Use of a substance targeting the mif-cd74 axis in the preparation of a medicament for treating male breast cancer
By blocking MIF-CD74 signaling with substances targeting the MIF-CD74 axis and inhibiting M2 polarization of TREM2+ macrophages, combined with a combination therapy regimen, the treatment of insufficient immune microenvironment in male breast cancer was addressed, anti-tumor immune response was restored, and treatment efficacy was improved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- WUHAN UNIV OF SCI & TECH
- Filing Date
- 2026-02-06
- Publication Date
- 2026-06-02
AI Technical Summary
Current treatment options lack effective means to target the specific immune microenvironment of male breast cancer, resulting in poor treatment outcomes, high recurrence rates, and poor prognosis.
By targeting substances along the MIF-CD74 axis, blocking the binding of MIF to CD74 or inhibiting its downstream signaling pathways, inhibiting M2 polarization of TREM2+ macrophages, restoring anti-tumor immune responses, and combining immune checkpoint inhibitors and chemotherapy drugs to form a combined treatment regimen.
It specifically inhibits M2 polarization of TREM2+ macrophages, reverses the immunosuppressive state of the tumor microenvironment, restores anti-tumor immune function, and improves treatment efficacy.
Smart Images
Figure CN122124245A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, specifically involving tumor immunology and molecular targeted therapy. It further elucidates the key role of the MIF and CD74 receptor signaling axis in mediating the polarization of TREM2-positive macrophages to the M2 type, as well as the specific function of this mechanism in the progression of male breast cancer. Background Technology
[0002] 1. Current Status of Research on Male Breast Cancer Male breast cancer (MBC) is a rare but distinctive clinicopathological malignancy. Although it accounts for only about 1% of all breast cancers, its incidence has been rising in recent years. Compared to female breast cancer (FBC), MBC patients are typically older at diagnosis and often at a more advanced stage, resulting in a mortality rate approximately 19% higher, lower survival rates, and a greater likelihood of distant metastasis.
[0003] Currently, most treatment options for breast cancer cystic leukemia (MBC) are based on inferences from clinical trials in women with breast cancer. However, due to differences in sex chromosomes, hormone levels (such as the androgen-to-estrogen ratio), and energy metabolism patterns, MBC exhibits significant biological differences from breast cancer cystic leukemia (FBC). These differences lead to greater resistance to conventional treatments (such as endocrine therapy and chemotherapy), higher recurrence rates, and worse prognoses in MBC. Therefore, there is an urgent need to develop specific treatment strategies based on the unique molecular mechanisms of MBC.
[0004] 2. Tumor microenvironment (TME) and immune escape The tumor microenvironment (TME) is a complex ecosystem on which tumor cells depend for survival and development, and is considered a key mechanism leading to targeted therapy resistance and immune escape. The TME includes not only tumor cells, but also the basement membrane, extracellular matrix, blood vessels, and various non-tumor cell components, such as immune cells (macrophages, T cells, etc.) and fibroblasts.
[0005] In the tumor microenvironment (TME), immunosuppression is a major reason why tumors evade immune surveillance. Tumor-associated macrophages (TAMs), in particular, are one of the most abundant immune cell populations infiltrating the TME and are closely associated with tumor initiation, progression, invasion, and metastasis. Macrophages are highly plastic and can be classified into two main subtypes through the process of "macrophage polarization": classically activated M1 macrophages (with pro-inflammatory and anti-tumor effects) and alternately activated M2 macrophages (with anti-inflammatory, tissue repair-promoting, and angiogenesis-promoting effects).
[0006] In most solid tumors, TAMs mainly exhibit an M2-like phenotype, creating an immunosuppressive microenvironment that promotes tumor growth by secreting anti-inflammatory cytokines (such as IL-10), growth factors, and pro-angiogenic factors, thereby inhibiting the activity of CD8+ T cells.
[0007] 3. Research progress on TREM2+ macrophages Trigger receptor expression in myeloid cells 2 (TREM2) is a protein specifically enriched in immunosuppressive macrophage populations. Studies have shown that TREM2+ macrophages possess unique transcriptional signatures associated with high expression of lipid metabolism, complement activation, and immune checkpoint molecules such as CD276 and SPP1.
