Application of MMP14 as a molecular marker and therapeutic target for leptomeningeal metastases

Abstract

The present invention belongs to the technical field of cell biology, specifically to the application of MMP14 as a molecular marker and therapeutic target for leptomeningeal metastases. Based on the discovery that macrophages and tumor cells in leptomeningeal metastases samples highly express MMP14 simultaneously, the present invention further discovers that MMP14 inhibitors can significantly inhibit the growth of tumor cells in a mouse model of leptomeningeal metastases and significantly prolong the overall survival of mice with leptomeningeal metastases. Therefore, MMP14, as a new molecular marker and therapeutic target for leptomeningeal metastases, is used to develop products for inhibiting leptomeningeal metastases, including preparing diagnostic and differential diagnostic reagents for leptomeningeal metastases targeting MMP14, and preparing targeted drugs for leptomeningeal metastases. The tumors include leptomeningeal metastases of lung cancer and breast cancer. Mouse model experiments show that compared with the non-drug administration group, the tumor burden in the MMP14 inhibitor administration group is reduced, and the overall survival time of mice with leptomeningeal metastases is prolonged.

Description

Technical Field

[0001] The present invention belongs to the technical field of cell biology, and particularly relates to the application of MMP14 as a new molecular marker, therapeutic target for leptomeningeal metastasis tumors, and preparation of products for inhibiting leptomeningeal metastasis tumors. Background Art

[0002] The process by which tumor cells spread to the arachnoid mater, subarachnoid space (cerebrospinal fluid), and pia mater of the brain or spinal cord through the blood or other routes is called leptomeningeal metastasis (LM), also known as leptomeningeal carcinomatosis, neoplastic meningitis, or carcinomatous meningitis. [1, 2] Leptomeningeal metastasis is a serious complication of tumors, with an average incidence rate of 5 - 15% and an increasing trend year by year. The primary tumors are mostly breast cancer (12 - 34%), lung cancer (10 - 26%), and melanoma (17 - 25%). The natural survival period of patients is only 4 - 6 weeks. [1, 3, 4] As a malignant central nervous system disease, LM patients often present with cerebral edema and are accompanied by various neurological symptoms such as headache, nausea, diplopia, limb paralysis, and vision loss.

[0003] Leptomeningeal metastasis tumor cells are diffusely distributed in the subarachnoid space and cannot be treated by surgical resection. Currently, the treatment options for leptomeningeal metastasis tumors are still traditional radiotherapy and chemotherapy, but the treatment effect is not good. [5-7] Radiotherapy and chemotherapy lack precision. While killing tumor cells, they damage normal tissue cells, have large toxic side effects, and can cause many adverse reactions such as nausea, loss of appetite, hair loss, and decreased immunity. Therefore, there is an urgent need to develop a precise treatment plan targeting leptomeningeal metastasis tumors to improve the survival period of patients and reduce adverse reactions. However, the pathogenesis of leptomeningeal metastasis tumors is not yet clear, and there is no effective report on molecular targets for leptomeningeal metastasis tumors. This research status restricts the development of targeted treatment strategies and related drugs for leptomeningeal metastasis tumors. Based on the field of leptomeningeal metastasis tumor treatment, the present invention develops new and efficient molecular targets for leptomeningeal metastasis tumors, which has great clinical significance and translational value.

[0004] The tumor microenvironment (TME) refers to the surrounding environment of tumor cells, including blood vessels, cells, signaling molecules, and the extracellular matrix. The interaction between tumor cells and the TME plays an important role in tumor cell survival, proliferation, metastasis, and drug resistance. [8] Immune cells and their secretions are important components of the tumor microenvironment. In addition, various immune cells infiltrate the cerebrospinal fluid of LM patients, and the content of inflammatory factors is significantly upregulated. However, the cerebrospinal fluid of LM usually presents an immunosuppressive TME, making tumor cells unable to be killed by the immune system. [7, 9, 10]。Gholamin et al. reported that melanoma cells in cerebrospinal fluid escape macrophage phagocytosis through the CD47-SIRPα anti-phagocytic axis

