Biomarker for predicting prognosis of lung cancer and application of biomarker
By detecting the HGF level in plasma, the problem of predicting the combined efficacy of immunotherapy and chemotherapy in patients with advanced NSCLC in the prior art is solved. HGF as a biomarker can effectively distinguish patient prognosis and improve the prediction accuracy of treatment effects.
Patent Information
- Application Number
- CN202411454746.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-17
- Publication Date
- 2024-12-13
AI Technical Summary
The prior art is difficult to effectively predict the efficacy of patients with advanced non-small cell lung cancer (NSCLC) after receiving combined treatment of immunotherapy and chemotherapy. Traditional biomarkers such as PD-L1 expression and tumor mutation burden have limitations in predicting effects, and dynamic monitoring is carried out. difficulty.
By detecting hepatocyte growth factor (HGF) levels in plasma before treatment, it was found that high levels of HGF were associated with shorter survival and progression times, and HGF as an independent predictive biomarker could predict the efficacy of patients on first-line chemoimmunotherapy.
A HGF level above 214.45pg/mL indicates a poor prognosis in the patient. Patients with low-level HGF survive longer, progress slowly, have good distinction ability, and independently predict immunotherapy response, which improves the accuracy of prediction of treatment effects.
Smart Images
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a biomarker for predicting the therapeutic effect of combined immunotherapy and chemotherapy for lung cancer and its application. Background Art
[0002] Lung cancer has the highest mortality rate among solid tumors worldwide, with a 5-year survival rate of less than 10% for advanced cases, and non-small cell lung cancer (NSCLC) accounts for 80%-85% of all cases (Bray F, Laversanne M, Sung H, et ai. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries [J]. CA Cancer J Clin 2024, 74 (3): 229-263.). Surgery is one of the main methods for treating lung cancer. If lung cancer is diagnosed early and radical surgery is performed, the 5-year survival rate can reach more than 90%. However, about 60% of lung cancer is in the locally advanced or metastatic stage when clinically diagnosed, missing the best time for surgical treatment. The treatment effects of radiotherapy (hereinafter referred to as radiotherapy) or chemotherapy (hereinafter referred to as chemotherapy) and combined radiotherapy and chemotherapy are not ideal, and the overall survival rate is no more than 50% (Qiu H, Cao S, Xu R. Cancer incidence, mortality, and burden in China: a time-trend analysis and comparison with the United States and United Kingdom based on the global epidemiological data released in 2020[J]. Cancer Commun (Lond), 2021, 41(10): 1037-1048; Shin S, Kim HK, Cho JH, et al. Adjuvant therapy in stage IIIA-N2 non-small cell lung cancer after neoadjuvant concurrent chemoradiotherapy followed by surgery[J]. J Thorac Dis, 2020, 12(5): 2602-2613.).
[0003] At present, in addition to surgical treatment, conventional chemotherapy, radiotherapy, immunotherapy, and molecular targeted drug therapy are the main treatments for NSCLC patients. Chemotherapy has always been an important first-line treatment for locally advanced and metastatic lung cancer (Hanna N, Johnson D, Temin S, et al. Systemic therapy for stage IV non-small-cell lung cancer: American Society of Clinical Oncology clinical practice guideline update [J]. J Clin Oncol, 2017, 35 (30): 3484-3515.). Preoperative chemotherapy has a certain effect on reducing tumor stage and eliminating tumor metastasis. However, during conventional chemotherapy, due to the poor specificity of the drug, it not only kills cancer cells but also damages normal tissues of the body, which can easily lead to more adverse reactions in patients and affect the treatment effect of patients (Yu Hongwei, Zhang Yibin, Zhang Lu, et al. Effect of Brucea javanica oil emulsion combined with GP regimen in the treatment of lymph node metastasis of non-small cell lung cancer [J]. China Practical Medicine, 2020, 15 (34): 108-110.). In recent years, programmed cell death protein-1 (PD-1) inhibitors have achieved good results in neoadjuvant immunotherapy for advanced lung cancer by activating T lymphocytes and promoting immune system response (Zhou Xiaodan, Ji Guofa, Liu Ya, et al. Analysis of the efficacy of tislelizumab combined with albumin-paclitaxel in the treatment of advanced lung cancer and its effect on T cell subsets [J]. Progress in Modern Biomedicine, 2022, 22(11): 2128-2132.). Immunotherapy using programmed cell death protein-1 (PD-1) or programmed death ligand 1 (PD-L1) has produced durable responses and prolonged survival in patients with advanced NSCLC, fundamentally changing the treatment strategy. In particular, immunotherapy combined with platinum chemotherapy has become the standard first-line treatment for advanced NSCLC. Although immunotherapy combined with chemotherapy provides a wider range of beneficiaries compared with monotherapy, the number of patients who achieve durable responses is still small. It is necessary to find suitable predictive biomarkers to help select NSCLC patients who benefit from the combination of immunotherapy and chemotherapy for long-term survival.
