US20230383352A1 - Microneedle devices and methods, and skin condition assays - Google Patents
Microneedle devices and methods, and skin condition assays Download PDFInfo
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Definitions
- autoimmune diseases can encompass a wide variety of different diseases and conditions that may impact a single tissue or organ, or many tissues or organs simultaneously. Some autoimmune diseases specifically target the skin. While the etiology of any particular autoimmune disease is often unknown, an imbalance in immune regulation is often involved.
- psoriasis is a chronic autoimmune disease characterized by raised areas of abnormal skin. It is a chronic condition that can cause thick, scaly patches, or plaques, to form on the skin. It is estimated that more than 8 million people in the United States have psoriasis. Currently, there is no cure for psoriasis, although there are treatments available to control the symptoms.
- Microneedle devices comprising arrays of relatively small structures, sometimes referred to as microneedles or micro-pins, can be used in connection with the delivery of therapeutic agents and other substances through the skin and other surfaces. While treatments exist for autoimmune diseases, patient response is variable. There is a need for new strategies for determining a likelihood that a subject with an autoimmune disease will respond to a therapeutic drug.
- a physician may prescribe a therapeutic aimed at inhibiting or blocking a particular immune system biochemical pathway to manage symptoms.
- a single therapeutic may not be an ideal candidate for two subjects with similar autoimmune diseases.
- This variability between subjects' genetic profiles leads to a trial and error methodology for prescribing therapeutics, often requiring several months of administering a therapeutic that may prove to be ineffective in treating a given subject.
- This approach is both costly to insurance payers and an ineffective route of managing an autoimmune disease that may have substantial impact on quality of life for the subject.
- An aspect of the present disclosure provides a microneedle device comprising: 1) a microneedle region comprising (i) a microneedle base substrate comprising a first base substrate surface and a second base substrate surface, wherein the first base substrate surface and the second base substrate surface are positioned on opposite sides of the microneedle base substrate; and (ii) a plurality of microneedles protruding from the first base substrate surface; and b) a support substrate adjacent to the microneedle base substrate, the support substrate connected to or integral with the microneedle base substrate and comprising a support substrate depth, wherein the support substrate depth is greater than the minimum distance between the first base substrate surface and the second base substrate surface.
- microneedle device comprising: 1) a microneedle region comprising a microneedle base substrate comprising a first base substrate surface with a plurality of microneedles protruding from the first base substrate surface, the microneedle base substrate also comprising a second base substrate surface on a side opposite from the first base substrate surface, the second base substrate surface comprising a recess aligned with at least a portion of the plurality of microneedles; and b) a support substrate adjacent to the microneedle base substrate, the support substrate connected to or integral with the microneedle base substrate.
- the minimum distance between the first base substrate surface and the second base substrate surface of the microneedle base substrate is between about 1 ⁇ m to about 500 ⁇ m less than the depth of the support substrate. In some embodiments, the minimum distance between the first base substrate surface and the second base substrate surface is between 150 ⁇ m to about 350 ⁇ m. In some embodiments, the ratio between (a) the minimum distance between the first base substrate surface and the second base substrate surface, and (b) the support substrate depth is at least 1:5. In some embodiments, the microneedle region comprises a perimeter and the support substrate are adjacent to at least half of the perimeter.
- the support substrate comprises a first support substrate surface proximal to the plurality of microneedles (at times, referred to herein as “the front surface of the support substrate surface”) and a second support substrate surface distal to the plurality of microneedles (at times, referred to herein as “the back surface of the support substrate surface”) and positioned opposite to the first support substrate surface, and wherein the second base substrate surface is not coplanar with the second support substrate surface distal to the plurality of microneedles.
- the first base substrate surface is not coplanar with a surface of the support substrate.
- the plurality of microneedles is plasma treated.
- a plurality of probes is coupled to a microneedle of the plurality of microneedles.
- the plurality of probes comprises a negative charge.
- the plurality of microneedles comprises a polyolefine resin.
- the polyolefine resin comprises one or both of Zeonor 1020R or Zeonor 690R.
- a microneedle of the plurality of microneedles is non-dissolvable.
- a microneedle of the plurality of microneedles is pyramidal.
- a microneedle of the plurality of microneedles is solid.
- an angle between a base of the microneedle and the microneedle base substrate is between about 60° and about 90°.
- the recess aligned with at least a portion of the plurality of microneedles comprises a width that is greater than a width of a mechanical applicator.
- kits comprising microneedle device comprising: 1) a microneedle region comprising a microneedle base substrate comprising a first base substrate surface with a plurality of microneedles protruding from the first base substrate surface, the microneedle base substrate also comprising a second base substrate surface on a side opposite from the first base substrate surface, the second base substrate surface comprising a recess aligned with at least a portion of the plurality of microneedles; and b) a support substrate adjacent to the microneedle base substrate, the support substrate connected to or integral with the microneedle base substrate, and (c) a mechanical applicator that fits within the recess aligned with at least a portion of the plurality of microneedles.
- the nucleic acid probe comprises a homopolymeric sequence. In some embodiments, the homopolymeric sequence comprises thymine or uracil. In some embodiments, the nucleic acid probe comprises DNA. In some embodiments, the nucleic acid probe comprises thymine. In some embodiments, the nucleic acid probe comprises thymidine. In some embodiments, the nucleic acid probe is covalently linked to the microneedle. In some embodiments, the support substrate comprises a fiducial marker. In some embodiments, no more than three microneedles of the plurality of microneedles are less than 600 ⁇ m in length or more than 1050 ⁇ m in length.
- Another aspect of the present disclosure provides a method of preparing a biological sample from a subject comprising: contacting skin of the subject with any one of the microneedle devices described herein, applying pressure to the microneedle device such that the microneedle device penetrates the skin of the subject; and allowing nucleic acids within the skin of the subject to contact the microneedle device.
- the method further comprises extracting nucleic acids, including, for example, RNA, mRNA.
- the method further comprises converting mRNA to cDNA.
- the subject is human.
- the subject has psoriasis or symptoms of psoriasis.
- the subject has a skin condition or symptoms of a skin condition.
- Another aspect of the present disclosure provides a method of preparing a biological sample from a subject comprising: a) contacting skin of the subject with a microneedle device, wherein the microneedle device comprises a plurality of nucleic acid probes coupled to a microneedle; b) applying pressure to the microneedle device such that the microneedle device penetrates the skin of the subject; c) allowing the microneedle device to penetrate the skin of the subject for no more than 10 minutes to obtain a ribonucleic acid (RNA) sample hybridized to the nucleic acid probes, wherein the RNA sample comprises a population of RNA fragments comprising about 700 bases or more and a population of RNA fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of RNA fragments comprising about 700 bases or more to the population of RNA fragments comprising about 150 bases to about 200 bases is greater than 1; and d) removing the microneedle device from the skin of the subject, the micro
- the RNA sample is substantially free of contaminants.
- the microneedle device is allowed to penetrate the skin of the subject for about 5 minutes. In some embodiments, the microneedle device penetrates the skin of the subject for less than 60 minutes, less than 45 minutes, less than 40 minutes, less than 30 minutes, less than 15 minutes, less than 10 minutes, less than 5 minutes, less than 3 minutes, less than 2 minutes, or less than 1 minute. In some embodiments, the microneedle device penetrates the skin of the subject for greater than 30 seconds, greater than 45 seconds, greater than 1 minutes, greater than 5 minutes, greater than 10 minutes, or greater than 30 minutes.
- Another aspect of the present disclosure comprises method of treating a subject with an autoimmune disease of the skin comprising: a) collecting a sample comprising RNA derived from skin from the subject, wherein the subject has not been administered the autoimmune therapeutic drug within 7 days prior to the collecting the sample comprising RNA; b) determining an expression level of at least one gene based on the RNA; c) predicting that a subject with the autoimmune disease of the skin will be responsive to the autoimmune therapeutic drug with a positive predictive value greater than 80% based on the expression level of the at least one gene; and d) based on the predicting in (c), treating the subject with the autoimmune therapeutic drug.
- the PPV is greater than 90%.
- the PPV is greater than 95% for a cohort that has greater than 100 patients.
- the autoimmune therapeutic drug is a biologic or comprises an antibody.
- the autoimmune therapeutic drug is an IL-17 mediated treatment, an IL-23 mediated treatment, or a TNF ⁇ mediated treatment.
- the autoimmune therapeutic drug is at least one drug selected from the group consisting of: etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, and risankizumab.
- the at least one gene comprises at least two genes, three genes, four genes, or five genes. In some embodiments, the at least one gene comprises at least two genes, three genes, four genes, five genes, six genes, seven genes, eight genes, nine genes, or ten genes from Table 6, Table 12, or Table 13. In some instances, the at least one gene comprises at least two genes that do not share a common upstream regulator.
- the autoimmune disorder is psoriasis. In some cases, the subject has a PASI greater than 8. In some cases, the subject has a PASI of at least 75 following the treating of the subject with the autoimmune therapeutic drug.
- the RNA comprises mRNA or microRNA, and is converted into cDNA, and then sequenced using next generation sequencing.
- collecting a sample comprising RNA derived from skin from the subject comprises penetrating the skin of the subject with a microneedle device, wherein the microneedle device comprises microneedles conjugated to nucleic acid probes.
- a trained algorithm is applied to data generated by the next generation sequence of the cDNA.
- the algorithm was trained using samples from patients that were administered a single type of drug selected from the group consisting of: IL-17 mediated treatment, TNF-alpha mediated treatment and IL-23 mediated treatment.
- the autoimmune therapeutic drug is an IL-17-mediated treatment and the at least one gene comprises at least one gene that is not involved in an IL-17 mediated pathway. In some embodiments, the autoimmune therapeutic drug is an IL-23-mediated treatment and the at least one gene comprises at least one gene that is not involved in an IL-23 mediated pathway. In some embodiments, the autoimmune therapeutic drug is a TNF- ⁇ -mediated treatment and the at least one gene comprises at least one gene that is not involved in a TNF- ⁇ -mediated pathway.
- Another aspect of the present disclosure comprises a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug comprising: collecting a sample comprising RNA derived from skin from the subject, wherein the subject has not been administered the autoimmune therapeutic drug within 7 days prior to the collecting the sample comprising RNA; converting the RNA into cDNA; determining expression of at least one gene based on the cDNA; and predicting whether the subject with the skin lesion will be responsive to the autoimmune therapeutic drug with a positive predictive value greater than 80%.
- the PPV is greater than 90%.
- the PPV is greater than 95% for a cohort that has greater than 100 patients.
- the autoimmune therapeutic drug is a biologic or comprises an antibody.
- the autoimmune therapeutic drug is an IL-17 mediated treatment, an IL-23 mediated treatment, or a TNF ⁇ mediated treatment.
- the autoimmune therapeutic drug is at least one drug selected from the group consisting of: etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, and risankizumab.
- the autoimmune therapeutic drug is at least one drug selected from the group consisting of: certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, and risankizumab.
- the at least one gene comprises at least two genes, three genes, four genes, or five genes.
- the at least one gene comprises at least two genes, three genes, four genes, five genes, six genes, seven genes, eight genes, nine genes, or ten genes from Table 6, Table 12, or Table 13.
- the at least one gene comprises at least two genes that do not share a common upstream regulator.
- the autoimmune disorder is psoriasis.
- the subject has a PASI greater than 8.
- the subject has a PASI of at least 75 following the treating of the subject with the autoimmune therapeutic drug.
- the RNA comprises mRNA or microRNA, and is converted into cDNA, and then sequenced using next generation sequencing.
- collecting a sample comprising RNA derived from skin from the subject comprises penetrating the skin of the subject with a microneedle device, wherein the microneedle device comprises microneedles conjugated to nucleic acid probes.
- a trained algorithm is applied to data generated by the next generation sequence of the cDNA.
- the algorithm was trained using samples from patients that were administered a single type of drug selected from the group consisting of: IL-17 mediated treatment, TNF-alpha mediated treatment and IL-23 mediated treatment.
- Another aspect of the present disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug, comprising: penetrating skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a microneedle; removing the microneedle device from the skin of the subject, thereby obtaining RNA molecules from the subject; performing high throughput sequencing on the RNA molecules to generate sequence reads; aligning the sequence reads with a signature of sequence reads associated with a positive response to an autoimmune disease therapeutic drug to obtain aligned sequence reads; and applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm has a greater than 80% positive predictive value for predicting response to the autoimmune disease therapeutic drug.
- Another aspect of the present disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug comprising: penetrating skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a solid microneedle; removing the microneedle device from the skin of the subject, thereby obtaining RNA molecules from the subject; performing high throughput sequencing on the RNA molecules to generate sequence reads; and aligning the sequence reads with a signature of sequence reads associated with a positive response to an autoimmune disease therapeutic drug to obtain aligned sequence reads; using the aligned sequence reads to determine an expression level of at least one RNA molecule; and applying a trained algorithm to the expression level of the at least one RNA molecule, wherein the trained algorithm predicts whether the subject with the skin lesion will be responsive to an IL-17 mediated treatment, an IL-23 mediated treatment, a TNF ⁇ mediated treatment, or any combination thereof.
- the subject will be responsive to the to the IL-17 mediated treatment, the IL-23 mediated treatment, and the TNF ⁇ -mediated treatment.
- the at least one gene comprises at least two genes, three genes, four genes, or five genes.
- the at least one gene comprises at least two genes, three genes, four genes, five genes, six genes, seven genes, eight genes, nine genes, or ten genes from Table 6, Table 12, or Table 13.
- the at least one gene comprises at least two genes that do not share a common upstream regulator.
- the autoimmune disorder is psoriasis.
- the subject has a PASI greater than 8.
- the subject has a PASI of at least 75 following the treating of the subject with the autoimmune therapeutic drug.
- the RNA comprises mRNA or microRNA, and is converted into cDNA, and then sequenced using next generation sequencing.
- the recommended treatment comprises one or more autoimmune therapeutic drugs for an autoimmune disease or condition.
- the recommended treatment comprises etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, risankizumab, or any combination thereof.
- the method described herein further comprises performing high throughput sequencing on the RNA biomarkers to generate one or more sequence reads for the subject; aligning the one or more sequence reads for the subject with a known signature of sequence reads, wherein the known signature of sequence reads is associated with a positive response to the recommended treatment, thereby obtaining aligned sequence reads; and classifying the subject as having a likelihood of positively responding to the recommended treatment by applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm comprises a greater than 50% positive predictive value for predicting a positive response to the recommended treatment. In some embodiments, the trained algorithm has greater than 50% negative predictive value.
- Another aspect of the present disclosure provides a method of determining whether a subject with an autoimmune disorder of the skin will be responsive to an autoimmune therapeutic drug comprising: extracting mRNA from skin of the subject; sequencing the mRNA from the skin of the subject; and predicting whether the subject with the autoimmune disorder will be responsive to etanercept, adalimumab, infliximab, certolizumab, secukinumab, ixekisumab, broadalumab, guselkumab, tildrakizumab, and risankizumab with a positive predictive value greater than 80%.
- Another aspect of the present disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an IL-23 mediated treatment comprising: contacting skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a solid microneedle such that the microneedle device penetrates the skin of the subject; removing the microneedle device from the skin of the subject, thereby obtaining RNA molecules from the subject; performing high throughput sequencing on the RNA molecules to generate sequence reads; and aligning the sequence reads with a signature of sequence reads associated with a positive response to an autoimmune disease therapeutic drug to obtain aligned sequence reads; and applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm predicts whether the subject with the skin lesion will be responsive to an IL-23 mediated treatment and the aligned sequence reads correspond to at least one gene from Table 13.
- Another aspect of the present disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an IL-17, IL-23, or TNF-alpha mediated treatment comprising: contacting skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a solid microneedle such that the microneedle device penetrates the skin of the subject; removing the microneedle device from the skin of the subject, thereby obtaining RNA molecules from the subject; performing high throughput sequencing on the RNA molecules to generate sequence reads; and aligning the sequence reads with a signature of sequence reads associated with a positive response to an autoimmune disease therapeutic drug to obtain aligned sequence reads; and applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm predicts whether the subject with the skin lesion will be responsive to an IL-17 mediated treatment, a TNF-alpha-mediated treatment, or IL-23 mediated treatment and the aligned sequence read
- subject is treated with IL-17 mediated treatment, wherein the aligned sequence reads correspond to at least one gene, two genes, three genes, four genes, five genes, or six genes from the table 12.
- subject is treated with TNF-alpha mediated treatment, wherein the aligned sequence reads correspond to at least one gene, two genes, three genes, four genes, five genes, or six genes from the table 6.
- subject is treated with IL-23 mediated treatment, wherein the aligned sequence reads correspond to at least one gene, two genes, three genes, four genes, five genes, or six genes from the table 13.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of CNFN, CTSC, GBAP1, CRABP2, PCDH7, PPIG, RAB31, C3, and EGR. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of CNFN, CTSC, GBAP1, CRABP2, PCDH7, and PPIG. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of PCDH7, PPIG, RAB31, C3, and EGR.
- the at least one gene is at least one gene, two genes, or three genes from the group consisting of CNFN, CTSC, GBAP1, and CRABP2. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PPIG, RAB31, C3, and EGR. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PCDH7, PPIG, RAB31, and C3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of GBAP1, CRABP2, PCDH7, PPIG.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of SERPINB3, SERPINB4, S100A7A, PI3, KRT6A, LCN2, DEFB4A, DEFB4B, SPRR1A, IL36G, MX1, IFI27, CD36, CD24, and IL4R.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of KRT6A, SPRR1A, CD36, IL4R, LCN2, and IFI27.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of CD36, IL4R, S100A7A, SERPINB4, MX1, and SERPINB3. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of LCN2, IFI27, DEFB4A, IL36G, CD24, and PI3.
- the at least one gene is at least one gene, two genes, or three genes from the group consisting of IL4R, LCN2, and IFI27. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PI3, IFI27, and SERPINB3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of IL4R, S100A7A, and MX1. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of CD36, LCN2, and SERPINB4.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of MTCO1P12, MTATP6P1, CLSTN1, PDPN, LDLRAD2, and GSTM3. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of AL158847.1, DAD1, LDLRAD2, ZNF395, MGMT, and AL136982.4. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of NREP, PPIF, PRIM1, AL136982.5, MTATP6P1, and SMPD3. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of PDPN, TXNRD1, GSTM3, GPSM1, GLRX, and USP2.
