoma, and the blood was drawn prior to any treatment for melanoma. The thickness of the tumors were 10 mm and 4 mm for patients #1 and #2, respectively. Ten healthy male blood donors were used as the control population. Five were Black, and five were White. Black males were included to determine if the blood level of TYRP1 is strongly associated with race.

[0023] A small volume of patient and control plasma was diluted, and identical volumes of each were spotted onto two strips of nitrocellulose. The other binding sites on the nitrocellulose were covered by incubating the nitrocellulose in a solution of bovine serum albumin.

[0024] A mouse monoclonal antibody to TYRP1 was purchased from GeneTex. This antibody had been shown to react with melanocytes, melanoma and nevi and that it does not react with carcinomas or sarcomas. See TRP1 antibody [Ta99] GTX11780 Datasheet. p. 1 GeneTex (Accessed January, 2012).

[0025] The antibody was biotinylated, then reacted with streptavidin-Alkaline-Phosphatase to produce an antibody to TYRP1 with the Alkaline-Phosphatase reporter attached. The experimental and control strips were put into incubation media with or without the anti-TYRP1-Alkaline Phosphatase complex, respectively. Both strips of nitrocellulose were incubated identically. Both were washed, then put into buffer containing the substrate for alkaline phosphatase for visualizing TYRP1.

[0026] The data in FIG. 1 show that the blood level of TYRP1 in both newly diagnosed melanoma patients is higher than the blood level of nine of the ten plasma samples from the healthy blood donors, whether Black or White.

[0027] These results indicate that the level of TYRP1 in human blood, or other human bodily fluids, could be used in diagnostic assays for distinguishing individuals who have melanoma from those who do not.

[0028] Thus, TYRP1 could be useful in diagnosing and monitoring of melanoma.

REFERENCES

[0029] Ando et al., Melanosomes Are Transferred from Melanocytes to Keratinocytes through the Processes of Packaging, Release, Uptake, and Dispersion. J. Invest. Dermatol. 132.4 (2012): 1222-1229. [0030] Architect System Total PSA. pp. 1-8 Abbott Laboratories (June, 2007). [0031] Cancer Facts and Figures 2015 pp. 1-56 American Cancer Society (Nov. 10, 2015). [0032] Hearing et al., Enzymatic Control of Pigmentation in Mammals. FASEB J. 5.14 (1991):2902-2909. [0033] Howlader et al., Melanoma of the Skin. SEER Cancer Statistics Review, 1975-2012, Section 16 pp. 1-27 National Cancer Institute. Bethesda, Md., http://seer.cancer.gov/csr/19752012/, based on November 2014 SEER data submission, posted to the SEER web site, April 2015. [0034] Howlader et al., Cancer of the Prostate (Invasive). SEER Cancer Statistics Review, 1975-2012, Section 23 pp. 1-22 National Cancer Institute. Bethesda, Md., http://seer.cancer.gov/csr/19752012/, based on November 2014 SEER data submission, posted to the SEER web site, April 2015. [0035] Howlader et al., Cancer of the Urinary Bladder Section (Invasive and In Situ). SEER Cancer Statistics Review, 1975-2012, Section 27 pp. 1-26 National Cancer Institute. Bethesda, Md., http://seer.cancer.gov/csr/19752012/, based on November 2014 SEER data submission, posted to the SEER web site, April 2015. [0036] Journe et al., TYRP1 mRNA Expression in Malignant Melanoma Metastases Correlates with Clinical Outcome. Br. J. Cancer. 105.11 (2011):1726-1732. [0037] Melanoma Skin Cancer. p. 62 American Cancer Society (Nov. 10, 2015). [0038] TRP1 antibody [Ta99] GTX11780 Datasheet. p. 1 GeneTex (Accessed January, 2012).