[0004] Tumor markers are often important examination indicators that reflect the growth and prognosis of malignant tumors, and their changes indicate the effect of tumor treatment to a certain extent. Biomarkers related to the prognosis of immunotherapy for advanced NSCLC have traditionally focused on tumor-related, including PD-L1 immunohistochemistry (IHC) and tumor mutation burden (TMB). Programmed cell death ligand 1 (PD-L1) expression is currently the most widely accepted predictive marker, but its expression status can change with the progress of treatment, and not all patients with high expression respond to immunotherapy. Therefore, the use of PD-L1 expression to predict the efficacy of immunotherapy still has certain limitations (Feng Hui, Xu Zihan, Nie Yuhui, et al. Clinical significance of prognostic score of immunotherapy for advanced non-small cell lung cancer in immunotherapy combined with chemotherapy for advanced non-small cell lung cancer [J]. Journal of Clinical Internal Medicine, 2023, 40(10): 677-680.). In addition, although PD-L1 expression and TMB have certain predictive value in single immunotherapy, their predictive value for the combination of chemotherapy and immunotherapy is limited. Although intracellular markers help to elucidate the tumorigenesis process, they usually require tissue biopsies, which are not easily available and hinder dynamic monitoring. The complex dynamic interactions between tumors and hosts may affect the efficacy of immunotherapy, and there is an urgent need to identify accurate and easily accessible dynamic clinical biomarkers to predict the efficacy of immunotherapy combined with chemotherapy for advanced NSCLC.
[0005] Hepatocyte growth factor (HGF) is a bioactive factor with multiple functions. It can act on the growth of multiple cells and participate in the regulation of angiogenesis and immune activity. HGF can promote the proliferation, metastasis and invasion of tumor cells and is a specific ligand for c-MET (Naldini L, Vigna E, Narsimhan RP, et al. Hepatocyte growth factor (HGF) stimulates the tyrosine kinase activity of the receptor encoded by the proto-oncogene c-MET [J]. Oncogene, 1991, 6 (4): 501-504.). Cellular-mesenchymal epithelial transition factor (c-MET) is a type of transmembrane receptor with autonomous phosphorylation activity encoded by the c-MET gene (Gherardi E, Birchmeier W, Birchmeier C, et al. Targeting MET in cancer: rationale and progress [J]. Nat Rev Cancer, 2012, 12 (2): 89-103.), and its overexpression has been shown to promote tumor growth in a variety of cancers. This signaling pathway is abnormally activated in NSCLC cell lines and tumor tissues, and c-Met has been validated as a potential therapeutic target by small interfering RNA targeting and small molecule inhibitors. In addition, MET amplification tends to promote PD-L1 expression in NSCLC, so the combination of MET inhibitors and ICIs may be an attractive therapeutic strategy. Summary of the invention
[0006] The present invention detects the level of soluble HGF in plasma before treatment and finds that high levels of HGF have shorter OS and PFS. Further analysis shows that HGF is an independent predictive biomarker for immunotherapy response in advanced NSCLC, which can predict the efficacy of patients after first-line chemoimmunotherapy and has a good ability to distinguish whether patients have progressed. Based on this, the present invention was completed.
