The use of CCL3 in the immunotherapy of multiple myeloma

Abstract

This disclosure belongs to the field of biomedical technology, specifically disclosing the use of CCL3 in the immunotherapy of multiple myeloma. On one hand, it provides the application of preparations for detecting CCL3 expression in the preparation of products analyzing the risk of drug resistance and relapse after multiple myeloma immunotherapy and in analyzing the severity of the immunosuppressive microenvironment in multiple myeloma. On the other hand, it provides the application of IMNP in the preparation of immunotherapy products for multiple myeloma. Furthermore, it provides the application of IMNP combined with PD-L1 antibodies in the preparation of immunotherapy products for multiple myeloma. This invention provides new observation and analysis methods for multiple myeloma immunotherapy, enabling better monitoring and evaluation of changes in immunotherapy efficacy during the treatment process. IMNP, by efficiently clearing CCL3 molecules, improves the immunosuppressive microenvironment of myeloma, reducing the risk of drug resistance and relapse during multiple myeloma immunotherapy. Furthermore, the combination of IMNP and PD-L1 antibodies enhances the efficacy of MM immunotherapy.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the use of CCL3 in the immunotherapy of multiple myeloma. Background Technology

[0002] Multiple myeloma (MM) is the second most common hematologic malignancy. It primarily invades the bone marrow, but can also form extramedullary lesions, such as in the liver, spleen, kidneys, lymph nodes, skin, lungs, and thyroid gland. The latter is a sign of rapid disease progression and indicates a poor prognosis. The emergence of new drugs for treating multiple myeloma (such as proteasome inhibitors, immunomodulators, and CD38 monoclonal antibodies) and the application of standard clinical treatment modalities, including induction therapy, consolidation therapy, and maintenance therapy, have significantly improved progression-free survival (PFS) and overall survival (OS) in MM patients. However, MM remains incurable, and relapse is inevitable. The key reason is that the bone marrow immunosuppressive microenvironment supports and protects against minimal residual disease (MRD) and induces drug resistance, making complete eradication of MRD difficult. Extensive clinical data show a close correlation between the degree of MRD clearance and PFS and OS benefits in MM patients. Effectively regulating the bone marrow immunosuppressive microenvironment to completely eradicate MRD and prolong PFS to strive for a cure for MM has significant clinical value.

[0003] Inflammatory factors are an important component of the immunosuppressive microenvironment in the bone marrow of multiple myeloma (MM), and the progression of MM is often accompanied by a significant increase in the levels of a large number of inflammatory factors. Chemokine ligand 3 (CCL3) is an important inflammatory chemokine, mainly derived from MM cells and the bone marrow microenvironment (such as osteoclasts). Previous studies on CCL3 have primarily focused on characterizing its chemotactic capacity and have expanded to its role in autoimmune inflammatory processes and hematologic malignancies (such as multiple myeloma), revealing a dual role for CCL3: as an effective immune activator and a promoter of inflammatory disease processes. However, no studies have revealed the role of CCL3 in the formation of the immunosuppressive microenvironment in myeloma.

[0004] Numerous studies have shown that the bone marrow microenvironment in MM patients is primarily characterized by an immunosuppressive phenotype that supports tumor growth (i.e., an immunosuppressive microenvironment). This includes abnormal dendritic cell function leading to antigen-presenting defects, an increase in the number of immunosuppressive M2 macrophages, regulatory T cells (Tregs), and myeloid-derived suppressor cells (MDSCs), a decrease in the number and dysfunction of cytotoxic T cells with MM-killing effects, and a significant increase in immunosuppressive soluble cytokines such as TGF-β, IL-10, and IL-8. Research indicates that the state of the bone marrow microenvironment in MM patients is correlated with the degree of minimal residual disease clearance; regulating the bone marrow microenvironment is beneficial for the complete clearance of minimal residual disease. Therapeutic drugs (such as immunomodulators and CD38 monoclonal antibodies) or treatment methods (such as autologous hematopoietic stem cell transplantation) with certain bone marrow microenvironment regulatory effects have achieved remarkable efficacy in the treatment of MM. However, drug resistance and relapse are still unavoidable in MM patients after the above treatments, suggesting that existing methods for reshaping the myeloma immunosuppressive microenvironment are insufficient to fully meet clinical needs in MM treatment. Furthermore, PD-L1 antibodies, which have shown remarkable efficacy in some solid tumors such as breast cancer and melanoma, have not performed as well in MM. Therefore, new methods for regulating the immunosuppressive microenvironment in myeloma urgently need to be developed. Summary of the Invention

[0005] To address the aforementioned issues, this invention provides the use of CCL3 in the immunotherapy of multiple myeloma, primarily to solve problems such as the need to supplement and improve new methods for detecting the severity of the bone marrow immunosuppressive microenvironment and the urgent need to develop methods for regulating the bone marrow immunosuppressive microenvironment.

