Compositions and methods for detecting oral neoplasm

Abstract

Methods and kits for assessing the presence of, and for detecting cancer, notably oral cancer, are disclosed. Such methods and kits use one or more lectins operably linked to a fluorophore, wherein the lectin binds differentially to cancerous and non-cancerous tissues, and wherein the fluorophore facilitates visualizing the differential binding. The lectins are applied to the oral mucosa of a subject, the fluorophore is exposed to light, and cancerous regions of the mucosa are visualized. In certain embodiments, the lectin specifically binds to one or more of a β-galactoside, an α- or β-N-acetylglucosamine, or a sialic acid moiety.

1 . A method for assessing the presence of possible oral cancer in a subject, the method comprising the steps of:
 applying a composition comprising one or more lectins operably linked to a detection moiety to the oral mucosa of a subject, wherein said lectins specifically bind to one or more mono and/or oligosaccharides;   exposing the oral mucosa of the subject to near infrared light, visible light, or ultraviolet light; and   visualizing the oral mucosa to detect possible oral cancer.   
   
   
       2 . The method of  claim 1 , wherein the one or more lectins are selected from the group consisting of Con A, LcH, VFA, PSA, GS-II, WGA, DSA, LEA, STA, LAA, OSA, PWM, PWA, UEA-II, UDA, PTA, HAA, VAA, Allo A, ABA, APA, PNA, CSA, TKA, RCA-I, RCA-II, ECA, CAA, SNA, MAA, LFA, LPA, HMA, and CCA. 
   
   
       3 . The method of  claim 2 , wherein the one or more lectins are selected from the group consisting of GS-II, WGA, PNA, VAA, Allo A, ABA, APA, CSA, TKA, and RCA-I. 
   
   
       4 . The method of  claim 3 , wherein the one or more lectins are selected from the group consisting of GS-II, WGA, and PNA. 
   
   
       5 . The method of  claim 1 , wherein the detection moiety is a dye, a chemiluminescent compound, an enzyme, a fluorescent compound, a metal complex, a biotin, a hapten, a radioluminescent compound, a radioactive-labeled biomolecule, a colored microparticle, a metallic nanoparticles, or a quantum dot. 
   
   
       6 . The method of  claim 5 , wherein the detection moiety is a fluorophore. 
   
   
       7 . The method of  claim 6 , wherein the fluorophore absorbs ultraviolet or visible light and emits visible light. 
   
   
       8 . The method of  claim 6 , wherein the composition further comprises a second lectin operably linked to a second fluorophore, wherein both the first and second fluorophore absorb light of substantially the same wavelength and emit light of substantially different wavelengths. 
   
   
       9 . The method of  claim 6 , wherein the composition further comprises a second lectin operably linked to a second fluorophore, wherein both the first and second fluorophore absorb light of substantially the same wavelength and emit light of similar wavelengths. 
   
   
       10 . The method of  claim 6 , wherein the composition further comprises a second lectin operably linked to a second fluorophore, wherein the first and second fluorophore absorb light of substantially the same wavelength and emit light of different wavelengths. 
   
   
       11 . The method of  claim 6 , wherein the composition further comprises a second lectin operably linked to a second fluorophore, wherein the first and second fluorophore absorb light of different wavelengths and emit light of similar wavelengths. 
   
   
       12 . The method of  claim 1 , wherein the light to which the oral mucosa of the subject is exposed is provided by a light source producing a narrow band of wavelengths. 
   
   
       13 . The method of  claim 1 , wherein the oral mucosa is visualized through one or more narrow band light filters. 
   
   
       14 . The method of  claim 1 , wherein the oral mucosa is visualized through a microscope. 
   
   
       15 . The method of  claim 1 , wherein the oral mucosa is visualized using a software analysis program. 
   
   
       16 . The method of  claim 1 , additionally comprising the step of visualizing the exogenous fluorescence of the oral mucosa. 
   
   
       17 . The method of  claim 1 , additionally comprising the step of applying a tissue dehydrating visual enhancer solution to the oral mucosa. 
   
   
       18 . The method of  claim 17 , wherein the visual enhancer solution comprises acetic acid. 
   
   
       19 . The method of  claim 1 , additionally comprising the step of applying a tissue penetrating agent to the oral mucosa. 
   
   
       20 . The method of  claim 19 , wherein the tissue penetrating agent is DMSO. 
   
