Tape stripping methods for analysis of skin disease and pathological skin state

• US 20070202540A1
• Filed: 02/23/2007
• Published: 08/30/2007
• Est. Priority Date: 03/31/2004
• Status: Active Grant

• Current Assignee: DermTech International
• Original Assignee: DermTech International
• Inventors: Benson, Nicholas
• Granted Patent: US 7,989,165 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 1/2813 Producing thin layers of samples on a substrate, e.g. smearing, spinning-on G01N1/30 takes precedence
• CPC Class Codes: G01N 2800/20 Dermatological disorders
• CPC Class Codes: G01N 2800/202 Dermatitis
• CPC Class Codes: G01N 2800/205 Scaling palpular diseases, e.g. psoriasis, pytiriasis
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/6881 from skin

Abstract

The present invention provides non-invasive methods for detecting, monitoring, and diagnosing skin disease and pathological skin states such as irritated skin and psoriasis. The methods include using tape stripping to analyze expression in epidermal samples, of one or more skin markers. In illustrative examples, the tape stripping is performed using pliable tape that has a rubber adhesive. Furthermore, the present invention provides methods for predicting and monitoring response to therapy for a skin disease, such as psoriasis or dermatitis. Finally, the methods can include the use of a microarray.

Claims

1. A non-invasive method for isolating or detecting a protein from an epidermal sample of a human subject, comprising:

a) applying an adhesive tape to a target area of the skin of the subject in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises cells from the stratum corneum of the subject;

b) lysing the cells to extract a protein; and

c) isolating or detecting a protein in the epidermal sample.

2. The method of claim 1, wherein the protein is selected from the group consisting of TNFα

, IFNγ

, CD2, IL-112B, Krt-16, Krt-17, IL-23A, GAPDH and β

-actin.

3. The method of claim 1, wherein the protein is an expression product of a gene listed in Table VI.

4. The method of claim 1, wherein the tape comprises a rubber adhesive on a polyurethane film.

5. The method of claim 1, wherein about one to ten adhesive tapes are applied and removed from the skin.

6. The method of claim 1, wherein about one to eight adhesive tapes are applied and removed from the skin.

7. The method of claim 1, wherein about one to five adhesive tapes are applied and removed from the skin.

8. A method for diagnosing melanoma in a human subject, comprising:

a) applying an adhesive tape to a target area suspected of being melanoma on the skin of the subject in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises cells from the stratum corneum;

b) lysing the cells to extract a protein; and

c) comparing the level of target protein to a normal or standard protein profile in similar tissue, wherein an altered level of the target protein is indicative of melanoma, thereby diagnosing melanoma in the subject.

9. The method of claim 8, wherein the protein is selected from the group consisting of MGSA and TNFα

.

10. The method of claim 8, wherein the tape comprises a rubber adhesive on a polyurethane film.

11. The method of claim 8, wherein between one and ten adhesive tapes are applied to the skin and removed from the skin.

12. The method of claim 8, wherein about one to eight adhesive tapes are applied and removed from the skin.

13. The method of claim 8, wherein about one to five adhesive tapes are applied and removed from the skin.

14. The method of claim 8, wherein the method further comprises taking a biopsy of the target area of the skin.

15. The method of claim 14, wherein protein is extracted from the biopsy sample, and the level of protein in the biopsy and the level of protein in the tape sample are analyzed.

16. The method of claim 8, further comprising obtaining a sample from uninvolved epidermal tissue of the human subject.

17. The method of claim 16, wherein the sample from uninvolved epidermal tissue is obtained by taking a biopsy of the uninvolved skin.

18. The method of claim 16, wherein the sample from uninvolved epidermal tissue is obtained by:

a) applying an adhesive tape to skin of the subject in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises cells from the stratum corneum and wherein the skin is unaffected by a disease to be tested;

b) lysing the cells to extract a protein; and

c) quantitating the extracted protein.

19. The method of claim 16, wherein the uninvolved skin is from the upper arm or the upper back.

20. A non-invasive method for predicting response to treatment for a disease or pathological state, comprising:

a) applying an adhesive tape to the skin of a subject afflicted with the disease or pathological state in a manner sufficient to isolate an epidermal sample comprising cells from the stratum corneum of the subject;

b) lysing the cells to extract a protein; and

c) detecting a target protein in the epidermal sample, wherein the presence of the target protein is indicative of a greater likelihood of response to treatment, thereby predicting response to treatment for the disease or pathological state.

21. The method of claim 20, wherein the disease or pathological state is melanoma.

22. The method of claim 20, wherein the treatment is Etanercept.

23. The method of claim 20, wherein the tape comprises a rubber adhesive on a polyurethane film.

24. The method of claim 20, wherein between one and ten adhesive tapes are applied to the skin and removed from the skin.

25. The method of claim 20, wherein about one to eight adhesive tapes are applied and removed from the skin.

26. The method of claim 20, wherein about one to five adhesive tapes are applied and removed from the skin.

27. A non-invasive method for identifying a predictive skin marker for response to treatment for a disease or pathological state, comprising:

a) applying an adhesive tape to the skin of a subject afflicted with the disease or pathological state at a first time point, in a manner sufficient to isolate an epidermal sample comprising cells from the stratum corneum;

b) treating the subject for the disease or pathological state;

c) determining whether the disease or pathological state has responded to the treatment;

d) lysing the cells to extract and quantify a target protein; and

e) determining whether the level of a target protein in the epidermal sample is predictive of response to treatment, thereby identifying a skin marker for response to treatment.

28. The method of claim 27, wherein the disease or pathological state is melanoma.

29. The method of claim 27, wherein the treatment is Etanercept.

30. The method of claim 27, wherein the tape comprises a rubber adhesive on a polyurethane film.

31. The method of claim 27, wherein between one and ten adhesive tapes are applied to the skin and removed from the skin.

32. The method of claim 27, wherein about one to eight adhesive tapes are applied and removed from the skin.

33. The method of claim 27, wherein about one to five adhesive tapes are applied and removed from the skin.