[0006] To solve the above problems, the present invention adopts the following technical solution:

[0007] The first aspect provides the application of formulations for detecting CCL3 expression in the preparation of products for detecting and analyzing the risk of drug resistance and relapse after immunotherapy for multiple myeloma. High CCL3 expression is associated with a high risk of drug resistance and relapse, and the CCL3 expression level is positively correlated with the risk of drug resistance and relapse after immunotherapy for multiple myeloma. In immunotherapy for multiple myeloma, high CCL3 expression indicates more severe immunosuppression in the bone marrow microenvironment, leading to a higher likelihood of drug resistance and relapse after immunotherapy. The risk of drug resistance and relapse after immunotherapy for multiple myeloma can be assessed and analyzed before and after immunotherapy. For example, the expected immunotherapy effect can be assessed based on CCL3 expression before immunotherapy, and the immunotherapy effect can also be assessed based on CCL3 expression after immunotherapy. Furthermore, a high risk of drug resistance and relapse indicates poor immunotherapy efficacy.

[0008] The level of CCL3 expression can be differentiated based on general clinical understanding, such as the expression level of CCL3 in the corresponding site or cell, and the CCL3 expression level can be used as a reference for treatment. CCL3 expression can be detected using conventional methods, such as IHC, WB, and ELISA, etc. The term "product" mentioned in this text can be interpreted as at least as detection reagents, kits, and detection equipment, without specific limitations on their exact form, the same applies below.

[0009] The second aspect provides the application of formulations for detecting CCL3 expression in the preparation of products for analyzing the severity of the immunosuppressive microenvironment in multiple myeloma. CCL3 expression levels are positively correlated with the severity of immunosuppression. The severity of the immunosuppressive microenvironment in myeloma can be determined by CCL3 expression; higher CCL3 expression indicates more severe immunosuppression in the myeloma microenvironment, providing guidance for clinical diagnosis and treatment. Besides clinical applications, it can also be used in research projects. When validating the therapeutic effects of different drugs on myeloma, monitoring CCL3 expression can reveal the extent of immune response suppression, thus clarifying whether the drug effectively treats myeloma. Existing technologies can be used to detect CCL3 expression, without specific limitations.

[0010] The third aspect provides a method for preparing IMNPs and the IMNPs obtained by this method.

[0011] The preparation method of IMNP includes the following steps: macrophage membranes obtained by co-incubating macrophages with IFN-γ and nanocarriers or nanoliposomes are mixed to prepare IMNP.

[0012] The purpose of co-incubating macrophages with IFN-γ is to optimize the stimulation of macrophages with IFN-γ, and then obtain IMNPs by coating nanocarriers with macrophage membranes or fusing them to the surface of nanoliposomes.

[0013] Regarding the preparation of IMNPs, any one or more of the conditions in the preparation process can be selected:

[0014] In reaction condition one, when preparing the membrane-coated nanocarrier: acetone containing a polyester material is injected into water, and then the acetone is removed to obtain the nanocarrier. The polyester material can be at least one of PLGA (polylactic acid-glycolic acid copolymer), PLA (polylactic acid), and PCL (polycaprolactone), but is not limited to these. The acetone can be removed by evaporation. In preparing the membrane-fused liposome: after the lipid material film is formed, the membrane material is added during hydration, mixed, sonicated, and extruded to form membrane-fused nanoliposomes. The nanoliposomes can be prepared using existing technology.

[0015] In reaction condition two, if the cell membrane is selected from any of the following: conventional RAW264.7 macrophage membrane, neutrophil membrane, hybrid membrane of different types of biological membranes, or cell membrane of gene-edited cells (not limited to these), then all such selections shall fall within the scope of the claims of this invention.

[0016] Reaction condition three: IMNPs were obtained by mixing macrophage membranes and nanocarriers and then sonicating them in an ice bath.

[0017] Reaction condition four, for the materials used in the preparation of IMNP, can also be limited depending on the circumstances:

[0018] One of them is that the stimulation concentration of IFN-γ is generally 10±0.5ng / ml. Of course, if it fluctuates outside this range but has a similar effect to the present invention, it should still be within the scope of the present invention.