[0008] In various cancers (such as triple-negative breast cancer and non-small cell lung cancer), the accumulation of TREM2+ TAMs is closely associated with resistance to immunotherapy (such as anti-PD-1 / PD-L1). These cells can inhibit T cell activation and proliferation and are associated with the accumulation of regulatory T cells (Tregs), leading to poor patient prognosis. However, the specific distribution, origin, and polarization mechanisms of TREM2+ macrophages in male breast cancer have not been reported in detail.
[0009] 4. Biological functions of the MIF-CD74 axis MIF (macrophage migration inhibitory factor) is a pleiotropic cytokine that acts as a key regulator of the innate immune response. It is highly expressed in various solid tumors, such as colon cancer, lung cancer, and breast cancer, and is associated with poor prognosis. MIF exerts its pro-cancer effect by binding to its receptor CD74 (a chaperone protein of major histocompatibility complex class II molecules) and activating downstream signaling pathways such as AKT.
[0010] Existing research has found that the MIF-CD74 axis plays a crucial role in regulating the immunosuppressive microenvironment and modulating the function of macrophages and dendritic cells. For example, in melanoma, blocking this axis can inhibit tumor proliferation by inducing apoptosis. However, there is currently no research or application regarding the mechanism by which the MIF-CD74 axis specifically mediates the M2 polarization of TREM2+ macrophages, thereby driving the progression of male breast cancer. Summary of the Invention
[0011] The invention aims to address the lack of effective therapeutic methods targeting the specific immune microenvironment of male breast cancer in existing technologies. Specifically, based on single-cell sequencing data analysis, this invention has discovered and confirmed that in male breast cancer, epithelial cells specifically induce M2-type polarization of macrophages by secreting MIFs that bind to CD74 receptors on the surface of TREM2+ macrophages, thereby constructing an immunosuppressive microenvironment and promoting tumor progression.
[0012] The technical solution of this invention is achieved as follows: the application of a substance targeting the MIF-CD74 axis in the preparation of a drug for treating male breast cancer, the application being based on the following pathological mechanism: In male breast cancer tissue, tumor epithelial cells highly express MIF, which binds to CD74 receptors on the surface of TREM2+ macrophages in the tumor microenvironment, mediating macrophage polarization to M2 type and thus promoting tumor immune escape. The substance inhibits M2 type polarization of TREM2+ macrophages and restores anti-tumor immune response by blocking the binding of MIF to CD74 or inhibiting its downstream signaling pathways.
[0013] The present invention provides a pharmaceutical composition for treating male breast cancer, the pharmaceutical composition comprising an effective amount of a MIF inhibitor, a CD74 inhibitor or a MIF-CD74 interaction blocker, and a pharmaceutically acceptable carrier.
[0014] Preferably, the MIF inhibitor is selected from anti-MIF antibodies, MIF small molecule antagonists (such as ISO-1), or MIF antisense oligonucleotides.
[0015] Preferably, the CD74 inhibitor is selected from anti-CD74 antibodies (such as Miltauzumab) or CD74 small molecule inhibitors.
[0016] Preferably, the pharmaceutical composition further comprises an immune checkpoint inhibitor (such as an anti-PD-1 or anti-PD-L1 antibody), a chemotherapy drug, or an endocrine therapy drug, forming a combination therapy regimen.
[0017] This invention provides biomarkers for the diagnosis or prognosis of male breast cancer, said biomarkers including the expression levels of MIF, CD74 and / or TREM2.
[0018] Using the above method, the expression levels of MIF, CD74, and TREM2 in male breast cancer tissues were significantly correlated, and patients with high expression of these three molecules had a worse prognosis. Therefore, detecting the expression levels of these three molecules can be used for molecular subtyping, efficacy prediction, and prognostic assessment of male breast cancer.
