Method for Detection of Melanoma

• US 20100279877A1
• Filed: 10/15/2009
• Published: 11/04/2010
• Est. Priority Date: 06/28/2001
• Status: Abandoned Application

• Current Assignee: DermTech International
• Original Assignee: DermTech International
• Inventors: Vogt, Thomas
• Application Number:
  US12/580,194
• Publication Number:
  US 20100279877A1
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/5743 of skin, e.g. melanoma
• CPC Class Codes: Y10S 435/912 Absidia

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.

Claims

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.

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 (β

A 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.

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.

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.

28. The method of claim 1, further comprising detecting RNA levels of the markers IGF Binding Protein 7, HLA-A0202 heavy chain, Activin A (β

A 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.

2. (canceled)

3. (canceled)

6. (canceled)

10. (canceled)

14-18. -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.

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.

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 (β

A 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 (β

A subunit), TNF RII, SPC4, and CNTF Rα

is indicative of late 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.

20. (canceled)

22-24. -24. (canceled)

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.