IA-stage lung adenocarcinoma detection system, kit and application of IA-stage lung adenocarcinoma detection system

Abstract

The invention belongs to the technical field of molecular biology detection, and particularly relates to an IA-stage lung adenocarcinoma detection system, a kit and application thereof. The detection system comprises a sample image analysis module, a classification evaluation module and an evaluation verification module, according to the detection system, on the basis of the PET molecular probe FAPI-04 of the targeted tumor fibroblast activation protein (FAP), the lung adenocarcinoma and metastatic focus with the diameter smaller than or equal to 3 cm in the IA stage can be taken for a higher time and longer time, and by means of the semi-quantitative index SUVmax value, the lung adenocarcinoma in the IA stage can be accurately detected in the early stage in a non-invasive mode. According to the embodiment of the invention, the expression condition of FAP is detected and analyzed by comparing < 18 > F-FDG PET / CT and < 18 > F-FAPI-04PET / CT imaging and combining an immunohistochemical method, and a surgical specimen is subjected to staining analysis; the lung adenocarcinoma can be noninvasively and accurately predicted in an early stage, and a scientific basis is provided for early intervention and treatment of the lung adenocarcinoma.

Description

Technical Field

[0001] The present invention belongs to the technical field of molecular biology testing, and particularly relates to a detection system, kit and application for stage IA lung adenocarcinoma. Background Art

[0002] Lung cancer is the leading cause of cancer-related deaths, with both high incidence and mortality rates. Adenocarcinoma (ADC) is the most common type of primary lung tumor, accounting for approximately 40% of all lung cancer cases. Unfortunately, most patients are diagnosed at an advanced stage, missing the optimal treatment opportunity and increasing the risk of death, especially in low- and middle-income countries. Early detection of lung cancer is crucial for high-risk patients, especially the identification of stage IA lung cancer. Timely intervention and treatment can improve the prognosis of patients and increase the success rate of treatment. The NCCN guidelines recommend 18 F-FDG PET / CT is widely used for the staging and restaging of lung cancer. However, false negatives may occur when evaluating nodules with a diameter less than 1.0 cm (T1a), adenocarcinoma in situ (AIS), or minimally invasive adenocarcinoma (MIA).

[0003] With the development of imaging, especially the increasing improvement and popularization of spiral CT, the detection rate of pulmonary nodules (diameter ≤ 3 cm) has increased significantly. However, the qualitative diagnosis of pulmonary nodular lesions remains difficult, which is a key and difficult problem in clinical diagnosis and treatment. Early and clear diagnosis of the disease is of great significance for clinical correct treatment and patient prognosis.

[0004] Non-invasive PET / CT imaging can integrate the functional metabolism and anatomical information of PET and CT imaging. 18 The application of F-FDG PET / CT in the diagnosis of lung cancer is currently very extensive. 18 F-FDG PET / CT shows high uptake in lung adenocarcinoma lesions and metastases larger than 3 cm. Because larger lesions have more solid components, which are composed of densely packed tumor cells and the tumor surrounding stroma, while stage IA adenocarcinoma (diameter ≤ 3 cm) shows relatively low uptake due to sparse tumor cells and less tumor surrounding stroma. 18 F-FDG PET / CT is difficult to accurately evaluate stage IA adenocarcinoma. Therefore, finding a method for early, non-invasive and accurate identification of stage IA lung adenocarcinoma is an important topic in the field of lung tumor diagnosis and treatment. Currently, clinically, 18 F-FDG PET / CT imaging is mainly used to evaluate stage IA lung adenocarcinoma, but 18 the sensitivity and specificity of F-FDG imaging agents are relatively low. Therefore, there is an urgent clinical expectation to develop new molecular probes for complementation.

[0005] Studies have shown that lung cancer is associated with fibroblasts at all stages of disease development, including tumorigenesis, proliferation, and metastasis. Fibroblasts play a subtle role in the regulation process in the tumor microenvironment and are considered cancer-associated fibroblasts (CAFs). The presence of CAFs is associated with various types of tumor entities, including non-small cell lung cancer (NSCLC). CAFs are identified by their marker, fibroblast activation protein (FAP). FAP belongs to the type II transmembrane cell surface serine protease of the dipeptidyl peptidase (DPP) family, consisting of 760 amino acids, with endopeptidase and DPP activities, and is considered the most promising target in cancer diagnosis and therapeutics. FAP is expressed during human development but rarely expressed in healthy adult tissues. FAP is the main component of the stroma around tumor epithelial cells, accounting for up to 90%. The amount of FAP expressed by CAFs is much higher than that in normal tissues, making FAP a good molecular target for tumors.

