Application of HER-2 combined three-level lymph structure in diagnosis and treatment of mixed hepatocyte-cholangiocarcinoma

Abstract

The invention discloses application of HER-2 combined with a three-level lymph structure in diagnosis and treatment of mixed hepatocyte-bile duct cell carcinoma. A prognosis evaluation system and a treatment decision mode constructed on the basis of HER-2 detection are beneficial to establishment of a standard cHCC-CCA prognosis system, and the HER-2 detection can adopt a clinical conventional immunohistochemical method and is directly in butt joint with an existing pathological examination process, so that the clinical transformation threshold is effectively reduced, and the clinical treatment efficiency is improved. The clinical practicability and operability are obvious. Meanwhile, the expression level of HER-2 can reflect whether the patient can directly benefit from adjuvant chemotherapy, so that treatment schemes can be quickly generated for different patients after pathological examination. Moreover, the invention also discloses the association between the HER-2 and the tumor immune microenvironment, and provides a new thought for developing HER-2 and TLS combined prognosis products and targeted drug combined immunotherapy strategies.

Description

Technical Field

[0001] This invention relates to the field of tumor molecular marker technology, specifically to the application of HER-2 combined with tertiary lymphoid structures in the diagnosis and treatment of mixed hepatocellular-cholangiocarcinoma. Background Technology

[0002] Mixed hepatocellular-cholangiocarcinoma (cHCC-CCA) is a rare primary malignant liver tumor characterized by histological features of both hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC). Currently, treatment strategies for cHCC-CCA largely draw on clinical experience with hepatocellular carcinoma or intrahepatic cholangiocarcinoma. Surgical resection is the only radical treatment, but the indications for surgery are stringent, limiting its application to a small number of patients.

[0003] The prognosis for cHCC-CCA is extremely poor, with a median overall survival of only 32 months after surgical resection and a 5-year overall survival rate ranging from 23.6% to 36.4%. Postoperative recurrence rates are also high. Currently, there is a lack of targeted prognostic assessment indicators. Traditional tumor prognostic assessment methods rely on indicators such as TNM staging, vascular invasion, tumor size, and lymph node metastasis. These indicators are not accurate enough for predicting the prognosis of cHCC-CCA patients and cannot accurately predict their postoperative survival.

[0004] Furthermore, although postoperative adjuvant chemotherapy has been proven to have survival benefits in various tumors such as gastric cancer, breast cancer, and intrahepatic cholangiocarcinoma, there is currently no clear clinical consensus on whether adjuvant chemotherapy should be used in cHCC-CCA patients. Some patients may suffer additional medical burdens and adverse reactions due to blindly accepting chemotherapy, while some potentially beneficial patients may not receive timely and effective treatment.

[0005] The inventors disclosed a hybrid hepatocellular carcinoma prognostic model in patent CN120766967A, which combines traditional clinicopathological features and immune microenvironment indicators to construct a nomogram. It has good prognostic predictive performance for cHCC-CCA and can provide decision-making for postoperative adjuvant chemotherapy for patients at different risk levels. It confirms the key role of CD8 and FOXP3, two tumor immune microenvironment indicators, in the prognosis of cHCC-CCA.

[0006] However, at present, there are no independent prognostic indicators for cHCC-CCA, and the basis for prognostic and adjuvant therapy decisions is very limited. Therefore, it is of great significance to actively explore specific cHCC-CCA biomarkers and establish and improve a standardized prognostic assessment system and treatment decision-making method for cHCC-CCA. Summary of the Invention

[0007] Human epidermal growth factor receptor 2 (HER-2) is a receptor tyrosine kinase located on the cell surface. In healthy cells, when growth factors (ligands) bind to other receptors in the HER family, HER-2 pairs with them (forming a dimer), transmitting signals into the cell to regulate normal cell growth, division, and repair. As a member of the epidermal growth factor receptor (EGFR) family, HER-2 has been established as a prognostic factor and therapeutic target in various solid tumors, including breast cancer and gastric cancer. However, the expression pattern, prognostic significance, and predictive value of HER-2 in cHCC-CCA remain unclear.

[0008] In this application, through the analysis of clinical samples from 87 cHCC-CCA patients, it was found that HER-2 expression is an independent protective factor for postoperative overall survival (OS) in cHCC-CCA patients. The overall survival of HER-2 positive patients was longer than that of HER-2 negative patients, indicating that HER-2 can effectively distinguish cHCC-CCA patients with different prognostic risks.

