Methods for capture and detection of micro-RNA molecules from the skin by non-invasive tape stripping
The present invention provides non-invasive methods for isolating an miRNA molecule from an epidermal sample, as well as detecting, monitoring, and diagnosing skin disease and pathological skin states such as irritated skin and psoriasis. The methods include using tape stripping to obtain the miRNA from epidermal samples, and may be used to further analyze expression 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.
1 . A method for isolating miRNA molecules from an epidermal sample from 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 miRNA molecules; and
b) isolating miRNA molecules from the epidermal sample.
2 . The method of claim 1 , wherein the tape is a rubber-based tape.
3 . The method of claim 2 , wherein the tape comprises a rubber adhesive on a polyurethane film.
4 . A method for isolating an miRNA sample from skin, comprising applying an adhesive tape to a target area of the skin in a manner sufficient to isolate an epidermal sample, wherein epidermal sample contains miRNA, wherein the tape comprises a rubber adhesive, and wherein the tape is pliable, thereby isolating an epidermal sample adhering to the adhesive tape.
5 . The method of claim 4 , wherein the tape is a rubber-based tape.
6 . The method of claim 4 , wherein the tape comprises a rubber adhesive on a polyurethane film.
7 . The method of claims 1 or 4 , wherein the tape is applied and removed from the skin about one to ten times.
8 . The method of claims 1 or 4 , wherein the tape is applied and removed from the skin about one to eight times.
9 . The method of claims 1 or 4 , wherein the tape is applied and removed from the skin about one to five times.
10 . A method for identifying an miRNA profile indicative of a disease, pathological or physiological state of a human subject, the method comprising:
a) applying an adhesive tape to an area of skin in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises miRNA molecules; and
b) determining miRNA levels from the sample, or an amplification product thereof, and comparing the levels with the miRNA levels of a control sample, wherein a difference in miRNA level is indicative of an miRNA profile of a disease, pathological or physiological state of a subject.
11 . The method of claim 10 , wherein the disease, pathological or physiological state is dermatitis.
12 . The method of claim 10 , wherein the disease, pathological or physiological state is psoriasis.
13 . The method of claim 10 , wherein the disease, pathological or physiological state is diabetes.
14 . The method of claim 10 , wherein the disease, pathological or physiological state is melanoma.
15 . The method of claim 10 , wherein the disease, pathological or physiological state is prostate cancer.
16 . The method of claim 10 , further comprising comparing the miRNA levels to expression levels of mRNA isolated from the same sample, thereby expression profiling the mRNA population in the sample.
17 . A method for predicting response to treatment for a disease, pathological or physiological state, comprising:
a) applying an adhesive tape to an area of skin in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises miRNA molecules; and
b) determining miRNA levels from the sample, or an amplification product thereof, and comparing the levels with the miRNA levels of an unaffected subject or sample, wherein a difference in miRNA level is indicative of a response to treatment of a disease, pathological or physiological state.
18 . The method of claim 17 , wherein the disease, pathological or physiological state is dermatitis.
19 . The method of claim 17 , wherein the disease, pathological or physiological state is psoriasis.
20 . The method of claim 17 , wherein the disease, pathological or physiological state is diabetes.
21 . The method of claim 17 , wherein the disease, pathological or physiological state is melanoma.
22 . The method of claim 17 , wherein the disease, pathological or physiological state is prostate cancer.