[0006] Compared with antibodies, fibroblast activation protein inhibitors (FAPIs) have a much smaller relative molecular mass, while retaining specific affinity for FAP, optimizing image quality. In particular, FAPI-04 exhibits better pharmacokinetic and biochemical properties, showing higher stability and lower clearance rate in tumor tissues, and higher and longer uptake in tumor lesions. Therefore, FAPI-04 is considered the most promising molecular probe for clinical application.

[0007] Currently, FAPI molecular probes have been applied to the diagnosis and staging of solid tumor lesions. This imaging agent has good pharmacokinetics and biological distribution, high sensitivity, and can clearly display tumor characteristics and a high tumor background ratio in common solid tumors. It can visualize the stroma in the tumor microenvironment and provide better image quality than FDG probes. Pang et al. found that FAPI PET / CT is more sensitive than FDG PET / CT in the examination of primary pancreatic tumors, metastatic lymph nodes, and distant metastases, and has obvious advantages in TNM staging. However, the accuracy of a single FAPI molecular probe in evaluating stage IA adenocarcinoma is not stable.

[0008] Therefore, there is an urgent need to develop a detection system that can efficiently and accurately evaluate stage IA lung adenocarcinoma. Summary of the Invention

[0009] The purpose of the present invention is to provide a detection system for stage IA lung adenocarcinoma, which can early, efficiently, and accurately evaluate stage IA lung adenocarcinoma.

[0010] To achieve the above purpose, the present invention provides the following technical solutions:

[0011] The present invention provides a detection system for stage IA lung adenocarcinoma, which includes a sample image analysis module, a classification and evaluation module, and an evaluation and verification module;

[0012] The sample image analysis module is configured to perform reconstruction analysis based on the PET / CT image of the sample to determine the maximum standardized uptake value of the sample;

[0013] The classification and evaluation module is configured to calculate the ratio of the tumor to the background of the sample based on the maximum standardized uptake value of the sample to distinguish between tumors and non-tumor lesions;

[0014] The evaluation and verification module is configured to evaluate the tumor stroma ratio based on the classification result and verify the expression of FAP in tumor tissues;

[0015] The detection system further includes a comprehensive evaluation of the expression of FAP, which is determined based on semi-quantitative immunohistochemical scoring.

[0016] The present invention also provides the specific operation steps of the above detection system as follows:

[0017] S1. The sample is subjected to PET / CT acquisition in the sample image analysis module. After PET imaging, image reconstruction analysis is performed to determine the maximum standardized uptake value of the sample;

[0018] S2. According to the maximum standardized uptake value described in S1, calculate the ratio of the tumor to the background of the sample to distinguish between tumors and non-tumor lesions;

[0019] S3. Evaluate the tumor stroma ratio within the tumor described in S2, and use immunohistochemical analysis to evaluate the expression of FAP in tumor tissues.

[0020] The present invention also provides a detection kit for stage IA lung adenocarcinoma, which is applied to the detection system for stage IA lung adenocarcinoma as described above and includes 18 F-FAPI-04, 5-micron hematoxylin-eosin, and FAP-α antibody.

[0021] Preferably, the 18 F-FAPI-04 is a PET molecular probe that targets fibroblast activation protein FAP in tumors.

[0022] The present invention also provides the application of the above detection kit for stage IA lung adenocarcinoma. The kit is configured in the detection system for lung adenocarcinoma as described in any one of claims 1-2 to assist in the detection and evaluation of stage IA lung adenocarcinoma.

