CA2328138A1 - A novel method of diagnosing, monitoring, and staging lung cancer - Google Patents

Abstract

The present invention provides a new method for detecting, diagnosing, monitoring, staging, and prognosticating lung cancer.

Description

I _ A NOVEL METHOD OF DIAGNOSING, MONITORING, AND STAGING LUNG CANCER
FIELD OF THE INVENTION
This invention relates, in part, to newly developed assays for detecting, diagnosing, monitoring, staging,. and prognosticating cance r;, particularly lung cancer.
BACKGROUND OF THE INVENTION
Primary lung cancer is divided into three main types including small cell lung cancer, non-small cell lung cancer, and mesothelioma. Smal:1 cell lung cancer is also called "Oat Cell" lung cancer because the cancer cells are a distinctive oat shape. There are three types of non-small cell lung cancer which are grouped together based upon similar behavior patterns and response to treatment which is different from small cell lung cancer. The three types of non-small cell lung cancer are squamous cell carcinoma, adenocarcinoma and large cell carcinoma. Squamous cell cancer is the most common type of lung cancer. It develops from the cells that line the airways. Adenocarcinonna also develops from the cells that line the airways, but it develops from a particular type of cell that produces mucus (phlegm). In large cell lung cancer, the cells appear large and rounded when viewed under a microscope. Mesothelioma is a rare type of cancer which affects the covering of the lung, the pleura. It is often caused by exposure to asbestos.
Secondary lung cancer is cancer that has started somewhere else in the body (for example, the breast or bowel) and spread to the lungs. The choice of treatment depends on where the cancer began. For example, cancer that has spread from the breast should respond to breast cancer treatments and cancer that has spread from the bowel should respond to bowel cancer treatments. The stage of a cancer provides information regarding how far a cancer has spread. Staging is important because treatment of the cancer is often decided based upon its stage. Staging i;s different for non-small cell versus small cell cancers of the lung.
Non-small cell cancer is divided into four stages.
Stage I is very localized cancer with no cancer in the lymph nodes. In stage II, cancer has spread to the lymph nodes at the top of the affected lung. In stage III, cancer has spread near to where the cancer started. This can be to the chest wall, the covering of the lung (pleura) , the middle of the chest (mediastinum) or other lymph nodes: Stage IV cancer has spread to another part of the body.
Small cell lung cancers are divided into two groups .
This is because small cell lung cancer often spreads quite early. Even if spreading of the cancer is not visible on scans, it is likely that some cancer cells will have broken away and traveled through the bloodstream or lymph system.
Accordingly, it is often preferred to treat small cell lung cancers as if they have spread, whether or not any secondary cancer is seen.
The two stages of small cell lung cancers are limited disease, that is canceo. that can only be seen in one lung and in nearby lymph nodes, and extensive disease, that is cancer that has spread outside the lung to the chest or to other parts of the body. Because surgery is not usually used to treat small cell cancer, except in very early cases, the staging is not as important as it is with some other types of cancer. Chemotherapy faith or without radiotherapy is usually preferred for treatment of small cell lung cancers. Initial scans and tests are used for comparison with later scans and test to see how well a patient is responding to treatment.
Procedures used for detecting, diagnosing, monitoring, staging and prognosticating lung cancer are of critical importance to the outcome of the patient. For example, patients diagnosed with early lung cancer generally have a much greater five-year survival rate as compared to the survival rate for patients diagnosed with distant metastasized lung cancer. New diagnostic methods which are more sensitive and specific for detecting early lung cancer are clearly needed.
Lung cancer patients are also closely monitored following initial therapy and during adjuvant therapy to determine response to therapy and to detect persistent or recurrent disease of metastasis. There is clearly a need for a lung cancer marker which is more sensitive and specific in detecting lung cancer recurrence.
Another important step in managing lung cancer is determination of the stage of the disease. Stage determination has potE~ntial prognostic value and provides criteria for design:ing optimal therapy. Generally, pathological staging of lung cancer is preferable over clinical staging because the former gives a more accurate prognosis. However, cl.i.nical staging would be preferred were it at least as accurate as pathological staging because it does not depend on an invasive procedure to obtain tissue for pathological evaluation. Staging of lung cancer would be improved by detecting new markers in cells, tissues or bodily fluids which could dif.Eerentiate between different stages of invasion.
In the present invention, methods are provided for detecting, diagnosing, monitoring, staging and prognosticating lung cancer via six (6') Lung Specific Genes (LSGs). The six LSGs refer, among other things, to native proteins expressed by the genes comprising the polynucleotide sequences of any of SEQ ID N0: 1, 2, 3, 4, 5 or 6. In the alternative, what is meant by the six hSGs as used herein, means the native mRNAs encoded by t:he genes comprising any of the polynucleotide sequences of SEQ ID N0: 1, 2, 3, 4, 5 or 6 or levels of the genes comprising any of the polynucleotide sequences of SEQ ID N0: 1, 2, 3, 4, 5 or 6.
Other objects, features, advantages and aspects of the present invention will. become apparent to those of skill in the art from the following description. It shou:Ld be understood, however, that the following description and the specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only. Various changes and modifications within the spirit and scope of the disclosed invention will become readily apparent to those skilled in the art from reading the following description and from reading the other parts of the present disclosure.
SUt~lARY OF THE INVENTION
Toward these ends, and others, it is an object of the present invention to provide a method for diagnosing the presence of lung cancer in a patient which comprises measuring levels of LSG in a sample of cells, tissue or bodily fluid from the patient and comparing the measured levels of LSG with levels of LSG in preferably the same cells, tissue, or bodily fluid type of a control, wherein an increase in the measured LSG levels in the patio=nt versus levels of LSG in the control is associated with lung cancer.
Another object of the present invention is to provide a method of diagnosing metastatic lung cancer in a patient which comprises measuring LSG levels in a sample of cells, tissue, or bodily fluid from the patient and comparing the measured LSG levels with levels of LSG in preferably the same cells, tissue, or bodily fluid type of a control, wherein an increase in measured LSG levels in the patient versus levels of LSG in the control is associated with a cancer which has metastasized.
Another object of the present invention is to provide a method of staging lung cancer in a patient which comprises identifying a patient :having lung cancer, measuring levels of LSG in a sample of cells, tissues, or bodily fluid obtained from the patient, and ~~omparing the measured LSG levels with levels of LSG in preferably the same cells, tissue or bodily fluid type of a control. An increase in measured LSG levels in the patient versus LSG levels in the control can be associated with a cancer which is progressing while a decrease or equivalent level of LSG measured in the patient versus the control can be associated with a cancer which is regressing or in remission.
Another object of the present invention is to provide a method of monitoring lung cancer in a patient for the onset of metastasis. The mE:thod comprises identifying a patient having lung cancer that is not known to have metastasized, periodically measuring levels of LSG in a sample of cells, tissues, or bodily fluid obtained from the patient, and comparing the measured LSG levels with levels of LSG in preferably the same cells, tissue, or bodily fluid type of a control, wherein an increase in measured LSG levels versus control LSG levels is associated with a cancer which has metastasized.
Yet another object of the present invention is to provide a method of monitoring the change in stage of lung cancer in a patient w'.hich comprises identifying a patient having lung cancer, periodically measuring levels of LSG in a sample of cells, tissue, or bodily fluid obtained from the patient, and comparing 'the measured LSG levels with levels of LSG in preferably the same cells, tissues, or bodily fluid type of a control wherein an increase in measured LSG levels versus the control LSf~ levels is associated with a cancer which is progressing and a decrease in the measured LSG levels versus the control LSG levels is associated with a cancer which is regressing or in remission.
Other objects, features, advantages and aspects of the present invention will become apparent to those of skill in the art from the following description. It should be understood, however, that the following description and the specific examples, whi=Le indicating preferred embodiments of the invention, are given by way of illustration only. Various changes and modifications within the spirit and scope of the disclosed invention will become readily apparent to those skilled in the art from reading the following description and from reading the other parts of the present disclosure.
DESCRIPTION OF THE INVENTION
The present invention relates to diagnostic assays and methods, both quantitative and qualitative for detecting, diagnosing, monitoring,. staging, and prognosticating cancers by comparing levels of :LSG with those of LSG in a normal human control. What is meant by "levels of LSG" as used herein, means levels of the native protein expressed by the gene comprising the polynucleotide sequence of any of SEQ ID NO:
1, 2, 3, 4, 5, or 6. In the alternative, what is meant by "levels of LSG" as used herein, means levels of the native mRNA encoded by the gene comprising any of the polynucleotide sequence of SEQ ID N0: 1, 2, 3, 4, 5, or 6 or levels of the gene comprising any of the polynucleotide sequence of SEQ ID
N0: 1, 2, 3, 4, 5, or 6. Such levels are preferably measured in at least one of, cells, tissues and/or bodily fluids, including determination of normal and abnormal levels. Thus, for instance, a diagnostic assay in accordance with the invention for diagno:>ing over-expression of LSG protein compared to normal control bodily fluids, cells, or tissue samples may be used i.o diagnose the presence of cancers, including lung cancer. Any of the six LSGs may be measured alone in the methods of- the invention, or all together or any combination of the six.
By "control" it :is meant a human patient without cancer and/or non cancerous samples from the patient, also referred to herein as a normal human control; in the methods for diagnosing or monitoring for metastasis, control may also include samples from a human patient that is determined by reliable methods to have lung cancer which has not metastasized.
All the methods of the present invention may optionally include measuring the .Levels of other cancer markers as well as LSG. Other cancer rnarkers, in addition to LSG, useful in the present invention will depend on the cancer being tested and are known to those of skill in the art.
Diagnostic Assays The present invention provides methods for diagnosing the presence of lung cancer by analyzing for changes in levels of LSG in cells, tissues or bodily fluids compared with levels of LSG in cells, tissues or bodily fluids of preferably the same type from a normal human control, wherein an increase in levels of LSG in the patient versus the normal human control is associated with the presence of lung cancer.
Without limiting the instant invention, typically, for a quantitative diagnostic assay a positive result indicating the patient being tested has cancer is one in which cells, tissues, or bodily fluid levels of the cancer marker, such as LSG, are at least two times higher, and most preferably are at least five times higher, than in preferably the same cells, tissues, or bodily fluid of a normal human control.
The present invention also provides a method of diagnosing metastatic, lung cancer in a patient having lung cancer which has not yet metastasized for the onset of metastasis. Ir. the mei~hod of the present invention, a human cancer patient suspected of having lung cancer which may have metastasized (but which was not previously known to have metastasized) is identified. This is accomplished by a variety of means known to those of skill in the art. For example, in the case of lung cancer, patients are typically diagnosed with lung cancer following traditional detection methods.

