MICROWAVE TREATMENT OF SKIN

Abstract

The present disclosure provides microwave-based methods for the modulation of certain genes and immunomodulatory factors. The various methods described herein may be used to modulate the expression of one or more genes thought to be beneficial in and/or associated with, the treatment and/or prevention of a disease and/or condition of the skin.

Claims

1. A method of modulating the expression of one or more genes, said method comprising administering microwave energy to a subject in need thereof.

2. The method of claim 1, wherein the microwave energy administered at a frequency of between about 900 MHz and about 200 GHz.

3. The method of claim 1, wherein the microwave energy is administered at about 915 MHz, at about 2.45 GHz, at about 5.8 GHz, at about 8.0 GHz, or at about 24.125 GHz.

4. The method of claim 1, wherein the microwave energy is administered at an input power of 0.5 W to 40 W.

5. The method of claim 1, wherein the microwave energy is administered for a duration of anywhere between about 0.1 s to 20 s.

6. The method of claim 1, wherein the microwave energy is administered at 5 W for 3s, 4 W for 3s or 3 W for 3s.

7. The method of claim 1, wherein the microwave energy is administered as a series of pulsed doses.

8. The method of claim 7, wherein each pulse dose in the series of pulses is separated from another pulsed dose in the series by a time gap of anywhere between about 1 s to about 60 s

9. The method of claim 1, wherein the microwave energy is administered as 3 doses with a 20 s-time gap between each administered dose.

10. The method of claim 1, wherein the administered microwave energy is non-ablative.

11. The method of claim 10, wherein the non-ablative microwave energy does not cause direct tissue or skin damage.

12. The method of claim 1, wherein one or more of the gene to be modulated encode or provide factors associated with the host immune system.

13. The method of claim 1, wherein one or more of the gene to be modulated encode or provide immunomodulatory factors.

14. The method of claim 1, wherein one or more of the genes to be modulated are oncogenes.

15. The method of claim 1, wherein the one or more genes are presented in Tables 1 or 3.

16. The method of claim 1, wherein the one or more genes are presented in Tables 2 or 4.

17. A method of treating or preventing a skin condition by modulating the expression of one or more genes, said method comprising administering a subject suffering from, or predisposed to, the skin condition, microwave energy.

18. The method of claim 17, wherein the one or more genes are associated with the skin condition to be treated or prevented.

19. The method of claim 17, wherein the gene or genes to be modulated may be directly or indirectly associated with a disease or condition affecting the skin and/or wherein one or more of the genes may be involved or linked with/to one or more pathways or mechanisms associated with a disease or condition of the skin.

20. The method of claim 17, wherein the microwave energy administered at a frequency of between about 900 MHz and about 200 GHz.

21. The method of claim 17, wherein the microwave energy is administered at about 915 MHz, at about 2.45 GHz, at about 5.8 GHz, at about 8.0 GHz, or at about 24.125 GHz.

22. The method of claim 17, wherein the microwave energy is administered at an input power of 0.5 W to 40 W.

23. The method of claim 17, wherein the microwave energy is administered for a duration of anywhere between about 0.1 s to 20 s.

24. The method of claim 17, wherein the microwave energy is administered at 5 W for 3s, 4 W for 3s or 3 W for 3s.

25. The method of claim 17, wherein the microwave energy is administered as a series of pulsed doses.

26. The method of claim 25, wherein each pulse dose in the series of pulses is separated from another pulsed dose in the series by a time gap of anywhere between about 1 s to about 60 s

27. The method of claim 17, wherein the microwave energy is administered as 3 doses with a 20 s-time gap between each administered dose.

28. The method of claim 17, wherein the administered microwave energy is non-ablative.

29. The method of claim 28, wherein the non-ablative microwave energy does not cause direct tissue or skin damage.

30. The method of claim 17, wherein the expression of one or more of the genes is/are either (i) downregulated, inhibited or reduced or (ii) upregulated, induced, promoted or stimulated.

31. The method of claim 17, wherein the microwave energy is administered to a diseased tissue.

32. The method of claim 17, wherein the microwave energy is administered to a tissue exhibiting the signs or symptoms characteristic of one or more diseases.

33. The method of claim 17, wherein the microwave energy is administered to the skin and/or diseased skin.

34. The method of claim 17, wherein the microwave energy is administered to: (i) aging skin; or (ii) skin which exhibits solar damage; or (iii) skin with one or more scars, erosion and/or lesions.

