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CN-121994953-A - Application of metabolic marker in preparation of head and neck squamous cell carcinoma diagnostic product, kit, screening method of head and neck squamous cell carcinoma metabolic marker, diagnostic model, construction method of diagnostic model and application of diagnostic model

CN121994953ACN 121994953 ACN121994953 ACN 121994953ACN-121994953-A

Abstract

The invention relates to the technical field of metabonomics analysis, in particular to application of a metabolic marker in preparing a head and neck squamous cell carcinoma diagnosis product, a kit, a screening method of the head and neck squamous cell carcinoma metabolic marker, a diagnosis model, a construction method and application of the diagnosis model. The metabolic marker comprises at least one of lactic acid, sphingosine, cadaverine, uridine diphosphate, allantoic acid, hydroxyproline, sphingosine-1-phosphate, indole-3-acetaldehyde, pantothenic acid, fumaric acid, malic acid, prostaglandin, or ornithine in erythrocytes and/or plasma. The red blood cells and plasma metabolic markers screened by the invention can be used for predicting or diagnosing head and neck squamous cell carcinoma respectively or independently, and especially the red blood cells metabolic markers can provide more stable tumor microenvironment information, have high metabolic stability and are not easily influenced by diet and circadian rhythm.

Inventors

  • XIA YANG
  • LI ZHENJIANG
  • HUANG WEILUN

Assignees

  • 中南大学湘雅医院

Dates

Publication Date
20260508
Application Date
20260128

Claims (10)

