Application of serum sTGFBR3 in diagnosis of lung cancer and evaluation of chemotherapy efficacy

Abstract

The application discloses application of serum sTGFBR3 in diagnosis of lung cancer and evaluation of chemotherapy effect, and belongs to the technical field of biotechnology.The application proves that serum sTGFBR3 can be used as a new marker for auxiliary diagnosis of lung cancer and dynamic monitoring of chemotherapy effect.The diagnostic efficiency of sTGFBR3 in distinguishing lung cancer patients from healthy people is better than that of traditional markers such as CEA, CYFRA21-1 and CA-125;sTGFBR3 can effectively identify lung cancer and pneumonia, and the baseline level of sTGFBR3 is not interfered by pneumonia, thus solving the clinical problem of false positive caused by inflammation.The serum sTGFBR3 level is also related to good chemotherapy response and is correlated with a carboplatin administration scheme.The application provides a simple and effective serological new index for early screening, differential diagnosis and early evaluation of chemotherapy effect of lung cancer.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to the application of serum sTGFBR3 in the diagnosis of lung cancer and the evaluation of chemotherapy efficacy. Background Technology

[0002] Lung cancer is one of the leading causes of cancer-related deaths worldwide, with an extremely low five-year survival rate for advanced-stage patients. Current treatment options for lung cancer mainly include surgical resection, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. Surgical resection is the preferred radical treatment for early-stage lung cancer, but most patients are diagnosed at an advanced stage and are no longer eligible for surgery. Radiotherapy is suitable for local control, but its effectiveness against distant metastases is limited. Targeted therapy benefits only those with specific mutation subtypes and inevitably leads to secondary drug resistance. Immunotherapy activates the host's anti-tumor immunity by blocking immune checkpoints such as PD-1 / PD-L1, which can prolong survival for some patients, but the overall response rate is only 20%–40%, and there are risks such as hyperprogression and immune-related adverse events. Chemotherapy, especially platinum-based dual-drug regimens (such as carboplatin and cisplatin), remains the core first-line treatment for advanced lung cancer (especially those without driver gene mutations or unsuitable for immunotherapy), and covers the broadest population.

[0003] Early diagnosis and real-time monitoring of chemotherapy efficacy are crucial for improving the prognosis of lung cancer patients. However, current clinical diagnosis and treatment face two major challenges: 1. Lack of highly specific serum diagnostic biomarkers: Currently used serum biomarkers for lung cancer include CEA, CYFRA21-1, and CA-125, but their sensitivity and specificity are not ideal. Especially in the differential diagnosis of lung cancer from inflammatory lung diseases (such as pneumonia), existing biomarkers are easily affected by inflammation, leading to false positives and unnecessary invasive examinations. 2. Lack of serum indicators for real-time dynamic monitoring of chemotherapy efficacy: There are significant individual differences in the sensitivity of advanced lung cancer patients to chemotherapy drugs. Clinically, it is usually necessary to complete four cycles of chemotherapy before efficacy can be assessed. This makes it difficult to identify ineffective patients early in the chemotherapy process, missing the optimal time for switching drugs or adjusting the treatment plan. Currently, there is a lack of serum dynamic monitoring indicators that can achieve early lung cancer screening and reflect the chemotherapy response process in real time. Summary of the Invention

[0004] The purpose of this invention is to provide the application of serum sTGFBR3 in the diagnosis of lung cancer and the evaluation of chemotherapy efficacy, thereby addressing the problems existing in the prior art. This invention demonstrates that serum sTGFBR3 can serve as a novel biomarker for the auxiliary diagnosis of lung cancer and dynamic monitoring of chemotherapy efficacy. The diagnostic efficacy of sTGFBR3 in distinguishing lung cancer patients from healthy individuals is superior to traditional biomarkers such as CEA, CYFRA21-1, and CA-125; it can also effectively differentiate between lung cancer and pneumonia, without interfering with baseline levels due to pneumonia, thus solving the clinical problem of false positives caused by inflammation. Serum sTGFBR3 levels are also associated with a good chemotherapy response and are correlated with carboplatin dosing regimens.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides the application of a reagent for detecting serum sTGFBR3 levels in the preparation of products for diagnosing lung cancer, where the sTGFBR3 level in the serum of lung cancer patients is higher than that in healthy individuals.

[0006] This invention also provides the application of a reagent for detecting serum sTGFBR3 levels in the preparation of products for differentiating between lung cancer and pneumonia, wherein the sTGFBR3 level in the serum of lung cancer patients is higher than that in pneumonia patients.

[0007] The present invention also provides the use of reagents for detecting serum sTGFBR3 levels in the preparation of products for evaluating the efficacy of chemotherapy for lung cancer.