[0009] In this application, the impact of adjuvant chemotherapy on overall survival was compared between different HER-2 expression groups, and PSM was used for verification. This demonstrated that HER-2 negative patients can benefit from adjuvant chemotherapy, but positive patients do not need to receive adjuvant chemotherapy.

[0010] Furthermore, this application analyzed the correlation between HER-2 and tertiary lymphoid structures (TLS), and found a significant positive correlation between HER-2 positivity and TLS scores, meaning that TLS was more enriched in tissues of HER-2 positive patients. Moreover, the accuracy of HER-2 positivity as an independent prognostic indicator was more pronounced in patients with higher TLS abundance. Therefore, combining HER-2 and tertiary lymphoid structures can further improve the prognostic ability of cHCC-CCA, and based on the correlation between the two, a treatment strategy combining HER-2 targeted drugs and immunotherapy can be constructed, which is beneficial for establishing a standard cHCC-CCA diagnosis and treatment protocol.

[0011] Based on the above experimental analysis, one objective of this invention is to provide the application of a HER-2 expression level detection reagent in the preparation of a prognostic assessment product for mixed hepatocellular-cholangiocarcinoma. Specifically, patients with HER-2 positivity have a better prognosis than those with HER-2 negativity. For example, HER-2 positivity patients have a longer overall survival than HER-2 negativity patients.

[0012] Another objective of this invention is to provide the application of a HER-2 expression level detection reagent combined with a tertiary lymphoid structure abundance detection reagent in the preparation of a prognostic assessment product for mixed hepatocellular-cholangiocarcinoma.

[0013] This invention also provides the application of HER-2 expression level detection reagent in the preparation of a product for predicting the benefit of adjuvant chemotherapy in mixed hepatocellular-cholangiocarcinoma. Patients with HER-2 negative results are suitable for adjuvant chemotherapy, while patients with HER-2 positive results are not suitable for adjuvant chemotherapy.

[0014] In some preferred embodiments, the adjuvant chemotherapy is preferably based on fluoropyrimidine, gemcitabine, or a combination of both. In one or more embodiments, adjuvant chemotherapy may also be other adjuvant treatments clinically used for mixed hepatocellular-cholangiocarcinoma.

[0015] The present invention also provides a pharmaceutical composition for treating mixed hepatocellular carcinoma, the pharmaceutical composition comprising a HER-2 targeted drug and an adjuvant chemotherapy drug, wherein the HER-2 targeted drug is used to treat patients with HER-2 positive mixed hepatocellular carcinoma, and the adjuvant chemotherapy drug is used to treat patients with HER-2 negative mixed hepatocellular carcinoma.

[0016] In one or more embodiments, the HER-2 targeted drug can be an existing HER-2 targeted drug, such as trastuzumab, pertuzumab, trastuzumab emtansine, trastuzumab dexamethasone, lapatinib, etc.

[0017] Furthermore, it also includes immunotherapy drugs for treating patients with mixed hepatocellular-cholangiocarcinoma who are HER-2 positive and have a high grade 3 lymphoid structure score.

[0018] This technical solution reveals the potential relationship between HER-2 and TLS, a key component of the tumor immune microenvironment. Based on the correlation study between HER-2 and the abundance of tertiary lymphoid structures, when HER-2 is positive, it further combines TLS-related immunotherapeutic drugs to improve the therapeutic effect.

[0019] In one or more embodiments, the TLS-related immunotherapeutic agent may be a PD-1 inhibitor, a PD-L1 inhibitor, or the like.

[0020] In one or more embodiments, the detection method for tertiary lymphoid structures may be immunohistochemistry, multiplex immunofluorescence, or other detection methods.

[0021] The present invention also provides a hybrid hepatocellular-cholangiocarcinoma prognostic device, comprising:

[0022] The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples;

[0023] The analysis unit is used to determine the prognosis of mixed hepatocellular-cholangiocarcinoma based on the expression level of HER-2, where HER-2 positivity corresponds to a better prognosis and HER-2 negativity corresponds to a worse prognosis.

[0024] The output unit is used to output the prognostic results.

[0025] In some preferred embodiments, the acquisition unit is also used to acquire the abundance of tertiary lymphoid structures in tumor tissue samples. If HER-2 is positive and tertiary lymphoid structures are present, the prognosis is better.