35. The method of claim 17, wherein the disease to be treated or prevented is a wart, eczema, psoriasis, acne, cherry angioma, hidradenitis suppurativa, rosacea, ichthyosis, keloid scars, seborrheic dermatitis, seborrheic keratosis, seborrheic hyperplasia, Sebaceous hyperplasia, basal cell carcinoma, actinic keratosis, syringoma, squamous cell carcinoma, nevus, lentigo maligna, Melasma, melanoma, milia, molluscum contagiosum, cervical intraepithelial neoplasia, vaginal intraepithelial neoplasia, vulvar intraepithelial neoplasia, Bowen's disease and/or erythroplasia of queyrat.

36. The method of claim 17, wherein the disease to be treated or prevented is gastric epithelial dysplasia, cardiovascular lesions, conditions involving oral cavity such as epithelial dysplasia, leukoplakia, hairy leukoplakia, erythroplakia, erythroleukoplakia, lichen planus, xerostomia, mucositis, pyogenic granuloma, angioma, nicotinic stomatitis, actinic cheilitis, keratoacantoma, hyperkeratosis, candidosis, erythema migrans and/or a canker sore.

37. A method of treating a disease or condition associated with the downregulation or inhibition of a particular gene, said method comprising administering a subject in need thereof microwave energy to upregulate the gene thereby restoring the level of expression, activity and/or function of the gene to treat or prevent the disease or condition.

38. The method of claim 37, wherein the disease or condition is a disease or condition of the skin.

39. A method of treating a disease or condition associated with the upregulation of a particular gene, said method comprising administering a subject in need thereof microwave energy to downregulate the gene thereby restoring the level of expression, activity and/or function of the gene to treat or prevent the disease or condition.

40. The method of claim 39, wherein the disease or condition is a disease or condition of the skin.

Description

DETAILED DESCRIPTION

[0078] The present invention will now be described with reference to the following figures which show:

[0079] FIG. 1: a schematic illustration of a microwave treatment system, in accordance with embodiments.

[0080] FIG. 2: PCA of significantly altered genes between microwave treated and untreated skin assessed on Immunology panel.

[0081] FIG. 3: a heatmap showing the significantly altered genes between microwave treated and untreated skin assessed on Immunology panel.

[0082] FIG. 4: Example of gene count in normal, microwave treated and diseased skin tissue on Immunology panel.

[0083] FIG. 5: PCA of significantly altered genes between microwave treated and untreated skin assessed on PanCancer panel.

[0084] FIG. 6: a heatmap of significantly altered genes between microwave treated and untreated skin assessed on PanCancer panel.

[0085] FIG. 7: Example of gene count in normal, microwave treated and diseased skin tissue on PanCancer panel.

MICROWAVE ENERGY SYSTEM

[0086] A microwave radiation delivery system 11, in accordance with embodiments, for treating a biological tissue, is illustrated in FIG. 1. The system comprises a microwave generator 12 for generating microwave radiation, a flexible or rigid interconnecting cable 13 and a microwave applicator assembly (also called microwave antenna) 14 for delivering microwave radiation to a biological tissue. Other variations of this arrangement are possible including integrated versions. The microwave radiation delivery system may further comprise a controller (not shown) which is configured to select an operational frequency or range of frequencies to be supplied by the generator. A frequency of the microwave radiation supplied by the microwave generator 12 may be between 900 MHz and 30 GHz, for example about 915 MHz, about 2.45 GHz, about 5.8 GHz, about 8.0 GHz, or about 24.125 GHz. In some embodiments, the microwave radiation supplied is pulsed meaning that the energy is delivered in precise and brief doses lasting a number of seconds.

Methods

[0087] A microwave system comprising a microwave generator; a controller configured to control the microwave generator to generate microwave energy having a selected operational frequency or range of frequencies; a microwave cable configured to deliver the microwave energy to a microwave antenna extending from or coupled to a distal end of the microwave cable; and a microwave antenna was used to apply microwave energy to skin tissue for example diseased skin tissue. This created thermal and non-thermal effects within the tissue and biopsies were taken for transcriptome studies.

