Intrahepatic cholangiocarcinoma targeted therapy molecular typing system based on FGFR2 (Fibroblast Growth Factor Receptor 2) gene fusion detection

Abstract

The invention provides an intrahepatic cholangiocarcinoma targeted therapy molecular typing system based on FGFR2 (Fibroblast Growth Factor Receptor 2) gene fusion detection. The intrahepatic cholangiocarcinoma targeted therapy molecular typing system based on FGFR2 gene fusion detection comprises a sample processing unit used for acquiring and processing an intrahepatic cholangiocarcinoma tissue sample according to a preset baseline sampling scheme; the FISH detection unit is provided with an FGFR2 gene specific probe and is used for detecting the fracture fusion state of the FGFR2 gene in the tumor tissue; an NGS detection unit; a histopathologic typing unit; and a data analysis and decision unit. The intrahepatic cholangiocarcinoma targeted therapy molecular typing system based on FGFR2 gene fusion detection has the advantages that by integrating fluorescence in situ hybridization and next-generation sequencing technologies and combining histopathologic characteristics, precise detection and molecular typing of the FGFR2 gene fusion state of ICC patients are achieved, and a decision basis is provided for clinical targeted therapy.

Description

Technical Field

[0001] This invention relates to the fields of tumor molecular diagnostics and precision medicine, and in particular to a molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection. Background Technology

[0002] Intrahepatic cholangiocarcinoma (ICC) is a malignant tumor originating from the lining epithelium of the secondary bile ducts to the smallest intrahepatic bile duct branches and the perichondral glands, accounting for 10%-20% of primary liver cancers. According to the "Expert Consensus on Pathological Diagnosis of Intrahepatic Cholangiocarcinoma (2022 Edition)," ICC can be divided into two main histological subtypes: large bile duct type and small bile duct type. Among them, small bile duct type ICC accounts for 41%-58%, and its characteristic molecular variations include IDH1 / 2 gene mutations and FGFR2 gene fusion / rearrangement.

[0003] Currently used targeted therapies such as pemigatinib and infigratinib have shown significant efficacy in patients with ICC due to FGFR2 fusions / rearrangements. However, existing technologies lack systematic molecular subtyping methods, making it impossible to comprehensively assess the correlation between FGFR2 gene fusion status and ICC histological subtypes, clinicopathological features, and prognosis, resulting in a lack of precise guidance for targeted therapy selection.

[0004] Therefore, it is necessary to provide a molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection to solve the above-mentioned technical problems. Summary of the Invention

[0005] The technical problem solved by this invention is to provide a molecular subtyping system for intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection, which integrates fluorescence in situ hybridization and next-generation sequencing technologies with histopathological features to achieve accurate detection and molecular subtyping of the FGFR2 gene fusion status in ICC patients, and provides a basis for decision-making in clinical targeted therapy.

[0006] To address the aforementioned technical problems, the present invention provides a molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection, comprising: a sample processing unit for acquiring and processing intrahepatic cholangiocarcinoma tissue samples according to a preset baseline sampling protocol; The FISH detection unit is equipped with an FGFR2 gene-specific probe to detect the breakage and fusion status of the FGFR2 gene in tumor tissue. The NGS detection unit is used to identify FGFR2 gene fusion breakpoints and partner genes using next-generation sequencing technology. Histopathological subtyping unit, used to determine the histological subtype of intrahepatic cholangiocarcinoma based on histomorphological and immunohistochemical characteristics; The data analysis and decision-making unit integrates FISH test results, NGS test results, and histopathological typing results to generate molecular typing reports and targeted therapy recommendations.

[0007] Preferably, the preset baseline sampling scheme is a "7-point baseline sampling" scheme, which includes: taking samples at the junction of the cancer and adjacent liver tissue at the 12 o'clock, 3 o'clock, 6 o'clock and 9 o'clock positions; taking at least one sample from the tumor in a location without bleeding or necrosis; and taking samples from liver tissue within ≤1cm and >1cm of the tumor edge.

[0008] Preferably, the interpretation criteria adopted by the FISH detection unit include: negative determination: red and green signals overlap and appear as yellow signals; positive determination: red and green signals are separated and the interval is greater than 2 signal point diameters, and one of the following conditions is met: single green signal ratio ≥15% of tumor cells; single red signal ratio ≥50% of tumor cells.