[0026] The present invention also provides a device for predicting the benefit of adjuvant chemotherapy in mixed hepatocellular-cholangiocarcinoma, comprising:

[0027] The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples;

[0028] The analysis unit is used to determine whether mixed hepatocellular-cholangiocarcinoma is suitable for adjuvant chemotherapy based on HER-2 expression levels. If HER-2 is negative, adjuvant chemotherapy is suitable; if HER-2 is positive, adjuvant chemotherapy is not suitable.

[0029] The output unit is used to output the predicted results of the benefits of adjuvant chemotherapy.

[0030] The present invention also provides a device for generating a hybrid hepatocellular-cholangiocarcinoma treatment regimen, comprising:

[0031] The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples;

[0032] The analysis unit is used to generate treatment plans based on HER-2 expression levels; if HER-2 is negative, adjuvant chemotherapy is recommended.

[0033] The output unit is used to output the treatment plan.

[0034] In one or more embodiments, if HER-2 is positive, the use of a HER-2 targeted drug is recommended.

[0035] In some preferred embodiments, the acquisition unit is also used to acquire the grade 3 lymphoid structure score in tumor tissue samples. When generating the treatment plan, the analysis unit recommends the use of HER-2 targeted drugs and / or immunotherapy drugs for HER-2 positive patients with a grade 3 lymphoid structure score of 1 to 3.

[0036] The present invention also provides a storage medium including a stored computer program, wherein, when the computer program is executed, the device where the storage medium is located is controlled to perform a method for generating a prognostic and / or treatment plan for mixed hepatocellular-cholangiocarcinoma, the method comprising the following steps:

[0037] Obtain HER-2 expression levels and grade III lymphoid structure scores from tumor tissue samples;

[0038] Prognosis is based on HER-2 expression levels and tertiary lymphoid structure scores, with a better prognosis for HER-2 positive expression and tertiary lymphoid structure, and a worse prognosis for HER-2 negative expression; and / or

[0039] Treatment plans are generated based on the HER-2 expression level and the tertiary lymphoid structure score. If the HER-2 expression level is negative, the treatment plan is adjuvant chemotherapy; if the HER-2 expression level is positive and the tertiary lymphoid structure score is 0, the treatment plan is HER-2 targeted therapy only; if the HER-2 expression level is positive and the tertiary lymphoid structure score is 1-3, the treatment plan is a combination of HER-2 targeted therapy and immunotherapy.

[0040] Output prognostic results and / or treatment plans.

[0041] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0042] 1. The prognostic assessment system and treatment decision-making method based on HER-2 detection in this invention are conducive to establishing a standard prognostic system for cHCC-CCA. Furthermore, HER-2 detection can be performed using routine clinical immunohistochemistry, directly connecting with existing pathological examination procedures, thereby effectively reducing the clinical translation threshold and demonstrating significant clinical practicality and operability.

[0043] 2. Through experiments, this invention has found that the expression level of HER-2 can reflect whether a patient can directly benefit from adjuvant chemotherapy, and thus treatment plans can be quickly generated for different patients after pathological examination.

[0044] 3. This invention also reveals the association between HER-2 and the tumor immune microenvironment, providing new ideas for developing HER-2 targeted drugs in combination with immunotherapy strategies. Attached Figure Description

[0045] The accompanying drawings, which are included to provide a further understanding of embodiments of the invention and form part of this application, do not constitute a limitation thereof. In the drawings:

[0046] Figure 1The following are the results of hematoxylin-eosin staining and HER-2 immunohistochemical detection in (A): HER-2 positive patients and (B): HER-2 negative patients after surgery, according to a specific embodiment of the present invention.

[0047] Figure 2 Kaplan-Meier curves for overall survival of HER-2 positive (HER-2(+)) and HER-2 negative (HER-2(-)) patients in a specific embodiment of the present invention;

[0048] Figure 3 The following is a comparison of Kaplan-Meier survival curves of overall survival in a propensity score-matched pre-cohort, where (A) represents HER-2 negative patients and (B) represents HER-2 positive patients; and a comparison of Kaplan-Meier survival curves of overall survival in a propensity score-matched post-cohort, where (C) represents HER-2 negative patients and (D) represents HER-2 positive patients.

[0049] Figure 4 The histopathological classification of the three-level lymphoid structures in a specific embodiment of the present invention is shown, wherein (A) is hematoxylin-eosin staining; (B) is CD20 immunohistochemical staining; (C) is CD21 immunohistochemical staining; (D) is CD4 immunohistochemical staining; and (E) is CD8 immunohistochemical staining.