[0088] The analysis of mRNA transcripts was performed using NanoString (NanoString Technologies Inc., Seattle, Wash. 98109 USA) nCounter gene expression system using, Immune pathway (nCounter Human Immunology V2 Panel, catalogue number: XT-CSO-HIM2-12) and Cancer pathway (nCounter PanCancer Pathways, catalogue number: XT-CSO-PATH1-12) to comprehend changes in the transcripts of number of genes (579 human genes in the Immunology V2 panel assay and 730 human genes in the PanCancer pathway assay) [20].

[0089] Gene expression analysis was performed using nSolver 4.0 (NanoString Technologies Inc., Seattle, Wash. 98109 USA) and open source Bioconductor DESeq2 in R studio (version 3.5.3). The results are presented in Tables 1-4 above and also 5-8 below.

[0090] Treated skin refers to microwave treated skin. Terms untreated skin/control/diseased skin are interchangeable.

TABLE-US-00005 TABLE 5 those genes found to be significantly upregulated between microwave treated and untreated skin assessed on Immunology panel. Base log2 Fold Gene Mean Change p-value p-adj CFH 1026.38 1.49 0.0000 0.0000 MSR1 157.37 1.44 0.0001 0.0015 CXCL12 2043.39 1.24 0.0000 0.0000 HLA-DPB1 2556.70 1.21 0.0000 0.0000 MRC1 296.88 1.17 0.0000 0.0002 FCER1A 370.56 1.12 0.0003 0.0053 C3 510.64 1.10 0.0000 0.0008 VCAM1 117.58 1.00 0.0000 0.0000 CFD 577.43 0.96 0.0012 0.0132 CCL13 291.62 0.87 0.0018 0.0184 LGALS3 2115.07 0.85 0.0001 0.0019 CDH5 139.47 0.83 0.0004 0.0062 KIT 129.87 0.82 0.0005 0.0072 CD209 41.49 0.82 0.0028 0.0247 LILRB4 65.57 0.79 0.0036 0.0298 CLEC5A 35.60 0.76 0.0045 0.0339 HLA-DRA 6024.43 0.76 0.0001 0.0024 SERPING1 1521.07 0.75 0.0000 0.0003 HLA-DPA1 2211.06 0.75 0.0001 0.0021 CD4 220.85 0.73 0.0000 0.0002 FCGRT 940.18 0.73 0.0000 0.0003 NT5E 301.10 0.72 0.0058 0.0384 C2 136.50 0.71 0.0000 0.0004 CSF1 88.71 0.69 0.0000 0.0011 HAVCR2 48.56 0.65 0.0035 0.0296 TNFSF12 183.38 0.65 0.0000 0.0001 LAIR1 158.32 0.65 0.0026 0.0245 B2M 22970.01 0.65 0.0008 0.0104 CMKLR1 135.93 0.65 0.0006 0.0081 PDCD1LG2 61.53 0.63 0.0000 0.0007 CD45R0 303.52 0.63 0.0009 0.0115 CD34 105.37 0.63 0.0047 0.0351 XCR1 45.12 0.62 0.0030 0.0261 CYBB 338.71 0.61 0.0022 0.0220 ITGAM 79.52 0.60 0.0013 0.0145 TGFBR2 634.57 0.55 0.0001 0.0024 HLA-DMB 393.95 0.53 0.0004 0.0066 CTSS 748.47 0.50 0.0056 0.0383 IKZF2 58.96 0.50 0.0028 0.0247 CD59 2503.76 0.49 0.0073 0.0443 TNFRSF11A 55.36 0.49 0.0027 0.0247 TNFSF13B 94.44 0.48 0.0068 0.0423 TLR3 53.11 0.46 0.0049 0.0363 STAT5A 163.79 0.45 0.0002 0.0042 IFITM1 1874.75 0.45 0.0068 0.0423 NFKB1 151.67 0.44 0.0010 0.0118 ITGB2 362.43 0.44 0.0078 0.0465 C1S 1402.07 0.43 0.0026 0.0245 SDHA 449.24 0.41 0.0016 0.0174 ETS1 539.67 0.40 0.0032 0.0278 CASP1 190.69 0.37 0.0057 0.0383 C1R 1843.39 0.37 0.0057 0.0383 HLA-DMA 489.96 0.36 0.0053 0.0383 CD74 7154.25 0.35 0.0067 0.0422 MAPK1 510.11 0.34 0.0055 0.0383 IL6ST 1035.06 0.32 0.0006 0.0084