[0023] Advantages of the present invention:

[0024] The PET molecular probe FAPI-04 targeting tumor fibroblasts shows higher and longer-lasting uptake in stage IA lung adenocarcinoma,18 The SUVmax value, a semi - quantitative index available for F - FAPI - 04 PET / CT, can be used for early, non - invasive evaluation and diagnosis of stage IA lung adenocarcinoma. Brief Description of the Drawings

[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0026] Figure 1 For 18 The correlation between F - FAPI - 04 uptake and histological FAP expression and 18 The correlation of F - FAPI - 04 ROC curve in diagnosing stage IA IAC; A. The bar chart shows the 18 significant difference in F - FAPI - 04 SUVmax between FAP IHC score 1' and 3' groups; B. The ROC curve represents the 18 diagnostic efficacy of F - FAPI - 04 SUVmax in differentiating IAC from AIS and MIA; C. The bar chart shows the 18 difference in F - FAPI - 04 SUVmax between the low stroma group and the high stroma group; D. The ROC curve represents the 18 diagnostic efficacy of F - FAPI - 04 TBR in differentiating IAC from AIS and MIA; *P < 0.05, **P < 0.01, ns, not significant.

[0027] Figure 2 It is a flow chart of the research process;

[0028] Figure 3 A - D are respectively 18 F - FAPI - 04 PET / CT MIP, PET, CT and axial fusion images; E - H are respectively 18 F - FDG PET / CT MIP, PET, CT and axial fusion images;

[0029] Figure 4 For 18 F - FAPI - 04 imaging, HE staining and FAP immunohistochemical images of different - differentiated lung adenocarcinomas;

[0030] Figure 5 For 18 F - FAPI - 04 PET / CT axial fusion images and FAP immunohistochemical staining images of lung adenocarcinomas with different FAP - IHC scores and different clinical stages. Detailed Embodiments

[0031] The present invention provides a lung adenocarcinoma detection system, characterized in that the system includes a sample image analysis module, a classification and evaluation module, and an evaluation and verification module.

[0032] The specific detection system of the present invention includes:

[0033] (1) The sample is collected by PET / CT in the sample image analysis module. After PET imaging, image reconstruction analysis is performed to determine the maximum standardized uptake value of the sample.

[0034] (2) According to the above maximum standardized uptake value, calculate the ratio of tumor to background of the sample to distinguish tumors from non-tumor lesions.

[0035] (3) Evaluate the tumor stroma ratio in the above tumor, and use immunohistochemical analysis to evaluate the expression of FAP in tumor tissues.

[0036] In the embodiment of the present invention, by comparing 18 F-FDG PET / CT and 18 F-FAPI-04 PET / CT imaging combined with immunohistochemical methods to detect and analyze the expression of FAP, and perform staining analysis on surgical specimens; the results show that the PET molecular probe FAPI-04 of the present invention can target fibroblast activation protein in tumors, showing higher and longer uptake in stage IA lung adenocarcinoma with a diameter ≤ 3 cm and metastases. Using the semi-quantitative index SUVmax value, it can detect stage IA lung adenocarcinoma early, accurately, and non-invasively, providing a scientific basis for early intervention and treatment of lung adenocarcinoma.

[0037] A PET molecular probe is a probe based on molecular imaging technology that can be localized and observed in a living body. Its basic principle is to label a radionuclide to a specific molecule, and these molecules can bind to specific molecules or tissues in a living body. When these radionuclides decay, they emit positrons, which collide with the negative electrons in the living body and are captured by the PET instrument, so that the molecules and tissues in the living body can be localized and observed.

[0038] The Spearman rank correlation coefficient (Spearman correlation coefficient) is a non-parametric statistical index that measures the correlation between two variables, proposed by Charles Edward Spearman. It evaluates the monotonic relationship between variables by converting the variables into rank data and calculating the correlation between ranks. The value range of the Spearman correlation coefficient is from -1 to 1, and the closer the absolute value is to 1, the stronger the correlation.

[0039] The Bayesian penalized likelihood algorithm (Q.Clear, GE Healthcare) is a statistical inference method that combines Bayesian methods and penalized likelihood techniques, mainly used for processing ultra-high dimensional data. This algorithm controls the complexity of the model by introducing a penalty term, thereby effectively preventing overfitting while maintaining the flexibility of the model.

[0040] Advantage Workstation (AW) is an advanced imaging workstation launched by GE Healthcare, aiming to provide advanced visualization and diagnostic functions to help clinicians access and process medical imaging data more quickly.

[0041] HRCT (High Resolution CT), namely high-resolution CT, is an examination technique for thin-layer (1-2 mm) scanning and high-resolution algorithm (usually bone algorithm) reconstructed images. Sometimes, the voltage and current need to be appropriately increased. It uses a traditional CT scanner, but some parameters will be precisely set during imaging to maximize the spatial resolution.