[0008] Optionally, the product may include reagents or kits.

[0009] The present invention also provides a product for diagnosing lung cancer, the product comprising a reagent for detecting serum sTGFBR3 levels.

[0010] The present invention discloses the following technical effects: This invention confirms that serum sTGFBR3 can serve as a novel biomarker for the auxiliary diagnosis of lung cancer and dynamic monitoring of chemotherapy efficacy. The AUC of sTGFBR3 distinguishing lung cancer patients from healthy individuals reaches 0.90, with both sensitivity and specificity at 90%, demonstrating superior diagnostic efficacy compared to traditional biomarkers such as CEA, CYFRA21-1, and CA-125. Furthermore, it effectively differentiates lung cancer from pneumonia (AUC=0.8595), while pneumonia does not interfere with baseline levels (AUC=0.5544), resolving the clinical challenge of false positives caused by inflammation. In addition, serum sTGFBR3 levels exhibit individualized dynamic changes during chemotherapy, with a sustained high level trend correlated with a good chemotherapy response and a relationship with carboplatin dosing regimens. This invention provides a simple and effective new serological indicator for early lung cancer screening, differential diagnosis, and early chemotherapy efficacy assessment. Attached Figure Description

[0011] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0012] Figure 1 A comparison chart of serum sTGFBR3 levels in healthy individuals, pneumonia patients, and lung cancer patients; Figure 2 A comparative graph of serum sTGFBR3 levels among normal individuals, pneumonia patients, newly diagnosed patients with advanced lung cancer, and lung cancer patients undergoing chemotherapy. Figure 3 ROC curves for serum sTGFBR3 and traditional tumor markers to differentiate lung cancer patients from normal individuals and pneumonia patients; Figure 4 Figure 1 shows the correlation between dynamic changes in serum sTGFBR3 and chemotherapy efficacy, and the validation of carboplatin induction mechanism. Specifically, A represents the dynamic changes in serum sTGFBR3 levels in each chemotherapy cycle for patients in the partial remission (PR) group; B represents the dynamic changes in serum sTGFBR3 levels in each chemotherapy cycle for patients in the stable disease (SD) group; C represents the dynamic changes in serum sTGFBR3 levels in each chemotherapy cycle for patients in the non-application (NA) group; D compares serum sTGFBR3 levels in PR patients before and after carboplatin treatment; and E shows the proliferation inhibition curves and IC50 values ​​of A549 cells after 24 hours of treatment with different concentrations of carboplatin. 50 Value; F is carboplatin (IC) 50 Western blot images of TGFBR3 protein expression in A549 cells before and after treatment with carboplatin (IC50) concentration; G represents carboplatin (IC50). 50 Bar chart showing the relative quantitative expression of TGFBR3 protein in A549 cells before and after treatment with (concentration); H represents carboplatin (IC50). 50 Bar chart comparing the secretion levels of sTGFBR3 in the supernatant of A549 cells before and after treatment (concentration). Detailed Implementation

[0013] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0014] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0015] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0016] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0017] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0018] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the instruments and equipment used in the following examples are all conventional laboratory instruments and equipment; unless otherwise specified, the experimental materials used in the following examples were all purchased from conventional biochemical reagent stores.

[0019] Example 1 From October 2023 to October 2024, 30 healthy individuals, 30 patients with pneumonia, and 110 patients with lung cancer were recruited. Baseline data for all subjects are shown in Table 1. The inclusion criteria for healthy individuals were no history of chronic diseases, no recent acute illness, and normal results in liver and kidney glucose and lipid tests and chest X-rays. Exclusion criteria included latent infection, pregnancy or lactation, and recent surgery or blood transfusion history. The inclusion criteria for pneumonia patients were clinical diagnosis of pneumonia (symptoms + new radiographic infiltrates + abnormal blood counts). Exclusion criteria included active pulmonary tuberculosis, fungal infection, pulmonary edema, or recent use of immunosuppressants. The inclusion criteria for lung cancer patients were not limited by stage or treatment history (including newly diagnosed and previously treated patients). Exclusion criteria included a second primary lung cancer.

[0020] Table 1 Baseline Data Blood samples were collected from healthy controls, pneumonia patients, and lung cancer patients. Serum was separated, and the level of sTGFBR3 in the serum was detected using ELISA. The ELISA kit was purchased from Huamei Biotechnology (catalog number CSB-EL023453HU). All steps were performed according to the manufacturer's instructions.

[0021] The results showed that, compared with the normal control group, the serum sTGFBR3 level in the lung cancer group was significantly increased (see [link to study]). Figure 1 ).