- the at least one gene is at least one gene, two genes, or three genes from the group consisting of MTCO1P12, CLSTN1, and GSTM3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of NREP, PPIF, and PRIM1. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of AL136982.5, MTATP6P1, and SMPD3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PDPN, TXNRD1, and GSTM3.
- microneedle devices, methods, and systems for characterizing a subject's autoimmune disease and the likelihood that a subject may respond positive to a given therapeutic to treat said subject's autoimmune disease are also provided herein.
- An aspect of the present disclosure comprises a microneedle device comprising: a microneedle region comprising (i) a plurality of microneedles protruding from a front surface of a microneedle base substrate, (ii) a back surface of said microneedle base substrate; and (iii) a minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate; and a support substrate adjacent to at least one side of said microneedle base substrate, said support substrate connected to or integral with said microneedle base substrate and comprising a support substrate depth, wherein said support substrate depth is greater than said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate.
- microneedle device comprising: a microneedle region comprising a plurality of microneedles protruding from a front surface of a microneedle base substrate, said microneedle base substrate also comprising a back surface, said back surface comprising a recess directly behind at least a portion of said microneedle region; and a support substrate adjacent to at least one side of said microneedle base substrate, said support substrate connected to or integral with said microneedle base substrate.
- said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate is between about 1 ⁇ m to about 500 ⁇ m less than said depth of said support substrate.
- said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate is between 150 ⁇ m to about 350 ⁇ m. In some embodiments, said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate and said support substrate depth comprise an about 1:5 ratio.
- said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate and said support substrate depth comprise at least about a 0.1:5 ratio, at least about a ratio, at least about 1:5 ratio, at least about 2:5 ratio, at least 3:5 ratio, at least 4:5 ratio, at least 1:1 ratio, at least 1:2 ratio, at least 1:3 ratio, at least 1:4 ratio, at least about a 1:10 ratio, at least about a 1:15 ratio, at least about a 1:20 ratio, at least about 1:25 ratio, or at least about 1:50 ratio.
- said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate and said support substrate depth comprise at most about a 0.1:5 ratio, at most about a 0.5:5 ratio, at most about 1:5 ratio, at most about 2:5 ratio, at most 3:5 ratio, at most 4:5 ratio, at most 1:1 ratio, at most 1:2 ratio, at most 1:3 ratio, at most about 1:4 ratio, at most about a 1:10 ratio, at most about a 1:15 ratio, at most about a 1:20 ratio, at most about 1:25 ratio, or at most about 1:50 ratio.
- said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate and said support substrate depth comprise at least about a 2:1 ratio, at least about a 3:1 ratio, at least about 3:2 ratio, at least about 4:1 ratio, at least 5:1 ratio, at least 5:3 ratio, at least 6:1 ratio, at least 7:1 ratio, at least 10:1 ratio, at least 15:1 ratio, at least about a 20:1 ratio, at least about a 25:1 ratio, or at least about a ratio.
- said minimum distance between said front surface of said microneedle base substrate and said back surface of said microneedle base substrate and said support substrate depth comprise at most about a 2:1 ratio, at most about a 3:1 ratio, at most about 3:2 ratio, at most about 4:1 ratio, at most 5:1 ratio, at most 5:3 ratio, at most about 6:1 ratio, at most 7:1 ratio, at most 10:1 ratio, at most 15:1 ratio, at most about a 20:1 ratio, at most about a 25:1 ratio, or at most about a 50:1 ratio.
- said microneedle region comprises a perimeter and said support substrate is adjacent to at least half of said perimeter.
- said back surface of said microneedle base substrate is not coplanar with a back surface of said support substrate.
- said front surface of said microneedle base substrate is not coplanar with a surface of said support substrate.
- said plurality of microneedles is plasma treated.
- a plurality of probes is coupled to a microneedle of said plurality of microneedles.
- said plurality of probes comprises a negative charge.
- said plurality of microneedles comprises a polyolefine resin.
- said polyolefine resin comprises one or both of Zeonor 1020R or Zeonor 690R.
- a microneedle of said plurality of microneedles is non-dissolvable. In some embodiments, a microneedle of said plurality of microneedles is pyramidal. In some embodiments, a microneedle of said plurality of microneedles is solid. In some embodiments, an angle between a base of said microneedle and said microneedle base substrate is between about 60° and about 90°. In some embodiments, said recess directly behind said at least a portion of said microneedle region comprises a width that is greater than a width of a mechanical applicator.
- kits comprising (a) said device of some aspects described herein; and (b) a mechanical applicator that fits within said recess directly behind said at least a portion of said microneedle region.
- at least one of said plurality of microneedles is coupled to a nucleic acid probe.
- said nucleic acid probe comprises a homopolymeric sequence.
- said homopolymeric sequence comprises thymine or uracil.
- said nucleic acid probe is covalently linked to said microneedle.
- said support substrate comprises a fiducial marker.
- no more than three microneedles of said plurality of microneedles are less than 600 ⁇ m in length or more than 1050 ⁇ m in length.
- Another aspect of the present disclosure comprises a method of preparing a biological sample from a subject comprising: contacting skin of said subject with a microneedle device, wherein said microneedle device comprises a plurality of nucleic acid probes coupled to a microneedle; applying pressure to said microneedle device such that said microneedle device penetrates said skin of said subject; allowing said microneedle device to penetrate said skin of said subject for no more than 10 minutes to obtain a ribonucleic acid (RNA) sample hybridized to said nucleic acid probes, wherein said RNA sample comprises a population of RNA fragments comprising about 700 bases or more and a population of RNA fragments comprising about 150 bases to about 200 bases, and wherein a ratio of said population of RNA fragments comprising about 700 bases or more to said population of RNA fragments comprising about 150 bases to about 200 bases is greater than 1; and removing said microneedle device from said skin of said subject, said microneedle device comprising said RNA sample hybridized
- Another aspect of the present disclosure comprises a method of predicting an individual patient's response to a class of biologic drugs, by the mechanism of action of that class of agent, to optimize treatment selection.
- a classifier is trained to predict the response of a patient to either the IL-17, IL-23, or TNF class of biologics using baseline transcriptomic biomarkers (e.g., certain genes) and with high positive predictive value, For example, by using a machine learning-derived classifier to predict response to the anti-IL-17 and anti-IL-23 biologics in patients with psoriasis with high level of positive predictive value.
- Another aspect of the present disclosure comprises an algorithm for the prediction of response to biologics (for example, anti-IL-17, anti-IL-23) used for the treatment of patients with psoriasis, by comparing baseline transcriptomes with clinical response to biologic drugs after initial treatment, for example, at 12 weeks after.
- biologics for example, anti-IL-17, anti-IL-23
- Another aspect of the present disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug, comprising: penetrating skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a microneedle; removing the microneedle device from the skin of the subject, thereby obtaining RNA molecules from the subject; performing high throughput sequencing on the RNA molecules to generate sequence reads; quantifying the RNA molecules comprising sequence reads associated with a positive response to an autoimmune disease therapeutic drug to determine an expression level of at least one gene; and applying a trained algorithm to the expression level, wherein the trained algorithm has a greater than 80% positive predictive value for predicting response to the autoimmune disease therapeutic drug.
- Another aspect of the present disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug comprising: penetrating skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a solid microneedle; removing the microneedle device from the skin of the subject, thereby obtaining RNA molecules from the subject; performing high throughput sequencing on the RNA molecules to generate sequence reads; and quantifying the RNA molecules comprising sequence reads associated with a positive response to an autoimmune disease therapeutic drug to determine an expression level of at least one gene; and applying a trained algorithm to the expression level of the at least one RNA molecule, wherein the trained algorithm predicts whether the subject with the skin lesion will be responsive to an IL-17 mediated treatment, an IL-23 mediated treatment, a TNF ⁇ mediated treatment, or any combination thereof.
- the subject will be responsive to the to the IL-17 mediated treatment, the IL-23 mediated treatment, and the TNF ⁇ -mediated treatment.
- the at least one gene comprises at least two genes, three genes, four genes, or five genes.
- the at least one gene comprises at least two genes, three genes, four genes, five genes, six genes, seven genes, eight genes, nine genes, or ten genes from Table 6, Table 12, or Table 13.
- the at least one gene comprises at least two genes that do not share a common upstream regulator.
- the autoimmune disorder is psoriasis.
- the subject has a PASI greater than 8.
- the subject has a PASI of at least 75 following the treating of the subject with the autoimmune therapeutic drug.
- the RNA comprises mRNA or microRNA, and is converted into cDNA, and then sequenced using next generation sequencing.
- the recommended treatment comprises one or more autoimmune therapeutic drugs for an autoimmune disease or condition.
- the recommended treatment comprises etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, risankizumab, or any combination thereof.
- the method described herein further comprises performing high throughput sequencing on the RNA biomarkers to generate one or more sequence reads for the subject; quantifying the RNA molecules comprising one or more sequence reads for the subject with a known signature of sequence reads, wherein the known signature of sequence reads is associated with a positive response to the recommended treatment, thereby obtaining aligned sequence reads; and classifying the subject as having a likelihood of positively responding to the recommended treatment by applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm comprises a greater than 50% positive predictive value for predicting a positive response to the recommended treatment. In some embodiments, the trained algorithm has greater than 50% negative predictive value.
- FIG. 1 illustrates a cross section of a microneedle device as described herein.
- FIG. 2 illustrates a detail from the cross section of FIG. 1 .
- FIG. 3 illustrates a front view of a microneedle device as described herein.
- FIG. 4 illustrates a microneedle of a microneedle device as described herein.
- FIGS. 5 A-C illustrate quality of different RNA samples collected with a microneedle device as described herein—measured in length of RNA fragments contained in the sample.
- FIG. 5 A illustrates an undegraded sample.
- FIGS. 5 B and 5 C illustrate degraded samples.
- FIG. 6 A illustrates an example flow diagram for a method of treating one or more subjects with a mild or severe form of a skin condition, as described in some embodiments herein.
- FIG. 6 B shows an example flow diagram for a method of preparing a biological sample and identifying one or more therapeutic drugs for one or more subjects, as described in some embodiments herein.
- FIG. 6 C shows an example flow diagram for identifying the activation of one or more pathways that may be associated with a disease or condition in a subject.
- FIG. 7 shows study patient sample enrollment and analysis flow chart.
- FIGS. 8 A-C show correlation between predicted response and patient week 12 PASI changes in psoriasis patients from independent validation data set.
- FIG. 8 A 43 patients treated with IL-23i;
- FIG. 8 B 31 patients treated with IL-17i;
- FIG. 8 C 11 patients treated with TNF ⁇ i.
- X-axis predicted responder or non-responder;
- Y-axis Week 12 PASI changes.
- Red dot median PASI change value.
- FIG. 9 shows predicted response prevalence in patients enrolled in the study.
- FIG. 10 shows overview of RNA-Seq Analysis.
- FIG. 11 Lesional versus non-lesional samples compared using Mindera patches versus punch biopsies from 66 patients in the Mindera database.
- FIG. 12 shows heatmap generated from expression data for the 17 genes identified as correlating to response to anti-IL-17 biologic treatment.
- microneedle devices comprising features that result in precise, patient-friendly sample collection, including microneedle devices comprising features that may minimize pain during the sample collection process.
- the devices described herein can be specifically designed such that they can be fabricated by scalable processes. Features such as the varying thickness of the distinct areas of the devices described herein can result in uniform and precise microneedle devices.
- the specific features of the microneedle devices described herein may further result in sharper microneedles that are not described in the current state of the art. These sharp microneedles may result in an enhanced user or patient (e.g. the subject) experience wherein the user may experience much less pain as compared to microneedle devices of the current state of the art. Combining the precision of the microneedle devices with the enhanced user experience can result in more precise and accurate analysis of biological samples (e.g. the user's skin).
- the microneedle devices provided herein may contain probes attached to the microneedle.
- these probes are configured to bind to one or more biomarkers within a sample or tissue (e.g., skin) of a subject in such a manner as to permit extraction of the biomarker for further analysis.
- the extracted biomarkers can be analyzed, alone or in combination (e.g. generating a genetic signature or gene expression profile) to provide a diagnosis or prediction of a response to a drug or other therapeutic treatment with respect to the subject.
- the gene expression profile is useful for determining an appropriate treatment regimen for an autoimmune disease.
- the methods of the disclosure are useful for personalized medicine, wherein treatment is tailored to a subject based on the particular characteristics of the disease in the subject.
- the present disclosure provides a microneedle device.
- the microneedle device comprises a microneedle region and a support substrate.
- the microneedle region may comprise a plurality of microneedles protruding from a front or first surface of a microneedle base substrate 110 , a back surface (or second base substrate surface) of the microneedle base substrate 160 ; and a minimum distance between the front surface of the microneedle base substrate and the back surface of the microneedle base substrate 170 .
- the minimum distance between the front surface of the microneedle base substrate and the back surface of the microneedle base substrate 170 configures the device such that during some methods of manufacturing the device, a molten medium (e.g. the polyolefin resins described herein) will fill a mold of the microneedles prior to filling the mold of other sections of the device.
- the minimum distance between the front surface of the microneedle base substrate and the back surface of the microneedle base substrate 170 configures the device such that the device comprises uniform and/or sharp microneedles.
- the uniform and/or sharp microneedles result in an enhanced user experience for a subject using the device.
- the enhanced user experience comprises lessened pain experienced by the subject using the device.
- microneedle devices for detection and acquisition of biomarkers from a subject in situ.
- the microneedle-based device may contain one or more microneedles that can be pierced into biological barriers such as the skin or mucous membrane. Often, the microneedles are non-invasive, or minimally-invasive.
- the device can also have a planar substrate which supports the microneedles.
- the substrate can be made of the same material as that of the microneedle. It can also be made of a different material.
- Microneedle devices of the present disclosure may comprise a base substrate comprising a microneedle region, and a support substrate.
- FIG. 1 illustrates a side view of an exemplary microneedle device 100 , comprising microneedles, 110 , on a microneedle base substrate, 120 .
- the microneedle base substrate comprises a front surface (used interchangeably herein with the term “first base substrate surface”), 140 , wherein the microneedles, 110 , protrude from the front surface (or first base substrate surface), and a back surface (used interchangeably herein with the term “second base substrate surface”), 160 .
- the device may comprise a support substrate, 130 , adjacent to at least one side of the interior microneedle base substrate, 120 , the support substrate connected to or integral with the microneedle base substrate. In some embodiments, an angle is formed between the back surface of the microneedle base substrate, 160 and the adjacent support substrate, 130 .
- the angle formed between the back surface of the microneedle base substrate, 160 , and the adjacent support substrate, 130 is a right angle or obtuse angle, e.g., about 90° to about 179°.
- the angle ( ⁇ ) formed between the back surface of the microneedle base substrate, 160 , and the adjacent support substrate, 130 generally refers to an angle that spans a region of space interior to the device; while the adjacent support substrate generally comprises a complementary angle ( ⁇ ) spanning the substrate and that is often an acute angle (e.g., between 1° and 90°).
- the angle ( ⁇ ) formed between the back surface of the microneedle base substrate and the adjacent support substrate is about 110°.
- the angle ( ⁇ ) formed between the back surface of the microneedle base substrate and the adjacent support substrate is about 90° to about 100°, about 90° to about 110°, about 90° to about 120°, about 90° to about 130°, about 90° to about 140°, about 90° to about 150°, about 90° to about 160°, about 90° to about 170°, about 90° to about 179°, about 100° to about 110°, about 100° to about 120°, about 100° to about 130°, about 100° to about 140°, about 100° to about 150°, about 100° to about 160°, about 100° to about 170°, about 100° to about 179°, about 110° to about 120°, about 110° to about 130°, about 110° to about 140°, about 110° to about 150°, about 110° to about 160°, about 110° to about 170°, about 110° to about 179°, about 120° to about 130°, about 120° to about 140°, about 110° to about 150°, about 110° to about 160°,
- the angle formed is between the back surface of the microneedle base substrate and the adjacent support substrate is about 90°, about 100°, about 110°, about 120°, about 130°, about 140°, about 150°, about 160°, about 170°, or about 179°. In some embodiments, the angle formed between the back surface of the microneedle base substrate and the adjacent support substrate is at least about 90°, about 100°, about 110°, about 120°, about 130°, about 140°, about 150°, about 160°, or about 170°.
- the angle formed between the back surface of the microneedle base substrate and the adjacent support substrate is at most about 100°, about 110°, about 120°, about 130°, about 140°, about 150°, about 160°, about 170°, or about 179°.
- the resulting shape of the support substrate, 130 may also influence the speed and uniformity of the flow of polymer and/or resin into the section mold resembling the structure of the device during manufacturing of the device.
- the resulting shape of the support substrate, 130 is circular.
- the resulting shape of the support substrate, 130 is linear.
- the arrangement of the microneedles is a circular pattern with a radius of the microneedle section being about 0.95 mm to about 4.15 mm.
- the radius is about 4.0 mm.
- the radius is about 0.95 mm to about 1.4 mm, about 0.95 mm to about 1.75 mm, about 0.95 mm to about 2.15 mm, about 0.95 mm to about 2.55 mm, about 0.95 mm to about 2.95 mm, about 0.95 mm to about 3.35 mm, about 0.95 mm to about 3.75 mm, about 0.95 mm to about 4.15 mm, about 1.4 mm to about 1.75 mm, about 1.4 mm to about 2.15 mm, about 1.4 mm to about 2.55 mm, about 1.4 mm to about 2.95 mm, about 1.4 mm to about 3.35 mm, about 1.4 mm to about 3.75 mm, about 1.4 mm to about 4.15 mm, about 1.75 mm, about 1.4 mm to about 2.15 mm, about 1.4
- the radius is about 0.95 mm, about 1.4 mm, about 1.75 mm, about 2.15 mm, about 2.55 mm, about 2.95 mm, about 3.35 mm, about 3.75 mm, or about 4.15 mm. In some embodiments, the radius is at least about 0.95 mm, about 1.4 mm, about 1.75 mm, about 2.15 mm, about 2.55 mm, about 2.95 mm, about 3.35 mm, or about 3.75 mm. In some embodiments, the radius is at most about 1.4 mm, about 1.75 mm, about 2.15 mm, about 2.55 mm, about 2.95 mm, about 3.35 mm, about 3.75 mm, or about 4.15 mm.
- the arrangement of the microneedles is a linear square pattern with total edge length of about 2.14 mm to about 9.34 mm.
