A biomarker for assessing the risk of progression of lung adenocarcinoma bone metastasis disease and application thereof

Abstract

The application discloses a biomarker for evaluating the risk of lung adenocarcinoma bone metastasis disease progression and application thereof, and particularly relates to three key aspects of early warning, risk stratification and prognosis evaluation. The application provides application of OLFML3 or a detection reagent thereof in preparation of a product for assisting in diagnosis of lung adenocarcinoma bone metastasis or evaluation of the risk of lung adenocarcinoma bone metastasis disease progression, and early warning of the disease progression can be realized by quantitatively detecting the OLFML3 protein concentration in serum of a lung adenocarcinoma patient, precise stratification of the metastasis risk of the patient is completed, and an objective basis is provided for prognosis evaluation. The method effectively solves the clinical bottleneck that existing imaging examination cannot be dynamically monitored and traditional serum markers have insufficient prediction specificity, has the significant advantages of non-invasiveness, convenience and repeatable monitoring, and provides a brand-new technical means for whole-process management of lung adenocarcinoma bone metastasis.

Description

Technical Field

[0001] This invention relates to a biomarker for assessing the risk of progression of lung adenocarcinoma bone metastasis and its application, belonging to the field of biomedical detection technology. Background Technology

[0002] Lung adenocarcinoma (LUAD) is the most common subtype of lung cancer, and its high mortality rate is mainly attributed to its high incidence of distant metastases. Bone is one of the most common sites of metastasis for LUAD. Bone metastases not only lead to severe skeletal-related events (such as severe pain, pathological fractures, and hypercalcemia), but more importantly, they often serve as a "stepping stone" for further tumor spread. The vast majority of LUAD patients with bone metastases ultimately die from multiple metastases mediated by tumor stem cells (such as liver, brain, and adrenal gland metastases), rather than from simple bone destruction itself. Therefore, the prognosis for patients with bone metastases is extremely poor, and clinical management faces significant challenges.

[0003] Currently, clinical prognostic assessment and monitoring of patients with bone metastases from lung adenocarcinoma mainly rely on two types of methods:

[0004] (1) Imaging examinations: including computed tomography (CT), bone scan (ECT), and positron emission tomography (PET-CT). Although these techniques can detect macroscopic metastases, they are not sensitive enough to micrometastases and have disadvantages such as radiation exposure, high cost, and inability to perform frequent dynamic monitoring, thus failing to achieve early warning. More importantly, they can only provide anatomical morphological information and cannot reflect the intrinsic molecular biological processes that drive tumor metastasis and stem enhancement.

[0005] (2) Conventional serum tumor markers (such as CEA, CYFRA 21-1): Although they have been widely used for auxiliary diagnosis, their concentration changes are weakly associated with multiple metastatic events, making it impossible to achieve accurate risk stratification and lacking specificity and sensitivity in predicting the risk of multiple metastases after bone metastasis.

[0006] The bone microenvironment plays a crucial role in tumor metastasis. The traditional "vicious cycle" theory emphasizes the supporting role of osteoblasts in tumor colonization and growth. However, the core function of osteoclasts in regulating tumor cell stemness and thus driving multiple metastases remains poorly understood. No clear core pivot molecule has yet been identified to link the two key events of "osteoclast activity" and "tumor stemness reprogramming."

[0007] Therefore, there is an urgent need in this field for a novel, specific biomarker to fill the above gap and provide a new tool for early warning, risk stratification and prognostic assessment of patients with lung adenocarcinoma bone metastases. Summary of the Invention

[0008] The purpose of this invention is to address the shortcomings of existing technologies by providing a biomarker for assessing the risk of progression of lung adenocarcinoma bone metastases and its application.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] In a first aspect, the present invention provides the use of OLFML3 or its detection reagent in the preparation of products for assisting in the diagnosis of bone metastases of lung adenocarcinoma or for assessing the risk of disease progression in patients with bone metastases of lung adenocarcinoma.

[0011] In some embodiments, the detection reagent includes reagents for detecting OLFML3 at the gene level and / or protein level.