WO 99/60160 PCT/tJS99/10344 _ g _ In the present invention, determining the presence of LSG level in cells, tissues, or bodily fluid, is particularly useful for discriminating between lung cancer which has not metastasized and lung cancer which has metastasized. Existing techniques have difficia.lty discriminating between lung cancer which has metastasizE~d and lung cancer which has not metastasized and proper treatment selection is often dependent upon such knowledge.
In the present. invention, the cancer marker levels measured in such cells, tissues, or bodily fluid is LSG, and are compared with levels of LSG in preferably the same cells, tissue, or bodily fluid type of a normal human control. That is, if the cancer marker being observed is just LSG in serum, this level is preferably compared with the level of LSG in serum of a normal human patient. An increase in the LSG in the patient versus the normal human control is associated with lung cancer which has nnetastasized.
Without limiting t:he instant invention, typically, for a quantitative diagnostic assay a positive result indicating the cancer in the patient being tested or monitored has metastasized is one in which cells, tissues, or bodily fluid levels of the cancer marker, such as LSG, are at least two times higher, and most: preferable are at least five times higher, than in preferably the same cells, tissues, or bodily fluid of a normal patient.
Staging The invention also provides a method of staging lung cancer in a human patiE:nt.
The method comprises identifying a human patient having such cancer; analyzing a sample of cells, tissues, or bodily fluid from such patient for LSG. Then, the method compares LSG levels in such cells, tissues, or bodily fluid with levels of LSG in preferably the same cells, tissues, or bodily fluid type of a normal human control sample, wherein an increase in LSG levels in the patient versus the normal human control is _ g _ associated with a cancer which is progressing and a decrease in the levels of LSG is associated with a cancer which is regressing or in remission.
Monitoring Further provided is a method of monitoring lung cancer in a human having such cancer for the onset of metastasis.
The method comprises identifying a human patient having such cancer that is not knc>wn to have met<~stasized; periodically analyzing a sample of cells, tissues, or bodily fluid from such patient for LSG; comparing the LSG levels in such cells, tissue, or bodily fluid with levels o:E LSG in preferably the same cells, tissues, or bodily fluid type of a normal human control sample, wherein an increase in LSG levels in the patient versus the normal human control is associated with a cancer which has metastasized.
Further provided by this inventions is a method of monitoring the change in stage of lung cancer in a human having such cancer. Tree method comprises identifying a human patient having such cancer; periodically analyzing a sample of cells, tissues, or ~>odily fluid from such patient for LSG;
comparing the LSG levels in such cells, tissue, or bodily fluid with levels of LSG in preferably the same cells, tissues, or bodily fluid type of a normal human control sample, wherein an increase in LSG levels in the patient versus the normal human control is associated with a cancer which is progressing in stage and a decrease in the levels of LSG is associated with a cancer which is regressing in stage or in remission.
Monitoring such patient for onset of metastasis is periodic and preferably done on a quarterly basis. However, this may be more or less frequent depending on the cancer, the particular patient, and the stage of the cancer.
Assay Techniques Assay techniques that can be used to determine levels of gene expression, such as LSG of the present invention, in a sample derived from a host are well-known to those of skill in the art. Such assay methods include radioimmunoassays, reverse transcriptase PCR (RT-PCR) assays, immunohistochemistry assays, in situ hybridization assays, competitive-binding assays, Western Blot analyses and ELISA
assays. Among these, ELISAs are frequently preferred to diagnose a gene's expressed protein in biological fluids.
An ELISA assay initially comprises preparing an antibody, if nct readily available from a commercial source, specific to LSG, pre:Eerably a monoclonal antibody. In addition a reporter antibody generally is prepared which binds specifically to LSG. The reporter antibody is attached to a detectable reagent such as radioactive, fluorescent or enzymatic reagent, for example horseradish peroxidase enzyme or alkaline phosphatase.
To carry out the ELISA, antibody specific to LSG is incubated on a solid support, e.g., a polystyrene dish, that binds the antibody. l~.ny free protein binding sites on the dish are then covered by incubating with a non-specific protein such as bovine serum albumin. Next, the sample to be analyzed is incubated in the dish, during which time LSG binds to the specific antibody attached to the polystyrene dish.
Unbound sample is washed out with buffer. A reporter antibody specifically directed to LSG and linked. to horseradish peroxidase is placed in the dish resulting in binding of the reporter antibody to any monoclonal antibody bound to LSG.
Unattached reporter ant_Lbody is then washed out. Reagents for peroxidase activity, including a colorimetric substrate are then added to the dish. Immobilized peroxidase, linked to LSG
antibodies, produces a colored reaction product. The amount of color developed in a, given time period is proportional to the amount of LSG protein present in the sample. Quantitative results typically are obtained by reference to a standard curve.