- the distance is about 4.0 mm. In some embodiments, the distance is about 2.14 mm to about 3.04 mm, about 2.14 mm to about 3.94 mm, about 2.14 mm to about 4.84 mm, about 2.14 mm to about 5.74 mm, about 2.14 mm to about 6.64 mm, about 2.14 mm to about 7.54 mm, about 2.14 mm to about 8.44 mm, about 2.14 mm to about 9.34 mm, about 3.04 mm to about 3.94 mm, about 3.04 mm to about 4.84 mm, about 3.04 mm to about 5.74 mm, about 3.04 mm to about 6.64 mm, about 3.04 mm to about 7.54 mm, about 3.04 mm to about 8.44 mm, about 3.04 mm to about 9.34 mm, about 3.94 mm to about 4.84 mm, about 3.94 mm to about 4.84 mm to
- the distance is about 2.14 mm, about 3.04 mm, about 3.94 mm, about 4.84 mm, about 5.74 mm, about 6.64 mm, about 7.54 mm, about 8.44 mm, or about 9.34 mm. In some embodiments, the distance is at least about 2.14 mm, about 3.04 mm, about 3.94 mm, about 4.84 mm, about 5.74 mm, about 6.64 mm, about 7.54 mm, or about 8.44 mm. In some embodiments, the distance is at most about 3.04 mm, about 3.94 mm, about 4.84 mm, about 5.74 mm, about 6.64 mm, about 7.54 mm, about 8.44 mm, or about 9.34 mm.
- the surface from which the microneedles protrude from, and which is applied to the tissue of interest may be termed the bottom surface or front surface.
- the surface that is opposite to the surface from which the microneedles protrude from, 160 may be termed the back or top surface.
- the microneedle base substrate, 120 is defined by the presence of microneedles.
- the microneedle base substrate, 120 may be thinner and/or narrower than the support substrate, 130 , e.g. the distance between the front surface of the microneedle base substrate, 140 , and back surface of the microneedle base substrate, 160 , termed D BS ( 170 in FIG. 1 ) is less than the distance between the respective, adjacent surfaces of the support substrate, 130 , termed D SS ( 180 in FIG. 1 ).
- D SS the distance between the respective, adjacent surfaces of the support substrate, 130 .
- the difference in depth between the microneedle base substrate, 120 , and the support substrate, 130 may create a recess in the device, the recess located adjacent to the microneedle base substrate, 120 .
- This structural feature is particularly advantageous because having a narrow microneedle base substrate, 120 , improves the flow and/or penetration of a polymer/resin used to fabricate the device into a mold resembling the structure of the device during manufacturing, specifically the flow and/or penetration of the polymer/resin into the section of the mold corresponding to the microneedles is increased due to this structural feature of a difference in depth between the microneedle base substrate, 120 , and the support substrate, 130 , resulting in more uniform and sharper microneedles. Sharper microneedles may cause less tissue damage when inserted, and therefore, may result in less pain to a subject. More uniform microneedles result in more standard and scalable manufacturing methods.
- the narrower microneedle base substrate may also concentrate pressure through the microneedles when the device is applied to skin or other tissues.
- the minimum distance between the front surface of the microneedle base substrate, 140 , and the back surface of the microneedle base substrate, 150 is between about 1 ⁇ m to about 500 ⁇ m less than the distance from the front surface of the support substrate to the back surface of the support substrate.
- the difference in depth between the microneedle base substrate (D BS ) and the support substrate (D SS ) is about 1 ⁇ m to about 550 ⁇ m. In some embodiments, the difference in depth between the microneedle base substrate (D BS ) and the support substrate (D SS ) is about 250 ⁇ m.
- the difference in depth between the microneedle base substrate (D BS ) and the support substrate (D SS ) is about 1 ⁇ m to about 50 ⁇ m, about 1 ⁇ m to about 100 ⁇ m, about 1 ⁇ m to about 150 ⁇ m, about 1 ⁇ m to about 200 ⁇ m, about 1 ⁇ m to about 250 ⁇ m, about 1 ⁇ m to about 300 ⁇ m, about 1 ⁇ m to about 350 ⁇ m, about 1 ⁇ m to about 400 ⁇ m, about 1 ⁇ m to about 450 ⁇ m, about 1 ⁇ m to about 500 ⁇ m, about 1 ⁇ m to about 550 ⁇ m, about 50 ⁇ m to about 100 ⁇ m, about 50 ⁇ m to about 150 ⁇ m, about 50 ⁇ m to about 200 ⁇ m, about 50 ⁇ m to about 250 ⁇ m, about 50 ⁇ m to about 300 ⁇ m, about 50 ⁇ m to about 350 ⁇ m, about 50 ⁇ m to about 400 ⁇ m, about 50 ⁇ m to about
- the difference in depth between the microneedle base substrate and the support substrate is about 1 ⁇ m, about 50 ⁇ m, about 100 ⁇ m, about 150 ⁇ m, about 200 ⁇ m, about 250 ⁇ m, about 300 ⁇ m, about 350 ⁇ m, about 400 ⁇ m, about 450 ⁇ m, about 500 ⁇ m, or about 550 ⁇ m.
- the difference in depth between the microneedle base substrate (D BS ) and the support substrate (D SS ) is at least about 1 ⁇ m, about 50 ⁇ m, about 100 ⁇ m, about 150 ⁇ m, about 200 ⁇ m, about 250 ⁇ m, about 300 ⁇ m, about 350 ⁇ m, about 400 ⁇ m, about 450 ⁇ m, or about 500 ⁇ m.
- the difference in depth between the microneedle base substrate and the support substrate is at most about 50 ⁇ m, about 100 ⁇ m, about 150 ⁇ m, about 200 ⁇ m, about 250 ⁇ m, about 300 ⁇ m, about 350 ⁇ m, about 400 ⁇ m, about 450 ⁇ m, about 500 ⁇ m, or about 550 ⁇ m.
- the minimum distance between the front surface of the microneedle base substrate and the back surface of the microneedle base substrate (D BS ) is between 150 ⁇ m to about 350 ⁇ m.
- the ratio of the distance between the front and back of the interior section and the front and back of the peripheral section is about a 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10 ratio.
- the microneedle section, or region comprises a perimeter and the peripheral region, or support substrate, is adjacent to at least half of the perimeter.
- the back surface of the microneedle base substrate is not coplanar with a back surface of the support substrate.
- the front surface of the microneedle base substrate is not coplanar with a surface of the support substrate.
- FIG. 2 provides a detail view of the circled region in FIG. 1 .
- the interior section may also have a ridge, 200 , which surrounds the microneedle region.
- the ridge may protrude from the surface from which the microneedles protrude from.
- the height of the ridge, H R ( 210 in FIG. 2 ), may be about 250 ⁇ m.
- the height of the ridge, H R may be about 50 ⁇ m, 100 ⁇ m, 150 ⁇ m, 200 ⁇ m, 250 ⁇ m, 254 ⁇ m, 300 ⁇ m, 350 ⁇ m, 400 ⁇ m, or 450 ⁇ m.
- the ridge surrounding the interior section may increase or maintain surface tension of a tissue, such as skin, when the device is applied to the tissue.
- the angle formed between the ridge and the bottom surface is about 5° to about 85°. In some cases, the angle formed between the ridge and the bottom surface is about 45°.
- the angle formed between the ridge and the bottom surface is about 5° to about 10°, about 5° to about 20°, about 5° to about 30°, about 5° to about 40°, about 5° to about 50°, about 5° to about 60°, about 5° to about 70°, about 5° to about 80°, about 5° to about 85°, about 10° to about 20°, about 10° to about 30°, about 10° to about 40°, about 10° to about 50°, about 10° to about 60°, about 10° to about 70°, about 10° to about 80°, about 10° to about 85°, about 20° to about 30°, about 20° to about 40°, about 20° to about 50°, about 20° to about 60°, about 20° to about 70°, about 20° to about 80°, about 20° to about 85°, about 30° to about 40°, about 30° to about 50°, about 30° to about 60°, about 30° to about 70°, about 30° to about 80°, about 30° to about 85°, about 30
- the angle formed between the ridge and the bottom surface is about 5°, about 10°, about 20°, about 30°, about 40°, about 50°, about 60°, about 70°, about 80°, or about 85°. In some cases, the angle formed between the ridge and the bottom surface is at least about 5°, about 10°, about 20°, about 30°, about 40°, about 50°, about 60°, about 70°, or about 80°. In some cases, the angle formed between the ridge and the bottom surface is at most about 10°, about 20°, about 30°, about 40°, about 50°, about 60°, about 70°, about 80°, or about 85°.
- the microneedle section may also have a bottom surface which is lower than the bottom surface of the support substrate, e.g., the bottom surface of the microneedle section may protrude farther from the device compared to the bottom surface of the support substrate.
- a bottom surface may overlap the base of the microneedles.
- the bottom surface of the interior section may protrude by about 250 ⁇ m.
- the bottom surface of the interior section may protrude by about 0 ⁇ m, 50 ⁇ m, 100 ⁇ m, 150 ⁇ m, 200 ⁇ m, 250 ⁇ m, 300 ⁇ m, 350 ⁇ m, 400 ⁇ m, or 450 ⁇ m.
- the protruding bottom surface of the interior section may increase or maintain surface tension of a tissue such as skin when the device is applied to the tissue.
- the support substrate, 130 may be formed of the same material as the microneedle section and the microneedles.
- the support substrate may be thicker than the interior section.
- the peripheral section comprises a fiducial marker.
- FIG. 3 provides a front view of microneedle device 100 , with a fiducial marker, 310 .
- a device of the present disclosure may be manufactured using a mold.
- a device may be manufactured by displacing a resin (e.g., polyolefin resin) into a mold for the device.
- the mold may comprise a plurality of cavities for forming a plurality of microneedles, cavity for forming an interior section comprising the plurality of microneedles; and one or more cavities for forming one or more peripheral sections adjacent to the interior section, wherein a width of the interior section is less than a width of the peripheral sections.
- the polyolefin resin may generate the plurality of microneedles prior to generating the interior section.
- the molded resin may be plasma treated to modify a surface.
- a device of the present disclosure may be manufactured using injection molding.
- a device may be manufactured by injecting heated (e.g. molten) material (e.g. a polyolefin resin described herein) into a mold of the device.
- the mold may comprise a plurality of cavities for forming a plurality of microneedles, cavity for forming an interior section comprising the plurality of microneedles, and one or more cavities for forming one or more peripheral sections adjacent to the interior section, wherein a width (e.g. depth and/or thickness) of the interior section is less than a width of the peripheral sections.
- the heated material may generate the plurality of microneedles prior to generating the interior or microneedle section.
- the entire device may be plasma treated, while in other cases a portion of the device, for example the microneedles, is plasma treated.
- the method of plasma treating the device or portion thereof comprises the steps of providing the device or portion thereof, placing the device or portion thereof into the plasma vacuum chamber, closing the plasma vacuum chamber creating a sufficient vacuum seal, evacuating all of the gas present in the chamber, pumping a plasma treatment gas into the plasma vacuum chamber to a desirable pressure, enabling the generation of gas plasma for a desirable period of time at a desirable power, evacuating the chamber of the plasma treated gas and removing the device or portion thereof.
- the entire device may be excimer laser treated, while in other cases a portion of the device, for example the microneedles, is excimer laser treated.
- the method of excimer laser treating the device or portion thereof comprises the steps of providing the device or portion thereof, placing the device or portion thereof into the excimer laser treating chamber, closing the excimer laser treating chamber, evacuating all of the gas present in the chamber, pumping a suitable excimer laser treatment gas into the excimer laser treating chamber to provide sufficient absorption of radiative laser energy by the device or portion thereof, enabling the radiative emission of an excimer laser for a desirable period of time at a desirable power, evacuating the excimer laser treatment gas and removing the device or portion thereof.
- one or more probes of the plurality of probes are attached to one or more microneedles of the plurality of microneedles.
- the probes may be covalently or non-covalently attached.
- the probes are attached via a linker or a spacer.
- the probes are attached to a microneedle as to achieve a maximum packing density on the surface of the microneedle.
- the microneedles comprise probes that can bind and extract biomarkers from a tissue.
- a plurality of probes can be attached to a microneedle of the disclosure.
- the probes comprise polynucleotides (e.g., DNA, RNA, cDNA, cRNA, etc.). Often the polynucleotide probes are designed to bind or hybridize a specific polynucleotide biomarker.
- This disclosure also provides methods and devices for detecting peptide or protein biomarkers.
- the probes attached to the microneedles can specifically recognize and bind to target peptides or proteins of interest.
- the probes can be any substance capable of binding to a specific peptide or protein biomarker.
- polynucleotide can be, e.g., a protein (e.g., an antibody, antigen, or fragment thereof), carbohydrate, or a polynucleotide.
- the polynucleotide may possess sequence specificity for the biomarker, e.g., by having a complementary sequence.
- the probes to be used often depend on the biomarker or biomarkers to be detected.
- the number of probes immobilized to a microneedle can be 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.
- the total number of probes immobilized to a microneedle can be from about 1 probe to about 1,000 probes, from about 1 probe to about 10,000 probes, from about 1 probe to about 100,000 probes, from about 1 probe to about 1,000,000 probes, from about 1 probe to about from about 1 probe to about 100,000,000 probes, from about 1,000 probes to about 100,000,000 probes, from about 10,000 probes to about 100,000,000 probes, or from about probes to about 100,000,000 probes.
- the total number of probes in a microneedle is at least about 10, about 20, about 50, about 100, about 500, about 1000, about 10000, about 100000, about 1,000,000, about 10,000,000, or about 100,000,000 probes. In some cases, the total number of probes in a microneedle is less than about 10, about 100, about 1000, about 10000, about 100000, about 1,000,000, about 10,000,000, or about 100,000,000 probes.
- a probe for the biomarker can be immobilized to multiple microneedles in the device of the disclosure, particularly for detection of a biomarker of low concentration, as described further herein.
- a single microneedle comprises multiple different probes capable of binding or detecting the same biomarker.
- a single microneedle comprises at least 2 different probes, at least 3 different probes, at least 4 different probes, at least 6 different probes, at least 8 different probes, at least 10 different probes, at least 15 different probes, at least 20 different probes, or at least 50 different probes.
- the same microneedle comprises more than 50 different types of probes for the same biomarker.
- the same microneedle comprises a plurality of different probes.
- the different probes can be specific for the same biomarker or for a different biomarker.
- a microneedle can comprise at least 2 different probes, at least 10 different probes, at least 100 different probes, at least 200 different probes at least 500 different probes, at least 1,000 different probes, at least 5,000 different probes, or at least 10,000 different probes.
- the individual probes of the plurality of probes are identical (e.g., identical copies of the same polynucleotide or antibody).
- a microneedle can be associated with numerous copies of the same probe (e.g., greater than 2, 5, 10, 50, 100, 1000, 5000, 7500, 10000, or 50000 copies of the same probe).
- a microneedle can comprise a plurality of copies of a polynucleotide probe designed to hybridize the same polymorphism or biomarker.
- a microneedle may comprise a plurality of copies of an antibody probe designed to bind the same epitope.
- a probe may bind to a class of biomarkers, for example mRNAs.
- a probe which binds to mRNAs may comprise a polynucleotide with a homopolymeric sequence or a polynucleotide that comprises a homopolymeric sequence.
- the probe comprises one or more thymine residues.
- the probe comprises at least 1, at least at least 10, at least 20, at least 25, at least 30, at least 40, at least 50, at least 100, or at least 200 thymine residues (e.g., a thymidine residue).
- the probe binds to a poly-A tail of an mRNA molecule.
- the probe comprises a homopolymeric sequence.
- the homopolymeric sequence is configured to bind to a complementary homopolymeric sequence.
- a probe comprises a sequence of thymines which bind to the poly A tails of the mRNAs.
- a probe may comprise any fragment of a sequence of consecutive thymines.
- the sequence of consecutive thymines comprises about 250 consecutive thymines.
- the sequence of consecutive thymines comprises about 200 consecutive thymines.
- the sequence of consecutive thymines comprises about 150 consecutive thymines.
- the sequence of consecutive thymines comprises about 100 consecutive thymines.
- the sequence of consecutive thymines comprises about 50 consecutive thymines. In some embodiments, the sequence of consecutive thymines comprises about 50 and 250 consecutive thymines. In some cases, a probe may comprise thymines interspersed with other bases.
- a microneedle comprises polynucleotide probes.
- the probes may be designed to detect different biomarkers (e.g. nucleic acid molecules, proteins) associated with the same disease, disorder or condition.
- a first probe may recognize a polymorphism (e.g., DNA polymorphism, RNA polymorphism) associated with a disease and a second probe may recognize a different polymorphism associated with the same disease.
- a first nucleic acid probe on a microneedle can be designed to detect a first polymorphism of an RNA biomarker associated with onchocerciasis, a skin condition.
- a second nucleic acid probe on a microneedle can be designed to detect a second polymorphism of an RNA biomarker associated with onchocerciasis.
- a polymorphism can be, for example, a single nucleotide polymorphism (SNP).
- SNP single nucleotide polymorphism
- Genetic and genomic variations can comprise a single SNP or a plurality of SNPs. SNPs can occur at a single locus, or at many loci. Individuals who carry a particular SNP on an allele at one locus can predictably carry additional SNPs at other loci.
- a correlation of SNPs can provide an association between alleles predisposing an individual to disease or condition.
- the different polynucleotide probes are designed to detect different biomarkers associated with different conditions.
- one probe may detect a biomarker of a disease (e.g. polynucleotides), while the other probe may detect a housekeeping gene or gene product (e.g. polypeptides).
- the microneedles are attached to polynucleotides, polypeptides, or a mixture of polynucleotides and polypeptides.
- the probes may comprise a negative charge.
- a plurality of probes may comprise a residual negative charge.
- the microneedles of the devices described herein are packed with probes at the maximum packing density for the microneedle. The residual negative charge generated from a maximum-packed microneedle can be strong enough to prevent the packed probes from non-specific binding to biomarkers that are not of interest.
- a microneedle can also be associated with a plurality of different protein or antibody probes.
- a first antibody probe on a microneedle can be designed to detect a first epitope of an antigen associated with, for example, a skin cancer.
- a second antibody probe can be designed to detect a second epitope associated with the antigen.
- the second antibody probe can detect an epitope associated with a different skin condition and/or different antigen.
- the present disclosure provides a microneedle device comprising a set of microneedles, wherein each microneedle in the set comprises identical probes or set of probes.
- an identical probe is attached to a plurality of microneedles of a device.
- An identical probe can be attached to, for example, about 1% of the microneedles, about 5% of the microneedles, about 10% of the microneedles, about 50% of the microneedles, about 90% of the microneedles, about 95% of the microneedles, or about 100% of the microneedles.