[11] 。Smalley et al. found that cerebrospinal fluid T cells in melanoma LM patients were in a state of exhaustion and lost their function of killing tumor cells

[10] 。

[0005] Macrophages are a type of highly migratory cells that can be chemotaxed by tumor cells to the TME and are the main participants and shapers of the TME [12-14] 。Tumor-associated macrophages (TAM) play an immunosuppressive function in the TME and contribute to tumor development

[15] 。TAM has poor antigen presentation ability and inhibits the immune response of T cells by releasing immunosuppressive factors IL-10 and TGF-β. At the same time, TAM produces chemokines CCL17 and CCL22, and recruits Treg cells to the TME through the receptor CCR4

[15] 。Treg can inhibit the activation and proliferation of effector T cells and also inhibit the killing function of NK cells against tumors

[16] 。In addition, TAM also directly inhibits the proliferation and cytotoxicity of T cells through the PD-L1 / PD-1 signaling axis [15, 17] 。The published work of the research group shows that a large number of macrophages infiltrate the subarachnoid cerebrospinal fluid of lung cancer LM patients, and these macrophages in cerebrospinal fluid secrete cytokines IL-6 / 8 to activate the LCN2 iron transport system of tumor cells and promote tumor survival [9] 。In addition, researchers performed single-cell transcriptome sequencing on cerebrospinal fluid of patients with leptomeningeal metastases of lung cancer and found that cerebrospinal fluid macrophages of patients presented an immunosuppressive phenotype [9, 18] 。Based on the above background, targeting macrophages in the cerebrospinal fluid of patients with leptomeningeal metastases of lung cancer has potential development prospects

[0006] The applicant's research found that in a mouse model of leptomeningeal metastases and patients, their cerebrospinal fluid macrophages and tumor cells simultaneously highly expressed matrix metalloproteinase 14 (MMP14). MMP14 is an important tumor microenvironment regulatory protein, mainly involved in the degradation and remodeling of the extracellular matrix, inflammation, and pro-angiogenic signaling pathways, and promotes tumor metastasis

[19] 。Literature-reported data show that MMP14 is involved in the execution of multiple tumor regulation-related functions of macrophages: (1) directly regulating the two-dimensional and three-dimensional migration of macrophages; (2) changing the expression of inflammatory factors; (3) inhibiting the lymphangiogenesis-promoting function of TAM [20-22]On the basis of discovering the high expression of MMP14 in cerebrospinal fluid macrophages and tumor cells, the applicant further found that the small molecule inhibitor of MMP14 can significantly inhibit the growth of tumor cells in the cerebrospinal fluid of the LM mouse model and significantly prolong the overall survival period of LM mice. Therefore, MMP14 can be used as a new molecular marker and therapeutic target for leptomeningeal metastases for the development of products that inhibit leptomeningeal metastases. Summary of the Invention

[0007] The object of the present invention is to provide a new molecular marker and therapeutic target for leptomeningeal metastases, as well as the application of MMP14 as a molecular marker for leptomeningeal metastases, and the application of MMP14 in the preparation of various diagnostic reagents and various targeted drugs for leptomeningeal metastases.

[0008] The present invention discovers for the first time that in the leptomeningeal metastases mouse model and patients, matrix metalloproteinase 14 is highly expressed in macrophages and tumor cells in their cerebrospinal fluid and tumor foci;

[0009] On this basis, it is further found that the small molecule inhibitor of MMP14 can significantly inhibit the growth of tumor cells in the LM mouse model and significantly prolong the overall survival period of LM mice. Therefore, MMP14 can be used as a new molecular marker and therapeutic target for leptomeningeal metastases for the development of products that inhibit leptomeningeal metastases.

[0010] On the one hand, the present invention provides the application of MMP14 as a molecular marker for leptomeningeal metastases, which can be used for the preparation of diagnostic and differential diagnostic reagents for leptomeningeal metastases targeting MMP14.