A competition assay may be employed wherein antibodies specific to LSG attached to a solid support and labeled LSG
and a sample derived from the host are passed over the solid support and the amount of label detected attached to the solid support can be correlated to a quantity of LSG in the sample.
Nucleic acid methods may be used to detect LSG mRNA as a marker for lung cancer. Polymerase chain reaction (PCR) and other nucleic acid mei~hods, such as ligase chain reaction (LCR) and nucleic acid sequence based amplification (NASABA), can be used to detec~~t malignant cells for diagnosis and monitoring of various malignancies. For example, reverse-transcriptase PCR (RT-fCR) is a powerful technique which can be used to detect the presence of a specific mRNA population in a complex mixture of thousands of other mRNA species. In RT-PCR, an mRNA species is first reverse transcribed to complementary DNA (cDIVA) with use of the enzyme reverse transcriptase; the cDNA :is then amplified as in a standard PCR
reaction. RT-PCR can thus reveal by amplification the presence of a single species of mRNA. Accordingly, if the mRNA is highly specific for the cell that produces it, RT-PCR
can be used to identify the presence of a specific type of cell.
Hybridization to clones or oligonucleotides arrayed on a solid support (i.e., gridding) can be used to both detect the expression of and quantitate the level of expression of that gene. In this approach, a cDNA encoding the LSG gene is fixed to a substrate. The substrate may be of any suitable type including but not :Limited to glass, nitrocellulose, nylon or plastic. At least a portion of the DNA encoding the LSG
gene is attached to the substrate and then incubated with the analyte, which may be RNA or a complementary DNA (cDNA) copy of the RNA, isolated from the tissue of interest.
Hybridization between the substrate bound DNA and the analyte can be detected and quantitated by several means including but not limited to radioactive labeling or fluorescence labeling of the analyte or a secondary molecule designed to detect the hybrid. Quantitation c>f the level of gene expression can be done by comparison of the intensity of the signal from the analyte compared with that determined from known standards.
The standards can be obi~ained by in vitro transcription of the target gene, quantitat:ing the yield, and then using that material to generate a standard curve.
The above tests can be carried out on samples derived from a variety of patients' cells, bodily fluids and/or tissue extracts (homogenates ar solubilized tissue) such as from tissue biopsy and autopsy material. Bodily fluids useful in the present invention include blood, urine, saliva, or_ any other bodily secretion or derivative thereof. Blood can include whole blood, plasma, serum, or any derivative of blood.
EXAMPLES
The present invention is further described by the following examples. 'The examples are provided solely to illustrate the invention by reference to specific embodiments.
These exemplifications, while illustrating certain specific aspects of the invention, do not portray the limitations or circumscribe the scope of the disclosed invention.
Example 1: LSGs Searches were carried out and LSGs identified using the following Search Tools as part of the LIFESEQ~ database available from Incyte Pharmaceuticals, Palo Alto, CA:
1. Library Comparison (compar.es one library to one other library) allows the identification of clones expressed in tumor and absent or expressed at a lower level in normal tissue.
2. Subsetting is similar to library comparison but allows the identification of clones expressed in a pool of libraries and absent or expressed at a lower level in a second pool of libraries.
3. Transcript Imaging lists all of the clones in a single library or a pool of libraries based on abundance.
Individual clones can then be examined using Electronic Northerns to determine the tissue sources of their component ESTs.
4. Protein Function: Incyte has identified subsets of ESTs with a potential protein function based on homologies to known proteins. Some examples in this database include Transcription Factors. and Proteases. Some lead were identified by searching in this database for clones whose component EST's showed disease specificity.
Electronic subtractions, transcript imaging and protein function searches were used to identify clones, whose component EST's were exclusively or more frequently found in libraries from specific tumors. Individual candidate clones were examined in detail. by checking where each EST originated.
TABLE 1: LSGs SEQ ID NO Clone ID Gene ID
1 126758 29997 Library Comparisons 2 2798946 26723 Library Comparisons 3 3107312 242842 Transcript Imaging 4 1472038 51968 Transcript Imaging 126263 221807 Transcript Imaging 6 58271 242745 Transcript Imaging The following example was carried out using standard techniques, which are well known and routine to those of skill in the art, except where otherwise described in detail.
Routine molecular biology techniques of the following example can be carried out as described in standard laboratory manuals, such as Sanbrook et al., MOLECULAR CLONING: A

LABORATORY MANUAL, 2noL Ed.: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (7.989).
Example 2: Relative Quantitation of Gene Expression Real-Time quantitative PCR with fluorescent Taqman probes is a quantitation detection system utilizing the 5'-3' nuclease activity of Taq DNA polymerase. The method uses an internal fluorescent oligonucleotide probe (Taqman) labeled with a 5' reporter dyE: and a downstream, 3' quencher dye.
During PCR, the 5'-3' nuclease activity of Taq DNA polymerase releases the reporter, whose fluorescence can then be detected by the laser detector of the Model 7700 Sequence Detection System (PE Applied Bio~;ystems, Foster City, CA, USA).
Amplification of an endogenous control is used to standardize the amount of sample RNA added to the reaction and normalize for Reverse franscriptase (RT) efficiency. Either cyclophilin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or 18S ribosomal RNA (rRNA) is used as this endogenous control. To calculate relative quanti.tation between all the samples studied, the target RNA levels for one sample were used as the basis far comparative results (calibrator).
Quantitation relative to the "calibrator" can be obtained using the standard curve method or the comparative method (User Bulletin #2: ABI PRISM 7700 Sequence Detection System).
The tissue distribution and the level of the target gene was evaluated for every example in normal and cancer tissue.
Total RNA was extracted from normal tissues, cancer tissues, and from cancers and the corresponding matched adjacent tissues. Subsequently, first strand cDNA was prepared with reverse transcriptase .and the polymerase chain reaction was done using primers and Taqman probe specific to each target gene. The results are analyzed using the ABI PRISM 7700 Sequence Detector. The absolute numbers are relative levels of expression of the target gene in a particular tissue compared to the calibrator tissue.

Comparative Examples For comparativE: examples similar mRNA expression analysis for genes coding for the diagnostic markers PSA
(Prostate Specific Antigen) and PLA2 (Phospholipase A2) was performed. PSA is the only cancer screening marker available in clinical laboratories. When the panel of normal pooled tissues was analyzed, PSA was expressed at very high levels in prostate, with a very low expression in breast and testis.
After we analyzed more than 55 matching samples from 14 different tissues, the data corroborated the tissue specificity seen with normal tissue samples. We compared PSA
expression in cancer and normal adjacent tissue for 12 matching samples of prostate tissue. The relative levels of PSA were higher in 10 cancer samples (83~). Clinical data recently obtained support the utilization of PLA2 as a staging marker for late stages of prostate cancer. Our mRNA
expression data showed overexpression of the mRNA in 8 out of the 12 prostate matching samples analyzed (660). The tissue specificity for PLA2 was not as good as the one described for PSA. In addition to prostate, also small intestine, liver, and pancreas showed high levels of mRNA expression for PLA2.
Measurement of SEQ ID NO:1; Clone ID 126758; Gene ID 29997 (Lng101) The absolute numbers as depicted in Table 2 are relative levels of expression of: LSG Lng101 (SEQ ID NO:1) in 12 normal different tissues. A:11 the values are compared to normal testis (calibrator). These RNA samples are commercially available pools, originated by pooling samples of a particular tissue from different :individuals.