[0022] This result provides direct evidence for sTGFBR3 as a serological diagnostic biomarker—although tissue-level TGFBR3 expression is downregulated in lung cancer, the level of its soluble form, sTGFBR3, is significantly elevated in serum, suggesting that tumor cells may release membrane-bound TGFBR3 into the bloodstream in a soluble form through mechanisms such as protease cleavage or exosome secretion. This consistent tissue-serum contrast (low tissue expression + high serum expression) further enhances the biological rationale and clinical feasibility of sTGFBR3 as a serum biomarker for lung cancer. Building on this, to further clarify the serological characteristics of newly diagnosed lung cancer patients, future studies will narrow the inclusion criteria to pathologically confirmed newly diagnosed lung cancer patients who have not received any anti-tumor treatment, in order to exclude the interference of previous treatments on serum biomarker levels and improve the reliability of the research results.

[0023] Example 2 From January 2024 to December 2025, 30 healthy individuals, 30 patients with pneumonia, and 170 newly diagnosed patients with advanced lung cancer were recruited. Baseline data for all participants are shown in Table 2. The inclusion criteria for healthy individuals were no history of chronic diseases, no recent acute illness, and normal results in liver and kidney glucose and lipid tests and chest X-rays. Exclusion criteria included latent infection, pregnancy or lactation, and recent surgery or blood transfusion history. The inclusion criteria for patients with pneumonia were a clinical diagnosis of pneumonia (symptoms + new radiographic infiltrates + abnormal blood counts). Exclusion criteria included active pulmonary tuberculosis, fungal infection, pulmonary edema, or recent use of immunosuppressants. The inclusion criteria for newly diagnosed patients with advanced lung cancer were pathologically confirmed stage IIIB-IV lung cancer and no prior history of any anti-tumor systemic treatment (including chemotherapy, targeted therapy, immunotherapy, and clinical trial drugs). Exclusion criteria included a second primary cancer or recent chest radiotherapy. Newly diagnosed patients with advanced lung cancer underwent chemotherapy after enrollment, and were followed up for four cycles.

[0024] Table 2 Baseline Data Blood was collected from each group of patients, serum was separated, and the level of sTGFBR3 in the serum was detected according to the method in Example 1. The diagnostic value of sTGFBR3 for lung cancer was evaluated using ROC curves, and the results are as follows: Figure 2 , Figure 3 As shown in Table 3. The results show: 1. Compared with healthy individuals and pneumonia patients, the serum sTGFBR3 level in newly diagnosed patients with advanced lung cancer was significantly increased. Further analysis revealed that after completing four cycles of chemotherapy, the serum sTGFBR3 level in these patients was higher than the baseline level in all newly diagnosed lung cancer patients. Figure 2 ).

[0025] 2. Overall diagnostic effectiveness The area under the curve (AUC) of sTGFBR3 in distinguishing between newly diagnosed lung cancer patients and normal individuals was as high as 0.9. With an optimal cutoff value of 42 ng / mL, its sensitivity was 90.0%, specificity was 90.0%, and Youden's index was 0.8, suggesting that this biomarker has certain application potential in the diagnosis of lung cancer. Figure 3 (Table 3).

[0026] 3. Its diagnostic efficacy is superior to that of currently used clinical indicators. In the same clinical sample, this invention compared the diagnostic efficacy of sTGFBR3 with commonly used clinical indicators, such as... Figure 3 As shown in Table 3, the AUC value of sTGFBR3 in distinguishing between newly diagnosed lung cancer patients and normal individuals was 0.9, while the AUC values ​​of traditional tumor markers such as CEA, CYFRA21-1, and CA-125 ranged from 0.8629 to 0.8841. The AUC value of sTGFBR3 was higher than that of the aforementioned traditional markers in this sample. This suggests that sTGFBR3 has certain research value and potential as an auxiliary diagnostic marker for lung cancer.

[0027] 4. Effectively differentiates between lung cancer and pneumonia (solving common clinical problems that cause confusion) Inflammatory lesions in the lungs (such as pneumonia and inflammatory pseudotumors) are among the most common interfering factors in the differential diagnosis of lung cancer. Clinically, some serological markers may produce false-positive results in the context of inflammation. Therefore, this invention further extends the analysis to a complex tri-category clinical scenario including normal individuals, pneumonia patients, and newly diagnosed lung cancer patients, evaluating the diagnostic efficacy of sTGFBR3 in differentiating between normal individuals and pneumonia patients, and between pneumonia patients and newly diagnosed lung cancer patients. As shown in Table 4, in differentiating between pneumonia patients and newly diagnosed lung cancer patients, the AUC of sTGFBR3 was 0.8595, indicating that this marker still has some discriminatory ability in the presence of inflammatory interference. However, in differentiating between normal individuals and pneumonia patients, the AUC of sTGFBR3 was 0.5544, indicating that this marker has no significant discriminatory ability between normal healthy individuals and pneumonia patients, and inflammation does not interfere with baseline levels.