[0011] Furthermore, the type of the tumor is any tumor type that can develop leptomeningeal metastases.

[0012] Preferably, the tumor includes lung cancer and breast cancer.

[0013] On the other hand, the present invention provides the application of MMP14 as a targeted therapeutic target for leptomeningeal metastases, which can be used for the preparation of various targeted drugs for leptomeningeal metastases.

[0014] Furthermore, the type of the tumor is any tumor type that can develop leptomeningeal metastases.

[0015] Preferably, the tumor includes lung cancer and breast cancer.

[0016] Furthermore, the targeted therapy includes targeting any cells with high expression of MMP14 in leptomeningeal metastases.

[0017] Preferably, the targeted therapy includes targeting tumor cells with high expression of MMP14, macrophages with high expression of MMP14, and the combination of both.

[0018] Furthermore, the drug includes traditional Chinese medicine, chemical medicine, biological medicine and their combinations, and the active ingredient of the drug or combination can inhibit the expression or function of MMP14. Such as small molecule inhibitors of MMP14.

[0019] Furthermore, the dosage form of the drug is any pharmaceutically acceptable dosage form, and the drug dosage is any pharmaceutically acceptable dosage.

[0020] Through bioinformatics analysis and experimental research, the present invention finds that in patient and mouse leptomeningeal metastasis samples, MMP14 is highly expressed in the tumor microenvironment of leptomeningeal metastases, mainly highly expressed in tumor cells and macrophages. The application of MMP14 inhibitors can significantly improve the health status of animal models of leptomeningeal metastases and prolong their survival time. Accordingly, the present invention provides the application of MMP14 as a molecular marker and therapeutic target for leptomeningeal metastases, and further provides the application of MMP14 in the preparation of products for inhibiting leptomeningeal metastases. By using products that inhibit the expression of MMP14, the effect of inhibiting leptomeningeal metastases can be achieved, and a reliable treatment plan can be designed based on this principle to prevent and treat leptomeningeal metastases. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is an immunofluorescence staining diagram of cerebrospinal fluid cultured cells of mice with lung cancer leptomeningeal metastases. The four diagrams are corresponding fluorescence diagrams under the same field of view. TdTomato autofluorescence labels tumor cells (excited by 561nm excitation light), GFP autofluorescence labels macrophages, MMP14 molecules are labeled with anti-MMP14 primary antibody and Rabbit-Alexa Fluor 647 secondary antibody (excited by 647nm excitation light), and DAPI labels cell nuclei (excited by 405nm excitation light). The figure shows that the expression of MMP14 in tumor cells and macrophages is significantly increased. Scale bar: 40μm.

[0022] Figure 2 It is an immunofluorescence staining diagram of a brain section (tumor focus site) of mice with lung cancer leptomeningeal metastases. The left diagram and the right diagram are corresponding fluorescence diagrams under the same field of view. The following diagram is an enlarged view of the above diagram. GFP autofluorescence labels macrophages, and MMP14 molecules are labeled with anti-MMP14 primary antibody and Rabbit-Alexa Fluor 647 secondary antibody (excited by 647nm excitation light). The figure shows that the expression of MMP14 in tumor cells and macrophages is significantly increased. Scale bar: 40μm for the above diagram and 10μm for the following diagram.

[0023] Figure 3Violin plot of MMP14 expression in cerebrospinal fluid cells of patients with lung cancer and breast leptomeningeal metastases. The abscissa represents different cell subsets clustered from single-cell sequencing data, and the ordinate represents MMP14 expression. Among them, macrophages (MΦ) and tumor cells have high expression of MMP14.

[0024] Figure 4 Effect diagram of targeting MMP14 in the treatment of leptomeningeal metastases in mice. A - B. Compared with the non - drug - administered group, the tumor burden in the MMP14 inhibitor - administered group decreased, indicating that the MMP14 inhibitor exerted an anti - leptomeningeal metastasis effect. C. The MMP14 inhibitor prolonged the overall survival time of LM mice. ** represents P < 0.01. Detailed implementation mode

[0025] The present invention provides the application of MMP14 as a molecular marker for leptomeningeal metastases.