[0042] The McNemar test is a statistical test method mainly used to compare the significance of the differences in classification results of two related samples in a binary classification problem.

[0043] The Spearman correlation coefficient is a non-parametric statistical method used to measure the dependence of two variables. Especially when the data does not conform to a linear relationship, it can provide a more robust evaluation.

[0044] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0045] 18 F-FDG and 18 F-FAPI-04 was provided by Dongcheng AMS (Guangdong, China) Pharmaceutical Co., Ltd., and the 20 MeV cyclotron was from Sumitomo, Japan, CYPRIS HM-20.

[0046] In the embodiments of the present invention, the production processes, experimental methods or detection methods involved, unless otherwise specified, are all conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the art and are very clear and definite in the relevant application fields. Those skilled in the art can understand the conventional process steps according to the name and apply the corresponding equipment, and implement them under conventional conditions or conditions recommended by the manufacturer.

[0047] In the embodiments of the present invention, there are no special restrictions on the sources of various instruments, equipment, raw materials or reagents used. They are all conventional products that can be obtained through regular commercial channels, and can also be prepared according to the conventional methods well-known to those skilled in the art.

[0048] Example 1

[0049] 1.1 Research design and population

[0050] Twenty patients were enrolled from February 2023 to October 2023 and underwent 18 F-FDG PET / CT examination at Maoming People's Hospital. All patients underwent 18 F-FAPI-04 PET / CT examination within one week, and were pathologically diagnosed with lung adenocarcinoma (stage IA) by surgery. None of them had received surgery or other anti-tumor treatments before the examination.

[0051] 1.2 Inclusion criteria

[0052] The criteria for patients to be included in this study were as follows: (1) age > 18 years; (2) Eastern Cooperative Oncology Group (ECOG) score ≤ 1; (3) patients with pulmonary nodules (≤ 3 cm) detected by CT scan and undergoing tumor marker examination; (4) obtaining informed consent; (5) only receiving symptomatic treatment, without any history of chemotherapy, radiotherapy or surgical resection before PET / CT scan.

[0053] 1.3 Exclusion criteria

[0054] Patients were excluded for the following reasons: (1) pulmonary nodules > 3 cm; (2) pregnant or suspected pregnant participants; (3) 18 The interval between 18 F-FDG PET / CT and

[0055] F-FAPI-04 examination exceeded one week; (4) more than two primary malignant tumors; (5) lack of final histological pathological diagnosis. 18 Preparation of 18 F-FDG and

[0056] 18 F-FDG and 18 F-FAPI-04 were prepared using a 20 MeV cyclotron. The synthesis of the radiotracer 18F-FAPI-04 strictly followed the protocol published by Wei et al., and the radiochemical purity exceeded 95%.

[0057] 1.5 Patient preparation and 18 F-FDG PET / CT protocol

[0058] According to the recommendations of the EANM in its version 2.0 tumor imaging guidelines, all patients undergoing examination should fast for at least 4 hours before injection of 18 the 18 F-FDG imaging agent, avoid strenuous exercise before PET / CT acquisition. After the patient arrives at the preparation room of the PET / CT examination room, they should change clothes and remove metal ornaments. Before injection of 18 F-FDG, the blood glucose level measured by a blood glucose meter should be ≤ 11.1 mmol / L (about 200 mg / dL). All patients received manual injection of 18 F-FDG at 4.44 MBq / kg (0.12 mCi / kg), and the patients were informed to lie flat and rest in a dimly lit and quiet lounge. 45 minutes after injection of the imaging agent, the patients were instructed to drink water (about 800 - 1000 ml), empty the bladder, and wait outside the PET / CT as prompted. At 18 60 ± 5 minutes after injection of 18 F-FDG, the patient lies on the examination table of the Discovery MI scanner (GE Healthcare, USA) as instructed, and PET / CT acquisition is started. Before PET imaging, low-dose CT scanning (120 kV, 80 mA, 3.75 mm thick) is used for anatomical localization and scanning. Subsequently, PET data acquisition is initiated. The PET acquisition range is from the skull vertex to the upper thigh, and each patient is acquired in 5 - 8 segments, with each segment having a length of 20 cm. During PET acquisition, the patient maintains shallow breathing, and all patients undergo breath-hold high-resolution CT scanning of the chest (120 kV, 150 mA, 1.25 mm thick). PET image reconstruction uses the Bayesian penalized likelihood algorithm with a penalty factor (b = 750) (Q.Clear, GE Healthcare). The low-dose CT scanning data is used for attenuation correction and anatomical localization.