[0009] Preferably, the classification criteria implemented by the histopathological typing unit include: large bile duct type: characterized by cuboidal or columnar tumor cells containing myxoid cytoplasm, and positive for S-100P protein and MUC5AC immunohistochemical staining; small bile duct type: characterized by cuboidal or low columnar tumor cells, and positive for CRP and N-cadherin immunohistochemical staining.

[0010] Preferably, the molecular typing performed by the data analysis and decision-making unit includes: FGFR2 fusion-positive small bile duct type; FGFR2 fusion-positive large bile duct type; FGFR2 fusion-negative small bile duct type; and FGFR2 fusion-negative large bile duct type.

[0011] Preferably, the targeted therapy recommendations include: FGFR inhibitor therapy for FGFR2 fusion-positive patients; IDH1 inhibitor therapy for FGFR2 fusion-negative patients with small bile ducts but with IDH1 / 2 mutations; and KRAS-targeted therapy for FGFR2 fusion-negative patients with large bile ducts but with KRAS mutations.

[0012] Preferably, the FGFR inhibitor is selected from pemitinib or inflistatinib.

[0013] Preferably, the multi-gene panel used in the NGS detection unit contains probes for at least the FGFR2, IDH1, IDH2, and KRAS genes.

[0014] Preferably, it also includes a report generation unit for automatically generating a molecular diagnostic report containing the following: FGFR2 gene fusion status (positive / negative); detected FGFR2 fusion partner gene (if applicable); histopathological typing results; molecular subtype classification; and recommended targeted therapy regimen.

[0015] Preferably, the system further includes a quality control unit for monitoring the following quality indicators: signal intensity and background noise level of FISH detection; coverage and uniformity of NGS sequencing; tumor cell content and DNA quality of tissue samples.

[0016] Compared with related technologies, the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by this invention has the following beneficial effects: This invention provides a molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection. By leveraging the complementary advantages of FISH and NGS technologies, it significantly improves the sensitivity and specificity of FGFR2 fusion detection, avoiding the limitations of single technologies. The "7-point baseline sampling" scheme ensures sample representativeness and improves the reliability of test results. Molecular subtyping combined with histopathological characteristics achieves organic integration of morphological and molecular features, more comprehensively reflecting the biological characteristics of the tumor. The systematic subtyping scheme and automated report generation provide precise decision-making basis for targeted therapy in clinical practice. The built-in quality control unit ensures the reliability of data at each testing stage, improving the overall stability of the testing system. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Figure 2 This is a detection diagram of FISH detection in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Figure 3 This is another detection diagram of FISH detection in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention; Figure 4 This is a detection diagram of NGS in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Figure 5 This is another detection diagram of NGS detection in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Detailed Implementation

[0018] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0019] Please refer to the following: Figure 1 , Figure 2 , Figure 3 , Figure 4 and Figure 5 ,in, Figure 1This is a schematic diagram of the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Figure 2 This is a detection diagram of FISH detection in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Figure 3 This is another detection diagram of FISH detection in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention; Figure 4 This is a detection diagram of NGS in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. Figure 5 This is another detection diagram of NGS detection in the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by the present invention. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection includes: a sample processing unit for acquiring and processing intrahepatic cholangiocarcinoma tissue samples according to a preset baseline sampling protocol; a FISH detection unit equipped with FGFR2 gene-specific probes for detecting the breakpoint and fusion status of the FGFR2 gene in tumor tissue; an NGS detection unit for identifying FGFR2 gene fusion breakpoints and partner genes using next-generation sequencing technology; a histopathological subtyping unit for determining the histological subtype of intrahepatic cholangiocarcinoma based on tissue morphology and immunohistochemical characteristics; and a data analysis and decision-making unit for integrating FISH detection results, NGS detection results, and histopathological subtyping results to generate a molecular subtyping report and targeted therapy recommendations.

[0020] In this embodiment, the preset baseline sampling scheme is a "7-point baseline sampling" scheme, which includes: taking samples at the junction of the cancer and adjacent liver tissue at the 12 o'clock, 3 o'clock, 6 o'clock and 9 o'clock positions; taking at least one sample from the tumor in a location without bleeding or necrosis; and taking samples from liver tissue within ≤1cm and >1cm of the tumor edge.

[0021] In this embodiment, the interpretation criteria adopted by the FISH detection unit include: negative determination: red and green signals overlap and appear as yellow signals; positive determination: red and green signals are separated and the interval is greater than 2 signal point diameters, and one of the following conditions is met: single green signal ratio ≥15% of tumor cells; single red signal ratio ≥50% of tumor cells.