Table 2: Relative levels of Lng101 Expression in Pooled Samples Tissue NORMAL

Brain 0 Heart 1.55 Kidney 0 Liver 0 Lung 72716 Mammary Gland 2 Prostate 0 Small Intestine 0 Spleen 0 Testis 1 Thymus 0 Uterus 0 The relative levels of expression in Table 2 show that mRNA expression of the LSG Lng101 (SEQ ID NO:1) is very high (72716) in lung compared with all the other normal tissues analyzed. Testis, the calibrator, with a relative expression level of 1, heart (1.55), and mammary gland (2) are the only tissues expressing the mRNA for Lng101. These results demonstrated that Lng101 mRNA expression is highly specific for lung.
The absolute numbers in Table 2 were obtained analyzing pools of samples of a particular tissue from different individuals. They can not be compared to the absolute numbers originated from RNA obtained from tissue samples of a single individual in Table 3.
The absolute numbers depicted in Table 3 are relative levels of expression of Lng101 in 44 pairs of matching samples. All the values are compared to normal testis (calibrator) . A matching pair is formed by mRNA from the cancer sample for a particular tissue and mRNA from the normal adjacent sample for that same tissue from the same individual.

Table 3: Relative Levels of Lng101 Expression in Individual Samples Sample Cancer Type Tissue Cancer Matching ID Normal Lng AC82 Adenocarcinoma Lung 1 17199 92042 Lng 60XL Adenocarcinoma Lung 2 4603 49971 Lng AC66 Adenocarcinoma Lung 3 7358 116907 Lng AC69 Adenocarcinoma Lung 4 82953 47649 Lng AC11 Adenocarcinoma Lung 5 37771 496008 Lng AC39 Adenocarcinoma Lung 6 2487 15771 Lng AC32 Adenocarcinoma Lung 7 12634 2042_54 Lng SQ9X Squamous cell Lung 8 90774 14462 carcinoma Lng SQ32 Squamous cell Lung 9 6677 677567 carcinoma Lng SQ80 Squamous ce7.l Lung 10 50711 47151 carcinoma Lng SQ16 Squamous cell Lung 11 396 41333 carcinoma Lng SQ79 Squamous cel_1 Lung 12 10261 354395 carcinoma Lng 47XQ Squamous ce~_1 Lung 13 2513 5293 carcinoma Lng SQ44 Squamous ce7_1 Lung 14 69033 72 carcinoma Lng 90X Squamous cell Lung 15 678 14715 carcinoma Lng LC71 Large cell Lung 16 155332 44762 carcinoma Lng Large cell Lung 17 10191 322737 LC109 carcinoma Lng 75XC Metastatic Lung 18 222033 165291 from bone cancer Lng MT67 Metastatic Lung 19 189 35982 from renal cell cancer Lng MT71 Metastatic Lung 20 122 4270 from melanoma Bld 32XK Bladder 1 0 0 Bld 46XK Bladder 2 0 0 Cln AS45 Colon 1 0 0 Cln C9XR Colon 2 0 0 Cvx KS52 Cervix 1 0 0 Cvx NK23 Cervix 2 0 0 End 28XA Endometrium 0 0 End 12XA Endometrium 0 0 Kid Kidney 1 0 0 Kid Kidney 2 0 0 Liv 94XA Liver 1 0 0 Liv 15XA Liver 2 0 0 Mam 82XI Mammary 1 0 0 Mam A06X Mammary 2 0 0 Pan 71XL Pancreas 1 0 0 Pan 77X Pancreas 2 0 0 Pro 20XB Prostate 1 0 0 Pro 12B Prostate 2 0 0 SmI 21XA Sm. Int. 1 0 0 SmI H89 Sm. Int. 2 0 0 Sto AC44 Stomach 13 0 Tst 39X Testis 4315 0 Utr Uterus 1 0 0 Utr Uterus 2 0 0 0= Negative In the analysis of matching samples, the higher levels of expression were in lung, showing a high degree of tissue specificity for this tissue. These results confirmed the tissue specificity results obtained with the panel of normal pooled samples (Table 2).
Furthermore, the level of mRNA expression in cancer samples and the isogeni.c normal adjacent tissue from the same individual were compared. This comparison provides an indication of specificity for the cancer stage (e. g. higher levels of mRNA expression in the cancer sample compared to the normal adjacent). Table 3 shows overexpression of LSG Lng101 in 6 lung cancer tissues compared with their respective normal adjacent (lung samples #4, 8, 10, 14, 16, and 18). There was overexpression in the cancer tissue for 30~ of the lung matching samples tested (total of 20 lung matching samples).
Altogether, the high level of tissue specificity, plus the mRNA overexpression in 300 of the lung matching samples tested are demonstrative of LSG Lng101 (SEQ ID N0:1) being a diagnostic marker for lung cancer. The amino acid sequence encoded by Lng101 (SEQ ID NO M) is depicted in SEQ ID N0: 7.
Measurement of SEQ ID Iht0:3~ Clone ID 3107312; Gene ID 242842 (Lng105 ) The absolute numbers depicted in Table 4 are relative levels of expression of LSG Lng105 (SEQ ID N0:3) in 12 normal different tissues. All the values are compared to normal kidney (calibrator). These RNA samples are commercially available pools, originated by pooling samples of a particular tissue from different individuals.

Table 4: Relative levels of Lng105 Expression in Pooled Samples Tissue NORMAL

Brain 1 Heart 1.11 Kidney 558 Liver 0 Lung 9248 Mammary Gland 6 Muscle 0 Prostate 0 Small Intestine 87 Testis 50 Thymus 6 Uterus 23 The relative levESls of expression in Table 4 show that mRNA expression of LSG Lng105 (SEQ ID N0:3) is more than 16 fold higher in the pool of normal lung (9248) compared with the next higher expres~sor (558 for kidney) . All the other pooled tissues samples analyzed showed a very low level of expression for Lng105 (SEQ ID N0:3). These results demonstrate that mRNA E=xpression of LSG Lng105 (SEQ ID N0:3) is highly specific for lung.
The absolute numk~ers in Table 4 were obtained analyzing pools of samples of a particular tissue from different individuals. They can not be compared to the absolute numbers originated from RNA obtained from tissue samples of a single individual in Table 5.
The absolute numbers depicted in Table 5 are relative levels of expression of Lng105 (SEQ ID N0:3) in 61 pairs of matching samples. All the values are compared to normal small intestine (calibrator). A matching pair is formed by mRNA
from the cancer sample for a particular tissue and mRNA from the normal adjacent sample for that same tissue from the same individual.

