Noninvasive detection of lung cancer using exhaled breath

Patent metadata

Field	Value
Publication number	US-9638695-B2
Application number	US-201414471793-A
Country	US
Kind code	B2
Filing date	Aug 28, 2014
Priority date	Aug 28, 2013
Publication date	May 2, 2017
Grant date	May 2, 2017

Abstract

A non-invasive method of detecting or screening for lung cancer in a subject specimen is provided. The method includes detecting elevated levels of one or more carbonyl-containing volatile organic compounds (VOCs) that are biomarkers for lung cancer in exhaled breath from the subject specimen. The method may further include obtaining exhaled breath from the subject specimen; forming adducts of the carbonyl-containing VOCs with a reactive chemical compound; quantifying the adducts of the carbonyl-containing VOCs to establish a subject value for each of the adducts; and comparing each subject value to a threshold healthy specimen value for each of the adducts of the carbonyl-containing VOCs. One or more subject values at quantities greater than threshold healthy specimen values are also useful for screening for lung cancer in the subject specimen.

First claim

What is claimed is:

1. A non-invasive method of detecting or screening for a lung cancer disease state in a subject specimen, the method comprising the steps of:
obtaining an exhaled breath sample from the subject specimen, wherein the exhaled breath sample includes one or more carbonyl-containing volatile organic compounds (VOCs), wherein the one or more carbonyl-containing VOCs is selected from the group consisting of 2-butanone,2-hydroxyacetaldehyde, 3-hydroxy-2-butanone, 4-hydroxy-2-hexenal (4-HHE), 4-hydroxy-2-nonenal (4-HNE), and a mixture of C 5 H 10 O compounds that includes 2-pentanone and pentanal;

passing the exhaled breath sample through a chemical preconcentrator comprising a reactive chemical compound and reacting the one or more carbonyl-containing VOCs in the exhaled breath sample with the reactive chemical compound to form charged adducts of the one or more carbonyl-containing VOCs, wherein the reactive chemical compound has a general formula (I) of:
H 2 N—Z-L-Y,  (I)

wherein Z is NH, NR or O; L is a Linking group; Y is a di-substituted or tri-substituted N or P moiety; R is selected from the group consisting of alkyls, aralkyls, aralkenyls, and aralkynyls, each of which may be substituted or unsubstituted, and optionally contain one or more heteroatoms, and wherein the plurality of carbonyl-containing VOCs react with the reactive chemical compound via a condensation reaction to form adducts thereof;
quantifying one or more of the charged adducts of the one or more carbonyl-containing VOCs to establish a subject value for the one or more of the charged adducts; and
comparing each subject value to a threshold healthy specimen value for the one or more of the charged adducts of the one or more carbonyl-containing VOCs, the threshold healthy specimen value corresponding to a value calculated from healthy specimens, in order to determine the presence of one or more subject values at quantities greater than their respective threshold healthy specimen values, thereby indicating a lung cancer disease state in the subject specimen.

2. The method of claim 1 , wherein the lung cancer disease state is selected from the group consisting of small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), squamous cell carcinoma, and adenocarcinoma.

3. The method of claim 1 , further comprising:
concentrating the one or more carbonyl-containing VOCs contained in exhaled breath samples obtained from a plurality of healthy specimens, wherein the one or more carbonyl-containing VOCs form charged adducts with the reactive chemical compound; and
quantifying the charged adducts of the one or more carbonyl-containing VOCs in the plurality of healthy specimens to establish the threshold healthy specimen values for each of the charged adducts of the one or more carbonyl-containing VOCs.

4. The method of claim 1 , wherein obtaining the exhaled breath sample from the subject specimen comprises collecting the exhaled breath sample in an inflatable, polymeric film device to provide an inflated device comprising a breath sample; and wherein forming adducts of the plurality of carbonyl-containing VOCs with a reactive chemical compound comprises passing the breath sample through a chemical preconcentrator comprising the reactive chemical.

5. The method of claim 4 , wherein passing the exhaled breath sample through a chemical preconcentrator comprises connecting the inflated device to an inlet of the chemical preconcentrator, and applying reduced pressure to an outlet of the chemical preconcentrator to induce flow of the exhaled breath sample from the inflated device through the chemical preconcentrator.