- an identical probe is attached to no greater than 5% of the microneedles, no greater than 10% of the microneedles, no greater than 25% of the microneedles, no greater than 50% of the microneedles, no greater than 95% of the microneedles, or no greater than 99% of the microneedles.
- a plurality of microneedles may comprise at least one microneedle attached to a first probe and at least one microneedle attached to a second probe that is different from the first probe.
- the first probe may be a polynucleotide or polypeptide (e.g., antibody, protein) that specifically binds a biomarker of a disease or disorder and the second probe may be a polynucleotide or polypeptide that specifically binds a different biomarker associated with the same disease or disorder.
- the first probe may be a polynucleotide or polypeptide (e.g., antibody, protein) that specifically binds a biomarker of a disease or disorder and the second probe may be a polynucleotide or polypeptide that specifically binds a different biomarker associated with a different disease, disorder, or condition.
- the different disease, condition, or disorder is associated with the same organ.
- the first probe may be associated with a first disease, disorder, or condition associated with skin; and the second probe may be associated with a second disease, disorder, or condition associated with skin or eye.
- the device may comprise an array of microneedles, wherein each microneedle comprises a probe that detects a biomarker associated with a different disease, disorder, or condition associated with the same organ.
- the array of microneedles may comprise greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 75, 100, 150, 200, 500, or 1000 microneedles associated with different diseases, disorders, or conditions.
- the different diseases, disorders, or conditions are associated with different organs (e.g., greater than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 organs).
- the microneedle device comprises a plurality of arrays of microneedles; often, the plurality of arrays of microneedles are suitable for multiplexed reactions. In some cases, the plurality of arrays comprises two or more arrays of microneedles, wherein the arrays are designed to detect a different biomarker. In some cases, a first array of microneedles may be designed to detect a biomarker associated with disease, disorder, or condition, and the second array of microneedles is designed to detect a different biomarker associated with the same disease, disorder, or condition.
- a first array of microneedles may be designed to detect a biomarker associated with disease, disorder, or condition
- a second array of microneedles may be designed to detect a different biomarker associated with a different disease, disorder, or condition.
- a first array of microneedles may be designed to detect a plurality of biomarkers associated with a disease, disorder, or condition
- a second array of microneedles may be designed to detect a plurality of biomarkers associated with a different disease, disorder, or condition.
- the second array of microneedles may be designed to detect control biomarkers (e.g., housekeeping genes), either positive controls or negative controls.
- the probes are covalently attached to the microneedles. In some embodiments, the probes are attached to the microneedles at about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% of a maximum packing density.
- a maximum packing density may be determined by the chemical structure of the microneedle surface.
- a high density of probes may decrease non-specific binding. For example, a high density of polynucleotide probes will discourage non-specific polynucleotide binding due to the residual negative charge.
- the microneedles may be treated to facilitate a high density of probe attachment.
- the microneedles may be plasma treated. The plasma treatment may comprise treatment with an oxygen, nitrogen, or carbon dioxide plasma to modify the surface.
- a microneedle can have a plurality of shapes, for example, a microneedle can be round, conical, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, pyramidal, or any other suitable shape.
- a microneedle can be a sharp microneedle, a blunt microneedle, or any combination thereof.
- a device of the present disclosure comprising a plurality of sharp microneedles can be used to penetrate the skin of a subject thereby contacting the probe(s) on the microneedle with, for example, a biomarker.
- a sharp microneedle can be used to disrupt tissue of a biological sample, such as a layer of cells or the outer membrane of a cell.
- a blunt microneedle can be used to touch the surface of the skin of a subject thereby contacting the microneedle with, for example, a cell-surface biomarker on the skin.
- the present disclosure provides microneedle devices that comprise microneedles with different shapes (e.g., sharp and blunt microneedles).
- the microneedles may be solid rather than hollow.
- the present disclosure provides microneedle devices in which all needles have a same shape.
- all microneedles on a device have the same shape and dimensions, within a 5%, 4%, 3%, 2%, 1.5%, 1%, 0.75%, 0.5%, or 0.25% error.
- no more than three microneedles of a plurality of microneedles do not meet the height tolerance. For example, if a desired microneedle height is 950 ⁇ m, then a height tolerance may be no less than 600 ⁇ m in length and no more than 1050 ⁇ m in length.
- a microneedle may have a shape that allows non-specifically bound material to be wiped off needle upon microneedles' exit from skin.
- the non-specifically bound material may be wiped off via shearing forces generated as the microneedle is drawn through the skin.
- the needle-shaped microneedles can be blunt-pointed objects, they are preferably sharp-pointed objects.
- the microneedles have a circular cone structure with a diameter of the base generally in the range of 10 ⁇ m to 500 ⁇ m, preferably in the range of 20 ⁇ m to 200 ⁇ m.
- FIG. 4 illustrates a surface of a device of the present disclosure with a plurality of microneedles wherein the diameter of the base is less than 10 mm in width.
- a microneedle of the disclosure can comprise a plurality of different diameters or base widths.
- the shape of the base of a microneedle can be, for example, round, rectangular, triangular, square, pentagonal, hexagonal, heptagonal, or other geometric shape.
- a microneedle of the disclosure can have a diameter or base width that is no greater than 500 ⁇ m, no greater than 400 ⁇ m, no greater than 300 ⁇ m, no greater than 200 ⁇ m, no greater than 100 ⁇ m, no greater than 50 ⁇ m, no greater than 40 ⁇ m, no greater than 30 ⁇ m, no greater than 20 pit, no greater than 10 pit, no greater than 1000 nm, no greater than 900 nm, no greater than 800 nm, no greater than 700 nm, no greater than 600 nm, no greater than 500 nm, no greater than 400 nm, no greater than 300 nm, no greater than 200 nm, or no greater than 100 nm.
- a microneedle with a circular cone structure may have an angle of the edge of the cone to the base of the cone from 70° to about 90°, from 71° to 89°, from 72° to 88°, from 73° to 86°, from 73° to 84°, from 73° to 80°, from 74° to 78°, from 74° to 76°, or from 75.5° to 76°.
- a microneedle with a circular cone structure may have an angle from the edge of the cone to the base of the cone of about 73°, 73.25°, 73.5°, 73.75°, 74°, 74.25°, 74.5°, 74.75°, 75°, 75.25°, 75.5°, 75.75°, 76°, 76.25°, 76.5°, 76.75°, or 77°.
- a microneedle with a circular cone structure may have an angle of the edge of the cone to the base of less than 73°, 73.25°, 73.5°, 73.75°, 74°, 74.25°, 74.5°, 74.75°, 75°, 75.25°, 75.5°, 75.75°, 76°, 76.25°, 76.5°, 76.75°, or 77°.
- the substrate and the microneedles of the arrays can be made of various biodegradable or non-biodegradable materials.
- the material for the microneedles or substrate include poly(methyl methacrylate), silicon, silicon dioxide, ceramic, metal (such as stainless steel, titanium, nickel, molybdenum, chromium, and cobalt), and synthetic or natural resin material.
- a biodegradable polymer such as polylactic acid, polyglycolide, polylactic acid-co-polyglycolide, pullulan, capronolactone, polyurethane or polyanhydride.
- a non-degradable material is employed to fabricate the microneedle array, e.g., a polymer polycarbonate, a synthetic or natural resin material such as polymethacrylic acid, ethylenevinylacetate, polytetrafluoroethylene, polysulfone, or polyoxymethylene.
- the microneedles may be non-dissolvable.
- the microneedle is fabricated with a thermoplastic polymer.
- the substrate and the microneedles of the arrays can be made of various thermoplastic polymers.
- thermoplastic polymers include acrylic polymers, such as poly(methyl methacrylate) (PMMA), nylon, polyethylene, polypropylene, polystyrene, polyvinyl chloride, or Teflon.
- PMMA poly(methyl methacrylate)
- nylon polyethylene
- polypropylene polystyrene
- polyvinyl chloride polyvinyl chloride
- Teflon Teflon
- a device of the present disclosure is fabricated with a thermoplastic polymer selected from the group consisting of polycarbonate, poly(methyl methacrylate), polyethylene and polypropylene.
- Non-limiting examples of non-degradable polymers include, for example, silicone, hydrogels such as crosslinked poly(vinyl alcohol) and poly(hydroxy ethylmethacrylate), ethylene-vivyl acetate, acyl substituted cellulose acetates and alkyl derivatives thereof, partially and completely hydrolyzed alkylene-vinyl acetate copolymers, unplasticized polyvinyl chloride, crosslinked homo- and copolymers of polyvinyl acetate, crosslinked polyesters of acrylic acid and/or methacrylic acid, polyvinyl alkyl ethers, polyvinyl fluoride, polycarbonate, polyurethane, polyamide, polysulphones, styrene acrylonitrile copolymers, crosslinked poly(ethylene oxide), poly(alkylenes), poly(vinyl imidazole), poly(esters), poly(ethylene terephthalate), polyphosphazenes, and chlorosulphonated polyolefines,
- biodegradable polymers include polyesters such as 3-hydroxypropionate, 3-hydroxybutyrate, 3-hydroxyvalerate, 3-hydroxycaproate, 3-hydroxyheptanoate, 3-hydroxyoctanoate, 3-hydroxynonanoate, 3-hydroxydecanoate, 3-hydroxyundecanoate, 3-hydroxydodecanoate, 4-hydroxybutyrate, 5-hydroxyvalerate, polylactide or polylactic acid including poly(d-lactic acid), poly(l-lactic acid), poly(d,l-lactic acid), polyglycolic acid and polyglycolide, poly(lactic-co-glycolic acid), poly(lactide-co-glycolide), poly( ⁇ -caprolactone) and polydioxanone.
- Polysaccharides including starch, glycogen, cellulose and chitin can also be used as biodegradable materials.
- the substrate and the microneedles of the arrays can be made of a polyolefin resin.
- polyolefin resins include thermoplastic polyolefins: polyethylene (PE), polypropylene (PP), polymethylpentene (PMP), and polybutene-1 (PB-1).
- Further examples of polyolefin resins include polyolefin elastomers (POE): polyisobutylene (PIB), ethylene propylene rubber (EPR), and ethylene propylene diene monomer (M-class) rubber (EPDM rubber).
- the polyolefin resin may be a Cyclo Olefin Polymer such as Zeonor or Zeonex. In some cases, the Zeonor is Zeonor 1020R, Zeonor 690R or Zeonor 1060R.
- the microneedles employed in the present disclosure have a length (height) that is typically in the range of 20 ⁇ m to 1 mm, preferably in the range of 50 ⁇ m to 500 ⁇ m.
- the height of the needles may range from about 400 ⁇ m to about 1000 ⁇ m.
- a microneedle device of the present disclosure may comprise any number of microneedles.
- a device of the present disclosure comprises at least 1 microneedle, at least 100 microneedles, at least 200 microneedles, at least 300 microneedles, at least 400 microneedles, at least 500 microneedles, at least 1000 microneedles, at least 3000 microneedles, or at least 5000 microneedles.
- a device of the present disclosure comprises at most 10000 microneedles, at most 5000 microneedles, at most 2500 microneedles, at most 2000 microneedles, at most 1000 microneedles, at most 500 microneedles, at most 400 microneedles, at most 300 microneedles, at most 100 microneedles, at most 90 microneedles, at most 50 microneedles, at most 40 microneedles, at most 30 microneedles, at most 20 microneedles, at most 15 microneedles, at most 10 microneedles, at most 9 microneedles, at most 8 microneedles, at most 7 microneedles, at most 6 microneedles, at most 5 microneedles, at most 4 microneedles, at most 3 microneedles, at most 2 microneedles, or 1 microneedle.
- a device of the disclosure comprises from about 1 microneedle to about 100 microneedles, from about 1 microneedle to about 200 microneedles, from about 1 microneedle to about 1000 microneedles, from about 10 microneedles to about 200 microneedles, from about 50 microneedles to about 150 microneedles, or from about 1 microneedle to about 5000 microneedles.
- a device of the disclosure comprises 100 microneedles. In some embodiments, a device of the disclosure comprises from 90 to 110 microneedles. In some embodiments, a device of the disclosure comprises from 80 to 120 microneedles. In some embodiments, a device of the disclosure comprises 50 to 150 microneedles.
- the microneedles can be present on the devices in rows.
- the rows can be spaced at virtually equal intervals to the space of the needles aligned in the row. In some embodiments, the rows can be spaced at irregular intervals.
- the density of the microneedles in the device may be at least about 10/cm 2 , about 15/cm 2 , about 20/cm 2 , about 25/cm 2 , about 30/cm 2 , about 35/cm 2 , about 2, about 45/cm 2 , about 50/cm 2 , about 55/cm 2 , about 60/cm 2 , about 65/cm 2 , about 70/cm 2 , about 75/cm 2 , about 80/cm 2 , about 85/cm 2 , about 90/cm 2 , about 95/cm 2 , about 100/cm 2 , about 110/cm 2 , about 120/cm 2 , about 130/cm 2 , about 140/cm 2 , about 150/cm 2 , or about 200/cm 2 .
- the density of the microneedles in the device may be less than about 10/cm 2 , about 15/cm 2 , about 20/cm 2 , about 25/cm 2 , about 30/cm 2 , about 35/cm 2 , about 40/cm 2 , about 2, about 50/cm 2 , about 55/cm 2 , about 60/cm 2 , about 65/cm 2 , about 70/cm 2 , about 75/cm 2 , about 80/cm 2 , about 85/cm 2 , about 90/cm 2 , about 95/cm 2 , about 100/cm 2 , about 110/cm 2 , about 120/cm 2 , about 130/cm 2 , about 140/cm 2 , about 150/cm 2 , or about 200/cm 2 .
- a distance between the center of two microneedles on a device of the disclosure can be calculated to determine a density of the microneedles in the device.
- the center-to-center distance between two microneedles can be less than 1000 ⁇ m, less than 900 ⁇ m, less than 800 ⁇ m, less than 700 ⁇ m, less than 600 ⁇ m, less than 500 ⁇ m, less than 400 ⁇ m, less than 300 ⁇ m, less than 200 ⁇ m, or less 100 ⁇ m.
- the center-to-center distance between two microneedles can be no greater than 100 ⁇ m, no greater than 200 ⁇ m, no greater than 300 ⁇ m, no greater than 400 ⁇ m, no greater than 500 ⁇ m, no greater than 600 ⁇ m, no greater than 700 ⁇ m, no greater than 800 ⁇ m, no greater than 900 ⁇ m, or no greater than 1000 ⁇ m.
- microneedles of the disclosure can comprise a plurality of different diameters or base widths.
- the shape of the base of a microneedle can be, for example, round, rectangular, triangular, square, pentagonal, hexagonal, heptagonal, or other geometric shape.
- a microneedle of the disclosure can have a diameter or base width that is no greater than 500 ⁇ m, no greater than 400 ⁇ m, no greater than 300 ⁇ m, no greater than 200 ⁇ m, no greater than 100 ⁇ m, no greater than 50 ⁇ m, no greater than 40 ⁇ m, no greater than 30 ⁇ m, no greater than 20 ⁇ m, no greater than 10 ⁇ m, no greater than 1000 nm, no greater than 900 nm, no greater than 800 nm, no greater than 700 nm, no greater than 600 nm, no greater than 500 nm, no greater than 400 nm, no greater than 300 nm, no greater than 200 nm, or no greater than 100 nm.
- kits comprising a microneedle device as described herein.
- a kit may comprise a microneedle device as described herein, wherein the microneedle device comprises a recess behind the microneedle region, and an applicator which fits within the recess.
- the applicator is a mechanical applicator.
- the microneedle device comprises a microneedle region comprising a plurality of microneedles protruding from a front surface of a microneedle base substrate, the microneedle base substrate also comprising a back surface, the back surface comprising a recess directly behind at least a portion of the microneedle region; and a support substrate adjacent to at least one side of the microneedle base substrate, the support substrate connected to or integral with the microneedle base substrate.
- the present disclosure provides a method of obtaining a biological sample from a tissue, such as the skin of a subject.
- a subject can be of any age.
- a subject can be an elderly adult, an adult, an adolescent, a pre-adolescent, a child, a toddler, or an infant.
- a subject can be a mammal, a bird, a fish, a reptile, or an amphibian.
- Non-limiting examples of a subject include humans, primates, dogs, cats, horses, pigs, and mice.
- a subject may be a patient. In some embodiments, the subject is a human.
- the method comprises contacting a device of the present disclosure with a tissue of a subject.
- the subject may contact the device with their tissue, for example by applying the device to their skin.
- the device may be applied to a subject's tissue by someone else.
- the tissue may be skin, and the skin may be cleaned prior to application of the device.
- the method comprises contacting a device of the present disclosure with a tissue, such as skin, of a subject.
- the tissue is an in situ tissue.
- the tissue is a biopsy.
- the tissue is skin tissue.
- Pressure may be applied to the device, thus causing the microneedles to penetrate the tissue.
- pressure is applied by hand, for example thumb pressure.
- pressure may be applied by an applicator.
- the pressure may be less than about 1 N/mm 2 , 5 N/mm 2 , 10 N/mm 2 , 50 N/mm 2 , 100 N/mm 2 , 200 N/mm 2 , 200 N/mm 2 , or within a range of about 1 N/mm 2 and about 200 N/mm 2 .
- the applicator may be a manual applicator, or a mechanical or electronic applicator which provides a uniform pressure. In some cases, one end of the applicator has a width that is less than the width of the recess behind the microneedle section.
- the surface of the device distal to the microneedles may be referred to as the back surface. The pressure may be applied to the entire back surface, or may be concentrated on the back surface of the interior section of the device 120 .
- the microneedle device may be retained in the tissue for up to 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 12 minutes, 14 minutes, 16 minutes, 18 minutes, 20 minutes, 22 minutes, 24 minutes, 26 minutes, 28 minutes, or 30 minutes.
- the residence time of the microneedles inside the tissue allows the probes to contact and bind to biomarkers, for example nucleic acid molecules, in the tissue.
- the residence time of microneedles inside the tissue may be optimized for a particular tissue, biomarker or probe.
- a biomarker such as an RNA biomarker may start to degrade after a given period of time, and thus a residence time long enough to allow binding but short enough to minimize degradation would be preferred.
- a residence time of about 1 minute to about 10 minutes may be preferred for RNA biomarkers.
- a residence time of less than 5 minutes may be preferred for RNA biomarkers.
- a residence time of about 5 minutes may be preferred for RNA biomarkers.