TABLE-US-00006 TABLE 6 those genes significantly downregulated between microwave treated and untreated skin assessed on Immunology panel. Base log2 Fold Gene Mean Change p-value p-adj IL8 1285.14 −3.51 0.0000 0.0008 IL1B 186.34 −2.93 0.0000 0.0001 IL6 20.73 −2.33 0.0000 0.0006 CD79A 227.05 −2.09 0.0003 0.0057 SOCS3 405.62 −2.06 0.0000 0.0000 CXCL13 252.65 −1.98 0.0000 0.0010 CXCL1 405.92 −1.98 0.0001 0.0025 PTGS2 53.91 −1.71 0.0001 0.0026 TNFRSF17 61.18 −1.42 0.0018 0.0183 EGR1 148.70 −1.41 0.0055 0.0383 CXCL2 159.48 −1.28 0.0002 0.0032 CCL20 49.70 −1.22 0.0039 0.0311 IL28A 6.95 −1.17 0.0037 0.0308 CD19 43.45 −1.12 0.0004 0.0061 LIF 48.54 −1.07 0.0001 0.0024 IL20 22.40 −0.99 0.0045 0.0339 XBP1 993.22 −0.94 0.0006 0.0081 BCL3 353.32 −0.91 0.0000 0.0004 CXCR4 660.62 −0.81 0.0000 0.0001 MIF 2064.06 −0.74 0.0004 0.0065 CD79B 77.02 −0.73 0.0071 0.0432 KLRG2 39.89 −0.73 0.0043 0.0334 LTB4R 597.46 −0.64 0.0040 0.0319 TNFRSF13C 98.61 −0.63 0.0054 0.0383 IRF3 72.05 −0.60 0.0008 0.0104 TNFAIP3 313.05 −0.59 0.0024 0.0231 BCL2L11 71.16 −0.55 0.0008 0.0104 MAPKAPK2 278.49 −0.51 0.0001 0.0027 HLA-C 3374.79 −0.44 0.0042 0.0325 IL1RAP 223.61 −0.43 0.0062 0.0403 TRAF3 245.66 −0.42 0.0026 0.0245 CASP2 316.72 −0.42 0.0076 0.0458 MCL1 1935.98 −0.40 0.0010 0.0118

TABLE-US-00007 TABLE 7 those genes that are significantly upregulated genes between microwave treated and untreated skin assessed on PanCancer panel; Base log2 Fold Gene Mean Change p-value p-adj THBS4 93.07 3.10 0.0000 0.0000 SFRP4 22.13 1.88 0.0001 0.0030 RELN 20.60 1.65 0.0001 0.0023 ETV1 34.49 1.55 0.0000 0.0000 TMPRSS2 17.24 1.31 0.0016 0.0220 MMP7 202.92 1.27 0.0007 0.0122 PPARGC1A 89.69 1.15 0.0000 0.0010 PLCB4 47.93 1.13 0.0000 0.0001 PRKAR2B 141.05 1.11 0.0000 0.0004 AR 72.39 1.10 0.0002 0.0047 FGF2 90.48 1.10 0.0000 0.0000 GHR 44.14 0.91 0.0000 0.0001 PPARG 61.65 0.87 0.0017 0.0220 PLA2G4A 58.95 0.79 0.0000 0.0008 BCL2 103.81 0.77 0.0000 0.0000 KIT 72.69 0.76 0.0002 0.0050 TGFB2 50.53 0.76 0.0001 0.0029 NTRK2 334.85 0.73 0.0030 0.0307 ID4 168.54 0.71 0.0030 0.0307 PDGFD 159.07 0.71 0.0050 0.0451 VEGFC 37.00 0.68 0.0027 0.0303 KITLG 309.93 0.67 0.0001 0.0036 NGFR 83.48 0.67 0.0007 0.0122 HDAC4 172.01 0.66 0.0000 0.0000 TSPAN7 119.23 0.65 0.0000 0.0013 LIFR 160.65 0.65 0.0007 0.0122 USP39 190.37 0.60 0.0000 0.0000 LIG4 107.93 0.58 0.0001 0.0024 FGFR1 346.35 0.54 0.0017 0.0220 B2M 24036.71 0.54 0.0009 0.0141 ID2 422.83 0.54 0.0028 0.0303 DDIT3 142.36 0.53 0.0000 0.0007 GPC4 53.80 0.53 0.0038 0.0369 TGFBR2 787.94 0.52 0.0001 0.0020 AKT3 183.97 0.49 0.0003 0.0057 ALKBH3 130.76 0.42 0.0029 0.0305 NOL7 409.12 0.35 0.0045 0.0415 MAPK1 436.49 0.35 0.0025 0.0283 PRKACB 103.76 0.33 0.0058 0.0497 MAPK9 192.79 0.32 0.0016 0.0220 SKP1 2449.63 0.32 0.0003 0.0064 NF1 276.22 0.29 0.0010 0.0160 SF3B1 740.39 0.29 0.0018 0.0220 RPS27A 23365.03 0.22 0.0037 0.0364

