Method for Detection of Melanoma

Abstract

The present invention provides non-invasive methods for detecting, monitoring, staging, and diagnosing malignant melanoma in a skin sample of a subject. The methods include analyzing expression in skin sample of one or more melanoma skin markers. The melanoma skin markers include IL-1 RI, endothelin-2, ephrin-A5, IGF Binding Protein 7, HLA-A0202 heavy chain, Activin A (βA subunit), TNF RII, SPC4, and CNTF Rα. The skin sample can include nucleic acids, and can be a human skin sample from a lesion suspected of being melanoma.

1 . A non-invasive method for detecting early stage melanoma in a skin sample of a human subject, comprising:
 (a) obtaining a skin sample by applying at least one application of an adhesive to the skin of a human subject and removing the adhesive from the skin to remove a skin sample comprising RNA; and   (b) detecting the level of RNA in the skin sample, wherein an increase in endothelin-2 RNA level, as compared to the RNA level in a control sample, is indicative of early stage melanoma, thereby detecting early stage melanoma in the skin sample.   
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , further comprising detecting the RNA levels of IL1 RI, and ephrin-A5, wherein a decrease in the level of IL-1 RI RNA, an increase in the level of endothelin-2 RNA or a decrease in the level of ephrin-A5 RNA is indicative of early stage melanoma. 
     
     
         5 . The method of  claim 1 , further comprising detecting RNA levels of one or more markers selected from IGF Binding Protein 7, HLA-A0202 heavy chain, Activin A (βA subunit), TNF RII, SPC4, and CNTF Rα, wherein an increase in the RNA levels of the one or more markers, as compared to the RNA levels in a control sample, is indicative of melanoma. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the sample is obtained in a manner such that residual skin nucleic acid after removing the adhesive is not affected. 
     
     
         8 . The method of  claim 1 , wherein the skin sample is isolated by applying the adhesive to the skin between one and two times to obtain the skin sample. 
     
     
         9 . The method of  claim 1 , wherein the adhesive comprises an adhesive tape and the skin sample is isolated from a lesion suspected to be melanoma. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 1 , further comprising analyzing expression of one or more of the markers. 
     
     
         12 . The method of  claim 11 , wherein the RNA comprises mRNA, and expression is analyzed by analyzing mRNA levels of one or more of the markers. 
     
     
         13 . The method of  claim 1 , wherein expression of the markers is analyzed by detecting polynucleotides encoding the markers. 
     
     
         14 - 18 . (canceled) 
     
     
         19 . A non-invasive method for monitoring the presence of melanoma in a skin sample from a human subject, comprising:
 (a) obtaining a first skin sample at a first time point by applying at least one application of an adhesive to the skin of a human subject and removing the adhesive from the skin to remove a skin sample comprising RNA;   (b) detecting RNA levels in the first skin sample of the marker endothelin-2;   (c) obtaining a second skin sample at a second time point by applying at least one application of an adhesive to the skin of said human subject and removing the adhesive from the skin to remove a skin sample comprising RNA,   (d) detecting RNA levels in the second skin sample of the marker endothelin-2; and   (e) comparing the RNA level of endothelin-2 at the first time point to the RNA level of endothelin-2 at the second time point to thereby monitor the presence of melanoma in the human subject, wherein an increase in the level of endothelin-2 RNA, as compared to the RNA level in a control sample, is indicative of early stage melanoma.   
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 19 , further comprising detecting the RNA levels of IL 1 RI, and ephrin-A5, wherein an increase in the level of endothelin-2 RNA, and a decrease in the level of IL-1 RI RNA or a decrease in the level of ephrin-A5 RNA is indicative of early stage melanoma. 
     
     
         22 - 24 . (canceled) 
     
     
         25 . The method of  claim 19 , further comprising detecting the RNA levels of IL-1 RI, ephrin-A5, IGF Binding Protein 7, HLA-A0202 heavy chain, Activin A (βA subunit), TNF RII, SPC4, and CNTF Rα in the nucleic acid sample, wherein an increase in the level of SPC4 RNA and a decrease in the level of ephrin-A5 RNA is indicative of early stage melanoma and an increase in the RNA levels of IGF Binding Protein 7, HLA-A0202 heavy chain, Activin A (βA subunit), TNF RII, SPC4, and CNTF Rα is indicative of late stage melanoma. 
     
     
         26 . A non-invasive method for staging malignant melanoma in a skin sample of a human subject, comprising:
 a) obtaining a skin sample by applying an adhesive surface to the skin of a human subject and removing the adhesive surface from the skin such that a skin sample comprising RNA in an amount sufficient for subsequent detection adheres to the adhesive surface after its removal; and   b) detecting the level of endothelin-2 RNA in the skin sample, wherein an increase in the level of endothelin-2 RNA, as compared to the RNA level in a control sample, is indicative of early stage melanoma.   
     
     
         27 . The method of  claim 26 , wherein the sample is obtained in a manner such that residual skin nucleic acid after removing the adhesive is not affected, thereby obtaining a skin sample for use in isolating or detecting RNA in a skin sample. 
     
     
         28 . The method of  claim 1 , further comprising detecting RNA levels of the markers IGF Binding Protein 7, HLA-A0202 heavy chain, Activin A (βA subunit), TNF RII, and CNTF Rα, wherein an increase in the RNA levels of these markers, as compared to the RNA levels in a control sample, is indicative of melanoma 
     
     
         29 . The method of  claim 5 , wherein the RNA levels of the markers are analyzed by applying the RNA to a microarray. 
     
     
         30 . The method of  claim 28 , further comprising detecting the RNA levels of the markers IL-1 RI and ephrin-A5, wherein a decrease in the level of IL-1RI RNA or the level of ephrin-A5, as compared to the RNA levels in a control sample, is indicative of early stage melanoma. 
     
     
         31 . The method of  claim 19 , wherein the skin sample is obtained in a manner such that residual skin nucleic acid after removing the adhesive is not affected, thereby obtaining a skin sample for use in isolating or detecting RNA in a skin sample.