[0050] Figure 5 This is a correlation comparison chart between TLS score and HER-2 expression in a specific embodiment of the present invention;

[0051] Figure 6 This paper presents a subgroup analysis and comparison of overall survival of HER-2 positive and negative patients under different clinicopathological factors in a specific embodiment of the present invention. Detailed Implementation

[0052] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the embodiments and accompanying drawings. The illustrative embodiments and descriptions of the present invention are only used to explain the present invention and are not intended to limit the present invention.

[0053] All raw materials used in this invention are not particularly limited in their source; they can be purchased commercially or prepared using conventional methods well-known to those skilled in the art. The purity of all raw materials used in this invention is not particularly limited; however, analytical grade or purity requirements conventional for biomarker research are preferred. All raw materials used in this invention have brand names and abbreviations that are conventional in the field, and each brand name and abbreviation is clearly defined within its relevant application. Those skilled in the art can obtain these materials from commercial sources or prepare them using conventional methods based on the brand name, abbreviation, and corresponding application.

[0054] In this application, clinicopathological characteristics of patients (age, gender, hepatitis B infection status, cirrhosis status, number of tumors, vascular invasion, maximum tumor diameter, AFP, CA19-9 concentration, etc.) and long-term follow-up data (median follow-up time 75.2 months) were systematically collected.

[0055]

Example 1

[0056] In this embodiment, immunohistochemistry (IHC) was used to detect HER-2 expression in tumor tissue samples from 87 patients with cHCC-CCA, and the overall survival of HER-2 positive and HER-2 negative patients was analyzed by survival curve analysis.

[0057] Specifically, tumor tissue samples surgically removed from cHCC-CCA patients were collected, fixed in formaldehyde, embedded in paraffin, and then prepared into 4 μm thick serial sections. These sections were routinely dewaxed to water for later use. Next, the sections were placed in citrate buffer (pH=6.0) and autoclaved for 15 minutes, then cooled to room temperature. 3% hydrogen peroxide solution was added and incubated at room temperature for 10 minutes to block endogenous peroxidase activity, followed by blocking buffer and incubation at room temperature for 30 minutes. Anti-HER-2 monoclonal antibody (1:100 dilution) was added and incubated overnight at 4°C. The next day, the sections were washed three times with PBS buffer for 5 minutes each time. Horseradish peroxidase-labeled secondary antibody was added and incubated at room temperature for 30 minutes, followed by three washes with PBS. Finally, DAB chromogenic solution was added and incubated at room temperature for 5 minutes. The staining was stopped by rinsing with water, counterstained with hematoxylin for 3 minutes, differentiated with hydrochloric acid ethanol, blued with ammonia, dehydrated and cleared, and mounted with neutral resin. The sections were interpreted by two pathologists in a double-blind manner. Figure 1 As shown, a brownish-yellow staining of the tumor cell membrane is considered positive, while no staining is considered negative. In case of inconsistent interpretation results, a consensus is reached through negotiation.

[0058] Based on the results of the double-blind interpretation, patients were divided into HER-2 positive and HER-2 negative groups. Survival curves were plotted using the Kaplan-Meier method, combined with clinical follow-up data (follow-up time ≥36 months). Figure 2 As shown, the results indicated that the 3-year overall survival (OS) rate was 62.5% in the HER-2 positive group and 31.5% in the negative group, with a statistically significant difference (P=0.0067), confirming that HER-2 can effectively distinguish cHCC-CCA patients with different prognostic risks.

[0059]

Example 2

[0060] In this embodiment, we investigated whether HER-2 positive and negative cHCC-CCA patients could benefit from adjuvant chemotherapy regimens and explored the role of HER-2 expression in treatment decisions.

[0061] Specifically, among the 87 patients enrolled in Example 1, 59 were HER-2 negative and 28 were HER-2 positive. All patients were pathologically confirmed and met the criteria for surgical resection, with a Child-Turcotte-Pugh classification of A or B. Postoperatively, based on multidisciplinary team evaluation and patient wishes, some patients received adjuvant chemotherapy, while the remaining patients did not.