[0026] The present invention also provides the application of MMP14 as a therapeutic target for leptomeningeal metastases. The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but it does not limit the protection scope of the present invention.

[0027] Example 1: Verification of the expression of MMP14 in the tumor microenvironment of a leptomeningeal metastases mouse model

[0028] 1. Cell culture

[0029] The LLC - Luc2 - tdTomato - LM cells used in the study were from the inventory resources of the applicant's research group. This cell line is a lung cancer leptomeningeal - specific metastatic cell line, which highly expresses firefly luciferase and fluorescent protein tdTomato, facilitating in - vivo observation. LLC - Luc2 - tdTomato - LM cells were cultured in a 10 - cm - diameter cell culture dish using DMEM complete medium (containing 10% FBS and 1% penicillin - streptomycin solution). The temperature of the cell culture incubator was 37 °C and the CO2 concentration was 5%.

[0030] 2. Establishment of a leptomeningeal metastases model

[0031] When the LLC - Luc2 - tdTomato - LM cells reached 70% confluence, they were digested with 1 mL of 0.25% trypsin - EDTA for 3 min, centrifuged at 200 g for 5 min, and the cells were resuspended with 1×PBS to a concentration of 2×10 5 cells / mL and placed on ice for later use. To conveniently label cerebrospinal fluid macrophages, we introduced the widely used Cx3cr1 GFP / WT transgenic mice in the field. These mice have high expression of green fluorescent protein in various macrophages including the meninges

[23] 。Select 6-week-old SPF-grade Cx3cr1 GFP / WT male mice. Inject Avertin into the mice for anesthesia, and carefully shave the hair on the dorsal side of the neck and the top of the head of the mice. Horizontally fix a 15 mL centrifuge tube on the operating table with tape. Place the mouse on its back, with the neck padded on the centrifuge tube, the long axis of the head perpendicular to the horizontal plane, forming an angle of about 90° with the neck, fully exposing the corresponding area of the cerebellomedullary cistern, and mark the needle insertion point in the middle of the cerebellomedullary cistern. Use a 1 mL syringe (30 G needle) to aspirate the LLC-Luc2-tdTomato-LM cell suspension, insert it into the cerebellomedullary cistern, and after feeling the loss of resistance when breaking through the dura mater, slowly inject 10 μL of the cell suspension into the cerebellomedullary cistern. After the injection is completed, stay for 1 min. Wait for the cells to fully enter, and then remove the syringe. Place the mouse on a heating pad and wait for it to wake up. On the 0th, 7th, and 14th days after modeling, use the IVIS Spectrum machine to monitor the tumor burden of the mice through bioluminescence imaging (BLI).

[0032] 3. Detect the expression of MMP14 in cerebrospinal fluid cells

[0033] When the tumor burden covers the entire surface of the central nervous system or the body weight of the mice significantly decreases, the mice are euthanized. The skull is opened, and the base of the skull and the brain surface are rinsed with 2 mL of sterile PBS. The rinsing solution is centrifuged at 500 g for 5 min, resuspended in DMEM medium containing 10% FBS, cultured, and inoculated into a 35 mm confocal dish pre-coated with polylysine. After culturing for 24 h, the medium is discarded, and the cells are fixed with 2 mL of 4% PFA for 15 min and incubated in a blocking solution (containing 0.3% Triton X-100 and 10% donkey serum) at room temperature for 1 h. Then, the cells are incubated with the anti-MMP14 primary antibody (ab51074, Abcam) overnight at 4 °C in 500 μL of staining buffer (containing 10% donkey serum). After the binding of the primary antibody, the cells are rinsed three times with PBS (containing 0.05% Tween-20), and then incubated with the Alexa Fluor 647 AffiniPure Donkey Anti-Rabbit IgG (H+L) (711-605-152, Jackson ImmunoResearch) secondary antibody in 1 mL of staining buffer (containing 10% donkey serum) at room temperature for 1 h. Subsequently, the cells are rinsed three times with PBS (containing 0.05% Tween-20). Before microscopic imaging, the cells are stained with 1 μg / mL DAPI for 2 min and thoroughly washed with PBS for 10 min. After mounting with Fluoromount-G anti-quenching mounting medium (Southern Biotech), the cells are imaged using an Olympus FV3000 confocal microscope.