- a residence time of no more than 10 minutes may be preferred for RNA biomarkers.
- a residence time of about 2 minutes to about 9 minutes may be preferred for RNA biomarkers.
- a residence time of about 3 minutes to about 8 minutes may be preferred for RNA biomarkers. In some cases, a residence time of about 4 minutes to about 7 minutes may be preferred for RNA biomarkers. In some cases, a residence time of about 5 minutes to about 6 minutes may be preferred for RNA biomarkers. In some cases, a residence time of about 5 minutes to about 7 minutes may be preferred for RNA biomarkers. In some cases, a residence time of about 5 minutes to about 8 minutes may be preferred for RNA biomarkers. In some cases, a residence time of about 5 minutes to about 9 minutes may be preferred for RNA biomarkers. In some cases, a residence time of about 5 minutes to about 10 minutes may be preferred for RNA biomarkers.
- the optimal residence time of the microneedles inside the tissue is determined by the cycle threshold (Ct) for detection of a target biomarker.
- Ct cycle threshold
- the Ct is lowest at about 5 minutes.
- the Ct of a sample collected after a residence time of 10 minutes is higher than the Ct of a sample collected after a residence time of 5 minutes.
- an increase of Ct from a minimum Ct indicates degradation of the sample may be occurring.
- the optimal residence time generates a high-quality biomarker sample.
- the quality of the RNA sample is determined by a ratio of larger RNA fragments (e.g. RNA fragments comprising about 700 bases or more) comprised in the RNA sample to smaller RNA fragments (e.g. fragments comprising about 150 bases to about 200 bases) comprised in the RNA sample.
- a residence time of no more than 10 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g.
- RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of large RNA fragments, e.g.
- RNA fragments comprising about 700 bases or more, and the small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases is greater than 1.
- a residence time of no more than 8 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and the small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, is greater than 1.
- a residence time of no more than 7 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and the small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, is greater than 1.
- a residence time of no more than 6 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g.
- a residence time of no more than 5 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g.
- RNA sample comprising a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of large RNA fragments, e.g.
- RNA fragments comprising about 700 bases or more, and the small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases is greater than 1.
- a residence time of no more than 3 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and the small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, is greater than 1.
- a residence time of about 5 minutes generates an RNA sample wherein the RNA sample comprises a population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and a population of small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, and wherein a ratio of the population of large RNA fragments, e.g. RNA fragments comprising about 700 bases or more, and the small RNA fragments, e.g. fragments comprising about 150 bases to about 200 bases, is greater than 1.
- the device after allowing a time for the probes to contact and bind the biomarkers, the device is removed from the tissue. Removal of the microneedles from the tissue may generate sheering pressures that remove non-specifically bound contaminants from the microneedles. In some cases, contaminants comprise any compound not intended for further genomic analysis.
- the device and extracted RNA biomarkers may be placed into a storage buffer, a transport buffer, or an analysis buffer.
- the device and extracted RNA biomarkers may be stored at ⁇ 80° C., ⁇ 20° C., ⁇ 4° C., 4° C. or room temperature.
- the device may be placed into a buffer to dissociate the extracted RNA biomarkers from the device, and the extracted RNA biomarkers may be stored at ⁇ 80° C., ⁇ ° C., ⁇ 4° C., 4° C. or room temperature.
- the extracted RNA biomarkers (with or without the device) may be sent to a laboratory for further analysis.
- the present disclosure provides a method for preparing a sample for genomic or transcriptomic analysis.
- the method may comprise, contacting a device of the present disclosure with skin of a subject, extracting nucleic acid biomarkers from the skin onto the device, washing the nucleic acid biomarkers from the device to produce a sample solution.
- the nucleic acids of the sample solution may be analyzed by polymerase chain reaction.
- the extracted nucleic acids of the sample solution may be amplified and analyzed by next generation sequencing (NGS). NGS of extracted mRNAs may allow determination of gene expression in the contacted skin of the subject.
- NGS next generation sequencing
- the present disclosure provides a method of determining whether a skin lesion on a subject suffering thereof will be responsive to an autoimmune therapeutic drug.
- the present disclosure provides a method of preparing a biological sample and identifying one or more autoimmune disease therapeutic drugs for a subject, the method comprising contacting skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a solid microneedle and applying pressure to the microneedle device such that the microneedle device penetrates the skin of the subject.
- Extracted ribonucleic acid (RNA) molecules from the subject may be obtained by removing the microneedle device from the skin of the subject.
- Performing high throughput sequencing on the extracted RNA molecules can generate one or more sequence reads for the subject, which may be aligned with a known signature of sequence reads, wherein the known signature of sequence reads is associated with a positive response to the autoimmune disease therapeutic drug, thereby obtaining aligned sequence reads; and used to classify the subject as having a greater than 50%, 60%, 70%, 80% or 90% likelihood of positively responding to the autoimmune disease therapeutic drug by applying a trained algorithm to the aligned sequence reads.
- the trained algorithm has a greater than 50%, 60%, 70%, 80% or 90% positive predictive value or greater than 50%, 60%, 70%, 80% or 90% negative predictive value.
- the autoimmune disease therapeutic drug may be an IL-17 mediated treatment, an IL-23 mediated treatment, an TNF ⁇ mediated treatment, or a combination thereof.
- the method may comprise the steps of extracting and sequencing mRNAs as described above, and comparing a gene signature of the lesion to known gene expression signatures using a trained algorithm, or analyzing the gene signature of the lesion using a trained algorithm.
- the trained algorithm may have a greater than 50% positive predictive value or greater than 50% negative predictive value for predicting a greater than 50% likelihood of positive response to the autoimmune disease therapeutic drug.
- Comparing the gene expression signatures may allow classification of the contacted skin lesion as likely to be responsive to a treatment, wherein the treatment is either IL-17 mediated treatment, IL-23 mediated treatment, TNF ⁇ mediated treatment, or a combination thereof.
- autoimmune disease therapeutics include etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, and risankizumab.
- the present disclosure provides a method for determining a likelihood that a subject with an autoimmune disease will respond to a therapeutic drug for an autoimmune disease the method comprising obtaining a biological sample from the subject; extracting one or more nucleic acid molecules from the biological sample; performing high throughput sequencing on the nucleic acid molecules to generate one or more sequence reads for the subject; and performing a genomic analysis on the sequence reads.
- the genomic analysis may comprise: determining an expression level of genes encoded by the sequence reads; comparing the expression level to a known positive-response genetic signature specific to a treatment for the autoimmune disease; and generating a treatment score from the comparison, wherein the treatment score is specific to a response to the treatment in the patient.
- the present disclosure provides a method of treating a skin lesion of a subject, the method comprising contacting the skin lesion with a device as described herein, extracting mRNAs from the skin lesion, using NGS to determine the gene expression signature, comparing the signature to known signatures or using a trained algorithm to select a treatment for the subject, and administering the selected treatment to the subject.
- selecting a treatment comprises comparing an expression level of a known positive-response genetic signature specific to a treatment for the autoimmune disease.
- This disclosure provides non-invasive and/or minimally invasive devices, methods, and systems that provide a subject specific dataset for disease diagnosis or guided clinical intervention. Further described herein are microneedle devices and methods and systems of using the same.
- the methods of the invention described herein may comprise, in some embodiments, determining a gene expression profile, precise classification of a disease or condition, prospective guided therapy and/or clinical intervention for the diseases or conditions, retrospective analysis and modeling of past treatments, identifying an activated pathway for a disease or condition, or any combination thereof for one or more subjects.
- the microneedle devices described herein may provide precise, minimally and/or non-invasive sample collection that may be used in downstream analysis as described elsewhere herein, particularly in the aforementioned methods.
- the methods of the disclosure are useful for providing a means for practicing personalized medicine, wherein treatment is tailored to a subject based on characteristics of the disease in the subject (e.g. the gene expression profile of the subject suffering from the disease).
- the microneedle devices may facilitate the collection of a subject's biological sample in situ for further analysis.
- the microneedle device may comprise one or more microneedle features configured to puncture the skin or mucosal membrane of the subject.
- the mucosal membrane may comprise oral mucosa, ophthalmic mucosal membranes, or any combination thereof.
- the device may comprise a planar substrate which supports the microneedles. The substrate can be made of the same material as that of the microneedle. It can also be made of a different material.
- the microneedles may further comprise probes adhered to the microneedle.
- These probes may be configured to bind to one or more biomarkers within a sample or tissue (e.g., skin) of a subject in such a manner as to permit extraction of the biomarker for further analysis.
- the extracted biomarkers may be analyzed, alone or in combination (e.g. generating a genetic signature or gene expression profile) to provide a diagnosis or prediction of a response to a drug or other therapeutic treatment with respect to the subject. Treating a
- known treatments for known diseases may not be efficacious against a particular form of a disease as compared to an alternative form of the same disease. Differences between forms of the disease may arise as a result of varying genetic signatures from each particular form.
- the methods described herein may provide targeted, specialized, personalized, or otherwise highly efficacious treatments or recommendation of treatments for subjects suffering from particular forms of known diseases.
- the present disclosure provides a method of determining whether a mild, moderate, or severe form of a skin condition of a subject suffering thereof will be responsive to a recommended treatment 600 , as seen in FIG. 6 .
- the method of determining whether a skin condition of a subject suffering thereof will be responsive to recommended treatment may comprise: (a) obtaining ribonucleic acid (RNA) biomarkers from a subject (as described elsewhere herein) 602 ; (b) determining the likelihood that the skin condition will respond to a recommended treatment, wherein the recommended treatment comprises a 50% or less aggregate efficacy rate against known forms of the skin condition 604 ; and (c) treating the subject with the recommended treatment 606 .
- RNA ribonucleic acid
- the step of determining may further comprise: (i) performing high throughput sequencing on the RNA biomarkers to generate one or more sequence reads for the subject; (ii) aligning the one or more sequence reads for the subject with a known signature of sequence reads, wherein the known signature of sequence reads is associated with a positive response to the recommended treatment, thereby obtaining aligned sequence reads; and (iii) classifying the subject as having a greater than 50% likelihood of positively responding to the recommended treatment by applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm comprises a greater than 50% positive predictive value for predicting a greater than 50% likelihood of positive response to the recommended treatment.
- the RNA biomarkers may comprise RNA molecules.
- the RNA biomarkers are transcribed from one or more of the follow genes listed in Table 6, Table 12, or Table 13.
- the trained algorithm may further comprise a greater than 50% negative predictive value for predicting a greater than 50% likelihood of positive response to the recommended treatment.
- the positive predictive value may be greater than 50%, 60%, 70%, 80% or 90%.
- the negative predictive value may be or greater than 50%, 60%, 70%, 80% or 90%.
- the recommended treatment may comprise one or more autoimmune therapeutic drugs for an autoimmune disease or condition.
- the autoimmune disease or condition may be psoriasis.
- the recommended treatment may comprise etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, risankizumab, or any combination thereof.
- one or more subject suffering from a similar disease or condition may respond to a treatment with varying efficacy.
- the efficacy may comprise a responder, non-responder, or adverse responder.
- genetic variability between subjects suffering from similar disease or condition may further stratify the varying efficacy of treatment intended to broadly treat a general form of the disease or condition.
- aspects of the disclosure provided herein comprise a method of preparing a biological sample and identifying one or more therapeutic drugs to effectively tailor treatment to the particular genetic variant of a subject's disease or condition 608 , as seen in FIG. 6 B .
- this disclosure provides a method of treating a subject with an autoimmune disease of the skin (e.g., psoriasis) or recommending a treatment to such subject comprising collecting a sample comprising RNA derived from skin from the subject.
- a sample comprising RNA derived from skin from the subject.
- Such collecting of a sample of RNA may involve use of a microneedle device or may involve lysis of a tissue biopsy or cell sample and extraction of nucleic acids (e.g., RNA) using an RNA extraction protocol.
- the subject has not been administered the autoimmune therapeutic drug prior to collecting the sample.
- the subject has not been administered the autoimmune therapeutic drug greater than 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 15 days or 20 days prior to collecting the sample from the subject.
- the method further comprises determining an expression level of at least one gene based on the RNA; predicting that a subject with the autoimmune disease of the skin will be responsive to the autoimmune therapeutic drug with a positive predictive value greater than 70%, a positive predictive value, a greater than 75% positive predictive value, a greater than 80% positive predictive value, a greater than 85% positive predictive value, a greater than 90% positive predictive value, or a greater than 95% positive predictive value based on the expression level of the at least one gene; and/or based on the predicting, treating the subject with the autoimmune therapeutic drug.
- the positive predictive value is determined using a cohort of a specific size.
- the method may have a positive predictive value provided herein when evaluated in a cohort of greater than 5, 10, 25, 50, 100
- this disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug comprising collecting a sample comprising RNA derived from skin from the subject.
- collecting of a sample of RNA may involve use of a microneedle device or may involve lysis of a tissue biopsy or cell sample and extraction of nucleic acids (e.g., RNA) using an RNA extraction protocol.
- the subject has not been administered the autoimmune therapeutic drug prior to collecting the sample.
- the subject has not been administered the autoimmune therapeutic drug greater than 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 15 days or 20 days prior to collecting the sample from the subject.
- the method further comprises converting the RNA into cDNA and determining an expression level of at least one gene based on the cDNA. In some cases, the method further comprises predicting that a subject with the skin lesion will be responsive to the autoimmune therapeutic drug with a positive predictive value greater than 70%, a positive predictive value, a greater than 75% positive predictive value, a greater than 80% positive predictive value, a greater than 85% positive predictive value, a greater than 90% positive predictive value, or a greater than 95% positive predictive value based on the expression level of the at least one gene; and/or based on the predicting, treating the subject with the autoimmune therapeutic drug.
- the positive predictive value is determined using a cohort of a specific size. For example, the method may have a positive predictive value provided herein when evaluated in a cohort of greater than 10, 25, 50, 100, 150, 200, 250, 500, or 1000 patients.
- this disclosure provides a method of determining whether a skin lesion of a subject will be responsive to an autoimmune therapeutic drug comprising penetrating skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a microneedle.
- RNA molecules are obtained from the subject by removing the microneedle device from the skin of the subject.
- high throughput sequencing on the RNA molecules is performed to generate sequence reads.
- the sequence reads are aligned with a signature of sequence reads associated with a positive response to an autoimmune disease therapeutic drug to obtain aligned sequence reads.
- a trained algorithm is applied to the aligned sequence reads, wherein the trained algorithm has a greater than 70%, 80%, 85%, 90% or 95% positive predictive value for predicting response to the autoimmune disease therapeutic drug.
- the aligned sequence reads is used to determine an expression level of at least one RNA molecule; and a trained algorithm is applied to the expression level of the at least one RNA molecule, wherein the trained algorithm predicts whether the subject with the skin lesion will be responsive to an IL-17 mediated treatment, an IL-23 mediated treatment, a TNF ⁇ mediated treatment, or any combination thereof.
- high throughput sequencing is performed on the RNA biomarkers to generate one or more sequence reads for the subject.
- the one or more sequence reads is aligned with a known signature of sequence reads, wherein the known signature of sequence reads is associated with a positive response to the recommended treatment, thereby obtaining aligned sequence reads.
- the subject is classified as having a likelihood of positively responding to the recommended treatment by applying a trained algorithm to the aligned sequence reads, wherein the trained algorithm comprises a greater than 50%, 60%, 70%, 80%, 85%, 90% or 95% positive predictive value for predicting a positive response to the recommended treatment.
- the trained algorithm also has a greater than 50%, 60%, 70%, 80%, 85%, 90% or 95% negative predictive value.
- the recommended treatment comprises one or more autoimmune therapeutic drugs for an autoimmune disease or condition, including, for example, etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, and risankizumab.
- autoimmune therapeutic drugs for an autoimmune disease or condition including, for example, etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, and risankizumab.
- the autoimmune disease or condition is psoriasis, acne, atopic dermatitis (i.e., eczema), Raynaud's phenomenon, rosacea, skin cancer (e.g., basal cell carcinoma, squamous cell carcinoma and melanoma), or vitiligo.
- this disclosure provides a method of determining whether a subject with an autoimmune disorder of the skin will be responsive to an autoimmune therapeutic drug comprising extracting mRNA from skin of the subject, sequencing the mRNA from the skin of the subject; and predicting whether the subject with the autoimmune disorder will be responsive to etanercept, adalimumab, infliximab, certolizumab, secukinumab, ixekisumab, broadalumab, guselkumab, tildrakizumab, and risankizumab with a positive predictive value greater than 50%, 60%, 70%, 80%, 85%, 90% or 95%.
- the at least one gene is at least one gene from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least two genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least three genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least four genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least five genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least six genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least ten genes from Table 6, Table 12, and/or Table 13.
- the at least one gene is at least fifteen genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least twenty genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least 25 genes from Table 6, Table 12, and/or Table 13. In some cases, the at least one gene is at least 50 genes from Table 6, Table 12, and/or Table 13.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of CNFN, CTSC, GBAP1, CRABP2, PCDH7, PPIG, RAB31, C3, and EGR. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of CNFN, CTSC, GBAP1, CRABP2, PCDH7, and PPIG. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of PCDH7, PPIG, RAB31, C3, and EGR.
- the at least one gene is at least one gene, two genes, or three genes from the group consisting of CNFN, CTSC, GBAP1, and CRABP2. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PPIG, RAB31, C3, and EGR. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PCDH7, PPIG, RAB31, and C3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of GBAP1, CRABP2, PCDH7, PPIG.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of SERPINB3, SERPINB4, S100A7A, PI3, KRT6A, LCN2, DEFB4A, DEFB4B, SPRR1A, IL36G, MX1, IFI27, CD36, CD24, and IL4R.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of KRT6A, SPRR1A, CD36, IL4R, LCN2, and IFI27.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of CD36, IL4R, S100A7A, SERPINB4, MX1, and SERPINB3. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of LCN2, IFI27, DEFB4A, IL36G, CD24, and PI3.
- the at least one gene is at least one gene, two genes, or three genes from the group consisting of IL4R, LCN2, and IFI27. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PI3, IFI27, and SERPINB3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of IL4R, S100A7A, and MX1. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of CD36, LCN2, and SERPINB4.
- the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of MTCO1P12, MTATP6P1, CLSTN1, PDPN, LDLRAD2, and GSTM3. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of AL158847.1, DAD1, LDLRAD2, ZNF395, MGMT, and AL136982.4. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of NREP, PPIF, PRIM1, AL136982.5, MTATP6P1, and SMPD3. In some cases, the at least one gene is at least one gene, two genes, three genes, four genes, five genes, or six genes from the group consisting of PDPN, TXNRD1, GSTM3, GPSM1, GLRX, and USP2.