  1. 1. Use of a metabolic marker comprising at least one of lactic acid, sphingosine, cadaverine, uridine diphosphate, allantoic acid, hydroxyproline, sphingosine-1-phosphate, indole-3-acetaldehyde, pantothenic acid, fumaric acid, malic acid, prostaglandin, or ornithine in erythrocytes and/or plasma for the preparation of a diagnostic product for head and neck squamous carcinoma.
  2. 2. The use according to claim 1, wherein said diagnostic product determines the risk of head and neck squamous cell carcinoma by detecting a significant increase or a significant decrease in the relative amount of metabolic markers in red blood cells and/or plasma in said red blood cells and/or plasma, said significant increase or decrease having a significant P value <0.05.
  3. 3. The use according to claim 1, wherein the metabolic markers each independently or together determine the risk of head and neck squamous cell carcinoma in red blood cells or plasma; In erythrocytes, the relative content of one or more of lactic acid, pantothenic acid, fumaric acid, malic acid, hydroxyproline or prostaglandin is significantly reduced, and the relative content of one or more of sphingosine-1-phosphate, sphingosine, cadaverine, uridine diphosphate, allantoin, indole-3-acetaldehyde or ornithine is significantly increased; In plasma, the relative content of one or more of sphingosine-1-phosphate, sphingosine, hydroxyproline, fumaric acid, malic acid, lactic acid, ornithine, allantoin or prostaglandin is significantly reduced, and the relative content of one or more of uridine diphosphate, cadaverine, pantothenic acid or indole-3-acetaldehyde is significantly increased.
  4. 4. The use according to claim 3, wherein, among said markers of erythrocyte metabolism, any one of lactic acid, sphingosine, cadaverine, uridine diphosphate, allantoin, hydroxyproline or sphingosine-1-phosphate is used as a single marker of metabolism for the independent judgment of the risk of head and neck squamous cell carcinoma; any one of indole-3-acetaldehyde, pantothenic acid, fumaric acid, malic acid, prostaglandin or ornithine is used as an auxiliary discrimination metabolic marker for judging the risk of head and neck squamous cell carcinoma; Among the plasma metabolic markers, any one of lactic acid, allantoic acid, hydroxyproline, sphingosine-1-phosphate, fumaric acid, malic acid, ornithine, cadaverine, uridine diphosphate, pantothenic acid or prostaglandin is used as a single metabolic marker for independently determining the risk of head and neck squamous cell carcinoma; indole-3-acetaldehyde is used as an auxiliary discrimination metabolic marker for judging the disease risk of head and neck squamous cell carcinoma.
  5. 5. A kit for the preparation of a diagnostic product for head and neck squamous carcinoma comprising the diagnostic product of any of claims 1-4.
  6. 6. A screening method of metabolic markers of head and neck squamous cell carcinoma, which is characterized by comprising the following steps: (1) Collecting red blood cell and plasma samples of a head and neck squamous cell carcinoma patient and a healthy control group, and performing LC-MS non-targeted metabonomics analysis on the red blood cell and plasma samples; (2) Performing single-partial least square discriminant analysis and univariate analysis on the obtained LC-MS non-targeted metabonomics analysis result, screening substances with VIP of more than 1.0 and P of less than 0.05 as obvious difference metabolites, performing ROC curve analysis on the screened obvious difference metabolites, and selecting metabolites with AUC of more than 0.65 as metabolic markers.
  7. 7. The method for constructing the head and neck squamous cell carcinoma diagnosis model is characterized by comprising the following steps of: s1, collecting red blood cell and plasma samples of a head and neck squamous carcinoma patient and a healthy control group, performing LC-MS targeted or non-targeted metabonomics analysis on the red blood cell and plasma samples, and screening to obtain a metabolic marker; S2, preprocessing analysis data, constructing a diagnosis model for the metabolic markers based on a Logistic regression model, and evaluating the diagnosis performance of the metabolic markers in single or different combinations through cross verification and independent verification sets.
  8. 8. A head and neck squamous cell carcinoma diagnostic model constructed by the construction method according to claim 7.
  9. 9. The diagnostic model of claim 8, wherein the diagnostic model uses sphingosine-1-phosphate, hydroxyproline, malic acid, and lactic acid in erythrocytes in plasma as a combination of metabolic markers for head and neck squamous cell carcinoma diagnosis, to construct a diagnostic model; The original detection value of the metabolic marker is a relative signal intensity value obtained based on mass spectrum detection, and the unit of the original detection value is a mass spectrum peak intensity value; Z-Score conversion is carried out, and Z-Score is calculated according to Z= (x-mu)/sigma, wherein x represents an actual measurement value of a certain metabolic marker in a single sample, mu represents an average value of the metabolic marker in a training sample overall, sigma represents a standard deviation of the metabolic marker in the training sample overall, and Z is a dimensionless standardized value; the scoring formula of the obtained diagnostic model is as follows: Diagnostic score = Z [ sphingosine-1-phosphate (plasma) ]x12.94 Z [ hydroxyproline (plasma) ]x5.47+z [ malic acid (plasma) ]x11.39+z [ lactic acid (red blood cells) ]x1.65+4.92; wherein, the meaning of each parameter is explained as follows: Z [ sphingosine-1-phosphate (plasma) ] represents the value of the concentration of sphingosine-1-phosphate in the plasma sample normalized by Z-Score; z [ hydroxyproline (plasma) ] represents the value of the hydroxyproline concentration in the plasma sample normalized by Z-Score; Z [ malic acid (plasma) ] represents the value of the malic acid concentration in the plasma sample normalized by Z-Score, Z [ lactic acid (red blood cells) ] represents the value of the lactic acid concentration in the red blood cell sample normalized by Z-Score; 12.94、 5.47, 11.39 and 1.65 are regression coefficients of the metabolic markers in the diagnosis model respectively, and reflect the relative weight and contribution direction of each metabolic marker to the diagnosis score, and are dimensionless coefficients; 4.92 is a constant term of the model for correcting the overall scoring baseline in dimensionless units; And judging the high risk of head and neck squamous cell carcinoma when the diagnosis score exceeds a preset Cut-off value of 0.37.
  10. 10. Use of a diagnostic model of head and neck squamous cell carcinoma, characterized in that the diagnostic model according to any of claims 8-9 or the diagnostic model constructed by the construction method according to claim 7 is used for constructing a clinical screening or auxiliary diagnostic system of head and neck squamous cell carcinoma.