[0034] As Figure 1 shown, by applying immunofluorescence staining, it was found that the expression of MMP14 in tumor cells (tdTomato positive) and macrophages (GFP positive) in the cerebrospinal fluid rinsing solution of mice with leptomeningeal metastases was significantly increased, indicating that MMP14 is a molecular marker for leptomeningeal metastases.

[0035] 4. Verification of the expression of MMP14 in the tumor microenvironment of the leptomeningeal layer

[0036] In step 3, mouse brains were collected synchronously, fixed with 10 mL of 4% PFA for 8 - 12 h (at 4 °C), dehydrated by soaking in 30% sucrose until the brains sank to the bottom, embedded in OCT, snap-frozen in dry ice, and 20-μm sections were prepared using a Leica cryostat. Brain sections were incubated in a blocking solution (containing 0.3% Triton X-100 and 10% donkey serum) for 1 h at room temperature. Then, they were incubated overnight at 4 °C with the primary antibody against MMP14 (ab51074, Abcam) in 300 μL of staining buffer (containing 10% donkey serum). After binding of the primary antibody, they were rinsed three times with PBS (containing 0.05% Tween-20), and then incubated for 1 h at room temperature with the secondary antibody Alexa Fluor 647 AffiniPure Donkey Anti-Rabbit IgG (H+L) (711-605-152, Jackson ImmunoResearch) in 300 μL of staining buffer (containing 10% donkey serum), followed by three rinses with PBS (containing 0.05% Tween-20). Before microscopic imaging, the cell nuclei were stained with 1 μg / mL DAPI for 2 min, thoroughly washed with PBS for 10 min, blotted dry, and mounted with Fluoromount-G anti-fade mounting medium (SouthernBiotech), and imaged using an Olympus FV3000 confocal microscope.

[0037] As Figure 2 shown, by applying immunofluorescence staining, it was found that the expression of MMP14 in macrophages in the tumor microenvironment was significantly increased at the leptomeningeal tumor foci in mice with leptomeningeal metastases, indicating that MMP14 is a molecular marker for leptomeningeal metastases in preclinical experiments.

[0038] Example 2. Verification of the expression of MMP14 in the tumor microenvironment of patients with leptomeningeal metastases

[0039] 1. Obtaining and normalizing sequencing data

[0040] Single-cell transcriptome sequencing of cerebrospinal fluid cells from four patients with lung cancer leptomeningeal metastases that had been completed in our laboratory was performed. Single-cell libraries were constructed based on the 10x genomics system, and the raw data were background-corrected and normalized using transcriptome data analysis software. Cells with mitochondrial gene expression exceeding 20%, total expressed genes less than 200 or more than 5218, and housekeeping gene expression less than 50 were removed to obtain a single-cell transcriptome expression matrix.

[0041] 2. Cell clustering and gene expression analysis

[0042] The PCA and UMAP algorithms are used to perform dimensionality reduction processing and information display on the data. After dimensionality reduction is completed, single-cell populations are divided through unsupervised clustering based on gene expression. A differential screening algorithm is used to calculate the population of marker genes for single-cell clusters, and the cell types to which each cell population belongs are inferred and identified through these marker genes. Using the VlnPlot function in the Seurat package of R language, a violin plot is made with cell types as the abscissa and the MMP14 expression level as the ordinate.

[0043] As Figure 3 shown, by applying single-cell transcriptome analysis, it is found that in the cerebrospinal fluid of leptomeningeal metastasis patients, macrophages and tumor cells have high expression of MMP14, indicating that in clinical patients, MMP14 can be used as a molecular marker for leptomeningeal metastasis.