[0007] In a first aspect, the present invention provides a biomarker for predicting the therapeutic effect of lung cancer patients after treatment, wherein the biomarker is HGF; when the HGF level in the patient's biological sample is higher than 214.45 pg / mL, it indicates that the patient has a poor prognosis.
[0008] Furthermore, the lung cancer includes non-small cell lung cancer and small cell lung cancer.
[0009] Preferably, the lung cancer is non-small cell lung cancer.
[0010] Furthermore, the biological sample includes one or more of whole blood, serum and / or plasma.
[0011] Preferably, the biological sample is plasma.
[0012] Furthermore, the treatment method includes one or more of immunotherapy, chemotherapy and / or surgical resection.
[0013] Preferably, the treatment method is immunotherapy combined with chemotherapy.
[0014] In a second aspect, the present invention provides an application of a biomarker in the preparation of a reagent for predicting the therapeutic effect of lung cancer patients after treatment, wherein the biomarker is HGF, and the reagent is a reagent containing HGF that can detect HGF in a biological sample of a patient.
[0015] Furthermore, when the HGF level in the patient's biological sample is higher than 214.45 pg / mL, it indicates a poor prognosis for the patient.
[0016] Furthermore, the lung cancer includes non-small cell lung cancer and small cell lung cancer.
[0017] Preferably, the lung cancer is non-small cell lung cancer.
[0018] Furthermore, the biological sample includes one or more of whole blood, serum and / or plasma.
[0019] Preferably, the biological sample is plasma.
[0020] Furthermore, the treatment method includes one or more of immunotherapy, chemotherapy and / or surgical resection.
[0021] Preferably, the treatment method is immunotherapy combined with chemotherapy.
[0022] In a third aspect, the present invention provides a kit for predicting the therapeutic effect of lung cancer patients after receiving treatment, wherein the kit comprises a reagent for detecting the level of HGF in a biological sample of the patient.
[0023] Furthermore, when the HGF level in the patient's biological sample is higher than 214.45 pg / mL, it indicates a poor prognosis for the patient.
[0024] Furthermore, the biological sample includes one or more of whole blood, serum and / or plasma.
[0025] Preferably, the biological sample is plasma.
[0026] Furthermore, the lung cancer includes non-small cell lung cancer and small cell lung cancer.
[0027] Preferably, the lung cancer is non-small cell lung cancer.
[0028] Furthermore, the treatment method includes one or more of immunotherapy, chemotherapy and / or surgical resection.
[0029] Preferably, the treatment method is immunotherapy combined with chemotherapy.
[0030] Furthermore, the kit may be one or more of an ELISA detection kit, a colloidal gold detection kit, an immunohistochemistry kit, an immunofluorescence kit and / or an in situ hybridization staining kit.
[0031] Furthermore, the diagnostic method of the kit includes one or more of a direct method, an indirect method, a double antibody sandwich method and / or a competitive method.
[0032] Beneficial Effects
[0033] 1. The biomarkers of the present invention can predict the therapeutic efficacy of lung cancer patients; compared with patients with higher HGF levels, patients with lower HGF levels have longer survival time; patients with low HGF levels progress more slowly than patients with higher HGF levels; in the analysis of OS and PFS of the screening cohort, the AUCs of baseline HGF levels for predicting patient survival were 0.725 and 0.751, respectively, showing good discrimination ability.
[0034] 2. In the univariate Cox regression analysis of clinical characteristics, it was found that poor physical status, bone metastasis and adrenal metastasis were associated with poor prognosis of patients; multivariate analysis showed that HGF was an independent predictive biomarker for immunotherapy response in advanced NSCLC. BRIEF DESCRIPTION OF THE DRAWINGS
[0035] Figure 1 Survival curves and ROC curves of overall survival in different HGF level groups.