[0059] 1.6 Patient preparation and 18 F-FAPI-04 PET / CT protocol

[0060] The same patient undergoes 18 F-FDG and 18 F-FAPI-04 PET / CT examinations at an interval of one week. It is recommended that the patient avoid strenuous physical activities before 18 F-FAPI-04 PET / CT, and no additional specific preparation is required. 18 F-FAPI-04 PET / CT examination is performed 1 hour after injection of 4.44 MBq / kg (0.12 mCi / kg). The operation steps and image analysis are the same as those of 18 F-FDG PET / CT. 18

[0061] 1.7 PET / CT image analysis

[0062] All the collected data were transferred to the Advantage Workstation (AW version 4.7, GE Healthcare), and the attention-corrected PET, CT, and fused PET / CT images (axial, coronal, and sagittal planes) were reviewed and analyzed.

[0063] The long diameter of the pulmonary nodules was measured on the HRCT images. 18 F-FDG or 18 Focal accumulations with higher F-FAPI-04 metabolism than the surrounding tissues were considered positive lesions, indicating suspected malignant lesions. Semi-automatic 3D delineation was used to measure 18 F-FDG or 18 the tumor lesions showing F-FAPI-04 uptake.

[0064] According to the EANM guidelines, a 3D isovolumetric volume (VOI) established with a 41% threshold of the maximum standardized uptake value (SUVmax) was applied, and SUVmax, mean standardized uptake value (SUVmean), and peak standardized uptake value (SUVPeak) were automatically generated within a 1-cm 3 sphere volume. 18 The metabolic tumor volume (MTV) and total lesion glycolysis (TLG) measured by F-FDG, 18 the corresponding values measured by F-FAPI-04: FAPI average tumor volume [FTV] and total lesion FAP expression [TLF] were recorded together.

[0065] A circular VOI with a diameter of 1 cm and a length of 2 cm (parallel to the descending aorta) was used to measure 18 F-FDG or 18 the uptake of F-FAPI-04 in the mediastinal blood pool, and the SUVmean of the mediastinal blood pool was recorded. The tumor-to-background ratio (TBR) was calculated by dividing the maximum standardized uptake of the tumor (SUVmax) by the SUV mean of the mediastinal blood pool. The calculation formula used was as follows:

[0066]

[0067] 1.8 Histological and immunohistochemical analysis

[0068] According to the 2021 World Health Organization (WHO) classification of lung tumors, invasive non-mucinous adenocarcinomas are classified into well-differentiated, moderately differentiated, or poorly differentiated types. The degree of differentiation, tumor size (long-axis diameter), pathological TNM stage (I, II, or III), and tumor stroma ratio (TSR) were recorded. The assessment of TSR was based on previous reports, and routine microscopic examination of 5-μm hematoxylin and eosin (H&E)-stained primary tumor sections was performed using a 5× microscope (total magnification of 50×). The most invasive part of the primary tumor was selected for further evaluation. This area usually consists of two components: the tumor cell component and the stromal component, and these two components are expressed as percentages.

[0069] The percentage of tumor cells in a single field of view was calculated by observing with a 100× microscope, and the remaining percentage was the TSR. For example, if the stained area of tumor cells accounted for 40% of the field of view, then the percentage of the stromal component was 60%, and the TSR was 60%. The TSR was calculated in a range of 10%, i.e., 30%, 40%, etc. Two to four different areas of the field of view were selected for evaluation, and the highest TSR value was used to determine the final value. For the evaluation of TSR within the tumor, tumor cells must be present around the stroma in the selected field of view, and areas with more stroma and no tumor cells around should not be selected for evaluation. According to the TSR evaluation results, cases were grouped with a cut-off value of 50%. Cases with a TSR value greater than 50% were defined as the high-stroma group, and cases with a TSR value less than or equal to 50% were defined as the low-stroma group.

[0070] Immunohistochemistry (IHC) staining confirmed the expression of FAP in tumor tissues. Surgical specimens were fixed and embedded, then sectioned and immunostained with an anti-FAP-α antibody (1:100; ab207178, Abcam). All surgical specimens were stained and the results analyzed to measure the intensity and percentage of FAP-positive cells. The staining intensity was divided into 4 grades, and image acquisition was performed using the Mantra multispectral imaging platform.