[0062] Some patients with cHCC-CCA received the same adjuvant chemotherapy regimen, specifically adjuvant chemotherapy based on fluoropyrimidine, gemcitabine, or a combination of both, with each cycle lasting 21 days, for a total of 4-6 cycles. Adverse reactions were closely monitored during chemotherapy, and the dosage was adjusted as needed. After chemotherapy, patients were followed up regularly (every 3 months for 2 years) to record relapse time and survival status. The Log-rank test was used to compare overall survival and disease-free survival between the two groups.

[0063] Figure 3 The figure shows the overall survival of HER-2 negative and HER-2 positive patients with and without adjuvant chemotherapy. As can be seen from the figure, the overall survival of HER-2 negative patients who received adjuvant chemotherapy was significantly better than that of HER-2 negative patients who did not receive adjuvant chemotherapy. However, there was no significant difference in overall survival between HER-2 positive patients who received adjuvant chemotherapy and those who did not.

[0064] Furthermore, propensity score matching (PSM) was used to match HER-2 positive patients with HER-2 negative patients in a 1:2 ratio, controlling for confounding biases in patient baseline characteristics such as tumor number and vascular invasion, making the baseline characteristics of patients in different HER-2 expression groups comparable and verifying the reliability of the difference in efficacy between HER-2 expression and adjuvant chemotherapy.

[0065] The test results are as follows Figure 3 As shown, the overall survival results of the propensity score-matched cohort were largely consistent with those of the pre-matched cohort. This further confirms that HER-2-negative patients can benefit from adjuvant chemotherapy, while HER-2-positive patients do not require adjuvant chemotherapy. Based on these results, adjuvant chemotherapy can be used for HER-2-negative patients according to their HER-2 expression, while HER-2-positive patients can be treated with HER-2 targeted drugs such as trastuzumab and pertuzumab.

[0066]

Example 3

[0067] In this embodiment, the correlation between HER-2 and tertiary lymphoid structures (TLS) was verified.

[0068] Specifically, serial sections were prepared from tumor tissue samples from 87 patients with cHCC-CCA in Example 1.

[0069] CD20, CD4, CD8, and CD21 specific antibodies (fluorescently labeled) were added respectively, incubated overnight at 4°C, washed with PBS, and observed and imaged under a fluorescence microscope. The imaging results are as follows: Figure 4 As shown.

[0070] Subsequently, scores were awarded based on the quantity, completeness, and maturity of TLS. A score of 0 represented no TLS, 1 represented a small number of immature TLS with a loose structure, 2 represented a moderate number of TLS with a relatively complete structure, and 3 represented a large number of mature TLS with a dense structure and orderly distribution of immune cells.

[0071] Finally, Spearman rank correlation analysis was used to associate HER-2 expression status with TLS score. The results showed r=0.426, P=0.003, confirming a significant positive correlation between HER-2 positivity and TLS score, meaning that TLS was more enriched in tumor tissues of HER-2 positive patients. Figure 5 As shown, the TLS score was significantly higher in HER-2 positive patients than in HER-2 negative patients.

[0072] Furthermore, such as Figure 6 As shown, in most subgroups (e.g., age, sex, HBV infection status, and tumor diameter), the hazard ratio (HR) was less than 1, indicating that under this pathological condition, the risk of death for HER-2 positive patients was significantly lower than that for HER-2 negative patients, strongly confirming that HER-2 is an independent and favorable prognostic marker for cHCC-CCA. The results showed that adjuvant chemotherapy significantly improved the survival rate of HER-2 negative patients, bringing it closer to that of HER-2 positive patients; in contrast, the marginal benefit of chemotherapy was not significant for HER-2 positive patients due to their better baseline prognosis. Furthermore, subgroup analysis showed that there was no significant difference in prognosis between the two groups when the tertiary lymphoid structure (TLS) score was 0; however, when the TLS score was 1–3, the prognostic advantage of the HER-2 positive group was statistically significant. This indicates that the HER-2-mediated survival benefit is highly dependent on TLS abundance, i.e., regulated by the immune status of the tumor microenvironment. In conclusion, HER2 can not only serve as an independent prognostic marker, but its prognostic predictive value is also more prominent in patients who have not received adjuvant chemotherapy and those with higher TLS abundance. Therefore, in clinical practice, HER-2 abundance alone or in combination with TLS can be used to construct prognostic assessment models to further improve the accuracy of prognostic judgment.

[0073] Meanwhile, based on the correlation between the two, it is reasonable to construct a treatment approach for mixed hepatocellular carcinoma-cholangiocarcinoma, which could be to use HER-2-positive cHCC-CCA patients with TLS structures in combination with TLS-related immunotherapy, such as PD-1 inhibitors.