[0022] In this embodiment, the classification criteria implemented by the histopathological typing unit include: large bile duct type: characterized by cuboidal or columnar tumor cells containing myxoid cytoplasm, and positive for S-100P protein and MUC5AC immunohistochemical staining; small bile duct type: characterized by cuboidal or low columnar tumor cells, and positive for CRP and N-cadherin immunohistochemical staining.

[0023] In this embodiment, the molecular typing performed by the data analysis and decision-making unit includes: FGFR2 fusion-positive small bile duct type; FGFR2 fusion-positive large bile duct type; FGFR2 fusion-negative small bile duct type; and FGFR2 fusion-negative large bile duct type.

[0024] In this embodiment, the targeted therapy recommendations include: FGFR inhibitor therapy for FGFR2 fusion-positive patients; IDH1 inhibitor therapy for FGFR2 fusion-negative patients with small bile ducts but with IDH1 / 2 mutations; and KRAS targeted therapy for FGFR2 fusion-negative patients with large bile ducts but with KRAS mutations.

[0025] In this embodiment, the FGFR inhibitor is selected from pemitinib or infligratinib.

[0026] In this embodiment, the multi-gene panel used by the NGS detection unit contains probes for at least the FGFR2, IDH1, IDH2 and KRAS genes.

[0027] In this embodiment, a report generation unit is also included to automatically generate a molecular diagnostic report containing the following information: FGFR2 gene fusion status (positive / negative); detected FGFR2 fusion partner gene (if applicable); histopathological typing results; molecular subtype classification; and recommended targeted therapy regimen.

[0028] In this embodiment, the system further includes a quality control unit for monitoring the following quality indicators: signal strength and background noise level of FISH detection; coverage and uniformity of NGS sequencing; tumor cell content and DNA quality of tissue samples.

[0029] The specific operation steps of the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by this invention are as follows: S1: Sample Collection and Processing This system employs a standardized "7-point baseline sampling" method for sample collection. The specific procedure is as follows: First, the fresh surgical specimen is examined, and a cross-section is taken every 1 cm along the maximum diameter of the tumor to ensure representativeness. Tissue samples are taken at the junction of cancerous and normal tissue at four locations: the 12 o'clock, 3 o'clock, 6 o'clock, and 9 o'clock positions (based on the tumor center). Simultaneously, at least one tissue sample is taken from the tumor parenchyma (avoiding necrotic areas), and two samples of liver tissue are taken as controls, one within 1 cm of the tumor edge and the other outside. The collected tissue samples require immediate processing: one portion is fixed in 10% neutral buffered formalin solution (stored at 4°C), while the other portion of fresh tissue is rapidly frozen in liquid nitrogen and then transferred to an ultra-low temperature freezer at -80°C for storage. If lymph nodes or other special sites are found, they need to be marked and treated separately.

[0030] S2: Pathological examination: Histological evaluation was performed according to standard procedures: First, tissue sections were prepared to a thickness of 4 micrometers and subjected to routine hematoxylin and eosin (HE) staining. Serial sections were also prepared for subsequent immunohistochemical analysis. Immunohistochemical analysis required a combination of two biomarkers: S-100P protein and MUC5AC for identifying large bile casts, and CRP and N-cadherin for identifying small bile casts. Positive and negative controls were included in each assay to ensure quality.

[0031] When observed under a microscope: the characteristics of the large bile cast are irregular glandular structure, columnar cells, and positive cytoplasmic mucus staining; the characteristics of the small bile cast are dense small glandular structure, cuboidal cells, and diffuse positive CRP staining.

[0032] S3: FISH detection: The detection is performed using a specially designed probe: the probe system contains two fluorescent labels: a red probe at the 5' end of the FGFR2 gene (547 / 565 nm wavelength) and a green probe at the 3' end (495 / 517 nm wavelength), while the cell nucleus is stained with DAPI (367 / 452 nm wavelength).

[0033] The testing steps are as follows: Pretreatment of slides: First, bake at 82℃ for 2 hours, then dewax with xylene, and then hydrate with graded alcohols; Proteinase K digestion: digest at 37℃ for 28 minutes; Probe hybridization: denature DNA and probe simultaneously at 73℃ for 5 minutes, and then incubate at 37℃ overnight; Elution: use a specific elution buffer (2×SSC / 0.3%NP-40) to elute at 72℃ for 2 minutes; use a professional digital slide scanning system to simultaneously acquire images of three channels at 80x magnification.