Description

Application of metabolic marker in preparation of head and neck squamous cell carcinoma diagnostic product, kit, screening method of head and neck squamous cell carcinoma metabolic marker, diagnostic model, construction method of diagnostic model and application of diagnostic model Technical Field The invention relates to the technical field of metabonomics analysis, in particular to application of a metabolic marker in preparing a head and neck squamous cell carcinoma diagnosis product, a kit, a screening method of the head and neck squamous cell carcinoma metabolic marker, a diagnosis model, a construction method and application of the diagnosis model. Background Cancer is the second leading cause of death worldwide, and its continuously rising morbidity and mortality have constituted a significant public health threat. GLOBOCAN data show that the global new cancer cases in 2020 reach 1930 thousands of cases and the death cases reach 1000 tens of thousands of cases. Head and Neck Squamous Cell Carcinoma (HNSCC) is the seventh most common cancer species worldwide, with nearly 90 tens of thousands of new cases and more than 45 tens of thousands of deaths, as malignant tumors originating in nasal, oral, laryngeal and pharyngeal epithelium. Although the existing treatment approaches (including surgery, chemoradiotherapy and emerging immunotherapy) are continuously developed, because early symptoms of the disease are hidden and an effective screening strategy is lacked, a higher proportion of patients already progress to late stage (stage III/IV) or metastasis occurs when diagnosis is confirmed, so that the total survival rate of 5 years is stopped for about 50% for a long time, and the poor prognosis is further aggravated by local high recurrence rate and frequent lymph node metastasis. In recent years, metabonomic diagnosis has become a new way of screening for early cancer symptoms. Compared with other early screening modes, the metabonomics analysis has the advantages of short screening time and high accuracy, and can exclude misdiagnosis caused by age, individual difference and comprehensive disease influence with great probability, and under the background, novel biomarkers are developed to realize early diagnosis, so that the method becomes a key path for breaking through the current clinical dilemma of head and neck squamous cell carcinoma. However, existing biomarker technologies still suffer from the following disadvantages: 1. mainly relies on a single class of biomarkers, such as: 1) The combination of circulating RNA markers or miRNAs can realize noninvasive detection, but is limited by RNA stability, individual expression difference and technical sensitivity, and the diagnosis accuracy is limited when the circulating RNA markers or miRNAs are used alone. 2) Although protein markers or metabolic enzymes are related to tumor progression, they are susceptible to non-tumor factors such as inflammation and infection, resulting in high false positive rates. Such single-dimensional markers are difficult to comprehensively capture complex pathological features of HNSCC, and cannot meet the requirement of high-precision diagnosis. 2. Multidimensional integration and collaborative analysis are lacking, although multiple studies propose different classes of markers (such as RNA, protein and metabolite), the prior art lacks an integration model for multi-source biological information, plasma markers (such as ctDNA and proteome) mainly reflect short-term change of tumor microenvironment, and tissue markers need invasive acquisition, and the two are difficult to dynamically associate. The systematic value of marker combinations is not exploited and reliable diagnostic decision models cannot be built. 3. The stability and repeatability of the marker are seriously interfered by external factors, the existing liquid biopsy marker only depends on a liquid biopsy signal of a single source, a stable reference system capable of reflecting the long-term pathological adaptation state of an organism is lacking, short-term physiological fluctuation and real disease signals are difficult to distinguish, the marker is easily influenced by temporary external factors, plasma metabolites are easily influenced by food and circadian rhythm fluctuation obviously, and circulating RNA is easily degraded in sample processing. 4. The prognosis prediction and treatment guidance values are limited, the existing markers (such as PD-L1 and tumor mutation load) are mainly used for predicting the lifetime of the late patients, but the early treatment optimization cannot be guided, and the treatment effect evaluation is delayed due to the lack of a dynamic monitoring model of the markers. Under the premise, red blood cells are taken as biological samples which are stable in function and can be easily obtained in a circulatory system, and are attracting attention of researchers, however, the role of red blood cell metabolites in head and ne