[0028] Table 3 Optimal cutoff values ​​and diagnostic efficacy indicators for different biomarkers Table 4 Diagnostic efficacy in normal population, pneumonia patients, and lung cancer patients Example 3 To further explore the relationship between serum sTGFBR3 levels and chemotherapy efficacy, this invention conducted long-term follow-up, measuring serum sTGFBR3 levels at each chemotherapy cycle in 23 lung cancer patients. Patients were grouped according to chemotherapy efficacy into four groups: partial remission (PR) group (n=15), stable disease (SD) group (n=2), progressive disease (PD) group (n=0), and non-target disease (NA) group (n=6). The PR group was characterized by a ≥30% reduction in the total diameter of target lesions from baseline after at least four cycles of first-line treatment, with no new lung or distant metastases and no clear progression of non-target lesions. The SD group was characterized by a change in the total diameter of target lesions between PR and PD (reduction <30% and increase <20%), with no new lesions and a stable state maintained for at least 6-8 weeks. The PD group was characterized by an increase in the total diameter of target lesions of at least 20%, or the appearance of new lesions. The NA group was characterized by patients tolerating chemotherapy, with chaotic tumor response on imaging, difficulty in measuring lesions, and efficacy that could not be categorized as PR / SD / PD. Serum sTGFBR3 levels in each group of patients were statistically analyzed.

[0029] The results showed that serum sTGFBR3 levels exhibited significant individual differences and a regular, inverse dynamic change. Patients were grouped according to chemotherapy efficacy, and the correlation between dynamic changes in sTGFBR3 and chemotherapy efficacy was observed and analyzed. The results are as follows: Figure 4 As shown in Table 5.

[0030] Table 5. Correlation between dynamic changes in sTGFBR3 and chemotherapy efficacy In particular, in-depth analysis of clinical medication records in PR patients revealed a clear correlation between the sTGFBR3 change pattern and the carboplatin dosing regimen: Seven patients discontinued or reduced carboplatin after four cycles, and their sTGFBR3 levels were significantly lower than at the time of no treatment. P =0.0481, see Figure 4 (D); In one patient, sTGFBR3 decreased after discontinuing carboplatin in the third cycle, and increased again after resuming medication in the fourth cycle, showing a dose-dependent fluctuation between medication use and discontinuation.

[0031] The sTGFBR3 level fluctuated dramatically across all patients, reaching a maximum of 158.9 ng / mL. Two patients showed a significant rebound increase in sTGFBR3 levels during the later stages of chemotherapy, suggesting that sTGFBR3 may serve as a potential indicator of chemotherapy efficacy and early warning of treatment resistance.

[0032] Example 4 Based on the clinical observation results of Example 3, this invention hypothesizes that carboplatin may induce the release of membrane-bound TGFBR3 into a soluble form (sTGFBR3), and conducts systematic in vitro cell experiments to verify this. The specific experimental procedure is as follows: 1. Experimental Design: Human lung adenocarcinoma A549 cell line was used, and cells were treated with different concentrations of carboplatin (0–400 µM). After 24 h, proliferation inhibition was detected by the CCK-8 assay, and the IC50 was calculated. 50 TGFBR3 protein expression was detected by Western blot, and sTGFBR3 secretion levels in cell supernatant were detected by ELISA. Specific experimental conditions and results are shown in Table 6. Figure 4 As shown in the figure. The results of this in vitro experiment corroborated the clinical observation results of Example 3, jointly confirming that carboplatin chemotherapy upregulates the expression of TGFBR3 in tumor cells and promotes its release into a soluble form, thereby leading to dynamic changes in serum sTGFBR3 levels in patients. This mechanism discovery provides a solid biological basis for sTGFBR3 as a biomarker for dynamic monitoring of chemotherapy efficacy.

[0033] Table 6 Experimental conditions and test results The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. The application of a reagent for detecting serum sTGFBR3 levels in the preparation of products for diagnosing lung cancer, characterized in that, The serum sTGFBR3 level in lung cancer patients is higher than that in healthy individuals.

2. The application of a reagent for detecting serum sTGFBR3 levels in the preparation of products for differentiating between lung cancer and pneumonia, characterized in that, The serum sTGFBR3 level in lung cancer patients is higher than that in pneumonia patients.

3. Application of reagents for detecting serum sTGFBR3 levels in the preparation of products for evaluating the efficacy of chemotherapy in lung cancer.

4. The application according to any one of claims 1-3, characterized in that, The products include reagents or kits.

5. A product for diagnosing lung cancer, characterized in that, The product contains reagents for detecting serum sTGFBR3 levels.

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