TABLE-US-00008 !TABLE 8 those genes that are significantly downregulated genes between microwave treated and untreated skin assessed on PanCancer panel; Base log2 Fold Gene Mean Change p-value p-adj OSM 88.91 −4.09 0.0000 0.0000 IL1B 202.15 −3.20 0.0000 0.0001 GNG4 15.32 −2.56 0.0000 0.0006 FOS 1526.49 −2.54 0.0000 0.0002 IL24 20.95 −2.49 0.0000 0.0001 IL6 19.49 −2.22 0.0000 0.0007 SOCS3 429.31 −2.08 0.0000 0.0000 NR4A1 94.69 −2.00 0.0026 0.0293 CD19 30.18 −1.94 0.0003 0.0064 PAX5 14.85 −1.29 0.0020 0.0240 DUSP2 59.50 −1.16 0.0002 0.0052 LIF 45.57 −1.16 0.0001 0.0027 BCL2A1 53.99 −1.06 0.0031 0.0311 MYC 889.67 −1.03 0.0000 0.0001 EPHA2 180.44 −0.89 0.0000 0.0003 FOSL1 106.32 −0.82 0.0057 0.0495 CACNB3 59.66 −0.70 0.0001 0.0029 ETS2 1508.69 −0.66 0.0003 0.0062 HSPB1 4060.38 −0.66 0.0019 0.0233 CDC7 120.29 −0.66 0.0009 0.0147 COL27A1 191.58 −0.62 0.0002 0.0047 PIM1 1315.32 −0.59 0.0022 0.0256 ID1 1020.68 −0.54 0.0030 0.0307 ALKBH2 67.57 −0.53 0.0017 0.0220 TNFAIP3 442.34 −0.52 0.0021 0.0255 CDKN2D 116.20 −0.51 0.0017 0.0220 MCM7 662.05 −0.48 0.0044 0.0407 VHL 478.69 −0.47 0.0048 0.0434 KRAS 289.73 −0.45 0.0028 0.0303 PIK3R2 292.17 −0.44 0.0016 0.0220 HSP90B1 721.15 −0.42 0.0007 0.0122 CIC 277.81 −0.34 0.0005 0.0098 PML 617.66 −0.32 0.0015 0.0220 MMP3 186.51 −0.23 0.0022 0.0261 TNFRSF100 53.31 −0.21 0.0043 0.0407 IL8 1724.97 −0.16 0.0000 0.0008

TABLE-US-00009 TABLE 9 shows significantly altered common genes between microwave treated and untreated skin assessed on Immunology and PanCancer panel Immunology Path Cancer Path Base log2 Fold Base log2 Fold Gene Mean Change pvalue padj Mean Change pvalue padj KIT 129.87 0.82 0.0005 0.0072 72.69 0.76 0.0002 0.0050 B2M 22970.01 0.65 0.0008 0.0104 24036.71 0.54 0.0009 0.0141 TGFBR2 634.57 0.55 0.0001 0.0024 787.94 0.52 0.0001 0.0020 MAPK1 510.11 0.34 0.0055 0.0383 436.49 0.35 0.0025 0.0283 IL8 1285.14 −3.51 0.0000 0.0008 1724.97 −0.16 0.0000 0.0008 TNFAIP3 313.05 −0.59 0.0024 0.0231 442.34 −0.52 0.0021 0.0255 LIF 48.54 −1.07 0.0001 0.0024 45.57 −1.16 0.0001 0.0027 CD19 43.45 −1.12 0.0004 0.0061 30.18 −1.94 0.0003 0.0064 SOCS3 405.62 −2.06 0.0000 0.0000 429.31 −2.08 0.0000 0.0000 IL6 20.73 −2.33 0.0000 0.0006 19.49 −2.22 0.0000 0.0007 IL1B 186.34 −2.93 0.0000 0.0001 202.15 −3.20 0.0000 0.0001