- the at least one gene is at least one gene, two genes, or three genes from the group consisting of MTCO1P12, CLSTN1, and GSTM3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of NREP, PPIF, and PRIM1. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of AL136982.5, MTATP6P1, and SMPD3. In some cases, the at least one gene is at least one gene, two genes, or three genes from the group consisting of PDPN, TXNRD1, and GSTM3.
- the gene or genes used to detect a response to treatment may, in some cases, engage in a common pathway, such as an IL-17-mediated pathway, TNF-alpha pathway, or IL-23 pathway.
- the at least one gene comprises at least 2, 3, 4, 5, 6, 7, 10, 15, or 20 genes that do not share a common upstream regulator or do not share a common pathway.
- the genes have some relationship with the recommended treatment, e.g., the genes may be involved with a particular pathway (e.g., IL-17, 11-23, or TNF-alpha mediated pathway).
- at least one gene does not have a known relationship with a recommended treatment.
- the gene used to evaluate a response to treatment for a particular therapeutic may not be involved in the pathway targeted by the therapeutic.
- the autoimmune therapeutic drug is an IL-17-mediated treatment and the at least one gene comprises at least one gene that is not involved in an IL-17 mediated pathway.
- the autoimmune therapeutic drug is an IL-23-mediated treatment and the at least one gene comprises at least one gene that is not involved in an IL-23 mediated pathway.
- the autoimmune therapeutic drug is a TNF-alpha-mediated treatment and the at least one gene comprises at least one gene that is not involved in a TNF-alpha-mediated pathway.
- a trained algorithm is applied to expression level of the at least one gene in order to predict the subject's likelihood of responding to the therapeutic.
- the algorithm is trained using samples from patients that were administered a single type of drug. For example, the patients may have been administered an IL-17 mediated treatment, TNF-alpha mediated treatment or an IL-23 mediated treatment.
- the methods comprise use of a microneedle device.
- the method of preparing a biological sample and identifying one or more therapeutic drugs may comprise: (a) contacting skin of a subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a microneedle, as described elsewhere herein 610 ; (b) applying pressure to the microneedle device such that the microneedle device penetrates the skin of the subject 612 ; (c) obtaining extracted RNA molecules from the subject by removing the microneedle device from the skin of the subject 614 ; (d) performing high throughput sequencing on the extracted RNA molecules to generate one or more sequence reads for the subject 616 ; (e) aligning the one or more sequence reads for the subject with a known signature of sequence reads, wherein the known signature of sequence reads is associated with a positive response to the one or more therapeutic drugs, thereby obtaining aligned sequence reads 618 ; and classifying the subject
- the subject has a PASI of at least 8, at least 9, at least 10, at least 15. In some cases, the subject has a PASI of at least 8 and at least a PASI 75, 80, 90, 95, or 100 after treatment with the recommended drug. In some cases, the subject has a PASI of at least 10 and a PASI 75, 80, 90, 95, or 100 after treatment.
- the trained algorithm may further comprise a greater than 50% negative predictive value for predicting a greater than 50% likelihood of positive response to the one or more therapeutic drugs. In some cases, the positive predictive value may be greater than 50%, 60%, 70%, 80% or 90%. In some instances, the negative predictive value may be or greater than 50%, 60%, 70%, 80% or 90%.
- the extracted RNA molecules may be transcribed from one or more of the following genes listed in Table 6, Table 12, or Table 13.
- the recommended treatment may comprise one or more autoimmune therapeutic drugs for an autoimmune disease or condition.
- the autoimmune disease or condition may be psoriasis.
- the recommended treatment may comprise etanercept, infliximab, adalimumab, certolizumab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, tildrakizumab, risankizumab, or any combination thereof.
- the variability of gene expression between one or more subjects may be predictive of the response of a particular treatment for a disease or condition, particularly psoriasis.
- aspects of the invention disclosed herein provide a method for determining the response to a treatment for a disease or condition in a subject afflicted with the disease or condition.
- the method may further comprise recommending an additional treatment specific to the score.
- the method may further comprise administering the treatment or the additional treatment to the subject.
- the one or more nucleic acid molecules may comprise deoxyribonucleic acid (DNA) molecules.
- the one or more nucleic acid molecules comprise RNA molecules.
- the RNA molecules may comprise messenger RNA (mRNA) molecules.
- the disease or condition may be an autoimmune disease.
- the autoimmune disease may be psoriasis.
- particular forms of a disease may comprise different gene signatures. Certain known treatments for known diseases or conditions are not as efficacious against a particular form of a disease as compared to an alternate form of the same disease that may comprise.
- the method may further comprise recommending a treatment specific to the score.
- the method may further comprise administering the treatment to the subject.
- the disease or condition may be an autoimmune disease.
- the autoimmune disease may be psoriasis.
- the one or more nucleic acid molecules may comprise DNA molecules.
- the one or more nucleic acid molecules may comprise RNA molecules.
- the RNA molecules may comprise mRNA molecules.
- the at least five genes from Table 6, Table 12, or Table 13 may comprise genes associated with one or more pathways. In some cases, the at least five genes from Table 6, Table 12, or Table 13 may comprise at least 2 genes associated with activation of the one or more pathways.
- the method for identifying the activation of one or more pathways that may be associated with a disease or condition in a subject may comprise the steps of: (a) contacting skin of the subject with a microneedle device, wherein the microneedle device comprises one or more nucleic acid probes coupled to a microneedle 644 ; (b) applying pressure to the microneedle device such that the microneedle device penetrates the skin of the subject 646 ; (c) obtaining extracted RNA molecules from the subject by removing the microneedle device form the skin of the subject 648 ; (d) performing high throughput sequencing on the extracted RNA molecules to generate one or more sequence reads for the subject 650 ; (e) aligning the one or more sequence reads for the subject with a known signature of sequence reads, where
- the method may further comprise recommending a treatment specific to the activation of the one or more pathways. In some cases, the method may further comprise administering the treatment specific to the activation of the one or more pathways to the subject.
- the microneedle may be a solid microneedle.
- the trained algorithm may further comprise a greater than 50% negative predictive value for predicting a greater than 50% likelihood of developing the disease or condition associated with activation of the one or more pathways.
- the positive predictive value may be greater than 50%, 60%, 70%, 80% or 90%.
- the negative predictive value may be or greater than 50%, 60%, 70%, 80% or 90%.
- the disease or condition may be an autoimmune disease or condition.
- the autoimmune disease or condition may be psoriasis.
- the extracted RNA molecules may be transcribed from one or more genes comprising at least one gene associated with activation of the one or more pathways.
- the one or more genes may comprise at least two genes associated with activation of the one or more pathways.
- the one or more genes may comprise any one of the genes listed in Table 6, Table 12 or Table 13.
- the term “subject” refers to any animal, including mammals and non-mammals), for example, humans, non-human primates (e.g., rhesus macaques, crab-eating macaques, stump-tailed macaques, pig-tailed macaques, squirrel monkeys, owl monkeys, baboons, chimpanzees, marmosets and spider monkeys, etc.), rodents (e.g., mice, rats, guinea pigs, etc.), dogs, cats, pigs, etc.
- the subject is a human subject.
- the subject may be experiencing a skin condition or have symptoms of a skin condition.
- skin conditions that may be detected and/or treated by the methods and devices provided herein include: psoriasis, acne, atopic dermatitis (i.e., eczema), Raynaud's phenomenon, rosacea, skin cancer (e.g., basal cell carcinoma, squamous cell carcinoma and melanoma), and vitiligo.
- the subject has, or has symptoms of, one or more skin conditions, for example, psoriasis and atopic dermatitis.
- the subject may be experiencing different degrees of skin conditions, for example, mild, moderate, severe, and very severe.
- Examples of symptoms of psoriasis include red patches of skin covered with thick, silvery scales; small scaling spots; dry, cracked skin that may bleed or itch; itching, burning or soreness; thickened, pitted or ridged nails; swollen and stiff joints.
- Psoriasis Area and Severity Index (PASI) Score Psoriasis Area and Severity Index (PASI) Score
- the Psoriasis Area and Severity Index (PASI) score is commonly used to measure discoloration, thickness, scaling, and coverage of these plaques.
- a caregiver e.g., a medical doctor, nurse, nurse practitioner, physician's assistant, etc.
- PASI score can be used to measure the severity and extent of psoriasis and observe the effectiveness of psoriasis treatments.
- Using the tool also allows a caregiver to monitor the progression of the condition and evaluate the effectiveness of treatment.
- the scoring involves rating the symptoms of psoriasis from none to very severe and estimating the percentage of the body that they affect.
- researchers also use PASI scores to determine the effectiveness of psoriasis medications in clinical trials.
- the range of absolute PASI scores is 0-72, with higher scores indicating a greater severity of psoriasis. A score of 0 indicates no psoriasis, while a score higher than 10 suggests severe psoriasis.
- the scoring system includes a section for intensity and another for body coverage. The intensity section of the score measures 1) discoloration (i.e., redness), 2) thickness, and 3) scaling. The area section shows the extent to which psoriasis affects the following: 1) head and neck, 2) upper limbs, 3) trunk, and 4) lower limbs.
- PASI scores can calculate PASI scores as follows:
- a scores Add together the three intensity scores (discoloration, thickness, and scaling) for each of the four body regions to get four A scores.
- Each intensity score can be given based on the severity of the symptoms: 0: absent symptoms; 1: mild symptoms; 2: moderate symptoms; 3: severe symptoms; 4: very severe symptoms.
- B scores A B score is assigned for each of the four body regions: 0: absence of symptoms, 1: 1- 9% coverage, 2: 10-29% coverage, 3: 30-49% coverage, 4: 50-69% coverage, 5: 70- 89% coverage, and 6: 90-100% coverage.
- C scores Multiply the A score by the B score for each body region to get four C scores
- D scores Multiply each C score by the amount of body surface area that the region represents. This amount is 0.1 for the head and neck, 0.2 for the arms, 0.3 for the trunk, and 0.4 for the legs. This gives four D scores.
- PASI Add together the four D scores. score
- PASI 75 means that a person's PASI score has decreased 75% from baseline, which indicates a significant improvement in the condition.
- Conventional goal of treatment has been to achieve PASI 75.
- a PASI 90 or PASI 100 i.e., a 90% or 100% reduction in PASI scores, respectively.
- a subject provided herein has a PASI of at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, or at least 15. In some cases, a subject has moderate-to-severe psoriasis and a baseline PASI of greater than or equal to 10. In some cases, the methods provided herein provide a prediction for whether a subject will respond to a particular treatment. In some cases, following being treated by the recommended treatment, the subject responds to the treatment and has as a decrease in PASI score.
- the subject's response to treatment may be a particular response at week 16 following treatment, such as week 16 PASI60; week 16 PASI75, week 16 PASI80, week 16 PASI80, week 16 PASI90, week 16 PASI95, week 16 PASI 99, or week 16 PASI 100.
- the subject's response to treatment may be a particular response at week 12 following treatment, such as week 12 PASI60; week 12 PASI75, week 12 PASI80, week 12 PASI80, week 12 PASI90, week 12 PASI95, week 12 PASI 99, or week 12 PASI 100.
- the subject's response to a treatment is a response at week 8 following treatment, such as week 8 PASI60; week 8 PASI75, week 8 PASI80, week 8 PASI80, week 8 PASI90, week 8 PASI95, week 8 PASI 99, or week 8 PASI 100.
- the subject's response to a treatment is a response at week 4 following treatment, such as week 4 PASI60; week 4 PASI75, week 4 PASI80, week 4 PASI80, week 4 PASI90, week 4 PASI95, week 4 PASI 99, or week 4 PASI 100.
- the subject's response to treatment is evaluated at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, or at least 24 weeks following treatment. In some embodiments, the subject's response to treatment is evaluated at most 2 weeks, at most 3 weeks, at most 4 weeks, at most 8 weeks, at most 12 weeks, at most 16 weeks, at most 20 weeks, at most 24 weeks following treatment, or at most 50 weeks following treatment. For example, the subject's response to treatment may, in some embodiments, be evaluated between 8 and 12 weeks following treatment, between 4 and 12 weeks following treatment, and/or between 8 and 16 weeks following treatment.
- the patient's PASI response may be at least PASI60, at least PASI70, at least PASI80, at least PASI90, at least PASI95, at least PASI99, or PASI100 for any of the periods following treatment.
- Nucleic acids can be extracted from a sample using the microneedle devices described herein, or extracted directly from a sample (e.g., biopsy sample) using an extraction method other than one that involves use of a microneedle device.
- the types of nucleic acids that can be extracted include any nucleic acid molecules that encode genetic information, including for example, messenger RNA (mRNA), microRNA, DNA (e.g., nuclear DNA or mitochondrial DNA), a mixture of mRNA and DNA, short RNA, isolated RNA, and isolated DNA.
- the biopsy samples that can be used for nucleic acids extraction can include any tissue removed from any part of the body, for example, from the surface of the body (e.g., skin, hair, scalp, surface skin, nails, skin debris, hair follicles); from inside the body (e.g., brain, heart, lungs, pancreas, kidney, liver, intestine, muscles, connective tissue, bone, cartilage, or blood).
- the biopsy sample can be obtained invasively, non-invasively, or minimally invasively. In some cases, the biopsy sample is obtained from one or more active lesions, dormant lesions, or normal tissues.
- the device and extracted RNA biomarkers may be placed into a storage buffer, a transport buffer, or an analysis buffer.
- the device and extracted RNA biomarkers may be stored at ⁇ 80° C., ⁇ 20° C., ⁇ 4° C., 4° C. or room temperature.
- the device may be placed into a buffer to dissociate the extracted RNA biomarkers from the device, and the extracted RNA biomarkers may be stored at ⁇ 80° C., ⁇ 20° C., ⁇ 4° C., 4° C. or room temperature.
- the extracted RNA biomarkers (with or without the device) may be sent to a laboratory for further analysis.
- DNA is sequenced using nanopore sequencing, in which a single molecule of DNA or RNA can be sequenced without the need for PCR amplification or chemical labeling of the sample.
- the extracted nucleic acids of the sample s may be amplified and analyzed by next generation sequencing (NGS), high-throughput sequencing, massively parallel sequencing, or sequencing-by-synthesis.
- NGS next generation sequencing
- Different sequencing approaches can include, for example, pyrosequencing, sequencing by reversible terminator chemistry, sequencing-by-ligation mediated by ligase enzymes, or phosphor-linked fluorescent nucleotides or real-time sequencing.
- Methods for performing genomic analyses may also include microarray methods. In some cases, genomic analysis may be performed in combination with any of the other methods herein.
- a sample may be obtained, tested for adequacy, and divided into aliquots.
- One or more aliquots may then be used for cytological analysis of the present invention, one or more may be used for gene expression profiling methods of the present invention, and one or more may be used for genomic analysis.
- the present invention anticipates that one skilled in the art may wish to perform other analyses on the biological sample that are not explicitly provided herein.
- the method further comprises converting the captured RNA to DNA (e.g., cDNA) that is readily available for sequencing (e.g. sequencing by synthesis, or nanopore sequencing).
- DNA e.g., cDNA
- RNeasy Mini kits Qiagen, Valencia, CA
- RNAase-Free DNase digestion Qiagen, Valencia, CA
- Taqman kits Applied Biosystems, Foster City, CA
- cDNA can be amplified and then sequenced by a commercial vendor (Psomagen, Inc., Rockville, MD) according to standard procedures. Library preparation can be accomplished using Illumina Nextera DNA Flex kits according to the manufacturer's instructions. Prepared indexed libraries can be loaded onto a NovaSeq6000 S4 with read length of 150PE for sequencing of 40 M reads per sample. During sequencing, the quality score (Q30) may be maintained at a certain level, e.g., over 75%. Upon completion of runs, FASTQ file quality may be checked with FASTQC and trimmed with the Trim_galore program. The trimmed FASTQ can be aligned and mapped to human reference genome (e.g., GRCh38) using hisat2 program.
- human reference genome e.g., GRCh38
- the number of reads is counted for each Ensemble gene ID using FeatureCounts program and Homo sapiens GRCH38.84.gtf.
- RNA expression analysis is further processed using Bioconductor package edgeR. Genes may be filtered using filterByExpr before logCPM (counts per million reads) were calculated. Plots may be made with ggplot2 and heatmap.2 functions in R. Percent of genes detected may be determined using the ratio of genes detected (with >0 count) over the total number of Ensemble genes.
- the general methods for determining gene expression product levels may include but are not limited to one or more of the following: additional cytological assays, assays for specific proteins or enzyme activities, assays for specific expression products including protein or RNA or specific RNA splice variants, in situ hybridization, whole or partial genome expression analysis, microarray hybridization assays, SAGE, enzyme linked immuno-absorbance assays, mass-spectrometry, immuno-histochemistry, or blotting.
- Gene expression product levels may be normalized to an internal standard such as total mRNA or the expression level of a particular gene including but not limited to glyceraldehyde 3 phosphate dehydrogenase, or tublin.
- microarray analysis begins with extracting and purifying nucleic acid from a biological sample, (e.g. a biopsy or fine needle aspirate) using methods known to the art.
- a biological sample e.g. a biopsy or fine needle aspirate
- it can be advantageous to extract and/or purify RNA from DNA. It can further be advantageous to extract and/or purify mRNA from other forms of RNA such as tRNA and rRNA.
- purified nucleic acid may further be labeled with a fluorescent, radionuclide, or chemical label such as biotin or digoxin for example by reverse transcription, PCR, ligation, chemical reaction or other techniques.
- the labeling can be direct or indirect which may further require a coupling stage.
- the coupling stage can occur before hybridization, for example, using aminoallyl-UTP and NHS amino-reactive dyes (like cyanine dyes) or after, for example, using biotin and labeled streptavidin.
- the modified nucleotides e.g.
- the aaDNA may then be purified with, for example, a column or a diafiltration device.
- the aminoallyl group is an amine group on a long linker attached to the nucleobase, which reacts with a reactive label (e.g. a fluorescent dye).
- the labeled samples may then be mixed with a hybridization solution which may contain SDS, SSC, dextran sulfate, a blocking agent (such as COT1 DNA, salmon sperm DNA, calf thymum DNA, PolyA or PolyT), Denhardt's solution, formamine, or a combination thereof.