[0044] Example 3: Verification of the therapeutic effect of MMP14-targeted drugs on a mouse model of leptomeningeal metastasis

[0045] 1. Group design

[0046] Set up ① the drug administration group (10 C57BL / 6J mice): Establish a leptomeningeal metastasis model and administer the MMP14 inhibitor NSC405020 (MCE Company); ② the non-drug administration group (10 C57BL / 6J mice): Establish a leptomeningeal metastasis model and administer the same amount of solvent DMSO as in ①.

[0047] 2. Administration of drugs to mice with leptomeningeal metastasis model

[0048] Construct a mouse model of lung cancer leptomeningeal metastasis according to the modeling method in Example 1, and prepare the drug in advance. Dissolve NSC405020 with DMSO to prepare a stock solution with a concentration of 40 mg / mL, aliquot and store at -80 °C, thaw on the day of administration, and dilute to a working concentration of 4 mg / mL. Mice in the drug administration group were intraperitoneally injected with NSC405020 at a dose of 20 g / kg (mouse body weight) every 3 days, and mice in the non-drug administration group were intraperitoneally injected with the same amount of solvent DMSO. Thereafter, the body weight and health status of the two groups of mice were observed every day, and the tumor burden of the mice was monitored by bioluminescence imaging (BLI). Once the mice showed obvious central nervous system tumors, significant weight loss, and unsteady gait and other symptoms, CO2 euthanasia was performed, and this was recorded as the end point of the survival curve of this individual.

[0049] As Figure 4 shown, by applying IVIS detection, it is found that compared with the non-drug administration group, the tumor burden in the MMP14 inhibitor drug administration group is reduced, indicating that the MMP14 inhibitor exerts an anti-leptomeningeal metastasis effect and prolongs the overall survival time of LM mice.

[0050] References

[0051] [1] LE RHUN E, TAILLIBERT S, CHAMBERLAIN M C. Carcinomatous meningitis: leptomeningeal metastases in solid tumors [J]. SurgNeurol Int, 2013, 4(Suppl4): S265-88.

[0052] [2] Liang Xiaohua, Huang Ruofan, Zhan Qiong. Shanghai Expert Consensus on the Diagnosis and Treatment of Brain Metastases in Patients with Driver Gene-Positive Non-Small Cell Lung Cancer (2019 Edition) [J]. China Oncology, 2019, 29(01): 71-80.

[0053] [3] PINA Y, GRAMATZKI D, FORSYTH P, et al. Leptomeningeal disease [J]. Hematol Oncol Clin North Am, 2022, 36(1): 189-215.

[0054] [4] SENER U, KUMTHEKAR P, BOIRE A. Advances in the diagnosis, evaluation, and management of leptomeningeal disease [J]. NeurooncolAdv, 2021, 3(Suppl 5): v86-v95.

[0055] [5] Zhou Lei, Zhan Renya. Research progress in the treatment of leptomeningeal metastases [J]. World Latest Medicine Information Digest, 2019, 19(59): 115-8.

[0056] [6] LE RHUN E, DEVOS P, WELLER J, et al. Prognostic validation and clinical implications of the EANO ESMO classification of leptomeningeal metastasis from solid tumors [J]. Neuro Oncol, 2021, 23(7): 1100-12.

[0057] [7]PRAKADAN S M, ALVAREZ-BRECKENRIDGE C A,MARKSON S C, et al. Genomicand transcriptomic correlates of immunotherapy response within the tumormicroenvironment of leptomeningeal metastases [J]. Nat Commun, 2021, 12(1):5955.

[0058] [8]BAYIK D, LATHIA J D. Cancer stem cell-immunecell crosstalk intumour progression [J]. Nat Rev Cancer, 2021, 21(8): 526-36.

[0059] [9]CHI Y, REMSIK J, KISELIOVAS V, et al. Cancercells deploylipocalin-2 to collect limiting iron in leptomeningeal metastasis [J].Science, 2020, 369(6501): 276-82.