[0012] In some embodiments, the detection reagent is a reagent used in one or more detection techniques or methods selected from the group consisting of: enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, radioimmunoassay, immunoprecipitation assay, immunoblotting, high performance liquid chromatography (HPLC), capillary gel electrophoresis, near-infrared spectroscopy, mass spectrometry, immunochemiluminescence assay, colloidal gold immunochromatography, fluorescence immunochromatography, surface plasmon resonance (SPR), immuno-PCR, or biotin-avidin assay.

[0013] In some implementations, the product includes at least one of reagents, kits, test strips, chips, detection systems, and devices.

[0014] In some embodiments, the product is a testing device, which includes a sample collection device, a sample testing device, and a diagnostic device; wherein:

[0015] The sample collection device is configured to collect blood samples from a subject who is a lung adenocarcinoma patient;

[0016] The sample detection device is a device capable of detecting the content or expression level of OLFML3 protein or its nucleic acid in the blood sample;

[0017] The diagnostic device includes a data acquisition module and a diagnostic module. The data acquisition module is configured to acquire data detected by the sample detection device, and the diagnostic module is configured to assess the risk of disease progression in the subject based on the data acquired by the data acquisition module.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] (1) Achieved early and non-invasive early warning of the progression of bone metastasis in lung adenocarcinoma: This invention provides a biomarker OLFML3 (interolaryngin-like protein 3) and related detection products (detection reagents, equipment, etc.) for assessing the risk of progression of bone metastasis in lung adenocarcinoma. Based on the detection scheme of this biomarker, the bottlenecks of insensitivity of imaging to micrometastases and inability to monitor frequently have been overcome, and the problem of early clinical warning has been solved.

[0020] (2) Accurate risk stratification of multiple metastases is achieved: The OLFML3 biomarker provided by this invention has a stronger correlation with multiple metastases than traditional biomarkers such as CEA, which significantly improves the specificity and sensitivity of risk stratification.

[0021] (3) It provides tools from diagnosis to mechanism exploration: This invention not only provides a basis for prognostic management, but also provides new key targets and research directions for revealing the core biological mechanism of "osteoclast-tumor stem reprogramming". Attached Figure Description

[0022] Figure 1 Serum OLFML3 levels in healthy individuals and patients with different types of lung adenocarcinoma.

[0023] Figure 2 Using healthy patients and patients with lung adenocarcinoma without metastasis as negative controls, this study evaluated the diagnostic efficacy of OLFML3 for bone metastasis.

[0024] Figure 3 Using patients with lung adenocarcinoma bone metastases (BM) as negative controls, this study evaluated the diagnostic efficacy of OLFML3 for multiple metastases. Detailed Implementation

[0025] To make the present invention more apparent and understandable, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0026] Experimental methods in the following examples, unless otherwise specified, are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0027] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0028] Example

[0029] Based on transcriptome sequencing (RNA-seq) and functional validation using a lung adenocarcinoma bone metastasis model, OLFML3 was found and confirmed to be specifically highly expressed in cells with high bone metastasis and strongly correlated with tumor stemness and metastatic phenotype. Therefore, it was established as a candidate biomarker for clinical validation.

[0030] This study included a retrospective cohort from the Lingang branch of the Sixth People's Hospital affiliated with Shanghai Jiao Tong University School of Medicine, comprising 24 healthy individuals undergoing routine checkups, 30 patients with lung adenocarcinoma without metastasis, 28 patients with isolated bone metastasis, and 37 patients with multiple metastases of lung adenocarcinoma. The multiple metastasis group was defined as including bone metastasis and at least one other distant metastasis. Although the metastatic lesions in this group were mostly discovered concurrently, making it impossible to determine the absolute chronological order, this group accurately defined a late-stage disease subgroup with "high tumor burden and high invasive potential." The core of this study is that OLFML3 can effectively distinguish between "isolated bone metastasis" and the "multiple metastases" status, which represents a higher risk of progression, providing direct clinical evidence for its use in risk assessment.