6. The method of claim 1 , wherein Z is O, and Y is a di-substituted or tri-substituted nitrogen to provide the reactive chemical compound having a general formula (II)

wherein R 1 and R 2 are independently selected from the group consisting of alkyls, aralkyls, aralkenyls, and aralkynyls, each of which may be substituted or unsubstituted, and optionally contain one or more heteroatoms, or wherein R 1 and R 2 in combination form a heterocyclic ring; R 3 is selected from the group consisting of H, alkyls, aralkyls, aralkenyls, and aralkynyls, each of which may be substituted or unsubstituted, and optionally contain one or more heteroatoms; and A is an anionic counter-ion; and wherein said Linking group L comprises a non-ionic segment selected from the group consisting of a substituted or unsubstituted alkyl, a substituted or unsubstituted aryl, and an ether.

7. The method of claim 1 , wherein Z is O, and Y is a di-substituted nitrogen to provide the reactive chemical compound having a general formula (III)

wherein R 1 and R 2 are independently selected from the group consisting of alkyls, aralkyls, aralkenyls, and aralkynyls, each of which may be substituted or unsubstituted, and optionally contain one or more heteroatoms, or wherein R 1 and R 2 in combination form a heterocyclic ring.

8. The method of claim 1 , wherein the reactive chemical compound is a 2-(aminooxy)-N,N,N-trimethylethanammonium salt.

9. The method of claim 1 , wherein quantifying the charged adducts of the one or more carbonyl-containing VOCs to establish the subject value for each member of the charged adducts of the one or more carbonyl-containing VOCs comprises analyzing the charged adducts of the one or more carbonyl-containing VOCs using a mass spectrometer.

10. The method of claim 9 , wherein the mass spectrometer uses Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS).

11. The method of claim 9 , wherein the mass spectrometer is coupled with a chromatography apparatus.

12. The method of claim 1 , wherein the one or more carbonyl-containing VOCs comprises 4-HNE and a mixture of C 5 H 10 O compounds that includes 2-pentanone and pentanal, the method further comprising:
distinguishing a SCLC disease state from a NSCLC disease state in the subject specimen by determining
a) a subject value of a 4-HNE adduct of the reactive chemical compound,
b) a subject value of C 5 H 10 O adducts of the reactive chemical compound, or
c) both a) and b);
and comparing the subject value of a), b), or c) with a concentration range of the respective adduct indicative of at least one of the SCLC disease state and the NSCLC disease state in the subject.

13. The method of claim 12 , further comprising:
d) identifying a concentration range for the 4-HNE adduct of the reactive chemical compound indicative of SCLC, and a concentration range for the 4-HNE adduct of the reactive chemical compound indicative of NSCLC;
e) identifying a concentration range for C 5 H 10 O adducts of the reactive chemical compound indicative of SCLC, and a concentration range for C 5 H 10 O adducts of the reactive chemical compound indicative of NSCLC; or
f) both d) and e).

14. The method of claim 1 , wherein the one or more carbonyl-containing VOCs comprises 4-HHE, the method further comprising:
distinguishing between squamous cell carcinoma, adenocarcinoma, and NSCLC disease states in the subject specimen by determining
a) a subject value of a 4-HHE adduct of the reactive chemical compound
and comparing the subject value of a) with a concentration range of the respective adduct indicative of at least one of the squamous cell carcinoma disease state, the adenocarcinoma disease state, and the NSCLC disease state in the subject.

15. The me

Assignees

• Univ Louisville Res Found Inc

Inventors

• Fu Xiaoan
• Nantz Michael
• Bousamra Michael

Classifications

• G01N33/64

  involving ketones · CPC title

• G01N33/543

  with an insoluble carrier for immobilising immunochemicals · CPC title

• G01N30/72 Primary

  Mass spectrometers {(mass spectrometers per se H01J49/00)} · CPC title

• G01N30/08

  using an enricher · CPC title

• G01N1/405

  by adsorption or absorption · CPC title