Claims for Patent: 7,183,057

Summary for Patent: 7,183,057

Title:	Tape stripping methods for analysis of skin disease and pathological skin state
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.
Inventor(s):	Benson; Nicholas R. (San Diego, CA)
Assignee:	Dermtech International (San Diego, CA)
Application Number:	10/816,457
Patent Claims:	1. A method of detecting expression of genes in the skin, comprising: a) applying an adhesive tape to a target area of the skin in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises nucleic acid molecules; and b) detecting expression of the nucleic acid molecules in the epidermal sample by determining a Ct value. 2. A method for detecting a response of a subject to treatment for dermatitis, comprising: a) treating the subject for dermatitis; b) applying an adhesive tape to irritated skin of the subject in a manner sufficient to isolate an epidermal sample, wherein the epidermal sample comprises nucleic acid molecules; and c) detecting expression of a keratin 10, keratin 16, or keratin 17 gene product, wherein an increase in expression is indicative of response of the subject to treatment for dermatitis, and wherein the method is performed prior to treatment and after treatment. 3. A method for detecting a response of a subject to treatment for dermatitis, comprising: a) treating the subject for dermatitis; b) applying an adhesive tape to irritated skin of the subject in a manner sufficient to isolate an epidermal sample, wherein the epidermal sample comprises nucleic acid molecules; and c) detecting expression of a keratin 16 or keratin 17 gene product, wherein an increase in expression is indicative of response of the subject to treatment for dermatitis, and wherein the method is performed prior to treatment and after treatment. 4. A non-invasive method for isolating or detecting nucleic acid molecules from an epidermal sample of a psoriatic lesion of a human subject, comprising: a) applying an adhesive tape to the psoriatic lesion of the subject in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises nucleic acid molecules; b) detecting expression of the nucleic acid molecules in the epidermal sample by determining a Ct value before and after treatment; c) detecting expression of the nucleic acid molecule by detecting a difference in a .DELTA.Ct value before and after treatment, wherein a .DELTA.Ct value is a difference in the number of amplification cycles required to reach a threshold signal level between the nucleic acid molecule and a control nucleic acid molecule. 5. A method for diagnosing psoriasis in a human subject, comprising: a) applying an adhesive tape to a lesion suspected of being a psoriatic lesion 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 a target nucleic acid molecule; b) detecting the target nucleic acid molecule; and c) comparing expression of the target nucleic acid molecule with expression of a control nucleic acid molecule in the same experiment using the same sample volumes and probes, wherein altered expression of the target nucleic acid molecule as compared with expression of the control nucleic acid molecule is determined by calculating a Ct value wherein altered expression is indicative of psoriasis, thereby diagnosing psoriasis in the subject. 6. The method of claim 1, wherein the tape comprises a rubber adhesive on a polyurethane film. 7. The method of claim 1, wherein about one to ten adhesive tapes are applied and removed from the skin. 8. The method of claim 1, wherein about one to eight adhesive tapes are applied and removed from the skin. 9. The method of claim 1, wherein about one to five adhesive tapes are applied and removed from the skin. 10. The method of claim 1, wherein the nucleic acid molecules are applied to a microarray to detect the nucleic acid molecules. 11. The method of claim 4, wherein the nucleic acid encodes for TNF.alpha., IFN.gamma., CD2, IL-12B, Krt-16 and IL-23A. 12. The method of claim 11, wherein the nucleic acid encodes a protein selected from CD2, TNF.alpha., and IFN.gamma.. 13. The method of claim 4, wherein between one and ten adhesive tapes are applied to the skin and removed from the skin. 14. The method of claim 4, wherein between one and eight adhesive tapes are applied to the skin and removed from the skin. 15. The method of claim 4, wherein between about one and four adhesive tapes are applied to the skin and removed from the skin. 16. The method of claim 4, wherein the method further comprises taking a biopsy of the psoriatic lesion. 