[0031] ELISA method for detecting OLFML3 levels in serum:

[0032] 1. Sample Collection: Draw venous blood from the subject into a blood collection tube containing separating gel, centrifuge at 3500 rpm for 15 minutes, collect the supernatant serum and centrifuge at 13000 rpm for 10 minutes, collect the supernatant. The sample can be stored at 4°C on the same day of testing, or stored long-term at -80°C. Before use, centrifuge again at 13000 rpm for 10 minutes and collect the supernatant.

[0033] 2. ELISA Procedure: Coat the wells with anti-OLFML3 antibody, add subject serum, incubate at 37°C for 1 hour, and wash the plate 3-5 times with deionized water for 3-5 minutes each time. Add OLFML3 enzyme-labeled antibody, incubate at 37°C for 30 minutes, and wash the plate 3-5 times with deionized water for 3-5 minutes each time. Add developing solution, incubate at 37°C for 15 minutes, add stop solution, and measure the absorbance at 450 nm using a microplate reader. A standard curve should be plotted using standards for each assay.

[0034] 3. Analysis of test results: The concentration of OLFML3 in the serum of the subjects was calculated based on the standard curve.

[0035] The test results showed that serum OLFML3 concentrations differed significantly among different clinical groups (p<0.01). Specifically, the serum OLFML3 concentration in the lung adenocarcinoma bone metastasis group (BM group) was significantly higher than that in the healthy group (HC group) and the non-metastasis group (NM group). The serum OLFML3 concentration in the lung adenocarcinoma multiple metastasis group (MM group) was significantly higher than that in other groups (including the HC group, NM group, and BM group). Figure 1 As shown.

[0036] Using healthy patients and those with non-metastatic lung adenocarcinoma as negative controls, the diagnostic efficacy of OLFML3 for lung adenocarcinoma metastasis was evaluated using ROC curves. The results are as follows: Figure 2As shown, OLFML3 exhibits excellent diagnostic efficacy for tumor metastasis tendency, with an AUC of 0.864. ROC analysis determined the optimal cutoff value to be 148.1 ng / mL. More importantly, ROC curve analysis, using patients with isolated bone metastases as negative controls, demonstrated that OLFML3 showed good diagnostic efficacy in differentiating between multiple metastatic states, as shown in the results. Figure 3 As shown, the AUC was 0.779, and the optimal cutoff value was determined to be 127.8 ng / mL by ROC analysis, providing a clear standard for the clinical application of this biomarker.

[0037] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any form or substance. It should be noted that those skilled in the art can make several improvements and additions without departing from the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention.

Claims

1. The use of OLFML3 or its detection reagent in the preparation of products for the auxiliary diagnosis of bone metastasis of lung adenocarcinoma or for assessing the progression of lung adenocarcinoma to multiple metastases in patients with bone metastasis, characterized in that, The detection reagent includes a reagent for detecting OLFML3 at the protein level.

2. The application according to claim 1, characterized in that, The detection reagent is a reagent selected from one or more detection techniques or methods from the group consisting of: enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, radioimmunoassay, immunoprecipitation assay, immunoblotting, high performance liquid chromatography (HPLC), capillary gel electrophoresis, near-infrared spectroscopy, mass spectrometry, immunochemiluminescence assay, colloidal gold immunoassay, surface plasmon resonance assay, immuno-PCR, or biotin-avidin assay.

3. The application according to claim 1 or 2, characterized in that, The product includes at least one of reagents, reagent kits, test strips, chips, detection systems, and detection equipment.

4. The application according to claim 3, characterized in that, The product is a testing device, which includes a sample collection device, a sample testing device, and a diagnostic device; wherein: The sample collection device is configured to collect blood samples from a subject who is a lung adenocarcinoma patient; The sample detection device is a device capable of detecting the content or expression level of OLFML3 protein in the blood sample; The diagnostic device includes a data acquisition module and a diagnostic module. The data acquisition module is configured to acquire data detected by the sample detection device, and the diagnostic module is configured to assess the risk of disease progression in the subject based on the data acquired by the data acquisition module.

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