[0019] Secondly, all materials can be directly subjected to ultrasonic reaction in pure water.

[0020] Taking PLGA nanoparticles as an example, IMNPs were prepared using RAW264.7 macrophages and the polyester material PLGA. PLGA nanoparticles can be prepared directly using existing materials or existing preparation methods. 500 μL of acetone containing 5 mg of PLGA polymer was rapidly injected into 1 mL of deionized water. After removing the acetone by vacuum evaporation, PLGA nanoparticles were obtained. 1 mg of IFN-γ-stimulated RAW264.7 macrophage membrane was added and mixed thoroughly. The mixture was then sonicated in an ice bath (200 W) for 5 minutes to obtain IMNPs. Large-scale preparation can be adjusted according to actual needs.

[0021] The IMNPs prepared by any of the aforementioned methods exhibit good biocompatibility. IMNPs generally have a particle size of around 100 nm, a potential of around -20 mV, and appear as regular spheres under an electron microscope. However, this is not limited to the aforementioned characteristics; any IMNPs that can be identified using conventional material identification methods as being identical to or having the same coating structure as those prepared by any of the aforementioned methods should be included within the scope of this invention.

[0022] The fourth aspect provides the application of IMNP in the preparation of adsorbent formulations for CCL3, wherein the CCL3 is present in the immunosuppressive microenvironment of multiple myeloma. IMNP can effectively remove CCL3 from the immunosuppressive microenvironment of myeloma. Of course, the removal mentioned here does not mean complete removal; it is a relative expression. The purpose is to reduce the content of CCL3 in the immunosuppressive microenvironment of multiple myeloma and to improve the immunosuppressive microenvironment of multiple myeloma, thereby reducing the inhibitory effect of the immune response. This type of adsorbent formulation has both pharmaceutical uses and can be used as an experimental reagent, such as in animal or cell models, where the amount of CCL3 can be controlled.

[0023] The fifth aspect provides the application of nanomedicines with CCL3 clearance effects in the preparation of products for immunotherapy of multiple myeloma; wherein, the nanomedicines with CCL3 clearance effects refer to any of the aforementioned IMNPs or nanomedicines formed by coating nanocarriers or fusing them to the surface of nanoliposomes with any of the following: RAW264.7 macrophage membranes, neutrophil membranes, hybrid membranes of different types of biological membranes, or cell membranes of gene-edited cells. In some specific examples, nanomedicines with CCL3 clearance effects are used to prepare products that inhibit drug resistance relapse after multiple myeloma immunotherapy. Specifically, the drug resistance relapse may refer to the inhibition of drug resistance relapse after multiple myeloma PD-L1 antibody immunotherapy, or nanomedicines with CCL3 clearance effects may be used to prepare products against drug resistance to multiple myeloma immunotherapy, thus resisting drug resistance.

[0024] The effects of IMNP are at least one of the following: IMNP improves the immunosuppressive microenvironment of bone marrow, and / or IMNP inhibits the osteoclast-inducing effect of CCL3, and / or IMNP inhibits myeloma cell proliferation and migration, and / or reduces the risk of drug resistance relapse after multiple myeloma immunotherapy, and / or improves the immunotherapy efficacy of myeloma PD-L1 antibody; satisfying any one of these conditions is within the scope of this invention, but is not limited to the aforementioned four effects. Through any of the aforementioned regulatory effects, it is shown that IMNP is beneficial to the immunotherapy of multiple myeloma, and can reduce the incidence of MM drug resistance relapse to a certain extent, improve the efficacy of immunotherapy, and has a significant synergistic effect. "At least one of the following effects" can be understood as at least one of the listed, but is not required to be one of the listed.

[0025] Regarding improving the immunosuppressive microenvironment of bone marrow: the aim is to reduce the number of immune cells supporting myeloma cell growth in the immunosuppressive microenvironment of myeloma, and increase the number of immune cells with myeloma cell-killing effects. This can also be used to eliminate minimal residual disease and reduce its drug resistance, thereby prolonging PFS and OS in MM patients. Nanomedicines with CCL3 clearance effects can be used in the preparation of drugs to inhibit resistance to immunotherapy in multiple myeloma. Nanomedicines with CCL3 clearance effects are used to inhibit resistance to PD-L1 antibody immunotherapy in multiple myeloma.