- a hybridization probe is a fragment of DNA or RNA of variable length, which is used to detect in DNA or RNA samples the presence of nucleotide sequences (the DNA target) that are complementary to the sequence in the probe. The probe thereby hybridizes to single-stranded nucleic acid (DNA or RNA) whose base sequence allows probe-target base pairing due to complementarity between the probe and target.
- the labeled probe is first denatured (by heating or under alkaline conditions) into single DNA strands and then hybridized to the target DNA.
- the probe is tagged (or labeled) with a molecular marker; commonly used markers are 32 P or Digoxigenin, which is non-radioactive antibody-based marker.
- DNA sequences or RNA transcripts that have moderate to high sequence similarity to the probe are then detected by visualizing the hybridized probe via autoradiography or other imaging techniques. Detection of sequences with moderate or high similarity depends on how stringent the hybridization conditions were applied—high stringency, such as high hybridization temperature and low salt in hybridization buffers, permits only hybridization between nucleic acid sequences that are highly similar, whereas low stringency, such as lower temperature and high salt, allows hybridization when the sequences are less similar.
- Hybridization probes used in DNA microarrays refer to DNA covalently attached to an inert surface, such as coated glass slides or gene chips, and to which a mobile cDNA target is hybridized.
- the mix may then be denatured by heat or chemical means and added to a port in a microarray.
- the holes may then be sealed, and the microarray hybridized, for example, in a hybridization oven, where the microarray is mixed by rotation, or in a mixer. After an overnight hybridization, non-specific binding may be washed off (e.g. with SDS and SSC).
- the microarray may then be dried and scanned in a special machine where a laser excites the dye and a detector measures its emission.
- the image may be overlaid with a template grid and the intensities of the features (several pixels make a feature) may be quantified.
- kits can be used for the amplification of nucleic acid and probe generation of the subject methods.
- kit that can be used in the present invention include but are not limited to Nugen WT-Ovation FFPE kit, cDNA amplification kit with Nugen Exon Module and Frag/Label module.
- the NuGEN WT-OvationTM FFPE System V2 is a whole transcriptome amplification system that enables conducting global gene expression analysis on the vast archives of small and degraded RNA derived from FFPE samples.
- the system is comprised of reagents and a protocol required for amplification of as little as 50 ng of total FFPE RNA.
- the protocol can be used for qPCR, sample archiving, fragmentation, and labeling.
- the amplified cDNA can be fragmented and labeled in less than two hours for GeneChip® 3′ expression array analysis using NuGEN's FL-OvationTM cDNA Biotin Module V2.
- the amplified cDNA can be used with the WT-Ovation Exon Module, then fragmented and labeled using the FL-OvationTM cDNA Biotin Module V2.
- the amplified cDNA can be fragmented and labeled using NuGEN's FL-OvationTM cDNA Fluorescent Module.
- Ambion WT-expression kit can be used.
- Ambion WT-expression kit allows amplification of total RNA directly without a separate ribosomal RNA (rRNA) depletion step.
- rRNA ribosomal RNA
- samples as small as 50 ng of total RNA can be analyzed on Affymetrix® GeneChip® Human, Mouse, and Rat Exon and Gene 1.0 ST Arrays.
- the Ambion® WT Expression Kit provides a significant increase in sensitivity.
- Ambion WT-expression kit may be used in combination with additional Affymetrix labeling kit.
- AmpTec Trinucleotide Nano mRNA Amplification kit (6299-A15) can be used in the subject methods.
- the ExpressArt® TRinucleotide mRNA amplification Nano kit is suitable for a wide range, from 1 ng to 700 ng of input total RNA. According to the amount of input total RNA and the required yields of aRNA, it can be used for 1-round (input >300 ng total RNA) or 2-rounds (minimal input amount 1 ng total RNA), with aRNA yields in the range of >10 ⁇ g.
- AmpTec's proprietary TRinucleotide priming technology results in preferential amplification of mRNAs (independent of the universal eukaryotic 3 ′-poly(A)-sequence), combined with selection against rRNAs.
- This kit can be used in combination with cDNA conversion kit and Affymetrix labeling kit.
- the raw data may then be normalized, for example, by subtracting the background intensity and then dividing the intensities making either the total intensity of the features on each channel equal or the intensities of a reference gene and then the t-value for all the intensities may be calculated. More sophisticated methods include z-ratio, loess and lowess regression and RMA (robust multichip analysis) for Affymetrix chips.
- PolyA tailed mRNA is captured on a support via hybridization to a polyT DNA capture probe or primer coupled to the surface of a support.
- the polyT strand is next extended with a reverse transcriptase polymerase to make a double stranded molecule comprising a DNA:RNA duplex.
- a transposome complex e.g., Tn5 transposase bound with an adaptor sequence and sequences complementary to surface amplification primers
- Tn5 transposase bound with an adaptor sequence and sequences complementary to surface amplification primers is added to the support, which undergoes a transposition reaction with and tagments the duplex, ligating a DNA adaptor oligo to the 5′ end of the RNA strand.
- a strand displacing polymerase (e.g., Bst polymerase) can then be used to extend the 3′ end of the DNA strand, displacing the non-transferred strand of the transposome complex and copying the RNA strand to its 5′ DNA chimeric end.
- the DNA adaptor oligo comprises a sequence complementary to an anchor primer on a substrate configured for sequencing.
- the DNA adaptor oligo comprises a sequence that is complementary to sequencing primer for sequencing.
- the double-stranded molecule can then be amplified (e.g., cluster amplification) and sequenced with a sequencing primer.
- the primer partially comprises the adaptor sequence and the upstream adaptor sequence.
- the other end of the molecule can be sequenced with a primer that anneals upstream of the polyT sequence and is extended with natural dATP nucleotides before commencing cycles of sequence by synthesis (SBS) chemistry.
- SBS sequence by synthesis
- RNA is fragmented and treated with a phosphatase.
- a single stranded adaptor molecule is ligated to the 3′end of each RNA fragment comprising the complement of a surface bound primer.
- the fragments are then added to a support and captured via hybridization.
- the hybridized RNA molecules are converted to a DNA:RNA duplex with a reverse transcriptase polymerase.
- a transposome complex or composition comprising a transposase and an adaptor duplex (i.e., transposon) of an ME with P5 is used to tagment the duplex.
- the molecules can be amplified (e.g., cluster amplification) and sequenced.
- PPV positive predictive value
- Mind.Px An algorithm (Mind.Px) is developed herein for the prediction of response to biologics (anti-IL-17, anti-IL-23) used for the treatment of patients with psoriasis, by comparing baseline transcriptomes with clinical response to biologic drugs at 12 weeks after initial treatment.
- Psoriasis Area and Severity Index PASI
- a sequence read from a subject is compared with a sequence read from a positive or negative control.
- a positive control is a subject diagnosed with the disease or condition.
- a negative control is a healthy person who does not have any symptoms or history of symptoms of the disease or condition.
- a negative control can be the baseline for a subject prior to treatment.
- biologicals means medicinal products obtained from living organisms (e.g., humans, animals, or microorganisms), including recombinant biologics, which are produced by genetic engineering technique.
- a biologic may contain proteins that control the action of other proteins and cellular processes, genes that control production of vital proteins, modified human hormones, or cells that produce substances that suppress or activate components of the immune system.
- a biologic is an antibody (e.g., a monoclonal antibody).
- the subject is treated with one or more of the following biologics: tumor necrosis factor inhibitors (e.g., adalimumab, etanercept, infliximab, golimumab and certolizumab pegol); IL-17 inhibitors (e.g., secukinumab, ixekizumab, and brodalumab); and the IL-23 inhibitors (e.g., tildrakizumab, guselkumab, ustekinumab, and risankizumab).
- tumor necrosis factor inhibitors e.g., adalimumab, etanercept, infliximab, golimumab and certolizumab pegol
- IL-17 inhibitors e.g., secukinumab, ixekizumab, and brodalumab
- the IL-23 inhibitors e.g., tildrakizum
- bases refers to nucleotides. In some cases, “bases” can refer to base pairs (“bp”), e.g. 1 base equals 1 base pair. As used herein, the terms, “bases,” and “base pairs,” are used interchangeably.
- the term “about” means within 10% above or below a given value. For example, “about 10”, would include values from 9 to 11, unless otherwise indicated by the context in which the term is used.
- a device of the present disclosure is manufactured using injection molding.
- the device is manufactured by injecting heated (e.g. molten) material, e.g. a polyolefin resin disclosed herein, into a mold of the device, according to the features disclosed herein.
- the mold comprises a plurality of cavities for forming a plurality of microneedles, cavity for forming an interior section comprising the plurality of microneedles, and one or more cavities for forming one or more peripheral sections adjacent to the interior section.
- the width of the interior section is less than the width of the peripheral section(s).
- the width of the intersection is about a 1:5 ratio in size as compared to the width of the peripheral section(s).
- the smaller width of the interior section (as compared to the peripheral section(s)) facilitates the heated materials movement to the one or more cavities to generate the plurality of microneedles prior to generating the interior section and the peripheral section(s).
- the result is a device as described herein comprising uniform, sharp microneedles. No more than 3% of the microneedles in the plurality of microneedles deviate (+/ ⁇ ). by more than about 50 ⁇ m from an average length of the microneedles of the plurality of microneedles.
- Example 2 Bonding Probes to the Device of Example 1
- the microneedles of the device of Example 1 are plasma treated as described herein.
- the device or a portion thereof is placed into a plasma vacuum chamber, the plasma vacuum chamber is closed, creating a sufficient vacuum seal, all of the pre-existing gas present in the chamber is evacuated and a plasma treatment gas is pumped into the plasma vacuum chamber to a defined pressure, enabling the generation of gas plasma.
- the gas plasma reacts with the heated material of the device (e.g. a polyolefin resin described herein) and renders the material more readily reactive to form covalent bonds with the probes described herein.
- the heated material of the device e.g. a polyolefin resin described herein
- the optimal time for mRNA extraction was determined by placing a microneedle device described herein on the skin of healthy subjects for varying periods of time, followed by removal of the patch from the skin and subsequent amplification, cDNA synthesis, and qPCR analysis.
- the devices were placed on subjects for 20 seconds, 1 minute, 5 minutes, and 10 minutes, and the cycle threshold (Ct) was measured for two biomarkers, GAPDH and KRT10, generated from the mRNA sample collected from the device at each residence time. All measurements were made in triplicate.
- bioanalyzer QC and qPCR analysis were performed with the mRNA extracted from a microneedle device described herein.
- the corrected area QC ratio was less than 1. Additionally, qPCR analysis was performed on these samples. Using a typical gene (B2M), the Ct value from degraded RNA sample increased to 22.70 from a Ct of 19.25 for the control sample (intact RNA); this increase of 3.45 Ct indicated approximately 11-fold decrease in the quantity of mRNA in the degraded sample.
- B2M typical gene
- Example 5 Prospective Prediction of Individual Patients Response to TNF ⁇ , IL-17 and IL-23 inhibitors
- RNA adhered on the Dermal Biomarker Patch was extracted and sequenced using next-generation sequencing.
- drug classes e.g., TNF ⁇ , IL-17i and IL-23i inhibitors
- RNA sequencing readout was then aligned and compared with a database of patients with a similar disease condition RNA gene profiles and their respective responsive or non-responsiveness to a particular drug class.
- the comparison between RNA sequencing readout and the database with a predictive linear classifier provided a score of the probability that a particular drug or drug class will provide an efficacious response to treating a disease condition.
- the predictive linear classifier's positive predictor value (PPV), sensitive, and balanced accuracy was assessed for TNF ⁇ i, (PASI75@W12), IL-17i (PASI75@W 12) and IL-23i (PASI150@W12) drug classes.
- the TNF ⁇ i inhibitor drug class was evaluated over a training set of 34 patients and a test set of 15.
- the resulting PPV, sensitivity and balanced accuracy for the linear classifier was 89%, 80%, and 80%, respectively.
- the IL-17i inhibitor drug class was evaluated over a training set of 75 patients and a test set of 6.
- the resulting PPV, sensitivity and balanced accuracy for the linear classifier was 100%, 100%, and 100%, respectively.
- the IL-23i inhibitor drug class was evaluated over a training set of 17 patients and a test set of 17.
- the resulting PPV, sensitivity and balanced accuracy for the linear classifier was 90%, 82%, and 83%, respectively.
- This study was designed to develop and prospectively validate a machine learning based algorithm that could predict patient response to the most common biologic drug classes used in the management of psoriasis patients. This type of tool would allow clinicians to have greater confidence that a given patient will respond to a specific drug class, which could lead to improved health outcomes and reduced wasted healthcare spend.
- DBPs dermal biomarker patches
- PASI measurements were made at baseline and 12 weeks to evaluate clinical response to a clinical phenotype. Responders were defined as those who reached PASI75 at 12 weeks.
- the transcriptomes obtained from the DBPs were sequenced and analyzed to derive and/or validate classifiers for each biologic class, which were then combined to yield predictive responses for all three biologic drug classes (IL-23i, IL-17i, and TNF ⁇ i).
- the IL-23i predictive classifier was developed from the earlier enrolled patients and independently validated with the latter enrolled patients.
- IL-17i and TNF ⁇ i predictive classifiers were developed using publicly available datasets and independently validated with patients from the STAMP studies. In the independent validation, positive predictive values for three classifiers (IL-23i, IL-17i, and TNF ⁇ i) were 93.1%, 92.3% and 85.7% respectively. Over the entire cohort, 99.5% of patients were predicted to respond to at least one drug class.
- Psoriasis is a T-cell mediated inflammatory skin disease characterized by discrete erythematous plaques and papules with micaceous scale. Worldwide, this is a common disease, with approximately 2.8% of the United States population, or 7.5 million people, diagnosed with psoriasis.
- Current treatment paradigms for psoriasis are distinguished by topical medications and/or phototherapy for mild to moderate patients, and systemic medications for patients who are classified as moderate to severe disease.
- the advent of biologic therapy as one of these systemic agents has revolutionized the management and treatment of psoriasis patients and is a direct result of the increased molecular understanding of the disease.
- biologic agents approved for use in the United States for the treatment of psoriasis, with more under development. However, it is not clear which biologic would be effective for a given patient.
- a novel biomarker capture platform that utilizes a dermal biomarker patch to capture the whole transcriptome including mRNA biomarkers from the epidermis and upper dermis.
- This platform showed excellent concordance with biopsy and provides a scalable method to access skin biomarkers in a minimally invasive manner. It is contemplated that the use of this platform in preliminary machine learning classifier builds for the prediction of response and non-response to IL-17 and IL-23 inhibitors. Also disclosed herein, is an extension of this preliminary study to the development and prospective validation of an actionable clinical test for predicting patient response to psoriasis biologics for all three drug classes.
- DBPs Dermal biomarker patches
- a customized spring-loaded applicator was used to standardize the application pressure across subjects and users.
- the loaded applicator was placed against the skin and the trigger pressed, applying the patch to the skin.
- the patch was then held in place against the skin for 5 minutes by a ring of medical tape. After this time, the patch was removed from the subject, immediately placed into storage buffer (LiCl, Triton X-100, Tris-EDTA), and stored at 4° C. until processing.
- Dermal transcriptomes were processed within 96 hours of collection from subjects.
- the applied DBPs were washed with chilled 1 ⁇ PBS and then dried under a stream of nitrogen.
- mRNA extraction from the patch was performed by applying PCR grade water (50 ⁇ L, 95° C.) to the DBP.
- the patch was then heated 1 minute at 95° C. to elute the bound mRNA from the DBP.
- This eluted mRNA was then converted to cDNA using the Takara SMART-Seq® Single Cell kit according to the manufacturer's instructions. Amplified cDNA samples were then stored at 4° C. until analysis.
- Amplified cDNA was sequenced by a commercial vendor (Psomagen, Inc., Rockville, MD) according to standard procedures. Library preparation was accomplished using Illumina Nextera DNA Flex kits according to the manufacturer's instructions. Prepared indexed libraries were then loaded onto a NovaSeq6000 S4 with read length of 150PE for sequencing of 40 M reads per sample. During sequencing, the quality score (Q30) was maintained over 75%. Upon completion of sequencing runs, FASTQ file quality was checked with FASTQC and trimmed with the Trim_galore program. The trimmed FASTQ files were aligned and mapped to human reference genome GRCh38 using the hisat2 program.
- RNA expression analysis was further processed using the Bioconductor package edgeR. Genes were filtered using filterByExpr before logCPM (log counts per million reads) were calculated as a measure of gene expression level. For downstream classifier builds, logCPM values were used.
- the selected classifiers have been frequently used in the medical field for exploring predictive or prognostic biomarkers and included glmnet (Lasso and Elastic-Net Regularized Generalized Linear Model), PAM (Nearest Shrunken Centroids), LM (Linear Regression Model), SVM (Support Vector Machine), and RF (Random Forest).
- glmnet Lasso and Elastic-Net Regularized Generalized Linear Model
- PAM Nearest Shrunken Centroids
- LM Linear Regression Model
- SVM Small Vector Machine
- RF Random Forest
- the five classifiers were compared for their predictive performance using the following experimental design: 1) the data set was split into ten stratified outer folds; 2) for each of the folds, the data were preprocessed for feature selection. The top 20, 50, or 200 differentially expressed genes (features) were selected using linear regression model; 3) The hyperparameters were tuned in the training set via a ten-fold cross-validation, and the process subsequently repeated five times; 4) Based on the selected hyperparameters, a model was derived from the training set and applied to the test set. Performance metrics on the test set were then calculated. This process was repeated five times for each classifier.
- IL-23i treated patients in STAMP studies were used for IL-23i classifier training.
- Baseline PASI filter (none, 6+, 8+, and 10+) were applied to explore the impact of disease severity on classifier performance.
- Classifier training were performed using the machine learning approaches stated above and test performance was assessed using 10-fold cross validation.
- IL-23i classifier were locked once a desired performance (>85% PPV and >85% sensitivity) were achieved.
- the IL-23i treatment patients enrolled after the classifier lock were used as the independent validation set.
- RNA profiling was performed using an Affymetrix microarray platform. The lesional samples collected at baseline were used in the predictive biomarker analysis.
- the 17 predictive genes were negatively correlating with patients' response to brodalumab.
- the 17 genes were mapped to 14 Ensemble gene IDs reported in RNASeq data from STAMP studies.
- the 14-gene classifier was validated in STAMP studies.
- Supervised predictive biomarker selection was applied to individual training data to filter genes based on the following assessment: 1) correlation between gene expression and patient response; 2) median gene expression level; 3) gene expression dynamic range; 4) difference between average gene expression of responder and non-responders. Ratios of genes down-regulated and gene up-regulated in TNF ⁇ i responders were used to develop a prediction of TNF ⁇ i treatment responses.
