Biomarkers for detection and diagnosis of head and neck squamous cell carcinoma

Abstract

Novel, sensitive and specific markers and methods for diagnostics and monitoring of head and neck squamous cell carcinoma (HNSCC) are provided. Kits and methods for the use of hyaluronic acid, hyaluronidase and CD44 to diagnose HNSCC are described.

1 . A method of diagnosing head and neck squamous cell carcinoma (HNSCC), or elevated risk of future occurrence thereof, in a subject, the method comprising: assaying a subject sample for the presence of at least one biomarker selected from the group consisting of hyaluronic acid (HA); hyaluronidase (HAase), and total protein; wherein the detection of said biomarker(s) is correlated with a diagnosis of HNSCC, or an elevated risk of future occurrence thereof, the correlation taking into account the presence and level of biomarker(s) in the subject sample as compared to normal subjects. 
     
     
         2 . The method of  claim 1 , further comprising assaying the subject sample for CD44 and comparing CD44 values in the subject sample with those from normal subjects, wherein elevated CD44 in the subject sample is correlated with a diagnosis of HNSCC or an elevated risk of future occurrence. 
     
     
         3 . The method of  claim 1 , wherein a plurality of the biomarkers are measured. 
     
     
         4 . The method of  claim 3  wherein HA, HAase, and CD44 are measured. 
     
     
         5 . The method of  claim 1 , wherein the subject sample is selected from the group consisting of oral rinse, saliva, sputum, breath condensate, blood, blood plasma, serum, urine, tissue, cells, and liver. 
     
     
         6 . The method of  claim 1 , wherein the subject sample is saliva or an oral rinse. 
     
     
         7 . The method of  claim 1 , wherein CD44 is soluble CD44 and isoforms thereof. 
     
     
         8 . The method of  claim 1 , wherein the biomarkers are detected using a binding assay. 
     
     
         9 . The method of  claim 8  wherein the binding assay is an immunoassay. 
     
     
         10 . The method of  claim 1 , wherein presence or increased level of HA, HAase, CD44, and/or total protein is indicative of tumor stage. 
     
     
         11 . A method of monitoring effectiveness of treatment of head and neck squamous cell carcinoma comprising measuring a plurality of biomarkers selected from the group consisting of hyaluronic acid (HA); hyaluronidase (HAase), CD44, and/or total protein in a biological sample obtained from said subject, wherein a decreased level of at least one of HA, HAase, CD44, and/or total protein compared to levels detected prior to treatment in the same patient, is indicative of effective treatment. 
     
     
         12 . The method of  claim 11  wherein HA, HAase, and CD44 are measured. 
     
     
         13 . A kit for diagnosing head and neck squamous cell carcinoma (HNSCC), the kit comprising a reference sample of at least one biomarker selected from the group consisting of hyaluronic acid (HA) and hyaluronidase (HAase), and means for detecting/measuring said biomarker(s). 
     
     
         14 . The kit of  claim 13 , additionally comprising a reference sample of CD44 and a means for detecting/measuring CD44. 
     
     
         15 . The kit of  claim 13  that additionally comprises means for measuring total protein. 
     
     
         16 . The kit of  claim 13  that additionally comprises a substrate or container for holding a biological sample from a subject. 
     
     
         17 . The kit of  claim 14  that comprises reference samples of HA, HAAse, and CD44 and means for measurement/detection thereof. 
     
     
         18 . The kit of  claim 17  that comprises at least one colorimetric agent for the detection of a biomarker. 
     
     
         19 . A kit for diagnosing head and neck squamous cell carcinoma (HNSCC) in a subject, the kit comprising: (a) a substrate for holding a saliva sample isolated from the subject, (b) a plurality of biomarkers selected from the group consisting of HA, HAase, and CD44; (c) agents for detection/measurement of said biomarkers; and, optionally (d) printed instructions for reacting the agents with the biological sample or a portion of the biological sample to detect the presence or amount of at least one biomarker in the biological sample. 
     
     
         20 . The kit of  claim 19  that additionally comprises means for measuring total protein in the saliva sample. 
     
     
         21 . The kit of  claim 19  wherein at least one agent is a calorimetric agent. 
     
     
         22 . The kit of  claim 19 , wherein the kit comprises antibodies specific for anyone or more biomarkers selected from the group consisting of hyaluronic acid (HA); hyaluronidase, and CD44.