[0060]

[10] SMALLEY I, CHEN Z, PHADKE M, et al.Single-cell characterizationof the immune microenvironment of melanoma brain and leptomeningealmetastases [J]. Clin Cancer Res, 2021, 27(14): 4109-25.

[0061]

[11] GHOLAMIN S, MITRA S S, FEROZE A H, et al. Disrupting the CD47-SIRPalpha anti-phagocytic axis by a humanized anti-CD47 antibody is an efficacious treatment for malignant pediatric brain tumors [J]. Sci TranslMed, 2017, 9(381): eaaf2968.

[0062]

[12] GUC E, POLLARD J W. Redefining macrophage and neutrophil biology in the metastatic cascade [J]. Immunity, 2021, 54(5): 885-902.

[0063]

[13] DUAN Z, LUO Y. Targeting macrophages in cancer immunotherapy [J]. Signal Transduct Target Ther, 2021, 6(1): 127.

[0064]

[14] KOHLI K, PILLARISETTY V G, KIM T S. Key chemokines direct migration of immune cells in solid tumors [J]. Cancer Gene Ther, 2022, 29(1): 10-21.

[0065]

[15] GARNER H, DE VISSER K E. Immune crosstalk in cancer progression and metastatic spread: a complex conversation [J]. Nat Rev Immunol, 2020, 20(8): 483-97.

[0066]

[16] TOGASHI Y, SHITARA K, NISHIKAWA H.Regulatory T cells in cancerimmunosuppression - implications for anticancer therapy [J]. Nat Rev ClinOncol, 2019, 16(6): 356-71.

[0067]

[17] DENARDO D G, RUFFELL B. Macrophages asregulators of tumourimmunity and immunotherapy [J]. Nat Rev Immunol, 2019,19(6): 369-82.

[0068]

[18] RUAN H, WANG Z, SUN Z, et al. Single-cellRNA sequencing revealsthe characteristics of cerebrospinal fluid tumour environment in breastcancer and lung cancer leptomeningeal metastases [J]. Clin Transl Med, 2022,12(6): e885.

[0069]

[19] CASTRO-CASTRO A, MARCHESIN V, MONTEIRO P,et al. Cellular andmolecular mechanisms of MT1-MMP-dependent cancer cell invasion [J]. Annu RevCell Dev Biol, 2016, 32: 555-76.

[0070]

[20] SHIMIZU-HIROTA R, XIONG W, BAXTER B T, etal. MT1-MMP regulatesthe PI3Kdelta.Mi-2 / NuRD-dependent control of macrophage immune function [J].Genes Dev, 2012, 26(4): 395-413.

[0071]

[21] WONG H L, JIN G, CAO R, et al. MT1-MMPsheds LYVE-1 on lymphaticendothelial cells and suppresses VEGF-C production to inhibitlymphangiogenesis [J]. Nat Commun, 2016, 7: 10824.

[0072]

[22] GIFFORD V, ITOH Y. MT1-MMP-dependent cellmigration: proteolyticand non-proteolytic mechanisms [J]. Biochem Soc Trans,2019, 47(3): 811-26.

[0073]

[23] GOLDMANN T, WIEGHOFER P, JORDAOM J, et al. Origin, fate anddynamics of macrophages at central nervous system interfaces [J]. NatImmunol, 2016, 17(7): 797-805。

Claims

1. Application of MMP14 as a target for targeted therapy of leptomeningeal metastases in the preparation of targeted drugs for leptomeningeal metastases.

2. The application according to claim 1, characterized in that, The leptomeningeal metastases are leptomeningeal metastases of lung cancer.

3. The application according to claim 1, characterized in that, The targeted therapy includes targeting cells with high expression of MMP14 in leptomeningeal metastases.

4. The application according to claim 3, characterized in that The targeted therapy includes targeting tumor cells with high expression of MMP14, tumor macrophages with high expression of MMP14, or a combination of both.

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