[0071] The expression of FAP was comprehensively evaluated in cross-sectional areas of tumors and adjacent non-malignant tissues. Semi-quantitative immunohistochemistry (IHC) scoring was performed to determine the percentage of FAP-positive cells and their staining intensity under an optical microscope at ×10 / 20 magnification.

[0072] The FAP intensity score was: 0 (none), 1 (weak), 2 (moderate), 3 (strong), and the semi - quantitative percentage of FAP - positive cells was 0 (0%), 1 (1 - 25%), 2 (26 - 50%), 3 (51 - 75%), 4 (76 - 100%) respectively. The final FAP IHC score ranged from 0 to 12, which was calculated by multiplying the intensity and percentage values. According to this scoring system, tumors were classified as: negative marked as "0" (IHC score = 0), mild marked as "1" (IHC score between 1 - 4), moderate marked as "2" (IHC score between 5 - 8), and severe marked as "3" (IHC score between 9 - 12).

[0073] 1.9 Statistical analysis

[0074] Statistical analysis was performed. Descriptive statistical methods were used to compare the general information of the participants and the standardized uptake values of FDG and FAPI. Normally distributed data were expressed as mean ± standard deviation, and non - normally distributed data were expressed as the median of IQR. The paired two - sample t - test was used to compare the normally distributed data between two groups (FAPI and FDG - TBR), and the McNemar test was used to compare the non - normally distributed data. The McNemar χ2 four - fold table test was used to compare 18 F - FDG PET / CT and 18 F - FAPI PET / CT for diagnostic efficacy, and the results were as Figure 1 shown; A. The bar chart showed a significant difference in 18 F - FAPI - 04 SUVmax between the FAP IHC score 1' and 3' groups; B. The ROC curve represented the 18 diagnostic efficacy of F - FAPI - 04 SUVmax in differentiating IAC from AIS and MIA; C. The bar chart showed a difference in 18 F - FAPI - 04 SUVmax between the low stroma group and the high stroma group; D. The ROC curve represented the 18 diagnostic efficacy of F - FAPI - 04 TBR in differentiating IAC from AIS and MIA; *P < 0.05, **P < 0.01, ns, not significant.

[0075] Categorical variables were expressed as numbers (percentages). The Spearman correlation coefficient was used to test the consistency between imaging findings and pathological results. The median was used as the cut - off point for grouping continuous variables. The independent - sample t - test was used to compare the differences between two groups. In the receiver operating characteristic (ROC) analysis, different 18The area under the curve (AUC), sensitivity, and specificity thresholds of the F-FAPI-04 parameters were used to distinguish IAS + MIA from IAC. A P value < 0.05 was considered statistically significant. All statistical analyses were performed using R (version: 4.3.2) and RStudio (version: 2023.12.1 + 402). As shown in Table 1, it was indicated that: in all tumors, 18 F-FDG and 18 there were significant differences in the SUVmax and TBR of F-FAPI-04 metabolism (P < 0.05). The SUVmax and TBR of FAPI-04 were both higher than those of FDG.

[0076] Table 1: Comparison of 18 F-FDG and 18 F-FAPI-04 uptake

[0077]

[0078]

[0079] Note: Data are presented as mean ± standard deviation. The maximum standardized uptake value is SUVmax. FAPI = fibroblast activation protein inhibitor, 18F = fluorine-18, FDG = fluorodeoxyglucose. P values were used for comparison of 18 F-FDG uptake and 18 F-FAPI uptake. AIS = adenocarcinoma in situ, MIA = minimally invasive adenocarcinoma, IAC = invasive adenocarcinoma.

[0080] Example 2 18 Comparative analysis of 18 F-FAPI-04 and

[0081] A total of 20 patients were included in this study. Among them, 2 patients had 2 pulmonary nodules each, 1 patient had 3 pulmonary nodules, and the remaining patients had 1 pulmonary nodule each, for a total of 24 pulmonary nodules. All pulmonary nodules were surgically resected and confirmed as lung adenocarcinoma by histopathology and immunohistochemistry. Among them, 6 patients (7 pulmonary nodules) had surgery at an external hospital and surgical specimens could not be obtained. FAP immunohistochemical staining was performed on the remaining 17 nodules. The flow chart of the research process is as Figure 2 shown.

