Compositions, methods and kits for diagnosis of lung cancer
Methods are provided for identifying biomarker proteins that exhibit differential expression in subjects with a first lung condition versus healthy subjects or subjects with a second lung condition. Also provided are compositions comprising these biomarker proteins and methods of using these biomarker proteins or panels thereof to diagnose, classify, and monitor various lung conditions. The methods and compositions provided herein may be used to diagnose or classify a subject as having lung cancer or a non-cancerous condition, and to distinguish between different types of cancer (e.g., malignant versus benign, SCLC versus NSCLC).
1. A method of determining the likelihood that a pulmonary nodule in a subject is not lung cancer, comprising:
(a) measuring an abundance of a plurality of transitions in a blood sample obtained from the subject, wherein each transition consists of a precursor ion m/z and a fragment ion m/z thereof, wherein said plurality of transitions comprise at least 3 transitions comprising a transition of ALQASALK (SEQ ID NO: 7) comprising m/z 401.25 and m/z 617.40, a transition of LGGPEAGLGEYLFER (SEQ ID NO: 24) comprising m/z 804.40 and m/z 913.40, and a transition of VEIFYR (SEQ ID NO: 25) comprising m/z 413.73 and m/z 598.30, and wherein said measuring is performed by selected reaction monitoring mass spectrometry;
(b) calculating a probability of lung cancer score based on the abundance of a plurality transitions of step (a); and
(c) ruling out lung cancer for the subject if the score in step (b) is lower than a pre-determined score.
2. The method of claim 1 , wherein said plurality of transitions further comprises at least one transition selected from the group consisting of a transition of AVGLAGTFR (SEQ ID NO: 11) comprising m/z 446.26 and m/z 721.40, a transition of GFLLLASLR (SEQ ID NO: 22) comprising m/z 495.31 and m/z 559.40, a transition of LTLLAPLNSVFK (SEQ ID NO: 8) comprising m/z 658.40 and m/z 804.50, a transition of NSLFEYQK (SEQ ID NO: 26) comprising m/z 514.76 and m/z 714.30, a transition of IIIQESALDYR (SEQ ID NO: 27) comprising m/z 660.86 and m/z 338.20, a transition of TWNDPSVQQDIK (SEQ ID NO: 23) comprising m/z 715.85 and m/z 260.20, a transition of LDTLAQEVALLK (SEQ ID NO: 20) comprising m/z 657.39 and m/z 330.20, a transition of QITVNDLPVGR (SEQ ID NO: 16) comprising m/z 606.30 and m/z 428.30, and a transition of ATVNPSAPR (SEQ ID NO: 33) comprising m/z 456.80 and m/z 527.30.
3. The method of claim 1 , wherein when lung cancer is ruled out the subject does not receive a treatment protocol.
4. The method of claim 3 , wherein said treatment protocol is a pulmonary function test (PFT), pulmonary imaging, a biopsy, a surgery, a chemotherapy, a radiotherapy, or any combination thereof.
5. The method of claim 4 , where said pulmonary imaging is an x-ray, a chest computed tomography (CT) scan, or a positron emission tomography (PET) scan.
6. The method of claim 1 , wherein said pulmonary nodule has a diameter of less than or equal to 3 cm.
7. The method of claim 1 , wherein said pulmonary nodule has a diameter of about 0.8 cm to 3.0 cm.
8. The method of claim 1 , wherein said score is calculated from a logistic regression model applied to the transition measurements.
9. The method of claim 1 , wherein said score is determined as P s =1/[1+exp(−α−Σ i=1 N β i *{hacek over (I)} i,s )], where {hacek over (I)} i,s is logarithmically transformed and normalized intensity of transition i in said sample (s), β i is the corresponding logistic regression coefficient, α was a panel-specific constant, and N was the total number of transitions in said panel.
10. The method of claim 8 , further comprising normalizing the transition measurements.
11. The method of claim 10 , wherein the transition measurements are normalized by one or more transitions selected from the group consisting of a transition of LQSLFDSPDFSK (SEQ ID NO:28) comprising m/z 692.34 and m/z 593.30, a transition of TGVITSPDFPNPYPK (SEQ ID NO:6) comprising m/z 816.92 and m/z 258.10, a transition of TASDFITK (SEQ ID NO:5) comprising m/z 441.73 and m/z 710.40, a transition of SLEDLQLTHNK (SEQ ID NO:29) comprising m/z 433.23 and m/z 499.30, a transition of INPASLDK (SEQ ID NO:30) comprising m/z 429.24 and m/z 630.30, and a transition of VITEPIPVSDLR (SEQ ID NO:31) comprising m/z 669.89 and m/z 896.50.
12. The method of claim 1 , wherein said blood sample is selected from the group consisting of plasma, serum, and whole blood.
13. The method of claim 1 , wherein the determining the likelihood that a pulmonary nodule in a subject is not lung cancer is determined by the sensitivity, specificity, negative predictive value or positive predictive value associated with the score.
14. The method of claim 1 , wherein said score determined in step (c) has a negative predictive value (NPV) that is at least about 80%.
15. A method of determining the likelihood that a pulmonary nodule in a subject is not lung cancer, comprising:
(a) measuring an abundance of a plurality of transitions in a blood sample obtained from the subject, wherein each transition consists of a precursor ion m/z and a fragment ion m/z thereof, wherein said plurality of transitions comprise a transition of ALQASALK (SEQ ID NO: 7) comprising m/z 401.25 and m/z 617.40, a transition of LGGPEAGLGEYLFER (SEQ ID NO: 24) comprising m/z 804.40 and m/z 913.40, a transition of VEIFYR (SEQ ID NO: 25) comprising m/z 413.73 and m/z 598.30, a transition of GFLLLASLR (SEQ ID NO: 22) comprising m/z 495.31 and m/z 559.40, and a transition of AVGLAGTFR (SEQ ID NO: 11) comprising m/z 446.26 and m/z 721.40, and wherein said measuring is performed by selected reaction monitoring mass spectrometry;
(b) calculating a probability of lung cancer score based on the abundance of a plurality of transitions of step (a); and
(c) ruling out lung cancer for the subject if the score in step (b) is lower than a pre-determined score.
16. The method of claim 15 , wherein said plurality of transitions further comprises at least one transition selected from the group consisting of a transition of LTLLAPLNSVFK (SEQ ID NO: 8) comprising m/z 658.40 and m/z 804.50, a transition of NSLFEYQK (SEQ ID NO: 26) comprising m/z 514.76 and m/z 714.30, a transition of IIIQESALDYR (SEQ ID NO: 27) comprising m/z 660.86 and m/z 338.20, a transition of TWNDPSVQQDIK (SEQ ID NO: 23) comprising m/z 715.85 and m/z 260.20, a transition of LDTLAQEVALLK (SEQ ID NO: 20) comprising m/z 657.39 and m/z 330.20, a transition of QITVNDLPVGR (SEQ ID NO: 16) comprising m/z 606.30 and m/z 428.30, and a transition of ATVNPSAPR (SEQ ID NO: 33) comprising m/z 456.80 and m/z 527.30.