Table 5: Relative Levels of Lng105 Expression in Individual Samples Sample Cancer Type Tissue Cancer Matching ID Normal Lng AC82 Adenocarcinama Lung 1 1278 792 Lng C17X Adenocarcinama Lung 2 1272 1948 Lng 60XL Adenocarcinama Lung 3 4345 2188 Lng AC66 Adenacarcinama Lung 4 1531 1558 Lng AC69 Adenocarcinoma Lung 5 7232 913 Lng AC88 Adenocarcinama Lung 6 7724 24749 Lng AC11 Adenocarcinoma Lung 7 690 21545 Lng AC39 Adenocarcinama Lung 8 16904 370 Lng AC90 Adenocarcinoma Lung 9 14614 34 Lng AC32 Adenocarcinama Lung 10 8720 5061 Lng SQ9X Squamous ce:Ll Lung 11 3603 659 carcinoma Lng SQ45 Squamous ce:l1 Lung 12 32998 1333 carcinoma Lng SQ56 Squamous ce:Ll Lung 13 829 15077 carcinoma Lng SQ14 Squamous ce:l1 Lung 14 7 6865 carcinoma Lng SQ32 Squamous cell Lung 15 976 10227 carcinoma Lng SQ80 Squamous ce:Ll Lung 16 2769 3554 carcinoma Lng SQ16 Squamous ce:Ll Lung 17 198 292 carcinoma Lng SQ79 Squamous ce:Ll Lung 18 1128 7777 carcinoma Lng C20X Squamous cell Lung 19 4 20 carcinoma Lng 47XQ Squamous ce:l1 Lung 20 276 117 carcinoma Lng SQ44 Squamous ce:l1 Lung 21 3126 1 carcinoma Lng BR94 Squamous ce:l1 Lung 22 709 6 carcinoma Lng 90X Squamous ce:Ll Lung 23 258 590 carcinoma Lng LC71 Large cell Lung 24 155332 44762 carcinoma Lng Large cell Lung 25 34280 33112 LC109 carcinoma Lng 75XC Metastatic Lung 26 749 902 from bone cancer Lng MT67 Metastatic Lung 27 70 6985 from renal cell cancer Lng MT71 Metastatic Lung 28 742 15992 from melanoma Bld 32XK Bladder 1 1 0 Bld 46XK Bladder 2 0 0 Cvx KS52 Cervix 1 4 0 Cvx NK23 Cervix 2 1 0 Cln AS45 Colon 1 0 1.

Cln C9XR Colon 2 2 1.

Cln CM67 Colon 3 0 0 End 28XA Endometrium 7 4 End 12XA Endometrium 0 0 Kid Kidney 1 0 186 Kid Kidney 2 82 458 Kid Kidney 3 169 438 Kid lOXD Kidney 4 21 186 Kid 11XD Kidney 5 586 110 Liv 94XA Liver 1 1 0 Liv 15XA Liver 2 1 0 Mam A06X Mammary 1 1 0 Mam Mammary 2 13 0 BOllX

Mam 12X Mammary 3 0 0 Mam 59X Mammary 4 0 C

Ovr 103X Ovary 1 15 2 Pan 71XL Pancreas 1 1 0 Pan 77X Pancreas 2 4 0 Pro 20XB Prostate 1 1 1 Pro 12B Prostate 2 8 0 SmI 21XA Sm. Int. 1 4 0 SmI H89 Sm. Int. 2 1 0 Sto AC49 Stomach 1 0 2 Sto AC99 Stomach 2 6 2 Tst 39X Testis 28 2 Utr 85XU Uterus 1 3 2 Utr Uterus 2 2 0 Utr Uterus 3 2 6 0= Negative In the analysis of matching samples, the higher levels of expression were in lung showing a high degree of tissue specificity for lung tissue. These results confirm the tissue specificity results obtained with normal pooled samples (Table 4) .
Furthermore, the level of mRNA expression in cancer samples and the isogenic normal adjacent tissue from the same individual were compared. This comparison provides an indication of specificity for the cancer stage (e. g. higher levels of mRNA expression in the cancer sample compared to the normal adjacent). Table 5 shows overexpression of LSG Lng105 (SEQ ID N0:3) in 13 lung cancer tissues compared with their respective normal adjacent (lung samples #1, 3, 5, 8, 9, 10, 11, 12, 20, 21, 22, 24, and 25). There is overexpression in the cancer tissue for ~16~ of the colon matching samples tested (total of 28 lung matching samples).
Altogether, the high level of tissue specificity, plus the mRNA overexpression in almost half of the lung matching samples tested are demonstrative of Lng105 (SEQ ID N0:3) being a diagnostic marker fo:r lung cancer. The amino acid sequence encoded by Lng105 (SEQ ID N0:3) is depicted as SEQ ID N0:8.
Measurement of SEQ ID N0:6; Clone ID 586271; Gene ID 242745 (Lng107) The absolute numbers depicted in Table 6 are relative levels of expression o:E LSG Lng107 (SEQ ID N0:6) in 12 normal different tissues. All the values are compared to normal mammary gland (calibrator). These RNA samples are commercially available pools, originated by pooling samples of a particular tissue from different individuals.
Table 6: Relative levels of Lng107 Expression in Pooled Samples Ti ue NO

Bladder 0 Heart 0 Kidney 0 Liver 0 Lung 23 Mammary Gland 1 Muscle 0 Prostate 0 Small Inte;>tine 0 Testis 0 Thymus 0 Uterus 0 The relative levels of expression in Table 6 show that mRNA expression oi= LSG Lng107 (SEQ ID N0:6) is 23 fold higher in the pool of normal lung (23) compared to the expression level in thE: calibrator mammary gland (1). All the other tissues analyzed were negative for Lng107 (SEQ ID
N0:6). These results demonstrate that Lng107 mRNA
expression is highly specific for lung.
The absolute numbers in Table 6 were obtained analyzing pools of samples of a particular tissue from different individuals. They can not be compared to the absolute numbers originated from RNA obtained from tissue samples of a single individual in Table 7.
The absolute numbers depicted in Table 7 are relative levels of expression of LSG Lng107 (SEQ ID N0:6) in 57.
pairs of matching samples. All the values are compared to normal prostate (calibrator). A matching pair is formed by mRNA from the cancer sample for a particular tissue and mRNA from the normal adjacent sample for that same tissue from the same individual.
Table 7: Relative Levels of Lng107 Expression in Individual Samples Sample Cancer Type Tissue Cancer Matching ID Normal Lng AC82 Adenocarcinoma Lung 1 6 2 Lng 60XL Adenocarcinoma Lung 2 1 4 Lng AC66 Adenocarcinoma Lung 3 1 0 Lng AC69 Adenocarcinoma Lung 4 117 6 Lng AC88 Adenocarcinoma Lung 5 12 6 Lng AC11 Adenocarcinoma Lung 6 1 18 Lng AC32 Adenocarcinoma Lung 7 9 2 Lng AC39 Adenocarcinoma Lung 8 2 1 Lng AC90 Adenocarcinoma Lung 9 1 0 Lng SQ9X Squamous cell Lung 10 7 0 Lng SQ45 Squamous cell Lung 1.1 45 7.

carcinoma Lng SQ56 Squamous cell Lung 12 1 23 carcinoma .