[0034] When interpreting the results, at least 100 tumor cells need to be counted. The positive criteria are: the diameter of more than 2 signal points that are separated from the red and green signals, and either of the following conditions is met: the proportion of cells with a single green signal is ≥15%, or the proportion of cells with a single red signal is ≥50%.

[0035] like Figure 2-3As shown, the "FGFR2 (10q26) Gene Breakage Probe Reagent (Fluorescence In Situ Hybridization)" produced by Wuhan Kanglu Biotechnology Co., Ltd. was used. The probe design principle is as follows: the 5' end of the FGFR2 gene carries a red probe, and the 3' end of the FGFR2 gene carries a green probe. When the DNA fragments containing the red and green probes are in normal positions, the red and green signals overlap, showing a yellow signal, indicating that the upstream and downstream genes of FGFR2 are not broken at this point. At this time, it is judged as FGFR2 breakage fusion negative. Normal diploid cells show two yellow signals. When the DNA fragments containing the red and green probes are abnormally broken, the red and green signals are separated, showing independent red and green signals. When the interval between the two signals is at least greater than the diameter of two signal points, it indicates that the upstream and downstream genes of FGFR2 have broken at this point. When the breakage ratio reaches 15% of the total number of tumor cells, it is judged as positive. Figure 2 In the middle, several cells can be seen to contain three colors: red, green, and yellow.

[0036] Figure 3 In the text, one can see forms such as two reds and two greens, two reds and one green, or one red and two greens.

[0037] S4: Next-Generation Sequencing (NGS) Detection: A specialized assay suite containing multiple genes is used: First, DNA (at least 50 ng) is extracted from the sample, and then a sequencing library is constructed using a specific kit. During quality control, the tumor cell content must be ≥20% (microdissection may be necessary), the library concentration ≥2 nM, and the insert size between 200-300 bp.

[0038] Sequencing was performed using the Illumina sequencing platform, requiring a coverage of ≥500-fold for the target region (≥1000-fold for important regions). Specialized software was used for data analysis, with a focus on breaks in the Intron 17 region of chromosome 10, setting a 5% threshold as a clinically significant variant frequency.

[0039] like Figure 4-5 As shown, the Burning Rock Multigene Mutation Detection Kit was used with an Illumina NextSeq 550Dx gene sequencer, employing NGS sequencing. The principle involves using RNA probe capture technology, including multiple FGFR2 probes, to capture and enrich target regions in the DNA sample library. High-throughput sequencing enables the simultaneous detection of multiple variants, including FGFR2 gene breaks and fusions. Bioinformatics analysis extracts gene variant information at this location, providing variant type and frequency. Using IGV software combined with a human genome reference sequence, the bioinformatics file is converted into an image, visually displaying the nucleotide sequences of relevant genes within the patient's genome.

[0040] Figure 4 In the study, the FGFR2 gene was broken at chr10 Intron17 and fused abnormally with the EEA1 gene at chr12 Intron20, resulting in the loss of downstream gene expression and affecting protein function. The deletion frequency was 32.68%.

[0041] Figure 5 In the study, the FGFR2 gene was broken at chr10 Intron17 and fused abnormally with the CCDC186 gene at chr10 Intron8, resulting in the loss of downstream gene expression and affecting protein function. The deletion frequency was 45.57%.

[0042] S5: Data Integration and Report Generation The system automatically receives and analyzes various test data: first, it integrates FISH image analysis data, NGS variant detection results, and histopathological diagnostic conclusions. The system's built-in intelligent typing algorithm automatically determines the molecular subtype based on this data, such as: FGFR2-positive small bile duct type; FGFR2-positive large bile duct type; IDH-mutant small bile duct type; KRAS-mutant large bile duct type, etc.

[0043] The final report will include all key information: FGFR2 gene status, identified fusion genes (if any), pathological classification, molecular subtype classification, and treatment recommendations. It will also include various quality control parameters, such as sequencing coverage and tumor purity. If any quality indicators are found to be abnormal (e.g., insufficient signal strength, inadequate coverage), the system will automatically prompt for a retest.

[0044] S6: Treatment Recommendations: The system will provide tiered treatment recommendations based on the test results: Pemitinib is the top priority for patients with FGFR2 fusion-positive disease (highest level of evidence). Evonib is recommended for patients with IDH1 mutations (second highest level of evidence). For patients with KRAS mutations, the corresponding targeted therapy is recommended (lower level of evidence). The system will also incorporate the latest clinical research data to provide prognostic assessments, including predicted 1-year, 3-year, and 5-year survival rates.