[0026] Regarding the inhibition of CCL3's osteoclast-inducing effect and myeloma cell proliferation and migration: the aim is to inhibit osteoclast differentiation and myeloma cell proliferation and migration, thereby inhibiting myeloma progression and improving the immunosuppressive microenvironment of myeloma. Of course, it is not required that CCL3 simultaneously inhibit all the aforementioned mechanisms to be within the scope of this invention. Meeting any one of these mechanisms is sufficient to be considered within the scope of this invention. For example, inhibiting myeloma cell proliferation and inhibiting myeloma cell migration are two separate mechanisms; meeting either one and being beneficial for the treatment of multiple myeloma should be within the scope of this invention.

[0027] The sixth aspect provides the application of IMNP combined with PD-L1 antibody in the preparation of immunotherapy products for multiple myeloma. The combination of IMNP and PD-L1 antibody demonstrates significant efficacy in the treatment of multiple myeloma, and the combined effect is significantly different from that of the IMNP group and the PD-L1 antibody group alone. This reflects the synergistic effect of IMNP and PD-L1 antibody in the treatment of multiple myeloma, which can more effectively prevent drug resistance and relapse.

[0028] Immunotherapy products for multiple myeloma are used to kill tumor cells in the bone marrow, and / or immunotherapy products for myeloma are used to inhibit extramedullary infiltration foci of myeloma in the spleen. Killing tumor cells in the bone marrow: Its main effect is to kill myeloma tumor cells while simultaneously reducing their migration. Inhibiting extramedullary infiltration foci of myeloma in the spleen: Specifically, IMNP combined with PD-L1 antibodies can effectively inhibit the formation of extramedullary infiltration foci (extramedullary invasion or extramedullary aggression: such as metastasis of multiple myeloma to the spleen) in multiple myeloma, thereby inhibiting the formation of infiltration foci in the spleen, thus alleviating the condition of patients with rapidly progressing or advanced multiple myeloma and prolonging their survival.

[0029] This disclosure provides a new observation and analysis method for immunotherapy of multiple myeloma, enabling better monitoring and evaluation of changes in immunotherapy efficacy during the treatment process. The invention also provides nanomedicines constructed using IFN-γ stimulation-optimized macrophage membrane-coated nanoparticles, exhibiting a stronger CCL3 adsorption effect than ordinary macrophage membrane-coated nanoparticles. By efficiently clearing CCL3 molecules, this improves the immunosuppressive microenvironment of myeloma, reduces the risk of drug resistance and relapse during multiple myeloma immunotherapy, and further enhances the efficacy of MM immunotherapy by combining IMNP with PD-L1 antibodies. Attached Figure Description

[0030] Figure 1 This is a flowchart of the research project.

[0031] Figure 2 Correlation analysis of CCL3 expression with overall prognosis and immunosuppression in multiple myeloma. CCL3 expression levels in myeloma patients and healthy individuals (A); Prognostic analysis of CCL3 expression levels in myeloma patients (B); Correlation analysis of CCL3 levels with immunosuppressive molecules in myeloma (C); Correlation analysis of CCL3 levels with different types of stromal cells in myeloma (D).

[0032] Figure 3 To optimize, prepare, and characterize MNPs, the effects of different concentrations of IFN-γ on the expression levels of CRR1 and CCR5 on macrophage membranes were analyzed by flow cytometry (A-D) and Western spectroscopy (E&F); the distribution of CCR1 and CCR5 on IMNPs (G&H); and the particle size (I), potential (J), electron microscopy (K), and stability (L) of IMNPs were determined.

[0033] Figure 4 The study investigated the scavenging effect of IMNP on CCL3 and its influence on myeloma cell proliferation and migration. Evaluation of the scavenging effect of IMNP and its control group on CCL3 (A); Evaluation of the scavenging effect of IMNP on CCL3 in different media (B); Evaluation of the scavenging effect of fixed IMNP concentration on different concentrations of CCL3 (C); Evaluation of the scavenging effect of different concentrations of IMNP on a fixed concentration of CCL3 (D); Time-dependent effect of IMNP on CCL3 scavenging (E); The scavenging effect of IMNP on CCL3 derived from the 5TGM1 myeloma cell line (F) and its influence on 5TGM1 cell proliferation (G&H) and migration (I).

[0034] Figure 5IMNP inhibits CCL3-induced osteoclast differentiation. Flow cytometry (A&B) and TRAP staining (C) were used to assess the effect of IMNP on CCL3-induced osteoclast differentiation; ELISA was used to detect the expression of the CCL3 effector molecule APRIL (D).