[0074]

Example 4

[0075] Based on the research results of the foregoing embodiments, this embodiment provides a hybrid hepatocellular-cholangiocarcinoma prognostic device, comprising:

[0076] The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples;

[0077] The analysis unit is used to determine the prognosis of mixed hepatocellular-cholangiocarcinoma based on the expression level of HER-2, where HER-2 positivity corresponds to a better prognosis and HER-2 negativity corresponds to a worse prognosis.

[0078] The output unit is used to output the prognostic results.

[0079] In this technical solution, the acquisition unit can be any existing device and method for detecting HER-2 expression levels, such as immunohistochemistry. Then, the HER-2 expression level is input to the analysis unit, which compares the input HER-2 expression level with a database. A positive HER-2 result indicates a better prognosis, such as longer overall survival, while a negative HER-2 result indicates a worse prognosis. Finally, the output unit outputs the prognostic results, facilitating physicians' assessment of the patient's prognosis.

[0080] In some preferred embodiments, the acquisition unit is also used to acquire the abundance of tertiary lymphoid structures in tumor tissue samples. If HER-2 is positive and tertiary lymphoid structures are present, it indicates a better prognosis.

[0081]

Example 5

[0082] Based on the research results of the foregoing embodiments, this embodiment provides a device for predicting the benefit of adjuvant chemotherapy in mixed hepatocellular-cholangiocarcinoma, comprising:

[0083] The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples;

[0084] The analysis unit is used to determine whether mixed hepatocellular-cholangiocarcinoma is suitable for adjuvant chemotherapy based on HER-2 expression levels. If HER-2 is negative, adjuvant chemotherapy is suitable; if HER-2 is positive, adjuvant chemotherapy is not suitable.

[0085] The output unit is used to output the predicted results of the benefits of adjuvant chemotherapy.

[0086]

Example 6

[0087] Based on the research results of the foregoing embodiments, this embodiment provides a device for generating a hybrid hepatocellular-cholangiocarcinoma treatment regimen, comprising:

[0088] The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples;

[0089] The analysis unit is used to generate treatment plans based on HER-2 expression levels; if HER-2 is negative, adjuvant chemotherapy is recommended.

[0090] The output unit is used to output the treatment plan.

[0091] In one or more embodiments, if HER-2 is positive, the use of a HER-2 targeted drug is recommended.

[0092] In some preferred embodiments, the acquisition unit further acquires a grade 3 lymphoid structure score from the tumor tissue sample. More preferably, the acquisition unit only acquires the TLS score of HER-2 positive patients. The analysis unit incorporates the TLS score into the analysis; for TLS scores that are not 0, it is recommended to combine immunotherapy with HER-2 targeted therapy.

[0093]

Example 7

[0094] Based on the research results of the foregoing embodiments, this embodiment also provides a storage medium, including a stored computer program, wherein, when the computer program is running, it controls the device where the storage medium is located to execute a method for generating a prognosis and / or treatment plan for mixed hepatocellular-cholangiocarcinoma, the method including the following steps:

[0095] Obtain HER-2 expression levels and grade III lymphoid structure scores from tumor tissue samples;

[0096] Prognosis is based on HER-2 expression levels and tertiary lymphoid structure scores, with a better prognosis for HER-2 positive expression and tertiary lymphoid structure, and a worse prognosis for HER-2 negative expression; and / or

[0097] Treatment plans are generated based on the HER-2 expression level and the tertiary lymphoid structure score. If the HER-2 expression level is negative, the treatment plan is adjuvant chemotherapy; if the HER-2 expression level is positive and the tertiary lymphoid structure score is 0, the treatment plan is HER-2 targeted therapy only; if the HER-2 expression level is positive and the tertiary lymphoid structure score is 1-3, the treatment plan is a combination of HER-2 targeted therapy and immunotherapy.

[0098] Output prognostic results and / or treatment plans.

[0099] In this embodiment, the aforementioned storage medium is a computer-readable storage medium. If the device or method for generating a mixed hepatocellular-cholangiocarcinoma treatment plan is implemented as a software functional unit and sold or used as an independent product, it can be stored in a computer-readable storage medium. Based on this understanding, all or part of the processes in the methods of the above embodiments can also be implemented by a computer program instructing related hardware. The computer program can be stored in a computer-readable storage medium, and when executed by a processor, it can implement the steps of the various method embodiments described above. The computer program includes computer program code, which can be in the form of source code, object code, executable files, or certain intermediate forms. The computer-readable medium can include: any entity or device capable of carrying the computer program code, a recording medium, a USB flash drive, a portable hard drive, a magnetic disk, an optical disk, a computer memory, a read-only memory (ROM), a random access memory (RAM), an electrical carrier signal, a telecommunication signal, and a software distribution medium, etc.