[0036] Figure 2 Survival curves and ROC curves of different HGF level groups and progression-free survival. DETAILED DESCRIPTION
[0037] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.
[0038] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.
[0039] Explanation of terms
[0040] Tumor mutation burden (TMB): refers to the density of non-synonymous mutations in protein coding regions, which is calculated by dividing the total number of non-synonymous mutation sites in the protein coding region by the total length of the protein coding region. The unit is mutation number / mb.
[0041] Overall survival (OS): The time from the start of treatment to the patient's death due to any cause.
[0042] Progression-free survival (PFS): The time from the start of treatment to disease progression or death from any cause. Generally, the longer the PFS, the longer the patient's quality of life and the better he or she lives.
[0043] Example 1 Clinical Data Collection and Analysis
[0044] 1.1 Case collection
[0045] Patients with NSCLC who received at least one immunotherapy combined with chemotherapy at Beijing Chest Hospital affiliated to Capital Medical University between December 2018 and January 2022 were retrospectively enrolled.
[0046] 1.2 Inclusion and exclusion standards
[0047] Patients were aged >18 years, with histopathologically diagnosed NSCLC, stage III / IV according to the eighth edition of the TNM staging system, Eastern Cooperative Oncology Group (ECOG) performance status score of 0-2, and adequate liver, kidney, and heart function.
[0048] At the same time, patients who did not meet the conditions for ICI administration, such as patients with active autoimmune diseases and active infections, were excluded.
[0049] 1.3 Study Groups
[0050] Patients who met the above inclusion criteria were divided into a screening cohort and a validation cohort, of which 55 were in the screening cohort and 24 were in the validation cohort.
[0051] 1.4 HGF determination
[0052] HGF MAP's Human Circulating Cancer Biomarker MagneticBead Panel 1 was used for detection. Standards, controls, samples, and Serum Matrix were added to the standard wells, control wells, and sample wells respectively. Antibody magnetic beads of the above cytokines were taken separately, mixed, and mixed magnetic beads were added to each well. The plate was sealed and shaken at room temperature in the dark. The plate was washed and the detection antibody was added to each well. The plate was sealed and shaken at room temperature in the dark for incubation. Streptavidin-phycoerythrin was added to each well. The plate was sealed and shaken at room temperature in the dark, the plate was washed, sheath fluid was added to each well, and the beads were resuspended. The detection was performed on the Luminex liquid phase chip instrument, and the data were analyzed using xPONENT software.
[0053] 1.5 Statistical analysis
[0054] The patient population was divided into a training set and a validation set by a 7:3 random split, and the baseline HGF cytokine level was divided into high and low groups according to the median. The median values of OS and PFS were calculated by the Kaplan-Meier method, and the survival differences between different cytokine groups were compared by the Log-Rank test. The Cox proportional hazard model was used to calculate the hazard ratio (HR) and 95% confidence interval (CI). All statistical analyses were performed using R, and P<0.05 was considered statistically significant.
[0055] Example 2 Correlation between baseline HGF levels and patient survival
[0056] The cutoff value was determined based on the median HGF. This grouping method can better reflect the correlation between HGF and patient prognosis, rather than selecting the best cutoff value that may achieve the best discrimination ability. The median baseline HGF level of 79 cases was 214.45pg / mL. According to this cutoff value, the patient population was divided into a high-HGF group and a low-HGF group, and survival analysis was performed. As of the last follow-up time, a total of 43 deaths and 52 progression events (including progression events and deaths) occurred in the patient cohort, with a median OS and median PFS of 24.4 (95% CI, 19.4-NR) months and 10.9 (95% CI, 7.9-16.2) months, respectively.
[0057] 2.1 Screening Queue
[0058] In the analysis of OS, patients with lower HGF levels had longer survival time compared with those with higher HGF levels (P=0.033) ( Figure 1 A), ROC curve of baseline HGF level predicting survival of patients in the screening cohort. Figure 1 As shown in C, the AUC is 0.725, showing good discrimination ability.