[0035] Figure 6 Qualitative (A) and quantitative (B&C) analysis of tumor burden in mice bearing the 5TGM1-Luc model under different treatments (PBS group, RNP group, MNP group, IMNP group, PD-L1 antibody group, and IMNP combined with PD-L1 antibody group).

[0036] Figure 7 Survival curves (A), peripheral blood globulin levels (B), bone marrow CCL3 levels (C&G top row), proportion of myeloma cells in peripheral blood (D), proportion of myeloma cells in bone marrow (E&G bottom row), and proportion of myeloma cells in spleen (F) are shown for different treatments (PBS group, RNP group, MNP group, IMNP group, PD-L1 antibody group, and IMNP combined with PD-L1 antibody group) in 5TGM1-Luc model mice.

[0037] Figure 8 To investigate the levels of bone MDSCs and CD8 in the spleen and bone marrow of 5TGM1-Luc model mice under different treatments (PBS group, RNP group, MNP group, IMNP group, PD-L1 antibody group, and IMNP combined with PD-L1 antibody group). + T cells, GranB + CD8 + T cells, CD4 + The ratio of T cells to Treg cells.

[0038] In the figure: * indicates a p-value less than 0.05, ** indicates a p-value less than 0.01, *** indicates a p-value less than 0.001, **** indicates a p-value less than 0.0001, and the smaller the p-value, the more significant the difference. Detailed Implementation

[0039] The invention will now be described in detail with reference to specific research projects.

[0040] I. Design and Fabrication of IMNPs

[0041] The preparation of IMNPs was as follows: S1, acetone containing dissolved PLGA was injected into water; S2, PLGA nanoparticles were obtained by evaporation to remove the acetone; S3, 10 ng / ml IFN-γ-stimulated RAW264.7 macrophage membranes were added and mixed, followed by sonication in an ice bath to obtain IMNPs. The ratio of macrophage membranes to PLGA nanoparticles was 1:2. The cell membrane content was 2.5 mg, and the PLGA content was 5 mg. Both were placed in pure water and sonicated.

[0042] The macrophage membrane can also be any of the following: conventional RAW264.7 macrophage membrane, neutrophil membrane, hybrid membrane of different types of biological membranes, or cell membrane of gene-edited cells.

[0043] II. CCL3 expression is associated with the immunosuppressive microenvironment of myeloma.

[0044] Analysis of data from public databases (two datasets: GSE47552 and MMRF) revealed that CCL3 expression was significantly higher in myeloma patients than in healthy individuals. Figure 2 In the middle A), high CCL3 expression suggests a poor overall prognosis for myeloma patients. Figure 2 In myeloma patients, the expression levels of CCL3 are positively correlated with the expression levels of various immunosuppressive molecules in myeloma, such as GEM, HACVR2, and CTLA4. Figure 2 The distribution of C cells was positively correlated with the distribution of immunosuppressive cells (such as Treg and M2 macrophages) and negatively correlated with the distribution of immune-activating cells such as osteoblasts and Th1 cells. Figure 2 The results suggest that CCL3 is associated with the severity of the immunosuppressive microenvironment in myeloma and may serve as an effective therapeutic target for reversing the immunosuppressive microenvironment in myeloma.

[0045] III. RAW264.7 cell membrane optimized by IFN-γ stimulation

[0046] IMNPs were prepared from macrophage membranes stimulated with IFN-γ and characterized. Figure 3 (AD) and Western ( Figure 3 The results of E&F studies suggest that 10 ng / ml IFN-γ is optimal for enhancing the expression of the natural receptors CCR1 and CCR5 for CCL3 on macrophage membranes. IMNPs prepared from macrophage membranes stimulated with 10 ng / ml IFN-γ effectively preserve the expression of CCR1 and CCR5 on the macrophage membrane surface. Figure 3 GH), IMNP particle size is about 100nm ( Figure 3 (I), potential around -20mV ( Figure 3 (J), under an electron microscope, it appears as a regular sphere. Figure 3After storage at 4℃ for one week, the particle size of IMNP showed no significant change, indicating that IMNP has good stability. Figure 3 (Middle L).