17. The method of claim 16, Wherein a nucleic acid sample is obtained from the biopsy, and the nucleic acid from the tape sample and the nucleic acid from the biopsy are analyzed. 18. The method of claim 4, wherein the adhesive tape comprises a rubber adhesive. 19. The method of claim 4, further comprising obtaining a nucleic acid sample from uninvolved epidermal tissue of the human subject. 20. The method of claim 19, wherein the nucleic acid sample is obtained by taking a biopsy of the uninvolved skin. 21. The method of claim 19, wherein the nucleic acid 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 nucleic acid and wherein the skin is unaffected by a disease to be tested; and b) isolating or detecting the nucleic acid from the epidermal sample of the unaffected skin. 22. The method of claim 19, wherein the uninvolved skin is from the upper arm or the upper back. 23. The method of claim 4, wherein the nucleic acid is deoxyribonucleic acid (DNA). 24. The method of claim 4, wherein the nucleic acid is ribonucleic acid (RNA). 25. The method of claim 5, wherein the target nucleic acid molecule encodes a protein selected from TNF.gamma., IFN.gamma., CD2, IL-12B, Krt-16 and IL-23A. 26. The method of claim 5, wherein two or more target nucleic acid molecules are detected. 27. The method of claim 26, wherein the two or more target nucleic acid molecules encode two or more proteins selected from CD2, TNF.alpha., or IFN.gamma.. 28. The method of claim 5, wherein a biopsy is taken at the site of the skin. 29. The method of claim 28, wherein a nucleic acid sample is obtained from the biopsy. 30. 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 nucleic acid molecules; b) treating the subject for the disease or pathological state; d) determining whether the disease or pathological state has responded to the treatment; and e) determining whether expression of a nucleic acid molecule in the epidermal sample is predictive of response to treatment, thereby identifying a skin marker for response to treatment. 31. The method of claim 30 wherein the disease or pathological state is psoriasis. 32. The method of claim 30 wherein the treatment is Etanercept, Clobetasol, Alefacept, or narrow band ultraviolet-B light. 33. 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 nucleic acid molecules; b) detecting a target nucleic acid molecule in the epidermal sample, wherein expression of the target nucleic acid molecule is indicative of a response to treatment, thereby predicting response to treatment for the disease or pathological state. 34. The method of claim 33 wherein the disease or pathological state is psoriasis. 35. The method of claim 33 wherein the treatment is Etanercept, Clobetasol, Alefacept, or narrow band ultraviolet-B light. 36. A non-invasive method for isolating or detecting a protein from an epidermal sample of a psoriatic lesion of a human subject, comprising: a) applying an adhesive tape to the psoriatic lesion 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) quantifying the protein, thereby isolating or detecting a protein in the epidermal sample. 37. The method of claim 36, wherein the protein is selected TNF.alpha., IFN.gamma., CD2, IL-12B, Krt-16 and IL-23A. 38. A method for diagnosing psoriasis in a human subject, comprising: a) applying an adhesive tape to a lesion suspected of being a psoriatic lesion 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 a target protein; b) detecting the target protein to determine the level of target protein in the sample; 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 psoriasis, thereby diagnosing psoriasis in the subject. 39. The method of claim 38, wherein the protein is selected TNF.alpha., IFN.gamma., CD2, IL-12B, Krt-16 and IL-23A.

Details for Patent 7,183,057

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Immunex Corporation	ENBREL	etanercept	For Injection	103795	11/02/1998		2039-02-26
Immunex Corporation	ENBREL	etanercept	For Injection	103795	05/27/1999		2039-02-26
Immunex Corporation	ENBREL	etanercept	Injection	103795	09/27/2004		2039-02-26
Immunex Corporation	ENBREL	etanercept	Injection	103795	02/01/2007		2039-02-26
Immunex Corporation	ENBREL MINI	etanercept	Injection	103795	09/14/2017		2039-02-26
Immunex Corporation	ENBREL	etanercept	Injection	103795			2039-02-26
Immunex Corporation	ENBREL	etanercept	Injection	103795	03/05/2020		2039-02-26