DETERMINING TUMOR ORIGIN
The disclosure provides methods for the use of gene expression measurements to classify or identify among 54 cancer types in samples obtained from a subject in a clinical setting, such as in cases of formalin fixed, paraffin embedded (FFPE) samples.
1 . A method of amplifying transcribed sequences for classifying a cell containing sample as containing tumor cells of a type of tissue, said method comprising
producing cDNA copies of 50 or more transcribed sequences from cells in a cell containing sample obtained from a human subject,
amplifying said cDNA copies to produce amplified molecules,
comparing said expression levels of 50 or more transcribed sequences to expression levels of the same 50 or more transcribed sequences in a plurality of known tumor types comprising one or more known tumor tissues selected from adrenal-cortical tumor, tumor of the salivary gland, squamous cell carcinoma, neuroendocrine-pancreas cancer, merkel cell carcinoma, lung carcinoid, primitive neuroectodermal tumor, sex cord stromal tumor, thymic carcinoma/thymoma, adenocarcinoma of urinary bladder, and squamous cell carcinoma of urinary bladder; and
classifying the sample as containing or not containing tumor cells of a tumor type or tissue in said plurality.
2 . The method of claim 1 wherein the plurality comprises adrenal-cortical tumor, tumor of the salivary gland, squamous cell carcinoma, neuroendocrine-pancreas cancer, merkel cell carcinoma, lung carcinoid, primitive neuroectodermal tumor, sex cord stromal tumor, thymic carcinoma/thymoma, adenocarcinoma of urinary bladder, and squamous cell carcinoma of urinary bladder, and
said classifying is of the sample as containing or not containing tumor cells of an adrenal-cortical tumor, tumor of the salivary gland, squamous cell carcinoma, neuroendocrine-pancreas cancer, merkel cell carcinoma, lung carcinoid, primitive neuroectodermal tumor, sex cord stromal tumor, thymic carcinoma/thymoma, adenocarcinoma of urinary bladder, and squamous cell carcinoma of urinary bladder.
3 . The method of claim 2 wherein the plurality comprises adrenal-cortical tumor, adrenal pheochromocytoma, tumor of the brain, adenocarcinoma of breast, cervical adenocarcinoma, cervical squamous cell carcinoma, cholangiocarcinoma, endometrial adenocarcinoma, esophageal squamous cell carcinoma, gastrointestinal stromal tumor, adenocarcinoma of gallbladder, gastro-esophageal adenocarcinoma, seminomatous germ cell tumor, nonseminomatous germ cell tumor, tumor of the salivary gland, squamous cell carcinoma, colorectal adenocarcinoma, small intestine adenocarcinoma, clear cell renal cell carcinoma, chromophobe renal cell carcinoma, papillary renal cell carcinoma, hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, lymphoma, melanoma, meningioma, mesothelioma, small/large cell neuroendocrine lung cancer, neuroendocrine-pancreas cancer, merkel cell carcinoma, gastrointestinal carcinoid, lung carcinoid, clear cell adenocarinoma, endometrioid adenocarcinoma, mucinous adenocarcinoma, serous adenocarcinoma, pancreatic adenocarcinoma, prostate adenocarcinoma, malignant fibrous histiocytoma, primitive neuroectodermal tumor, leiomyosarcoma, liposarcoma, osteosarcoma, synovial sarcoma, sex cord stromal tumor, basal cell carcinoma, skin squamous cell carcinoma, thymic carcinoma/thymoma, follicular/papillary carcinoma, medullary carcinoma, transitional cell carcinoma, adenocarcinoma of bladder, and squamous cell carcinoma of bladder, and
said classifying is of the sample as containing or not containing tumor cells of an adrenal-cortical tumor, adrenal pheochromocytoma, tumor of the brain, adenocarcinoma of breast, cervical adenocarcinoma, cervical squamous cell carcinoma, cholangiocarcinoma, endometrial adenocarcinoma, esophageal squamous cell carcinoma, gastrointestinal stromal tumor, adenocarcinoma of gallbladder, gastro-esophageal adenocarcinoma, seminomatous germ cell tumor, nonseminomatous germ cell tumor, tumor of the salivary gland, squamous cell carcinoma, colorectal adenocarcinoma, small intestine adenocarcinoma, clear cell renal cell carcinoma, chromophobe renal cell carcinoma, papillary renal cell carcinoma, hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, lymphoma, melanoma, meningioma, mesothelioma, small/large cell neuroendocrine lung cancer, neuroendocrine-pancreas cancer, merkel cell carcinoma, gastrointestinal carcinoid, lung carcinoid, clear cell adenocarinoma, endometrioid adenocarcinoma, mucinous adenocarcinoma, serous adenocarcinoma, pancreatic adenocarcinoma, prostate adenocarcinoma, malignant fibrous histiocytoma, primitive neuroectodermal tumor, leiomyosarcoma, liposarcoma, osteosarcoma, synovial sarcoma, sex cord stromal tumor, basal cell carcinoma, skin squamous cell carcinoma, thymic carcinoma/thymoma, follicular/papillary carcinoma, medullary carcinoma, transitional cell carcinoma, adenocarcinoma of bladder, and squamous cell carcinoma of bladder.
4 . The method of claim 1 wherein said 50 or more transcribed sequences comprise the disclosed 87 gene sequences.
5 . The method of claim 1 wherein all said expression levels are determined by use of a microarray.
6 . The method of claim 1 wherein said classifying is with an accuracy of 60% or higher.
7 . The method of claim 1 wherein said amplifying comprises amplification of all or part of the transcribed sequences, or reverse transcription and labeling RNA corresponding to said transcribed sequences.
8 . The method of claim 7 wherein said amplification comprises linear RNA amplification or quantitative PCR.
9 . The method of claim 8 wherein said amplification is quantitative PCR amplification of at least 50 nucleotides of the transcribed sequences.
10 . The method of claim 1 , wherein cDNA copies of 50 to 100 transcribed sequences are produced.
11 . The method of claim 1 , wherein said sample is a formalin fixed, paraffin embedded (FFPE) sample.
12 . The method of claim 1 , wherein a majority of the expression levels of 50 or more transcribed sequences overlap in said plurality.
13 . The method of claim 1 , wherein 30 or more of the expression levels of 50 or more transcribed sequences overlap in said plurality.
14 . The method of claim 1 , wherein 35 or more of the expression levels of 50 or more transcribed sequences overlap in said plurality.
15 . The method of claim 1 , wherein 40 or more of the expression levels of 50 or more transcribed sequences overlap in said plurality.
16 . The method of claim 10 , wherein 55 or more of the expression levels of 50 or more transcribed sequences overlap in said plurality.
17 . The method of claim 10 , wherein 60 or more of the expression levels of 60 or more transcribed sequences overlap in said plurality.