[0045] Compared with related technologies, the molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection provided by this invention has the following beneficial effects: This invention provides a molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection. A standardized "7-point baseline sampling" protocol ensures sample representativeness. Combined with FISH testing to visually display FGFR2 gene breakage status and NGS technology to accurately identify fusion partner genes and accompanying mutations, it achieves dual verification at the molecular level and histopathological level. The system automatically generates a structured report containing molecular subtyping and targeted therapy recommendations by integrating multi-dimensional detection data. This provides clinicians with precise treatment decisions, such as using FGFR inhibitors for FGFR2 fusion-positive patients and IDH1 inhibitors for IDH1 / 2 mutation patients, significantly improving detection accuracy and treatment effectiveness. Simultaneously, a comprehensive quality control system and intelligent data analysis module ensure the reliability of the test results, enabling the system to provide precise management of intrahepatic cholangiocarcinoma patients from diagnosis to treatment, effectively improving patient prognosis.

[0046] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention specification and drawings, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection, characterized in that, include: The sample processing unit is used to acquire and process intrahepatic cholangiocarcinoma tissue samples according to a preset baseline sampling plan. The FISH detection unit is equipped with an FGFR2 gene-specific probe to detect the breakage and fusion status of the FGFR2 gene in tumor tissue. The NGS detection unit is used to identify FGFR2 gene fusion breakpoints and partner genes using next-generation sequencing technology. Histopathological subtyping unit, used to determine the histological subtype of intrahepatic cholangiocarcinoma based on histomorphological and immunohistochemical characteristics; The data analysis and decision-making unit integrates FISH test results, NGS test results, and histopathological typing results to generate molecular typing reports and targeted therapy recommendations.

2. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The preset baseline sampling scheme is a "7-point baseline sampling" scheme, including: Samples were taken at the junction of the cancerous and adjacent liver tissue at the 12 o'clock, 3 o'clock, 6 o'clock and 9 o'clock positions; At least one sample should be taken from a site where the tumor has no bleeding or necrosis; Liver tissue samples were taken from areas ≤1cm and >1cm from the tumor margin.

3. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The interpretation criteria adopted by the FISH detection unit include: Negative result determination: Overlapping red and green signals appear as a yellow signal; Positive result determination: Red and green signals are separated and the interval is greater than the diameter of two signal points, and one of the following conditions is met: Tumor cells with a single green signal ratio ≥15%; The proportion of single red signal cells is ≥50% in tumor cells.

4. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The classification criteria applied by the histopathological typing unit include: Bold casts: Characterized by cuboidal or columnar tumor cells containing myxoid cytoplasm, and positive for S-100P protein and MUC5AC immunohistochemical staining; Small bile duct type: Characterized by cuboidal or low columnar tumor cells, with positive CRP and N-cadherin immunohistochemical staining.

5. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The molecular subtyping performed by the data analysis and decision-making unit includes: FGFR2 fusion-positive small bile duct type; FGFR2 fusion positive bold tubular type; FGFR2 fusion-negative small bile duct type; FGFR2 fusion negative bold tubular type.

6. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The targeted therapy recommendations include: FGFR inhibitor therapy is recommended for patients with positive FGFR2 fusions; IDH1 inhibitor therapy is recommended for patients with small bile duct type who are FGFR2 fusion negative but have IDH1 / 2 mutations; KRAS-targeted therapy is recommended for patients with bold casts who are FGFR2 fusion-negative but have KRAS mutations.

7. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 6, characterized in that, The FGFR inhibitor is selected from pemitinib or infligratinib.

8. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The multi-gene panel used in the NGS detection unit contains probes for at least the FGFR2, IDH1, IDH2, and KRAS genes.

9. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, It also includes a report generation unit for automatically generating molecular diagnostic reports containing the following: FGFR2 gene fusion status; Detected FGFR2 fusion chaperone gene; Histopathological typing results; Molecular subtype classification; Recommended targeted therapy regimen.

10. The molecular subtyping system for targeted therapy of intrahepatic cholangiocarcinoma based on FGFR2 gene fusion detection according to claim 1, characterized in that, The system also includes a quality control unit for monitoring the following quality indicators: FISH detection measures signal strength and background noise levels; NGS sequencing coverage and uniformity; The tumor cell content and DNA quality of the tissue sample.

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