Lng SQ16 Squamous cell Lung 13 0 0 carcinoma Lng SQ32 Squamous cell Lung 14 9 5 carcinoma Lng SQ80 Squamous ce:Ll Lung 15 2 0 carcinoma Lng SQ79 Squamous ce:Ll Lung 16 5 11 carcinoma Lng C20X Squamous ce:Ll Lung 17 0 0 carcinoma Lng 47XQ Squamous ce:L:1 Lung 18 1 0 carcinoma Lng SQ44 Squamous cell Lung 19 1 0 carcinoma Lng BR94 Squamous ce:Ll Lung 20 1 0 carcinoma Lng 90X Squamous cell Lung 21 0 13 carcinoma Lng LC71 Large cell Lung 22 31 12 carcinoma Lng Large cell Lung 23 1 83 LC109 carcinoma Lng 75XC Metastatic Lung 24 2 9 from bone cancer Lng MT67 Metastatic Lung 25 0 1.

from renal cell cancer Lng MT71 Metastatic Lung 26 0 24 from melanoma Bld 32XK Bladder 1 0 0 Bld 46XK Bladder 2 0 0 Cln AS45 Colon 1 0 0 Cln C9XR Colon 2 0 0 Cvx KS52 Cervix 1 0 0 Cvx NK23 Cervix 2 0 0 End 28XA Endometrium 7 0 End 12XA Endometrium 0 0 End 68X Endometrium 3 2 End 8XA Endometrium 0 0 Kid Kidney 1 0 0 Kid Kidney 2 0 0 Liv 94XA Liver 1 0 0 Liv 15XA Liver 2 0 0 Mam A06X Mammary 1 0 0 Mam Mammary 2 116 C

Mam 47XP Mammary 3 0 0 Mam 59X Mammary 4 1 0 Ovr 103X Ovary 1 0 0 Pan 71XL Pancreas 1 0 0 Pan 77X Pancreas 2 0 0 Pro 20XB Prostate 1 0 0 Pro 12B Prostate 2 0 0 SmI 21XA Sm. Int. 1 0 0 SmI H89 Sm. Int. 2 0 0 Sto AC44 Stomach 1 0 0 Sto MT59 Stomach 2 0 0 Sto TA73 Stomach 3 1 ?

Tst 39X Testis 0 0 Utr Uterus 1 0 0 Utr Uterus 2 0 0 0= Negative In the analysis of matching samples, the higher level of expression was in lung, showing a high degree of tissue specificity for this tissue. These results confirm the tissue specificity results obtained with normal pooled samples (Table 6).
Furthermore, the level of mRNA expression in cancer samples and the isogenic normal adjacent tissue from the same individual were compared. This comparison provides an indication of specific_~ty for the cancer stage (e. g. higher levels of mRNA expression in the cancer sample compared to the normal adjacent). Table 7 shows overexpression of LSG
Lng107 (SEQ ID N0:6) in 15 lung cancer tissues compared with their respective normal adjacent (lung samples #1, 3, 4, 5, 7, 8, 9, 10, 11, 14, 15, 18, 19, 20, and 22) . There is overexpression in the cancer tissue for 570 of the lung matching samples tested (total of 26 :lung matching samples).
Altogether, the high level of tissue specificity, plus the mRNA overexprE:ssion in more than half of the lung matching samples tested are demonstrative of Lng107 being a diagnostic marker for .Lung cancer. The amino acid sequence encoded by Lng107 is depicted in SEQ ID N0:9.

SEQUENCE LISTING
<110> Yang, Fei Macina, Roberto A.
Sun, Yongming <120> A Novel Method of Diagnosing, Monitoring and Staging Lung Cancer <130> DEX-0036 <140>
<141>
<150> 60/086,212 <151> 1998-05-21 <160> 9 <170> PatentIn Ver. 2.0 <210> 1 <211> 507 <212> DNA
<213> Homo Sapiens <900> 1 ggcaagtgga accactggct tggtggattt tgctagattt ttctgatttt taaactcctg 60 aaaaatatcc cagataactg tcatgaa~gct ggtaactatc ttcctgctgg tgaccatcag 120 cctttgtagt tactctgcta ctgcctt.cct catcaacaaa gtgccccttc ctgttgacaa 180 gttggcacct ttacctctgg acaacat.tct tccctttatg gatccattaa agcttcttct 240 gaaaactctg ggcatttctg ttgagca~cct tgtggagggg ctaaggaagt gtgtaaatga 300 gctgggacca gaggcttctg aagctgt.gaa gaaactgctg gaggcgctat cacacttggt 360 gtgacatcaa gataaagagc ggaggtggat ggggatggaa gatgatgctc ctatcctccc 420 tgcctgaaac ctgttctacc aattata.gat caaatgccct aaaatgtagt gacccgtgaa 980 aaggacaaat aaagcaatga atacatt 507 <210>2 <211>1680 <212>DNA