[0019] After adopting the above technical solution, the beneficial effects of the present invention are: 1. High specificity: This invention reveals for the first time the core role of the MIF-CD74 axis in the specific immune microenvironment of male breast cancer. The intervention strategy targeting this target has high gender specificity and is expected to solve the problem of poor efficacy of existing therapies for MBC.
[0020] 2. Mechanism is clear: By blocking MIF-CD74 signaling and specifically inhibiting M2 polarization of TREM2+ macrophages, the immunosuppressive state of the tumor microenvironment can be effectively reversed and the body's anti-tumor immune function can be restored.
[0021] 3. Broad application prospects: This invention not only provides new drug targets, but also diagnostic biomarkers and combination drug strategies, which have important clinical translational value and commercial potential. Attached Figure Description
[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.
[0023] Figure 1 Annotated diagram of cell subpopulations provided for embodiments of the present invention; Figure 2 A cell proportion composition diagram provided for an embodiment of the present invention; Figure 3 Cell scoring diagram provided in an embodiment of the present invention; Figure 4 A detailed annotation diagram of myeloid cell subdivision provided in an embodiment of the present invention; Figure 5 A statistical chart of cell percentage provided for an embodiment of the present invention; Figure 6 This is a macrophage polarization score map provided in an embodiment of the present invention; Figure 7 Cell communication analysis diagram provided for embodiments of the present invention; Figure 8 A diagram illustrating the mechanism intervention strategy provided in this embodiment of the invention. Detailed Implementation
[0024] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Example
[0025] Single-cell atlas construction and differential analysis of tumor microenvironments in male and female breast cancer To gain a deeper understanding of the unique biological characteristics of male breast cancer (MBC), this embodiment utilizes single-cell RNA sequencing technology to construct high-resolution atlases of tumor tissues from MBC and FBC patients.
[0026] 1. Sample collection and processing: Fresh tumor tissue resected from 5 newly diagnosed, untreated male breast cancer patients (MBC group) and 5 female breast cancer patients (FBC group) was collected. The tissue was minced and digested using collagenase and DNase to prepare a single-cell suspension. Flow cytometry was used to sort and remove dead cells and double cells, obtaining high-quality single-cell samples.
[0027] 2. Single-cell transcriptome sequencing: Single cells were captured, reverse transcribed, and libraries were constructed using the 10x Genomics Chromium system. Sequencing was performed on an Llumina NovaSeq 6000 platform at a depth of 50,000 reads per cell.
[0028] 3. Data Analysis and Cell Subpopulation Annotation: The raw sequencing data were aligned and quantified using CellRanger software to generate a gene expression matrix. Data standardization, dimensionality reduction (PCA), and cluster analysis were performed using the Seurat software package.
[0029] Based on known marker genes, we annotated all cells into six major lineages: epithelial cells (marker gene EPCAM), endothelial cells (marker gene PECAM1), fibroblasts (marker gene COL1A1), T cells (marker gene CD3D), B cells (marker gene CD19), and myeloid cells (marker genes CD14 / CD68). 4. Results Analysis: For example... Figure 3 As shown, by comparing the cell proportions of the MBC and FBC groups, we found that the proportion of myeloid cells and epithelial cells in the MBC sample was significantly higher than that in the FBC sample (myeloid cells accounted for approximately 25% in the MBC group and approximately 15% in the FBC group). Further subdivision of myeloid cells revealed nine different macrophage subpopulations, including: LYVE1+ Mac, TREM2+ Mac, FOLR2+ Mac, NLRP3+ Mac, ISG15+ Mac, TOP2A+ Mac, and dendritic cells (DCs).
[0030] Further investigation revealed that the proportion of TREM2+ macrophages (TREM2+Mac) was highest in MBC samples and significantly higher than in FBC samples (e.g., Figure 3 As shown in the bar chart, TREM2+Mac accounts for up to 12% in MBC, while it is only about 4% in FBC. Example
[0031] Phenotypic characteristics and polarization status analysis of TREM2+ macrophages To determine the functional phenotype of TREM2+ macrophages enriched in MBC, this embodiment conducted an in-depth analysis of their transcriptomic characteristics.