[0091] Results (see tables 1-9 above) comparing the level of genes expression in (microwave) untreated and (microwave) treated skin tissues (tested using Wald test) revealed:

89 genes in the Immunology V2 panel and 80 genes in the PanCancer pathway were significantly modulated (up and/or down regulated). All achieve significance at p<0.05.

Immunology V2 Panel Results:

[0092] Total number of significantly altered genes between treated and untreated skin=89

[0093] Significantly upregulated genes (n=56) in the treated skin (see Tables 1 and 5):

B2M, C1R, C1S, C2, C3, CASP1, CCL13, CD209, CD34, CD4, CD59, CD74, CFH, CDH5, CFD, CXCL12, CLEC5A, CMKLR1, CSF1, CTSS, FCER1A, HLA-DPB1, MRC1, MSR1, CYBB, ETS1, FCGRT, HAVCR2, HLA-DMA, HLA-DMB, HLA-DPA1, HLA-DRA, IFITM1, IKZF2, IL6ST, ITGAM, ITGB2, KIT, LAIR1, LGALS3, LILRB4, MAPK1, NFKB1, NT5E, PDCD1LG2, CD45R0, SDHA, SERPING1, STAT5A, TGFBR2, TLR3, TNFRSF11A, TNFSF12, TNFSF13B, VCAM1, XCR1.

[0094] Significantly downregulated genes (n=33) in the treated skin (see Tables 2 and 6):

BCL2L11, BCL3, CASP2, CCL20, CD19, CD79B, CXCR4, CD79A, CXCL1, HLA-C, CXCL13, IL1B, IL1RAP, IL20, IL28A, CXCL2, IL8, IRF3, KLRG2, EGR1, LTB4R, MAPKAPK2, MCL1, MIF, IL6, LIF, PTGS2, SOCS3, TNFAIP3, TNFRSF13C, TNFRSF17, TRAF3, XBP1.

[0095] In total forty-four genes were found to be significantly upregulated between treated and control skin. On the other hand, thirty-six genes were observed as significantly downregulated between treated and control skin.

PanCancer Pathway Results:

[0096] Total number of significantly altered genes between treated and untreated skin=80 (see Tables 3 and 7)

[0097] Significantly upregulated genes (n=44) in the treated skin:

AKT3, ALKBH3, AR, B2M, BCL2, DDIT3, ETV1, FGF2, FGFR1, GHR, GPC4, HDAC4, ID2, ID4, KIT, KITLG, LIFR, LIG4, MAPK1, MAPK9, MMP7, NF1, NGFR, NOL7, NTRK2, PDGFD, PLA2G4A, PLCB4, PPARG, PPARGC1A, PRKACB, PRKAR2B, RELN, RPS27A, SF3B1, SFRP4, SKP1, TGFB2, TGFBR2, THBS4, TMPRSS2, TSPAN7, USP39, VEGFC.

[0098] Significantly downregulated genes (n=36) in the treated skin (see Tables 4 and 8):

ALKBH2, BCL2A1, CACNB3, CD19, CDCl7, CDKN2D, CIC, COL27A1, DUSP2, EPHA2, ETS2, FOS, FOSL1, GNG4, HSP90B1, HSPB1, ID1, IL1B, IL24, IL6, IL8, KRAS, LIF, MCM7, MMP3, MYC, NR4A1, OSM, PAX5, PIK3R2, PIM1, PML, SOCS3, TNFAIP3, TNFRSF10C, VHL.

Principal Component Analysis (PCA)

[0099] The analysis comprises principal component (PC) axes that elucidate distinguished distribution of control and treated skin samples. In FIG. 2, PCA 32 on the Immunology panel comprising first principal axis PC1 33 and second principal axis PC2 34 show variation of 38% and 16.2% respectively depicting 54.2% of variation in total between control and treated skin. Control skin data cells are distinctly clustered together in the region designated numeral 35 whereas treated data cells are prominently clustered together in the region designated 36.