[0082] By comparing the MIP, PET, CT, and axial fusion images of 18 F-FAPI-04 PET / CT and 18 F-FDG PET / CT in the same patient, the results are as Figure 3 shown.

[0083] Figure 3One of the 20 patients, a 58-year-old female with moderately differentiated IAC (1.8 cm in diameter, stage IA2) in the apical segment of the right upper lobe of the lung, as Figure 3 shown in A-3D are 18 F-FAPI-04 PET / CT MIP, PET, CT, and axial fusion images, showing significantly increased metabolism of the lesion (red arrow) (SUVmax = 6.0, P < 0.05; tumor-to-background ratio (TBR) = 5.5, total lesion expression of fibroblast activation protein (TLF) = 14.4 cm 3 ); Figure 3 Shown in E-3H are 18 F-FDG PET / CT MIP, PET, CT, and axial fusion images, showing slightly increased metabolism of the lesion (red arrow) (SUVmax = 1.6, TBR = 0.6, TLF = 4.0 cm 3 ); Among them, Figure 3 the black arrows in A, 3E indicate radioactive uptake in the right rib fracture.

[0084] The results show that: in stage IA LUAD, 18 the SUVmax, TBR, and TLF of F-FAPI-04 are all significantly higher than 18 those of F-FDG (P < 0.01).

[0085] Example 3 Verification by Different Methods 18 Trial of F-FAPI-04 in the Detection of Stage IA Lung Adenocarcinoma

[0086] Randomly select one male and one female from the 20 patients in Example 2.

[0087] The results are as Figure 4 shown, being the 18 F-FAPI-04 imaging, HE staining, and FAP immunohistochemical images of different differentiated lung adenocarcinomas. Figure 4 A-4E is a 76-year-old female patient with well-differentiated MIA (1.5 cm in diameter, stage IA2) in the apical posterior segment of the left upper lobe of the lung; Figure 4 A-4C is 18 F-FAPI-04 PET, CT, and axial fusion images showing slightly increased metabolism of the lesion (SUVmax = 2.5, TBR = 2.1, TLF = 1.3 cm 3 ); Figure 4 D-4E shows rich tumor stroma by HE staining (TSR > 50%), and mild expression of tumor stroma by FAP-IHC (IHC score = 1', ×200).

[0088] Figure 4F-4J is a 51-year-old male patient among 20 patients, with moderately differentiated IAC in the apical segment of the right upper lobe (1.7 cm in diameter, stage IA2); Figure 4 F-4H is 18 F-FAPI-04 PET, CT, and axial fusion images show increased metabolic activity in the lesion (SUVmax = 2.9, TBR = 3.6, TLF = 2.4 cm 3 ); Figure 4 I-4J shows less abundant tumor stroma by HE staining (TSR < 50%), and moderate expression of tumor stroma by FAP-IHC (IHC score = 2', ×200).

[0089] The results show that the IHC FAP expression level is associated with histopathological characteristics in LUAD specimens.

[0090] Example 4 verifies 18 the accuracy of F-FAPI-04

[0091] As Figure 5 shown, the F-FAPI-04 PET / CT axial fusion images and FAP immunohistochemical staining images of lung adenocarcinoma with different FAP-IHC scores and different clinical stages 18 are presented.

[0092] Figure 5 A-5B is a 50-year-old male patient among 20 patients, with moderately differentiated IAC in the apical-posterior segment of the left upper lobe (1.5 cm in diameter, stage IA2), ( Figure 5 A, arrow) 18 The F-FAPI-04 PET / CT axial fusion image shows increased metabolic activity in the lesion (SUVmax = 2.3, TBR = 1.9, TLF = 2.8 cm 3 ); Figure 5 B shows mild expression of tumor stroma by FAP-IHC (FAP-IHC score = 1', ×20).

[0093] Figure 5 C-5D is a 52-year-old female patient among 20 patients, with well-differentiated IAS in the posterior basal segment of the right lower lobe (1.1 cm in diameter, stage IA2), ( Figure 5 C, arrow) 18 The F-FAPI-04 PET / CT axial fusion image shows increased metabolic activity in the lesion (SUVmax = 4.5, TBR = 4.5, TLF = 1.3 cm 3 ); Figure 5 D shows moderate expression of tumor stroma by FAP-IHC (FAP-IHC score = 2', ×20).