[0100] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above description is only a specific embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. Application of HER-2 expression level detection reagent in the preparation of prognostic assessment products for mixed hepatocellular-cholangiocarcinoma.

2. Application of HER-2 expression level detection reagent combined with tertiary lymphoid structure abundance detection reagent in the preparation of prognostic assessment products for mixed hepatocellular-cholangiocarcinoma.

3. The application of HER-2 expression level detection reagent in the preparation of products for predicting the benefit of adjuvant chemotherapy in mixed hepatocellular-cholangiocarcinoma, characterized in that... Patients who are HER-2 negative are suitable for adjuvant chemotherapy, while patients who are HER-2 positive are not suitable for adjuvant chemotherapy.

4. A pharmaceutical composition for treating mixed hepatocellular-cholangiocarcinoma, characterized in that, The pharmaceutical composition includes a HER-2 targeted drug and an adjuvant chemotherapy drug, wherein the HER-2 targeted drug is used to treat patients with HER-2 positive mixed hepatocellular carcinoma, and the adjuvant chemotherapy drug is used to treat patients with HER-2 negative mixed hepatocellular carcinoma.

5. The pharmaceutical composition according to claim 4, characterized in that, It also includes immunotherapy drugs for treating patients with mixed hepatocellular-cholangiocarcinoma who are HER-2 positive and have a high grade 3 lymphoid structure score.

6. A hybrid hepatocellular-cholangiocarcinoma prognostic device, characterized in that, include: The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples; The analysis unit is used to determine the prognosis of mixed hepatocellular-cholangiocarcinoma based on the expression level of HER-2, where HER-2 positivity corresponds to a better prognosis and HER-2 negativity corresponds to a worse prognosis. The output unit is used to output the prognostic results.

7. A hybrid hepatocellular-cholangiocarcinoma prognostic device according to claim 6, characterized in that, The acquisition unit is also used to acquire the abundance of tertiary lymphoid structures in tumor tissue samples. If HER-2 is positive and tertiary lymphoid structures are present, the prognosis is better.

8. A device for predicting the benefit of adjuvant chemotherapy in mixed hepatocellular-cholangiocarcinoma, characterized in that, include: The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples; The analysis unit is used to determine whether mixed hepatocellular-cholangiocarcinoma is suitable for adjuvant chemotherapy based on HER-2 expression levels. If HER-2 is negative, adjuvant chemotherapy is suitable; if HER-2 is positive, adjuvant chemotherapy is not suitable. The output unit is used to output the predicted results of the benefits of adjuvant chemotherapy.

9. A device for generating a hybrid hepatocellular-cholangiocarcinoma treatment regimen, characterized in that, include: The acquisition unit is used to acquire the expression level of HER-2 in tumor tissue samples; The analysis unit is used to generate treatment plans based on HER-2 expression levels; if HER-2 is negative, adjuvant chemotherapy is recommended. The output unit is used to output the treatment plan.

10. A storage medium, characterized in that, The system includes a stored computer program, wherein, when the computer program is executed, the device containing the storage medium is controlled to perform a method for generating a prognostic and / or treatment plan for mixed hepatocellular-cholangiocarcinoma, the method comprising the following steps: Obtain HER-2 expression levels and grade III lymphoid structure scores from tumor tissue samples; Prognosis is based on HER-2 expression levels and tertiary lymphoid structure scores, with a better prognosis for HER-2 positive expression and tertiary lymphoid structure, and a worse prognosis for HER-2 negative expression; and / or Treatment plans are generated based on the HER-2 expression level and the tertiary lymphoid structure score. If the HER-2 expression level is negative, the treatment plan is adjuvant chemotherapy; if the HER-2 expression level is positive and the tertiary lymphoid structure score is 0, the treatment plan is HER-2 targeted therapy only; if the HER-2 expression level is positive and the tertiary lymphoid structure score is 1-3, the treatment plan is a combination of HER-2 targeted therapy and immunotherapy. Output prognostic results and / or treatment plans.

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