[0032] 1. Differentially expressed gene analysis: Gene expression data of TREM2+ macrophages were extracted and differentially expressed. The results showed that this subset highly expressed immunosuppression-related genes (such as CD206 / MRC1, ARG1), lipid metabolism-related genes (such as APOE, FABP5), and immune checkpoint molecules (such as SPP1, CD276), which is highly consistent with the characteristics of typical M2 macrophages.
[0033] 2. M1 / M2 polarization scoring: The ssGSEA (single-sample gene set enrichment analysis) algorithm was used to score the M1 (pro-inflammatory) and M2 (anti-inflammatory) polarization of different macrophage subsets. Results are as follows: Figure 4 As shown, TREM2+ macrophages had significantly higher M2 scores than other macrophage subsets (such as LYVE1+Mac or FOLR2+Mac), while their M1 scores were extremely low. This indicates that TREM2+ macrophages enriched in male breast cancer are mainly in an M2 polarized state and possess strong immunosuppressive functions. Example
[0034] Cell communication analysis reveals that the MIF-CD74 axis is a key pathway driving TREM2+ macrophage polarization. To investigate what signals lead to the specific enrichment and polarization of TREM2+ macrophages in MBC, this embodiment uses CellChat software to perform intercellular communication analysis.
[0035] 1. Inference of ligand-receptor interactions: Based on single-cell expression data, the strength of ligand-receptor interactions between epithelial cells and TREM2+ macrophages was calculated.
[0036] The analysis results showed that among numerous signaling pathways, the interaction weight of the MIF-CD74 pathway was significantly higher in the MBC group than in the FBC group (e.g., ...). Figure 5 As shown, the thicker and darker-colored lines connecting the MBC groups indicate a stronger effect.
[0037] 2. Mechanism Deduction: Data shows that epithelial cells of male breast cancer highly express the ligand MIF, while TREM2+ macrophages highly express the receptor CD74. This specific ligand-receptor dialogue suggests that MBC tumor cells "educate" macrophages to polarize towards the TREM2+M2 type by secreting MIF, which binds to CD74 on the surface of macrophages. Example
[0038] In vitro validation of the effect of the MIF-CD74 axis on macrophage polarization To verify the hypotheses of the above bioinformatics analysis, in vitro cell experiments were conducted in this embodiment.
[0039] 1. Cell culture: Human peripheral blood mononuclear cells were isolated and induced to differentiate into MO macrophages using GM-CSF. Male breast cancer cell lines (such as MCF7 or ZR-75-1, although derived from females, are often used for model construction) or primary MBC cells were cultured.
[0040] 2. Co-culture experiment: M0 macrophages were co-cultured with male breast cancer cells using Transwell assays. Experimental groups: Control group: M0 macrophages were cultured alone.
[0041] Model group: M0 macrophages + male breast cancer cells (simulating TME).
[0042] Intervention group: MO macrophages, male breast cancer cells, anti-MIF neutralizing antibody (or CD74 blocking antibody).
[0043] 3. Detection indicators: The expression ratios of macrophage surface markers CD206 (M2 marker) and CD86 (M1 marker) were detected by flow cytometry; the mRNA expression levels of TREM2, ARG1, IL-10 (M2 gene) and iNOS and TNF-α (M1 gene) were detected by gRT-PCR.
[0044] 4. Experimental Results: Model group: After co-culturing with male breast cancer cells, the proportion of CD206+ in macrophages increased significantly and TREM2 expression was upregulated, indicating that tumor cells induced macrophages to polarize towards TREM2+M2 type.
[0045] In the intervention group, the addition of anti-MIF or anti-CD74 antibodies significantly reduced the proportion of CD200+ macrophages, decreased TREM2 expression, and simultaneously increased the expression of M1-related genes such as iNOS. This directly demonstrates that blocking the MIF-CD74 axis can inhibit the polarization of TREM2+M2 macrophages. Example
[0046] Preparation and application of the pharmaceutical compositions of the present invention Based on the above findings, this embodiment provides several specific drug formulation schemes.