[0059] In the analysis of PFS, patients with low HGF levels progressed more slowly than those with high HGF levels (P = 0.0036) ( Figure 2 A), and its AUC was 0.751, which showed good ability to distinguish whether the patient progressed ( Figure 2 C).
[0060] 2.2 Verification Queue
[0061] In the analysis of OS, patients with lower HGF levels survived longer compared with those with higher HGF levels ( Figure 1 B) ROC curve of baseline HGF level predicting survival of patients in the validation cohort Figure 1 As shown in D, the AUC is 0.590, which also shows good discrimination ability.
[0062] In the analysis of PFS, patients with low HGF levels progressed more slowly than those with high HGF levels (P = 0.007) ( Figure 2 B), and its AUC was 0.682, which has a certain ability to distinguish whether the patient has progressed ( Figure 2 D).
[0063] Example 3 Univariate and multivariate Cox regression analysis
[0064] In univariate Cox regression analysis, high levels of HGF (HR: 1.96, 95% CI: 1.08-3.57, P = 0.029) had a shorter OS (Table 1). This correlation was also observed in PFS (HR: 3.13, 95% CI: 1.72-5.88, P < 0.001) (Table 2). In addition, poor performance status, bone metastasis, and adrenal metastasis were found to be associated with poor prognosis in clinical characteristics.
[0065] Further multivariate analysis showed that HGF was an independent predictive biomarker for immunotherapy response in advanced NSCLC, among which high HGF levels predicted worse OS (HR: 2.04, 95% CI: 1.09-3.85, P = 0.028) (Table 1) and PFS (HR: 3.33, 95% CI: 1.85-6.25, P < 0.001) (Table 2).
[0066] Table 1 Univariate and multivariate Cox regression analysis of overall survival
[0067]
[0068]
[0069] Table 2 Univariate and multivariate Cox regression analysis of progression-free survival
[0070]
[0071]
Claims
1. A biomarker for predicting the therapeutic effect of lung cancer patients after treatment, wherein the biomarker is HGF; when the HGF level in the patient's biological sample is higher than 214.45 pg / mL, it indicates that the patient has a poor prognosis.
2. The biomarker according to claim 1, wherein the lung cancer includes non-small cell lung cancer and small cell lung cancer; preferably non-small cell lung cancer.
3. The biomarker according to any one of claims 1 or 2, wherein the biological sample comprises one or more of whole blood, serum and / or plasma; preferably plasma.
4. The biomarker according to any one of claims 1 to 3, wherein the treatment method comprises one or more of immunotherapy, chemotherapy and / or surgical resection; preferably immunotherapy combined with chemotherapy.
5. Use of a biomarker in the preparation of a reagent for predicting the therapeutic effect of lung cancer patients after treatment, wherein the biomarker is HGF, and the reagent is a reagent that can detect HGF in a patient's biological sample; when the HGF level in the patient's biological sample is higher than 214.45 pg / mL, it indicates that the patient has a poor prognosis.
6. A kit for predicting the therapeutic effect of lung cancer patients after treatment, the kit comprising a reagent for detecting the HGF level in the patient's biological sample; when the HGF level in the patient's biological sample is higher than 214.45 pg / mL, it indicates that the patient has a poor prognosis.
7. The kit according to claim 6, wherein the lung cancer includes non-small cell lung cancer and small cell lung cancer; preferably non-small cell lung cancer.
8. The kit according to any one of claims 6 or 7, wherein the treatment method comprises one or more of immunotherapy, chemotherapy and / or surgical resection; preferably immunotherapy combined with chemotherapy.
9. The kit according to any one of claims 6 to 8, wherein the kit can be one or more of an ELISA detection kit, a colloidal gold detection kit, an immunohistochemistry kit, an immunofluorescence kit and / or an in situ hybridization staining kit.
10. The kit according to any one of claims 6 to 9, wherein the diagnostic method of the kit comprises one or more of a direct method, an indirect method, a double antibody sandwich method and / or a competitive method.