<213>Homo Sapiens <900> 2 ggtgtgcagg atataaggtt ggacttc:cag acccactgcc cgggagagga grggagcggg 60 ccgaggactc cagcgtgccc aggtctg~gca tcctgcactt gctgccctct gacacctggg 120 aagatggccg gcccgtggac cttcacc:ctt ctctgtggtt tgctggcagc caccttgatc 180 caagccaccc tcagtcccac tgcagttctc atcctcggcc caaaagtcat caaagaaaag 240 ctgacacagg agctgaagga ccacaac:gcc accagcatcc tgcagcagct gccgctgctc 300 WO 99/601b0 PCT/US99/10344 agtgccatgc gggaaaagcc agccggagga tccctgtgct gggcagcctg gtgaacaccg 360 tcctgaagca catcatctgg ctgaaggtca tcacagctaa catcctccag ctgcaggtga 920 agccctcggc caatgaccag gagctgctag tcaagatccc cctggacatg gtggctggat 480 tcaacacgcc cctggtcaag accatcgtgg agttccacat gacgactgag gcccaagcca 540 ccatccgcat ggacaccagt gcaagtggcc ccacccgcct ggtcctcagt gactgtgcca 600 ccagccatgg gagcctgcgc atccaactgc tgcataagct ctccttcctg gtgaacgcct 660 tagctaagca ggtcatgaac ctcctagtgc catccctgcc caatctagtg aaaaaccagc 720 tgtgtcccgt gatcgaggct tccttcaatg gcatgtatgc agacctcctg cagctggtga 780 aggtgcccat ttccctcagc attgaccgtc tggagtttga ccttctgtat cctgccatca 840 agggtgacac cattcagctc tacctggggg ccaagttgtt ggactcacag ggaaaggtga 900 ccaagtggtt caataactct gcagcttccc tgacaatgcc caccctggac aacatcccgt 960 tcagcctcat cgtgagtcag gacgtggtga aagctgcagt ggctgctgtg ctctctccag 1020 aagaattcat ggtcctgttg gactctgtgc ttcctgagag tgcccatcgg ctgaagtcaa 1080 gcatcgggct gatcaatgaa aaggctgcag ataagctggg atctacccag atcgtgaaga 1140 tcctaactca ggacactccc gagtttttta tagaccaagg ccatgccaag gtggcccaac 1200 tgatcgtgct ggaagtgttt ccctccagtg aagccctccg ccctttgttc accctgggca 1260 tcgaagccag ctcggaagct cagttttaca ccaaaggtga ccaacttata ctcaacttga 1320 ataacatcag ctctgatcgg atccagctga tgaactctgg gattggctgg ttccaacctg 1380 atgttctgaa aaacatcatc actgagatca tccactccat cctgctgccg aaccagaatg 1440 gcaaattaag atctggggtc ccagtgtcat tggtgaaggc cttgggattc gaggcagctg 1500 agtcctcact gaccaaggat gcccttgtgc ttactccagc ctccttgtgg aaacccagct 1560 ctcctgtctc ccagtgaaga cttggatggc agccatcagg gaaggctggg tcccagctgg 1620 gagtatgggt gtgagctcta tagaccatcc ctctctgcaa tcaataaaca cttgcctgtg 1680 <210> 3 <211> 2060 <212> DNA
<213> Homo Sapiens <400> 3 cttgagagct ctcaaatact tggtcatgga tgaagccgac cgaatactga atatggattt 60 tgagacagag gttgacaagc ctcgagatcg gaaaacattc ctcttctctg ccaccatgac 120 caagaaggtt caaaaacttc agcgagcagc tctgaagaat cctgtgaaat gtgccgtttc 180 ctctaaatac cagacagttg aaaaattaca gcaatattat atttttattc cctctaaatt 290 caaggatacc tacctggttt atattctaaa tgaattggct ggaaactcct ttatgatatt 300 ctgcagcacc tgtaataata cccagagaac agctttgcta ctgcgaaatc ttggcttcac 360 tgccatcccc ctccatggac aaatgagtca gagtaagcgc ctaggatccc ttaataagtt 420 taaggccaag gcccgttcca ttcttctagc aactgacgtt gccagccgag gtttggacat 480 acctcatgta gatgtggttg tcaactttga cattcctacc cattccaagg attacatcca 540 tcgagtaggt cgaacagcta gagctgggcg ctccggaaag gctattactt ttgtcacaca 600 gtatgatgtg gaactcttcc agcgcataga acacttaatt gggaagaaac taccaggttt 660 tccaacacag gatgatgagg ttatgatgct gacagaacgc gtccccagcg atgtctccac 720 caccgctgct gcaacccctg ctgctgctgc tgcctctgct gaatgtggag ccttccgggg 780 ccacactgat ccgcatccct cttcatcgag tccaacctgg acgcaggacc ctgaacctac 840 tgaggggatg gagagaacca gcagagctcc ccaagttggg ggccccatcc cctggggaca 900 agcccatctt cgtacctctc tcgaactaca gggatgtgca gt:attttggg gaaattgggc 960 tgggaacgcc tccacaaaac ttcar_tgttg cctttgacac tggctcctcc aatctctggg 1020 tcccgtccag gagatgccac ttcttcagtg tgccctgctg gttacaccac cgatttgatc 1080 ccaaagcctc tagctccttc caggcc;aatg ggaccaagtt tgccattcaa tatggaactg 1190 ggcgggtaga tggaatcctg agcgaggaca agctgactat tggtggaatc aagggtgcat 1200 cagtgatttt cggggaggct ctctgc~gagc ccagcctggt cttcgctttt gcccattttg 1260 atgggatatt gggcctcggt tttccc:attc tgtctgtgga aggagttcgg cccccgatgg 1320 atgtactggt ggagcagggg ctattc~gata agcctgtctt ctccttttac ctcaacaggg 1380 accctgaaga gcctgatgga ggagac~ctgg tcctgggggg ct:cggacccg gcacactaca 1940 tcccacccct caccttcgtg ccagtc:acgg tccctgccta ctggcagatc cacatggagc 1500 gtgtgaaggt gggcccaggg ctgactctct gtgccaaggg ctgtgctgcc atcctggata 1560 cgggcacgtc cctcatcaca ggaccc:actg aggagatccg ggccctgcat gcagccattg 1620 ggggaatccc cttgctggct ggggacrtaca tcatcctgtg ctcggaaatc ccaaagctcc 1680 ccgcagtctc cttccttctt gggggggtct ggtttaacct cacggcccat gattacgtca 1790 tccagactac tcgaaatggc gtccgc:ctct gcttgtccgg tttccaggcc ctggatgtcc 1800 ctccgcctgc agggcccttc tggatc:ctcg gtgacgtctt cttggggacg tatgtggccg 1860 tcttcgaccg cggggacatg aagagc:agcg cccgggtggg cctggcgcgc gctcgcactc 1920 gcggagcgga cctcggatgg ggagagactg cgcaggcgca gttccccggg tgacgcccaa 1980 gtgaagcgca tgcgcagcgg gtggtcgcgg aggtcctgct acccagtaaa aatccactat 2040 ttccattgaa aaaaaaaaaa 2060 <210> 9 <211> 315 <212> DNA
<213> Homo sapiens <400> 9 taaacactga ctcagatttt aagaaataac ttttgagaaa tagaacaaat gaaatcagtt 60 tctccaccac ttaagtatat ctcttagaga tctacagcct ccctttaggg gacatacaaa 1.20 gtcagttgtg ttgcctttgt tgagtcccac cttatattca agtaggtatg actacaaatt 1.80 ttgaaaatag attgtcacac aataaactgg agtttatgga aacatcagta gaaggaaata 240 caacattcca tccctttaca gagatcattt acttgcaact caggataatt tgtcatgtgt 300 attatctact tatgc 315 <210> 5 <211> 895 <212> DNA
<213> Homo sapiens <900> 5 ctaatctgtt acgtaacagc aagacagcgt cacctcacct gttctcgccc tcaaatggga 60 acgctggcct gggactaaag catagaccac caggctgagt atcctgacct gagtcatccc 120 cagggatcag gagcctccag cagggaacct tccattatat tcttcaagca acttacagct 180 gcaccgacag ttgcgatgaa agttctaatc tcttccctcc tcctgttgct gccactaatg 240 ctgatgtcca tggtctctag cagcctgaat ccaggggtcg ccagaggcca cagggaccga 300 ggccaggctt ctaggagatg gctccaggaa ggcggccaag aatgtgagtg caaagattgg 360 ttcctgagag ccccgagaag aaaattcatg acagtgtctg ggctgccaaa gaagcagtgc 420 ccctgtgatc atttcaaggg caatgtgaag aaaacaagac accaaaggca ccacagaaag 480 ccaaacaagc attccagagc ctgccagcaa tttctcaaac aatgtcagct aagaagcttt 540 gctctgcctt tgtaggagct ctgagcgccc actcttccaa ttaaacattc tcagccaaga 600 agacagtgag cacacctacc agacactctt cttctcccac ct:cactctcc cactgtaccc 660 WO 99/60160 PC1'/US99/10344 acccctaaat cattccagtg ctctcaaaaa gcatgttttt caagatcatt ttgtttgttg 720 ctctctctag tgtcttcttc tctcgtcagt cttagcctgt gccctcccct tacccaggct 780 taggcttaat tacctgaaag attccaggaa actgtagctt cctagctagt gtcatttaac 840 cttaaatgca atcaggaaag tagcaaacag aagtcaataa atatttttaa atgtc 895 <210> 6 <211> 543 <212> DNA
<213> Homo Sapiens <400> 6 ccggcgctgg aggggcgagg accgggt ata agaagcctcg tggccttgcc cgggcagccg 60 caggttcccc gcgcgccccg agcccccgcg ccatgaagct, cgccgccctc ctggggctct 120 gcgtggccct gtcctgcagc tccgc;tgctg ctttcttagt gggctcggcc aagcctgtgg 180 cccagcctgt cgctgcgctg gagtcggcgg cggaggccgg ggccgggacc ctggccaacc 240 ccctcggcac cctcaacccg ctgaag~~tcc tgctgagcag cctgggcatc cccgtgaacc 300 acctcataga gggctcccag aagtgtgtgg ctgagctggg tccccaggcc gtgggggccg 360 tgaaggccct gaaggccctg ctgggggccc tgacagtgtt tggctgagcc gagactggag 920 catctacacc tgaggacaag acgctgccca cccgcgaggg ctgaaaaccc cgccgcgggg 480 aggaccgtcc atccccttcc cccggcccct ctcaataaac gtggttaaga gcaaaaaaaa 540 aaa 543 <210> 7 <211> 93 <212> PRT
<213> Homo Sapiens <400> 7 Met Lys Leu Val Thr Ile Phe :Geu Leu Val Thr Ile Ser Leu Cys Ser Tyr Ser Ala Thr Ala Phe Leu :Ile Asn Lys Val Pro Leu Pro Val Asp Lys Leu Ala Pro Leu Pro Leu i~sp Asn Ile Leu Pro Phe Met Asp Pro Leu Lys Leu Leu Leu Lys Thr '.Leu Gly Ile Ser Val Glu His Leu Val Glu Gly Leu Arg Lys Cys Val i9sn Glu Leu Gly Pro Glu Ala Ser Glu Ala Val Lys Lys Leu Leu Glu A:La Leu Ser His Leu Val <210> B