[0100] Correspondingly, FIG. 5 illustrates PCA of the significantly transformed genes between treated and control skin on PanCancer panel. The first principal axis PC1 54 and second principal axes PC2 55 represent variation of 35.2% and 11.9% respectively totaling 47.1% variation between treated and control skin clustered distinctly in the regions designated 56 and 57 respectively.

Heatmaps

[0101] Further, significantly altered genes between control and microwave treated skin are visualised based on hierarchical clustering and are demonstrated using heatmaps.

[0102] The columns/rows of the data matrix are re-ordered according to the hierarchical clustering result, putting similar observations close to each other.

[0103] Heatmaps in this document are shown on greyscale depicting blocks of ‘high’ and ‘low’ values on a scale.

[0104] In FIG. 3, the heatmap illustrates distinct distribution of significantly altered genes between untreated and microwave treated skin datasets on Immunology panel. Clustered gene dataset values are predominantly positive whereas gene values in the regions designated 47 and 49 are largely negative on the scale. Concurrently, FIG. 6 shows heatmap clustering of significantly altered genes on the PanCancer panel between untreated and microwave treated skin. Clustered gene dataset values between 66 and 68 are mainly positive whereas gene values in the regions designated 67 and 69 are largely negative on the scale.

Gene Count Graphs

[0105] Example of gene count comparison between normal (healthy), treated (microwave treated) and control (diseased) skin tissue is depicted in FIG. 4 (Immunology panel) and FIG. 7 (PanCancer pathway).

[0106] In FIG. 4, IL6 (interleukin 6, interferon, beta 2) count in the tissue, for example a skin tissue, before 73 and after microwave treatment 72 is shown and is compared with a normal tissue 71 for example healthy skin. IL6 participates in numerous important immunomodulatory pathways such as Cytokine Signaling, Host-pathogen Interaction, Lymphocyte Activation, NLR signalling, Oxidative Stress, Th17 Differentiation, Th2 Differentiation, TNF Family Signaling and TLR Signaling. Microwave treated skin has shown to restore the abnormally upregulated expression of the IL6 gene from the diseased skin and downregulate it to be equivalent to the normal healthy skin.

[0107] Similarly, in FIG. 7, gene count of MYC (v-myc avian myelocytomatosis viral oncogene homolog) a proto-oncogene that plays a vital role in cell cycle progression, apoptosis and cellular transformation is shown. MYC also participates in key cancer pathways such as Wnt, TXmisReg, TGF-B, MAPK, JAK-STAT and PI3K. MYC read count is aberrantly upregulated in the diseased skin 83 as compared to the normal skin 81. Upon microwave treatment, the gene count is downregulated and restored 82.

Commonly Affected Genes

[0108] Furthermore, as shown in Table 9, eleven common genes in Immunology and PanCancer panels were found to be significantly altered between microwave treated and control skin.

Common Significantly Altered Genes in PanCancer and Immunology V2 Panel (See Table 9: n=11):

[0109] Significantly upregulated (n=4): TGFBR2, KIT B2M, MAPK1

[0110] Significantly downregulated (n=7): SOCS3, IL1B, IL6, IL8, LIF, CD19, TNFAIP3

[0111] The significantly transformed genes in the Human Immunology V2 Panel participate in and modulate various cellular pathways and contribute to vital aspects of the immune system such as adaptive immune system, apoptosis, autophagy, B cell receptor signalling, cell adhesion, chemokine signalling, complement system, cytokine signalling, haemostasis, host-pathogen interaction, immunometabolism, inflammasomes, innate immune system, lymphocyte activation, lymphocyte trafficking, MHC Class I Antigen Presentation, MHC Class II Antigen Presentation, NF-kB signalling, NLR signalling, oxidative stress, phagocytosis and degradation, T cell receptor signalling, TGF-b signalling, Th1 differentiation, Th17 differentiation, Th2 differentiation, TNF family signalling, TLR signalling, Transcriptional regulation, Treg differentiation, Type I Interferon signalling and Type II Interferon signalling.

[0112] Significantly dysregulated genes on the PanCancer pathway participate in several key cancer pathways such as Notch, APC (Wnt), Hedgehog, Chromatin Modification, Transcriptional Regulation, DNA Damage Control, TGF-B, MAPK, JAK-STAT, PI3K, RAS, cell cycle and apoptosis [20].

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