MICRONEEDLE DEVICES, AND USES THEREOF
The present disclosure provides microneedle devices and kits for extracting biomarkers from a tissue. The present disclosure provides microneedle-device-mediated extraction methods for extracting biomarkers from a lesional skin of a subject. The present disclosure also provides computer-implemented methods for predicting a patient's response to an autoimmune therapeutic drug. The present disclosure provides methods selecting a treatment for an autoimmune disease. The present disclosure also provides methods of nucleic acid analysis to prospectively predict a patient's response to a therapeutic.
1 . A microneedle-device-mediated extraction method comprising:
a. obtaining an injection-molded microneedle device, wherein microneedles in the microneedle device are coupled to probes for an analyte and wherein the injection-molded microneedle device is coupled to a spring-loaded applicator;
b. contacting the injection-molded microneedle device into lesional skin of a subject, thereby contacting the probes with the analyte;
c. extracting the injection-molded microneedle device from the lesional skin of the subject; and
d. eluting the analyte from the microneedle device by exposing the microneedles in the injection-molded microneedle device to heat.
2 . The method of claim 1 , wherein a height of the microneedles is in the range of 400 μm to 1000 μm.
3 . The method of claim 1 , wherein a width of the microneedles is less than 500 μm.
4 . The method of claim 1 , wherein the injection-molded microneedle device comprises at least 100 microneedles.
5 . The method of claim 1 , wherein the analyte comprises RNA.
6 . The method of claim 5 , wherein the RNA comprises mRNA.
7 . The method of claim 6 , wherein the mRNA is intact mRNA.
8 . The method of claim 6 , wherein the mRNA is greater than 700 base pairs.
9 . The method of claim 6 , further comprising quantifying the mRNA.
10 . The method of claim 9 , wherein the quantifying the mRNA comprises performing a PCR reaction.
11 . The method of claim 1 , wherein the probes comprise a polynucleotide with a homopolymeric sequence.
12 . The method of claim 11 , wherein the homopolymeric sequence comprises thymine or uracil.
13 . The method of claim 12 , wherein the homopolymeric sequence comprises two or more consecutive thymine residues.
14 . The method of claim 13 , wherein the homopolymeric sequence is capable of binding to a poly-A tail of the mRNA.
15 . The method of claim 10 , wherein the mRNA is associated with an autoimmune disease.
16 . The method of claim 15 , wherein the autoimmune disease is psoriasis, acne, atopic dermatitis, Raynaud's phenomenon, rosacea, skin cancer, or vitiligo.
17 . The method of claim 16 , further comprising sequencing the mRNA by high-throughput sequencing to obtain sequence reads; and applying a trained algorithm to the sequence reads.
18 . The method of claim 17 , wherein the trained algorithm predicts whether the subject will be responsive to an autoimmune therapeutic drug.
19 . The method of claim 18 , wherein the autoimmune therapeutic drug is an IL-17 mediated treatment, an IL-23 mediated treatment, a TNFα mediated treatment, or any combination thereof.
20 . The method of claim 19 , further comprising administering the autoimmune therapeutic drug to the subject.
21 . The method of claim 1 , wherein the subject is human.
22 . A computer-implemented method for determining a skin lesion of a subject will be responsive to an autoimmune therapeutic drug, comprising:
a. providing a sample comprising mRNA derived from the skin lesion of the subject;
b. converting the mRNA to cDNA;
c. performing next generation sequencing of the cDNA; and
d. applying a trained algorithm to data generated by the next generation sequencing of the cDNA wherein the trained algorithm predicts whether a subject will be responsive to the autoimmune therapeutic drug.
23 . The method of claim 22 , wherein the skin lesion is a psoriatic lesion.
24 . The method of claim 23 , wherein the subject has a PASI score of at least 5.
25 . The method of claim 23 , wherein the subject has a PASI score of at least 8.
26 . A microneedle device wherein:
a. the microneedle device is injection-molded;
b. the microneedle device comprises a first face and a second face;
c. the microneedle device is coupled to an applicator; and
d. microneedles in the microneedle device comprise probes for mRNA.