   
       21 . A method for assessing the presence of possible oral cancer in a subject, the method comprising the steps of:
 applying one or more lectins to the oral mucosa of a subject, wherein said lectins specifically bind to one or more mono and/or oligosaccharides;   applying an antibody or an avidin protein to the oral mucosa of the subject, wherein the antibody or avidin protein is linked to a fluorophore;   exposing the oral mucosa of the subject to near infrared light, visible light, or ultraviolet light; and   visualizing the oral mucosa to detect possible oral cancer.   
   
   
       22 . The method of  claim 21 , wherein the one or more lectins are selected from the group consisting of Con A, LcH, VFA, PSA, GS-II, WGA, DSA, LEA, STA, LAA, OSA, PWM, PWA, UEA-II, UDA, PTA, HAA, VAA, Allo A, ABA, APA, PNA, CSA, TKA, RCA-I, RCA-II, ECA, CAA, SNA, MAA, LFA, LPA, HMA, and CCA. 
   
   
       23 . The method of  claim 22 , wherein the one or more lectins are selected from the group consisting of GS-II, WGA, PNA, VAA, Allo A, ABA, APA, CSA, TKA, and RCA-I. 
   
   
       24 . The method of  claim 23 , wherein the one or more lectins are selected from the group consisting of GS-II, WGA, and PNA. 
   
   
       25 . The method of  claim 21 , wherein avidin protein is applied to the oral mucosa of the subject, and additionally comprising the step of applying biotin to the oral mucosa of the subject. 
   
   
       26 . The method of  claim 25 , further comprising the step of applying one or more antibodies that are not linked to a fluorophore to the oral mucosa of the subject. 
   
   
       27 . The method of  claim 26 , wherein at least one of the antibodies that are not linked to a fluorophore is an anti-lectin antibody. 
   
   
       28 . The method of  claim 21 , wherein an antibody is applied to the oral mucosa of the subject. 
   
   
       29 . The method of  claim 28 , wherein the antibody is an anti-lectin antibody. 
   
   
       30 . The method of  claim 28 , further comprising the step of applying a second antibody to the oral mucosa of the subject, wherein the second antibody is not linked to a fluorophore. 
   
   
       31 . A kit for assessing the presence of oral cancer, the kit comprising:
 One or more lectins operably linked to a fluorophore, wherein said lectins specifically bind to one or more mono and/or oligosaccharides; and   a light source emitting light in the ultraviolet, visible, or near infrared spectrum.   
   
   
       32 . The kit of  claim 31 , wherein the one or more lectins are selected from the group consisting of Con A, LcH, VFA, PSA, GS-II, WGA, DSA, LEA, STA, LAA, OSA, PWM, PWA, UEA-II, UDA, PTA, HAA, VAA, Allo A, ABA, APA, PNA, CSA, TKA, RCA-I, RCA-II, ECA, CAA, SNA, MAA, LFA, LPA, HMA, and CCA. 
   
   
       33 . The kit of  claim 32 , wherein the one or more lectins are selected from the group consisting of GS-II, WGA, PNA, VAA, Allo A, ABA, APA, CSA, TKA, and RCA-I. 
   
   
       34 . The kit of  claim 33 , wherein the one or more lectins are selected from the group consisting of GS-II, WGA, and PNA. 
   
   
       35 . The kit of  claim 31 , further comprising an optical device to facilitate visualizing the oral cancer. 
   
   
       36 . The kit of  claim 35 , wherein the optical device comprises one or more optical filters. 
   
   
       37 . The kit of  claim 36 , wherein the optical filters are narrow band filters, broadpass filters, or both. 
   
   
       38 . The kit of  claim 31 , wherein the one or more lectins are in the form of lyophilized powder. 
   
   
       39 . The kit of  claim 38 , further comprising a buffer solution for reconstituting the lectins. 
   
   
       40 . The kit of  claim 31 , wherein the one or more lectins are dissolved in a buffer solution. 
   
   
       41 . The kit of  claim 31 , further comprising one or more solution applicators. 
   
   
       42 . The kit of  claim 31 , further comprising a punch biopsy device. 
   
   
       43 . The kit of  claim 31 , further comprising a dehydrating visual enhancer solution. 
   
   
       44 . The kit of  claim 43 , wherein the visual enhancer solution comprises acetic acid. 
   
   
       45 . The kit of  claim 31 , further comprising a tissue penetrating agent. 
   
   
       46 . The kit of  claim 45 , wherein the tissue penetrating agent is DMSO. 
   
   
       47 . The kit of  claim 31 , further comprising a container to collect the patient's saliva or serum for further confirmatory diagnosis.