[0094] Figure 5E-5F is a 66-year-old male patient among 20 patients, with moderately differentiated IAC in the dorsal segment of the lower lobe of the left lung (diameter 1.2 cm, stage IA2), ( Figure 5 E, arrow) 18 The axial fused image of F-FAPI-04 PET / CT shows increased metabolism of the lesion (SUVmax = 4.3, TBR = 2.9, TLF = 3.5 cm 3 ); Figure 5 F shows obvious expression in the tumor stroma by FAP-IHC (FAP-IHC score = 3’, ×20);

[0095] Figure 5 G-5H is a 73-year-old female patient among 20 patients, with moderately differentiated IAC in the posterior segment of the upper lobe of the right lung (diameter 1.0 cm, stage IA1), ( Figure 5 G, arrow) 18 The axial fused image of F-FAPI-04 PET / CT shows increased metabolism of the lesion (SUVmax = 3.5, TBR = 4.4, TLF = 2.2 cm 3 ); ( Figure 5 H) FAP-IHC shows moderate expression in the tumor stroma (FAP-IHC score = 2’, ×20).

[0096] Figure 5 I-5J is a 63-year-old female patient among 20 patients, with moderately differentiated IAC in the posterior basal segment of the lower lobe of the left lung (diameter 1.2 cm, stage IA2), ( Figure 5 I, arrow) 18 The axial fused image of F-FAPI-04 PET / CT shows significantly increased metabolism of the lesion (SUVmax = 6.1, TBR = 6.1, TLF = 18.6 cm 3 ); Figure 5 J shows obvious expression in the tumor stroma by FAP-IHC (FAP-IHC score = 3’, ×20);

[0097] Figure 5 K-5L is a 63-year-old male patient among 20 patients, with moderately differentiated IAC in the posterior basal segment of the lower lobe of the left lung (diameter 2.5 cm, stage IA3), ( Figure 5 K, arrow) 18 The axial fused image of F-FAPI-04 PET / CT shows significantly increased metabolism of the lesion (SUVmax = 4.1, TBR = 5.1, TLF = 4.6 cm 3 ); ( Figure 5 L) FAP-IHC shows strong expression in the tumor stroma (FAP-IHC score = 3’, ×20).

[0098] The results show that the lesions of stage IA LUAD18 F-FAPI-04 uptake is associated with histological FAP expression.

[0099] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A stage IA lung adenocarcinoma detection system, characterized in that: The system includes a sample image analysis module, a classification evaluation module and an evaluation verification module; The sample image analysis module is configured to perform reconstruction analysis based on the PET / CT image of the sample to determine the maximum standard uptake value of the sample; The classification evaluation module is configured to calculate the ratio of the sample tumor to the background based on the maximum standard uptake value of the sample to distinguish between tumors and non-tumor lesions; The evaluation and verification module is configured to evaluate the tumor stroma ratio based on the classification result and verify the expression of FAP in the tumor tissue; The detection system also includes a comprehensive assessment of the expression of FAP, which is determined based on a semi-quantitative immunohistochemical score.

2. The detection system according to claim 1, characterized in that: The following steps are involved: S1. The sample is collected by PET / CT in the sample image analysis module, and image reconstruction analysis is performed after PET imaging to determine the maximum standard uptake value of the sample; S2. Calculate the ratio of sample tumor to background based on the maximum standard uptake value described in S1 to distinguish tumor from non-neoplastic lesions; S3. Evaluate the tumor-stroma ratio within the tumor described in S2, and evaluate the expression of FAP in tumor tissue using immunohistochemical analysis.

3. A stage IA lung adenocarcinoma detection kit, characterized in that: The detection kit is applied to the stage IA lung adenocarcinoma detection system as claimed in claim 1, comprising 18 F-FAPI-04, 5 μM hematoxylin-eosin, and FAP-α antibody.

4. The kit according to claim 3, characterized in that: Said 18 F-FAPI-04 is a PET molecular probe that targets tumor fibroblast activation protein FAP.

5. Application of a stage IA lung adenocarcinoma detection kit, characterized in that: The kit is configured in the lung adenocarcinoma detection system as described in any one of claims 1-2, and is used to assist in the detection and evaluation of stage IA lung adenocarcinoma.

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