Methods for typing of lung cancer
Methods and compositions are provided for the molecular subtyping of lung cancer samples. Specifically, a method of assessing whether a patient's lung cancer subtype is adenocarcinoma, squamous cell carcinoma, or a neuroendocrine (encompassing both small cell carcinoma and carcinoid) is provided herein. A method for assessing whether a patient's lung cancer subtype is adenocarcinoma, squamous cell carcinoma, small cell carcinoma or carcinoid lung cancer is also provided. The methods provided herein entail probing the levels of the classifier biomarkers of Table 1-Table 6 or a subset thereof at the nucleic acid level, in a lung cancer sample obtained from the patient. Based in part on the levels of the classifier biomarkers, the lung cancer sample is classified as a particular lung cancer subtype.
1. A method of detecting biomarkers in a lung tissue sample obtained from a human patient, the method comprising measuring nucleic acid expression levels for every classifier biomarker in a group of classifier biomarkers consisting of only (a) CDH5, PECAM1, PAICS, PAK1, TFAP2A, CDKN2C, INSM1, STMN1, ACVR1, CIB1, LRP10, CHGA, MAPRE3, SNAP91, CAPG, LGALS3, SFN, PSMD14, CBX1, NFIL3, SCD5, HOXD1, ICAM5, FOXH1, CACNB1, TCF2, FEN1, TUBA1, CYB5B, PIK3C2A, ANTXR1, LIPE, MYBPH, DOK1, SIAH2, ITGA6, ICA1, TTF1, HPN, TRIM29, DSC3, MGRN1, HYAL2, MYO7A, ABCC5, GJB5, PLEKHA6, ME3, and ALDH3B1 or (b) CDH5, CLEC3B, PAICS, PAK1, TFAP2A, CDKN2C, INSM1, STMN1, ACVR1, LRP10, CHGA, MAPRE3, SNAP91, CAPG, LGALS3, SFN, PSMD14, CBX1, NFIL3, SCD5, HOXD1, ICAM5, CACNB1, TCF2, TCP1, FEN1, CYB5B, PIK3C2A, ANTXR1, MYBPH, DOK1, SIAH2, ITGA6, ICA1, TTF1, HPN, TRIM29, DSC3, MGRN1, HYAL2, MYO7A, ABCC5, GJB5, PLEKHA6, ME3 and ALDH3B1 in the lung tissue sample, wherein the measuring is performed using an amplification, hybridization and/or sequencing assay.
2. The method of claim 1 , wherein the lung tissue sample was previously diagnosed as lung cancer by histological examination.
3. The method of claim 1 , wherein the measuring is performed using an amplification assay, wherein the amplification assay is qRT-PCR.
4. The method of claim 3 , wherein the measuring of the nucleic acid expression levels comprises using at least one pair of oligonucleotide primers per each of the classifier biomarkers of only (a) or (b).
5. The method of claim 4 , further comprising comparing the nucleic acid expression levels of every classifier biomarker in a group of classifier biomarkers consisting of only (a) or (b) to the expression levels of every classifier biomarker in a group of classifier biomarkers consisting of only (a) or (b) from a reference sample.
6. The method of claim 2 , wherein the lung cancer is adenocarcinoma, squamous cell carcinoma or neuroendocrine.
7. The method of claim 2 , wherein the lung cancer is adenocarcinoma, squamous cell carcinoma, small cell carcinoma or carcinoid.
8. The method of claim 5 , wherein the reference sample is a normal sample from an individual that is healthy.
9. The method of claim 5 , wherein the reference sample is from an individual known to have a lung cancer subtype.
10. The method of claim 1 , wherein the measuring is performed using a hybridization assay, wherein the hybridization assay comprises:
(a) probing the nucleic acid expression levels of the classifier biomarkers in the group of classifier biomarkers consisting of only (a) or (b) in the lung cancer sample obtained from the patient, wherein the probing comprises:
(i) mixing the sample with oligonucleotides that are complementary to portions of nucleic acid molecules of the classifier biomarkers of only (a) or (b) under conditions suitable for hybridization of the oligonucleotides to their complements;
(ii) detecting whether hybridization occurs between the oligonucleotides to their complements;
(iii) obtaining hybridization values of the classifier biomarkers based on the detecting; and
(b) comparing the hybridization values of the classifier biomarkers to reference hybridization value(s) from at least one sample training set, wherein the at least one sample training set comprises hybridization values from a reference adenocarcinoma sample, hybridization values from a reference squamous cell carcinoma sample, hybridization values from a reference neuroendocrine sample, hybridization values from a reference small cell carcinoma sample, hybridization values from a reference carcinoid sample, or a combination thereof.
11. The method of claim 10 , wherein the comparing comprises determining a correlation between the hybridization values of the classifier biomarkers and the reference hybridization values.
12. The method of claim 10 , wherein the comparing further comprises determining an average expression ratio of the biomarkers and comparing the average expression ratio to an average expression ratio of the biomarkers obtained from the references values in the sample training set.
13. The method of claim 10 , wherein the hybridization comprises hybridization of a cDNA probe to a cDNA biomarker, thereby forming a non-natural complex.
14. The method of claim 10 , wherein the hybridization comprises hybridization of a cDNA probe to an mRNA biomarker, thereby forming a non-natural complex.
15. The method of claim 1 , wherein the lung tissue sample is selected from a formalin-fixed, paraffin-embedded (FFPE) lung tissue sample, fresh and a frozen tissue sample.