- IL-23i, IL-17i and TNF ⁇ i classifiers were independently validated using the patients enrolled in STAMP studies. Each classifier discretely predicted a patient as either a responder or non-responder for biologic class. Response was defined as achieving PASI75 at week 12. The cross-tabulation of observed and predicted classes with associated statistics was calculated with the confusionMatrix function of the R caret package.
- STAMP is an actively recruiting study designed to continue enrolling new patients to support psoriasis biomarker research. Varied demographics and clinical features of the study subjects were observed (Table 3). With regard to drug class, 49.6% patients were treated with IL-23i, 33.2% were treated with IL-17i, 14.7% were treated with TNF ⁇ i, and 2.5% were treated with IL-12/23i.
- the patient response rate was 64.1% for the whole cohort, and ranged from 47.6% to 72.5% for different biologics (Table 4).
- PASI ⁇ 8 PASI patients were used for predictive classifier development and validation.
- the IL-23i treated patient population was divided into two subsets, 17 IL-23i treated patients were used for training an IL-23i predictive classifier, and the remaining 43 patients were used for prospective validation. All high PASI IL-17i and TNF ⁇ i patients were used for the classifier validation.
- IL-23i predictive classifier training A subset of 17 IL-23i treated high PASI ( ⁇ 8) patients were used for IL-23i predictive classifier training, including 9 responders and 8 non-responders.
- the best performing model was built on glmnet using the top 50 features selected with linear regression model. Test performance was assessed with ten-fold cross validation and the positive predictive value (PPV), sensitivity and balanced accuracy were 89.7%, 96.3%, and 91.9%, respectively.
- PPV positive predictive value
- Table 5 Four publicly available datasets (Table 5) were identified and used for the TNF ⁇ i response classifier development. A total of 73 patients were included in these datasets, out of which 58 patients had both transcriptome data and outcome assessment data for predictive biomarker discovery. Patient outcome was assessed with PASI75 at week 12 or 16, or histological response.
- Supervised predictive biomarkers selection was applied to the four training data sets.
- Nine genes were determined as predictive of TNF ⁇ i response in at least two datasets (Table 6).
- the output of the classifier was a TNF ⁇ i response prediction score; in this scoring system, the lower the prediction score, the higher chance the patient will respond to TNF ⁇ i treatment.
- the classifier performance showed PPV, sensitivity and balanced accuracy of 78.9%, 43.1%, and 63.7%, respectively with this training set (Table 7).
- the predicted response prevalence of patients by all three classifiers was assessed using 195 patients who had baseline DBP samples and completed RNASeq sequencing data ( FIG. 9 and Table 11). Individually, the predicted response prevalence was 72.3%, 51.7% and 67.1% for IL-23i, IL-17i and TNF ⁇ i classifiers, respectively. Critically, 99.5% (194/195) patients were predicted as to responder to at least one of the three drug classes.
- the final IL-23i classifier was developed and validated using patients enrolled in the STAMP studies. A subset of the total IL-23i enrolled patients were used as the training set and the remaining patients in the cohort were used for classifier validation. Since the training and test set were from the same study with the sample and data processed in the same manner, the classifier developed with the training set can be applied to the test set without the need for additional normalization.
- the strategy for the development of the IL-17i and TNF ⁇ i classifiers was different from the IL-23i classifier.
- the IL-17i and TNF ⁇ i patient sample sizes in STAMP studies were smaller than IL-23i patients and were not sufficient to divide into separate training and test sets, so publicly available datasets were used for the training of these two classifiers.
- the training sets from the public data differed significantly than the test sets in some aspects, including sample collection method (punch biopsy vs. dermal patch), RNA preparation protocol, transcriptome profiling method (array vs. sequencing based). Due to these different natures of the training sets, the training sets primarily were used only for feature selection.
- BMI and age have been reported as having clinical prognostic value in assessing biologic treatment response.
- the predictive significance of BMI and age as a possible orthogonal input variable in the classifier is tested. However, adding either variable as a covariate when exploring the predictive models, no improvement was observed in the predictive accuracy or positive predictive value.
- an actionable machine learning-based precision medicine test that can predict psoriasis patient response to biologics (TNF ⁇ i, IL17i, or IL23i) with high positive predictive value, by combining dermal biomarker patch platform with machine learning methods.
- biologics TNF ⁇ i, IL17i, or IL23i
- Using baseline biomarkers combined with machine learning algorithm development the proper biologic for a given patient can be prescribed the first time. This test could lead to improved patient outcomes while also translating into tremendous cost savings for healthcare systems. It is contemplated that this test can effectively minimize the trial-and-error approach to the biologic treatment of psoriasis, and provide physicians, patients, and payers with a powerful tool to bring personalized medicine to the management of psoriasis patients.
- Example 7 Efficient Prediction of Response to Psoriasis Biologics Using a Machine Learning Classifier
- psoriasis affects upward of 3% of the population and treatment can cost from $87,585 to $366,645 annually with the most frequently used drugs. It generally takes 12 to 16 weeks for clinical response to treatment to be meaningful, and efficacy of presently available therapies ranges from 30% to 80%, due at least in part to the inability to predict response to a specific treatment regimen.
- Mind.Px An algorithm (Mind.Px) is developed herein for the prediction of response to biologics (anti-IL-17, anti-IL-23) used for the treatment of patients with psoriasis, by comparing baseline transcriptomes with clinical response to biologic drugs at 12 weeks after initial treatment.
- Psoriasis Area and Severity Index PASI
- a total of 17 genes were finalized as correlating with patient response to anti-IL-17 biologics.
- a total of 27 genes were finalized for use in this classifier and the positive predictive value (PPV) across both classifiers was 100%.
- Classifiers for biologic prediction largely utilized orthogonal biomarkers and achieved high PPV.
- the use of Mind.Px results in better outcomes in patients with psoriasis and significantly reduced costs to the healthcare system.
- a customized spring-loaded applicator was used to standardize the application pressure across subjects and across users.
- the loaded applicator was placed against the skin and the trigger pressed, applying the patch to the skin.
- the patch was then held in place against the skin for 5 minutes by a ring of medical tape. After this time, the patch was removed from the subject, immediately placed into storage buffer (LiCl, Triton X-100, Tris-EDTA), and stored at 4° C. until processing.
- Dermal transcriptomes were processed within 72 hours of collection from subjects. Samples were prepared by washing the applied DBP with chilled 1 ⁇ PBS and then drying the patch under a stream of nitrogen. The dried DBP was then place on a heat block preset to 95° C. for 1 minute, at which time, 50 ⁇ L of PCR grade water previously heated to 95° C. was then applied to the microneedle array for 1 minute to elute the bound mRNA from the DBP. This eluted mRNA was then converted to cDNA using Takara SMART-Seq® Single Cell kit according to the manufacturer's instructions. Amplified cDNA samples were then stored at 4° C. until qPCR or NGS analysis.
- Amplified cDNA was sequenced by a commercial vendor (Psomagen, Inc., Rockville, MD) according to standard procedures. Library preparation was accomplished using Illumina Nextera DNA Flex kits according to the manufacturer's instructions. Prepared indexed libraries were then loaded onto a NovaSeq6000 S4 with read length of 150PE for sequencing of 40 M reads per sample. During sequencing, the quality score (Q30) was maintained over 75%. Upon completion of runs, FASTQ file quality was checked with FASTQC and trimmed with the Trim_galore program. The trimmed FASTQ were aligned and mapped to human reference genome GRCh38 using hisat2 program.
- RNA expression analysis was further processed using Bioconductor package edgeR. Genes were filtered using filterByExpr before logCPM (counts per million reads) were calculated. Plots were made with ggplot2 and heatmap.2 functions in R. Percent of genes detected was determined using the ratio of genes detected (with >0 count) over the total number of Ensemble genes.
- the Mind.Px algorithm has been implemented as an R-script on the DNAnexus® system (Mountain View, CA), a cloud-based data analysis and management platform for DNA sequencing data. Cutoff points for anti-IL-17 and anti-IL-23 response prediction have been defined and implemented in the R-script.
- RNA-Seq FASTQ files were trimmed with the Trim_galore Program, which was followed by mapping to human reference genome GRCh38 using the HISAT2 Program. The number of reads was counted for each Ensemble gene ID using the FeatureCounts Program, and human GRCh38.84.gtf.
- RNA expression analysis was further processed using bioconductor package edgeR. Genes were filtered using filterByExpr before logCPM (counts per million reads) was calculated. Plots were made with ggplot2 and heatmap.2 functions compared with the standard R function. The percent of genes detected was determined using the ratio of genes detected (with >0 count) divided by the total number of Ensemble genes.
- FIG. 12 The overview of RNA-Seq analysis is summarized in FIG. 12 .
- RNA from the skin using a dermal biomarker patch.
- Subsequent next-generation sequencing of the extracted RNA has allowed us to take a genetic and transcriptomic snapshot of the skin at the exact moment of the test.
- This rich patient-specific data set can then be analyzed by machine-learning algorithms to ask sophisticated questions of the data (e.g., predicting the appropriate biologic drug for a patient prior to therapeutic selection and treatment).
- Also disclosed herein is an algorithm for the prediction of response to biologics (anti-IL-17, anti-IL-23) treatment in patients with psoriasis.
- the data set from Tomalin et al (2020) was used for the identification of genes whose RNA expression levels correlate with the responses to treatment with anti-IL-17 biologics in patients with psoriasis.
- the Akaike information criterion is an estimator of out-of-sample prediction error and thereby relative quality of statistical models for a given set of data. Akaike information criterion estimates the relative amount of information lost by a given model: the less information a model loses, the higher the quality of that model.
- a heatmap was generated from expression data for these 17 genes ( FIG. 14 ).
- Each column is a baseline sample collected from a patient. From left to right the patients were sorted by low to high PASI changes. The blue bar on the top shows the 12 non-responders (W12 PASI changes ⁇ 0.75); the yellow bar shows the 62 responders (W12 PASI changes >0.75).
- W12 PASI changes ⁇ 0.75 the blue bar
- the yellow bar shows the 62 responders (W12 PASI changes >0.75).
- a subset of responder patients near right had strong down-regulation (blue) of most of genes, suggesting the potential of using these 17 genes to predict patient response to anti-IL-17 treatment.
- the down-regulation of these 17 genes indicated the better responses to treatment, whereas the up-regulation indicated the worse responses.
- Disclosed herein is a platform that utilizes a simple, minimally invasive patch that takes a painless biomarker sample from a patient with psoriasis in a matter of minutes.
- the Mind.Px test has the ability to collect patient data at scale and, when combined with high-precision molecular testing, results in a powerful platform with excellent sensitivity and specificity.
- Use of this platform can potentially translate into huge cost savings for healthcare systems, particularly when applied to the prediction of response to expensive treatments—effectively eliminating the present trial-and-error approach to the treatment of psoriasis.
- Biomarkers captured using Mind.Px include DNA, RNA, proteins, and small molecules.
- RNA Ribonucleic acid
- next-generation sequencing is used to evaluate more than 7000 biomarkers per test sample.
- the results of this biomarker analysis can be used by healthcare providers and payers to predict an individual patient's response to a class of biologic drugs, by the mechanism of action of that class of agent, to optimize treatment selection.
- the use of Mind.Px could result in better outcomes and significantly reduced costs to the healthcare system.
- the methods disclosed herein directed to determining whether a subject suffering from psoriasis will positively respond to an IL-17, IL-23, or TNF ⁇ mediated treatment further comprise determining the presence, absence, or expression level of at least 5 genes selected from Table 14. In some embodiments, the methods further comprise determining the presence, absence, or expression level of at least 6 genes selected from Table 14 to 17 genes selected from Table 14.
- the methods further comprise determining the presence, absence, or expression level of at least 6 genes selected from Table 14 to 7 genes selected from Table 14, 6 genes selected from Table 14 to 8 genes selected from Table 14, 6 genes selected from Table 14 to 9 genes selected from Table 14, 6 genes selected from Table 14 to 10 genes selected from Table 14, 6 genes selected from Table 14 to 11 genes selected from Table 14, 6 genes selected from Table 14 to 12 genes selected from Table 14, 6 genes selected from Table 14 to 13 genes selected from Table 14, 6 genes selected from Table 14 to 14 genes selected from Table 14, 6 genes selected from Table 14 to 15 genes selected from Table 14, 6 genes selected from Table 14 to 16 genes selected from Table 14, 6 genes selected from Table 14 to 17 genes selected from Table 14, 7 genes selected from Table 14 to 8 genes selected from Table 14, 7 genes selected from Table 14 to 9 genes selected from Table 14, 7 genes selected from Table 14 to 10 genes selected from Table 14, 7 genes selected from Table 14 to 11 genes selected from Table 14, 7 genes selected from Table 14 to 12 genes selected from Table 14, 7 genes selected from Table 14 to 13 genes selected from Table 14, 7 genes selected from Table 14 to 14 genes selected from Table 14 from Table 14
- the methods further comprise determining the presence, absence, or expression level of at least 6 genes selected from Table 14, 7 genes selected from Table 14, 8 genes selected from Table 14, 9 genes selected from Table 14, 10 genes selected from Table 14, 11 genes selected from Table 14, 12 genes selected from Table 14, 13 genes selected from Table 14, 14 genes selected from Table 14, 15 genes selected from Table 14, 16 genes selected from Table 14, or 17 genes selected from Table 14.
- the methods further comprise determining the presence, absence, or expression level of at least at least 6 genes selected from Table 14, 7 genes selected from Table 14, 8 genes selected from Table 14, 9 genes selected from Table 14, 10 genes selected from Table 14, 11 genes selected from Table 14, 12 genes selected from Table 14, 13 genes selected from Table 14, 14 genes selected from Table 14, 15 genes selected from Table 14, or 16 genes selected from Table 14.
- the methods further comprise determining the presence, absence, or expression level of at least at most 7 genes selected from Table 14, 8 genes selected from Table 14, 9 genes selected from Table 14, 10 genes selected from Table 14, 11 genes selected from Table 14, 12 genes selected from Table 14, 13 genes selected from Table 14, 14 genes selected from Table 14, 15 genes selected from Table 14, 16 genes selected from Table 14, or 17 genes selected from Table 14. In some embodiments, the methods further comprise determining the presence, absence, or expression level of at least 20 genes selected from Table 14 to 50 genes selected from Table 14.
- the methods further comprise determining the presence, absence, or expression level of at least 20 genes selected from Table 14 to 25 genes selected from Table 14, 20 genes selected from Table 14 to 30 genes selected from Table 14, 20 genes selected from Table 14 to 35 genes selected from Table 14, 20 genes selected from Table 14 to 40 genes selected from Table 14, 20 genes selected from Table 14 to 45 genes selected from Table 14, 20 genes selected from Table 14 to 50 genes selected from Table 14, 25 genes selected from Table 14 to 30 genes selected from Table 14, 25 genes selected from Table 14 to 35 genes selected from Table 14, 25 genes selected from Table 14 to 40 genes selected from Table 14, 25 genes selected from Table 14 to 45 genes selected from Table 14, 25 genes selected from Table 14 to 50 genes selected from Table 14, 30 genes selected from Table 14 to 35 genes selected from Table 14, 30 genes selected from Table 14 to 40 genes selected from Table 14, 30 genes selected from Table 14 to 45 genes selected from Table 14, 30 genes selected from Table 14 to 50 genes selected from Table 14, 35 genes selected from Table 14 to 40 genes selected from Table 14, 35 genes selected from Table 14 to 45 genes selected from Table 14, 35 genes selected from Table 14 to 45 genes selected from Table 14, 35 genes selected
- the methods further comprise determining the presence, absence, or expression level of at least 20 genes selected from Table 14, 25 genes selected from Table 14, 30 genes selected from Table 14, 35 genes selected from Table 14, 40 genes selected from Table 14, 45 genes selected from Table 14, or 50 genes selected from Table 14. In some embodiments, the methods further comprise determining the presence, absence, or expression level of at least at least 20 genes selected from Table 14, 25 genes selected from Table 14, 30 genes selected from Table 14, 35 genes selected from Table 14, 40 genes selected from Table 14, or 45 genes selected from Table 14. In some embodiments, the methods further comprise determining the presence, absence, or expression level of at least at most 25 genes selected from Table 14, 30 genes selected from Table 14, 35 genes selected from Table 14, 40 genes selected from Table 14, 45 genes selected from Table 14, or 50 genes selected from Table 14.
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US6663820B2 (en) * | 2001-03-14 | 2003-12-16 | The Procter & Gamble Company | Method of manufacturing microneedle structures using soft lithography and photolithography |
ES2650188T3 (es) * | 2004-06-10 | 2018-01-17 | 3M Innovative Properties Company | Dispositivo y kit de aplicación de parches |
US7132054B1 (en) * | 2004-09-08 | 2006-11-07 | Sandia Corporation | Method to fabricate hollow microneedle arrays |
DE602005024038D1 (de) * | 2004-12-10 | 2010-11-18 | 3M Innovative Properties Co | Medizinische vorrichtung |
EP2052088A2 (fr) * | 2006-08-02 | 2009-04-29 | Genizon Biosciences | Carte génique des gènes humains associés au psoriaris |
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US20190184366A1 (en) * | 2016-08-03 | 2019-06-20 | Verndari, Inc. | Microarrays and methods |
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---|---|---|---|---|
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Non-Patent Citations (8)
Title |
---|
Bagel et al Skin. 05 Nov 2021. 5(6): 621-638 (Year: 2021) * |
Coleman, R. Drug Discovery Today. 2003. 8: 233-235 (Year: 2003) * |
dCorrea da Rosa et al. J Invest Dermatology. 2017. 137: 305-312 and Supplemental Materials and Methods, 35 pages total (Year: 2017) * |
Ibrahim et al 2017. J Invest Dermatology. "Non-invasive transcriptome extraction from the skin using MiNDERA microneedle technology" available via URL <wp-content/uploads/2019/03/MiNDERA-SID-2017-Poster-.pdf (Year: 2017) * |
Liu et al Clinical Immunology. 2004. 112: 225-230 (Year: 2004) * |
Shaheen et al Int J Molec Sciences. 19 June 2018. 19, 1810, p. 1-13 (Year: 2018) * |
Sulaiman et al ACS Nano. 27 August 2019. 13(8), p. 1-16 (Year: 2019) * |
Visvanathan et al J Allergy Clin Immunol. 2019. 143: 2158-2169 (Year: 2019) * |
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