NOVEL GENE CLASSIFIERS AND USES THEREOF IN NON-MELANOMA SKIN CANCERS
Described herein are methods, systems, and compositions for non-invasively diagnosing or detecting a skin disease or disorder. Diagnosing or detecting a non-melanoma skin cancer as provided herein comprises detecting gene expression levels of a set of identified genes and in some instances further detecting mutations in a gene of interest.
1 . A method of detecting gene expression levels of at least two of IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1 in a subject in need thereof, comprising:
(a) isolating nucleic acids from a biological sample obtained from the subject; and
(b) detecting the expression levels of the at least two of IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1, by contacting the isolated nucleic acids with a set of probes that recognizes the at least two of IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1, and detects binding between the at least two of IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1 and the set of probes.
2 . The method of claim 1 , wherein the set of probes recognizes:
either IGFL1 or MMP1 in combination with COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, or SPP1, or a combination thereof;
IGFL1 and MMP1 in combination with COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, or SPP1, or a combination thereof; or
IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1.
3 . The method of claim 1 , further comprising detecting a mutational change of TERT, CDKN2A, TP53, or PTCH1, or a combination thereof.
4 . The method of claim 3 , wherein a mutation in TP53 translates to amino acid positions in TP53 selected from: R175, S240, G245, R248, R249, R273, R282, or T284, wherein the numberings of the amino acid residues correspond to SEQ ID NO: 1.
5 . The method of claim 3 , wherein a mutation in TP53 is in exon 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or a combination thereof.
6 . The method of claim 3 , wherein a mutation in TP53 is in exon 5, 7, 8, or a combination thereof.
7 . The method of claim 3 , wherein a mutation in PTCH1 is in exon 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or a combination thereof of PTCH1.
8 . The method of claim 3 , wherein a mutation in PTCH1 is in exon 14, 15, 17, 23, or a combination thereof.
9 . The method of claim 3 , wherein a mutation in CDKN2A is in exon 1, 2, 3, 4, 5, 6, 7, 8, or a combination thereof.
10 . The method of claim 3 , wherein a mutation in CDKN2A translates to amino acid positions in CDKN2A selected from V51, M53, R58, E61, G67, E69, or R80, wherein the numbering of the amino acid residues correspond to SEQ ID NO: 5.
11 . The method of claim 3 , wherein a mutation in TERT is in exon 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or a combination thereof.
12 . The method of claim 3 , wherein a mutation in TERT is in a promoter region of TERT.
13 . The method of claim 3 , wherein the mutational change comprises at least 1.5×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, 10×, 11×, or 12× more mutations in TERT CDKN2A, TP53, PTCH1, or a combination thereof, compared to a normal biological sample.
14 . The method of claim 3 , wherein the mutational change comprises at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or 80% more mutations in TERT, CDKN2A, TP53, PTCH1, or a combination thereof, compared to a normal biological sample.
15 . The method of claim 1 , wherein the subject is suspected of having a non-melanoma skin cancer, optionally BCC or SCC.
16 . The method of claim 1 , wherein the biological sample comprises a skin sample, optionally comprising keratinocytes, melanocytes, basal cells, T-cells, or dendritic cells.
17 . The method of claim 16 , wherein the skin sample is obtained by applying a plurality of adhesive patches to the skin sample in a manner sufficient to adhere the skin sample to the adhesive patch, and removing the adhesive patch from the skin in a manner sufficient to retain the adhered skin sample to the adhesive patch.
18 . The method of claim 1 , wherein isolating the nucleic acids comprises using a plurality of silica-coated beads, optionally a plurality of silica-coated magnetic beads.
19 . The method of claim 1 , wherein the nucleic acids comprise RNA, DNA, or a combination thereof.
20 . A method of detecting gene expression levels and mutational changes from a skin sample, comprising:
a) isolating nucleic acids from the skin sample; and
b) detecting the expression levels of one or more genes selected from: IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1; and a mutational change of TERT, CDKN2A, TP53, or PTCH1, or a combination thereof;
wherein the gene expression levels are detected by contacting the isolated nucleic acids with a set of probes that recognizes at least one but no more than nine genes selected from IGFL1, MMP1, COL5A2, IL24, AADACL2, PTCH1, CD68, PRKACA, and SPP1, and detect binding between the genes and the set of probes.