<211> 920 <212> PRT
<213> Homo Sapiens <400> 8 Met Ser Pro Pro Pro Leu Leu Gln Pro Leu Leu Leu Leu Leu Pro Leu Leu Asn Val Glu Pro Ser Gly Ala Thr Leu Ile Arg Ile Pro Leu His Arg Val Gln Pro Gly Arg Arg Thr Leu Asn Leu Leu Arg Gly Trp Arg Glu Pro Ala Glu Leu Pro Lys heu Gly Ala Pro Ser Pro Gly Asp Lys Pro Ile Phe Val Pro Leu Ser P,sn Tyr Arg Asp Val Gln Tyr Phe Gly Glu Ile Gly Leu Gly Thr Pro Pro Gln Asn Phe Thr Val Ala Phe Asp Thr Gly Ser Ser Asn Leu Trp Val Pro Ser Arg Arg Cys His Phe Phe Ser Val Pro Cys Trp Leu His His Arg Phe Asp Pro Lys Ala Ser Ser Ser Phe Gln Ala Asn Gly Thr Lys Phe Ala Ile Gln Tyr Gly Thr Gly Arg Val Asp Gly Ile Leu Ser Glu Asp Lys Leu Thr Ile Gly Gly Ile Lys Gly Ala Ser Val Ile Phe Gly Glu Ala Leu Trp Glu Pro Ser Leu Val Phe Ala Phe Ala His Phe Asp Gly Ile Leu Gly Leu Gly Phe Pro Ile Leu Ser Val Glu Gly Val Arg Pro Pro Met Asp Val Leu Val Glu Gln Gly Leu Leu Asp Lys Pro Val Phe Ser Phe Tyr Leu Asn Arg Asp Pro Glu Glu Pro Asp Gly Gly Glu Leu Val Leu Gly Gly Ser Asp Pro Ala His Tyr Ile Pro Pro Leu Thr Phe Val Pro Val Thr Val Pro Ala Tyr Trp Gln Ile His Met Glu Arg Val Lys Val Gly Pro Gly Leu Thr Leu Cys Ala Lys Gly Cys Ala Ala Ile Leu Asp Thr Gly Thr Ser Leu Ile Thr Gly Pro Thr Glu Glu Ile Arg Ala Leu His Ala Ala Ile Gly Gly Ile Pro Leu Leu Ala Gly Glu Tyr Ile Ile Leu Cys Ser Glu Ile Pro Lys Leu Pro Ala Val Ser Phe Leu Leu Gly Gly Val Trp Phe Asn Leu Thr Ala His Asp Tyr Val. Ile Gln Thr Thr Arg Asn Gly Val Arg Leu Cys Leu Ser Gly Phe Gln ;?~:la Leu Asp Val Pro Pro Pro Ala Gly Pro Phe Trp Ile Leu Gly Asp 'Val Phe Leu Gly Thr Tyr Val Ala Val Phe Asp Arg Gly Asp Met Lys Ser Ser Ala Arg Val Gly Leu Ala Arg Ala Arg Thr Arg Gly Ala Asp lGeu Gly Trp Gly Glu Thr Ala Gln Ala Gln Phe Pro Gly <210> 9 <211> 109 <212> PRT
<213> Homo sapiens <400> 9 Met Lys Leu Ala Ala Leu Leu Gly Leu Cys Val Ala Leu Ser Cys Ser Ser Ala Ala Ala Phe Leu Val Gly Ser Ala Lys Pro Val Ala Gln Pro Val Ala Ala Leu Glu Ser Ala .Ala Glu Ala Gly Ala Gly Thr Leu Ala Asn Pro Leu Gly Thr Leu Asn Pro Leu Lys Leu Leu Leu Ser Ser Leu 50 5r> 60 Gly Ile Pro Val Asn His Leu Ile Glu Gly Ser Gln Lys Cys Val Ala Glu Leu Gly Pro Gln Ala Val Gly Ala Val Lys Ala Leu Lys Ala Leu Leu Gly Ala Leu Thr Val Phe G1y

Claims (6)

What is Claimed is:

1. A method for diagnosing the presence of lung cancer in a patient comprising:
(a) measuring levels of LSG in a sample of cells, tissue or bodily fluid. obtained from the patient; and (b) comparing the measured levels of LSG with levels of LSG in a sample of cells, tissue or bodily fluid obtained from a control, wherein an increase in measured levels of LSG in the patient versus the LSG levels in the control is associated with the presence of lung cancer.

2. A method of diagnosing metastatic lung cancer in a patient comprising:
(a) measuring levels of LSG in a sample of cells, tissue, or bodily fluid obtained from the patient; and (b) comparing the measured levels of LSG with levels of LSG in a sample of cells, tissue, or bodily fluid obtained from a control, wherein an increase in measured LSG levels in the patient versus the LSG levels in the control is associated with a cancer which has metastasized.

3. A method of staging lung cancer in a patient comprising:
(a) identifying a patient suffering from lung cancer;
(b) measuring levels of LSG in a sample of cells, tissue, or bodily fluid obtained from the patient; and (c) comparing the measured levels of LSG with levels of LSG in a sample of cells, tissue, or bodily fluid obtained from a control, wherein an increase in the measured levels of LSG versus the levels of LSG in the control is associated with a cancer which is progressing and a decrease in the :measured levels of LSG versus the levels of LSG in the control is associated with a cancer which is regressing or in remission.

4. A method of monitoring lung cancer in a patient for the onset of metastasis comprising:
(a) identifying a patient having lung cancer that is not known to have metastasized;
(b) periodically measuring LSG levels in samples of cells, tissue, or bodily fluid obtained from the patient;
and (c) comparing the periodically measured levels of LSG
with levels of LSG in cells, tissue, or bodily fluid obtained from a control, wherein an increase in any one of the periodically measured levels of LSG in the patient versus the levels of LSG in the control is associated with a cancer which has metastasized.

5. A method of monitoring changes in a stage of lung cancer in a patient comprising:
(a) identifying a patient having lung cancer;
(b) periodically measuring levels of LSG in samples of cells, tissue, or bodily fluid obtained from the patient; and (c) comparing the measured levels of LSG with levels of LSG in a sample of the same cells, tissue, or bodily fluid of a control, wherein an increase in any one of the periodically measured levels of LSG versus levels of LSG in the control is associated with a cancer which is progressing in stage and a decrease in any one of the periodically measured levels of LSG versus the levels of LSG in the control is associated with a cancer which is regressing in stage or in remission.

6. The method of claim 1, 2, 3, 4, or 5 wherein the LSG comprises SEQ ID NO: 1, 3 or 6.