[0047] Option A: Anti-MIF monoclonal antibody injection Active ingredient: Recombinant anti-human MIF monoclonal antibody, concentration 20 mg / mL.
[0048] Pharmaceutical excipients: histidine-histidine hydrochloride buffer (pH 6.0), sodium chloride, polysorbate 20.
[0049] Preparation method: Dissolve the antibody in buffer solution, adjust the pH value, filter sterilely, and dispense.
[0050] Application: By binding to MIF in the tumor microenvironment and blocking its binding to CD74 on macrophages, TREM2+ macrophages are inhibited, thereby reducing their polarization and making them suitable for the treatment of male breast cancer.
[0051] Option B: CD74-targeted small molecule inhibitor capsules Active ingredient: Compound X, an inhibitor of CD74 intracellular signal transduction, at a concentration of 50 mg / capsule.
[0052] Excipients: Microcrystalline cellulose, lactose, sodium carboxymethyl starch, magnesium stearate.
[0053] Preparation method: conventional wet granulation, tableting, and coating.
[0054] Uses: Oral administration, interferes with CD74-mediated downstream signaling, and reverses the immunosuppressive microenvironment.
[0055] Option C: Combination therapy strategy Combination: Anti-PD-1 antibody + MIF inhibitor.
[0056] Mechanism: MIF inhibitors are responsible for "reprogramming" the tumor microenvironment, reducing TREM2+M2 macrophages, and relieving immunosuppression; anti-PD-1 antibodies are responsible for activating T cells. The combination of these two can produce a synergistic effect, addressing the issue of MBC's insensitivity to immunotherapy. Example
[0057] Construction of diagnostic kits This embodiment provides a kit for detecting the prognosis of male breast cancer patients.
[0058] Contents include: specific primers and probes targeting the MIF, CD74, and TREM2 genes; or immunohistochemical antibody reagents targeting these three proteins. Interpretation criteria: If MIF, CD74, and TREM2 are all highly expressed in the patient's tumor tissue (High-High-High pattern), it is classified as "MIF-CD74 axis activated" MBC, with a poor prognosis, but the patient is highly likely to benefit from the targeted drug of this invention.
[0059] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several improvements and additions without departing from the method of the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention.
Claims
1. The application of a substance targeting the MIF-CD74 axis in the preparation of a drug for treating male breast cancer, characterized in that, The application is based on the following pathological mechanism: In male breast cancer tissue, tumor epithelial cells highly express MIF, which binds to CD74 receptors on the surface of TREM2+ macrophages in the tumor microenvironment, mediating macrophage polarization to M2 type and thus promoting tumor immune escape. The substance inhibits M2 type polarization of TREM2+ macrophages and restores anti-tumor immune response by blocking the binding of MIF to CD74 or inhibiting its downstream signaling pathways.
2. The application of the substance targeting the MIF-CD74 axis according to claim 1 in the preparation of a drug for treating male breast cancer, characterized in that, The substance is selected from any one or more of the following: anti-MIF antibody, MIF small molecule antagonist, anti-CD74 antibody, CD74 small molecule inhibitor, MIF antisense nucleotide, or siRNA.
3. A pharmaceutical composition for treating male breast cancer, characterized in that, It comprises an effective amount of the substance as described in claim 1 or 2, and a pharmaceutically acceptable carrier.
4. The pharmaceutical composition according to claim 3, characterized in that, The dosage form of the pharmaceutical composition is an injection, a lyophilized powder for injection, a tablet, or a capsule.
5. A combination therapy regimen, characterized in that, The substance described in claim 1 or 2 may be used in combination with an immune checkpoint inhibitor, a chemotherapy drug, or an endocrine therapy drug.
6. A biomarker for the diagnosis or prognosis of male breast cancer, characterized in that, The biomarkers include the gene or protein expression levels of MIF, CD74, and / or TREM2.
7. The biomarker according to claim 6, characterized in that, The expression levels of MIF, CD74, and TREM2 in samples were detected to screen suitable targets for receiving MIF-CD74 axis.