[0047] IV. The scavenging effect of IMNP on CCL3

[0048] After mixing nanomedicines with CCL3 for a period of time, the supernatant was collected by ultracentrifugation, and the residual CCL3 was determined by enzyme-linked immunosorbent assay (ELISA) to evaluate the scavenging effect of IMNPs on CCL3. The results showed that IMNPs had better CCL3 adsorption capacity compared with MNPs (conventional macrophage membrane-coated nanoparticles), RNPs (erythrocyte membrane-coated nanoparticles), and NPs (non-membrane-coated nanoparticles). Figure 4 The adsorption effect of IMNPs on CCL3 (A) may be related to the significantly higher levels of CCL3-corresponding acceptors such as CCR1 and CCR5 on the IMNP surface compared to MNPs, allowing for more efficient adsorption and scavenging of CCL3. The adsorption effect of IMNPs on CCL3 varies in different media. Figure 4 In the middle B), IMNP clearance of CCL3 showed a concentration-dependent effect. Figure 4 C&D) and time-dependent methods ( Figure 4 IMNP also has a certain scavenging effect on CCL3 derived from the 5TGM1 myeloma cell line. Figure 4 Treatment with IMNP can inhibit the proliferation of 5TGM1 cells by CCL3 (F). Figure 4 G&H) and migration ( Figure 4 The promoting effect of I).

[0049] 5. IMNP can inhibit the osteoclast-inducing effect of CCL3 and suppress APRIL expression.

[0050] CLL3 and IMNP were mixed or not mixed, and the supernatant was collected by ultracentrifugation for inducing osteoclast differentiation. Flow cytometry was used to analyze the supernatant. Figure 5 A&B) and TRAP staining ( Figure 5 The results showed that IMNP could inhibit the osteoclast-inducing effect of CCL3, and the ELISA results further showed that the expression of the CCL3 effector molecule APRIL was decreased in the IMNP-treated group. Figure 5 (D).

[0051] VI. IMNP combined with PD-L1 significantly enhances the immunotherapy effect against myeloma.

[0052] A mouse model of myeloma bearing 5TGM1-Luc was established. After treatment with different groups (PBS group, RNP group, MNP group, IMNP group, PD-L1 antibody group, and IMNP combined with PD-L1 antibody group), the tumor burden of the model mice was monitored periodically using small animal in vivo imaging for qualitative analysis. Figure 6(A) and semi-quantitative ( Figure 6 Both the B and C results showed that IMNP combined with PD-L1 antibody treatment could significantly reduce the tumor burden in myeloma-bearing mice.

[0053] The combination therapy of IMNP and PD-L1 antibody significantly prolonged the survival curve of model mice. Figure 7 (A) significantly reduced the peripheral blood globulin level in model mice. Figure 7 B), CCL3 level in bone marrow ( Figure 7 C& Figure 7 Middle G (top row), peripheral blood ( Figure 7 D), bone marrow tissue ( Figure 7 China E& Figure 7 (lower part of G) and spleen tissue ( Figure 7 Flow cytometry and immunohistochemical results from the combined treatment group (IMNP) showed that the combined treatment group significantly reduced the myeloma cell burden in peripheral blood, bone marrow, and spleen compared to other treatment groups, and the combined IMNP and PD-L1 antibody group showed a significantly different effect compared to other groups. It also reduced the proportion of immunosuppressive MDSCs and Tregs, and increased the proportion of GranB1 cells with tumor-killing effects. + The proportion of T cells ( Figure 8 ), and pharmacodynamic results ( Figure 6 & Figure 7 (To maintain consistency)

[0054] Those skilled in the art will appreciate that various modifications to the above embodiments can be made without departing from the overall spirit and concept of the present invention, and existing technical means can be used for any aspects not detailed herein. All such modifications fall within the protection scope of the present invention. The protection scheme of the present invention is defined by the appended claims.

Claims

1. Application of IMNP in the preparation of products that reduce the risk of drug resistance and relapse after immunotherapy for multiple myeloma; among which, The IMNP is prepared by the following method: macrophage membrane obtained by co-incubating macrophages with IFN-γ is mixed with a nanocarrier or nanoliposomes to prepare IMNP, wherein the macrophage membrane is coated with the nanocarrier or the macrophage membrane is fused to the surface of the nanoliposomes.

2. The application according to claim 1; wherein, When preparing membrane-coated nanocarriers: acetone containing polyester materials is injected into water, and the acetone is then removed to obtain nanocarriers. The polyester materials are at least any one of PLGA, PLA, and PCL. Alternatively, when preparing membrane-fused liposomes: after the lipid material film is formed, the membrane material is added during hydration, mixed, sonicated, and extruded to form membrane-fused nanoliposomes.

3. The application according to claim 2; wherein, The macrophage membrane is the cell membrane of the RAW264.7 cell line.

4. The application according to claim 1; wherein, The stimulating concentration of IFN-γ was 10 ± 0.5 ng / ml.

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