Abstract: Background Multiple Myeloma (MM) is a common hematological malignancy characterized by clonal proliferation of plasma cells in the bone marrow, accompanied by Hypercalcemia (C), Lytic Renal Insufficiency (R), Anemia (R), and Bone Lesions (R). Although new treatment strategies such as proteasome inhibitors, immunomodulatory agents, anti-CD38 monoclonal antibody, and CAR-T cell therapy have shown certain efficacy, patients still have recurrence or disease progression due to drug resistance and other reasons. Objective In recent years, with the in-depth study of the molecular mechanism of MM, it has been found that multiple tumor microenvironment abnormalities are involved in genetic abnormalities, metabolic reprogramming and immune escape, but the specific mechanism has not been fully elucidated. Leukocyte Immunoglobulin-like Receptor B1 (LILRB1) and Cluster of Differentiation 36 (CD36) is associated with metabolic abnormalities and immune escape in humans. Lymphoid Enhancer-binding Factor-1 (LEF-1) is expressed on T cells and pre-B cells and is involved in early lymphocyte development. As a key transcription factor of Wnt/β-catenin signaling pathway, its elevated level is closely related to tumorigenesis, disease progression and drug resistance. The Hepatocyte Nuclear Factor 1Beta (HNF1β) gene is located in the human chromosome 17q12 region, and its deletion leads to β-catenin activation and increased expression of its downstream effector LEF-1. In skin tumors, mutations in LEF-1 can lead to a significant reduction in P53 activity, and the close relationship between LEF-1 and P53 can be observed in human tumor cells and LEF-1 mutant mouse models. Methods The diagnosis, staging and treatment response of MM patients were evaluated according to the 2014 and 2016 International Myeloma Working Group (IMWG) criteria. A total of 76 MM patients and 33 healthy controls (HC) were collected from December 2023 to January 2025 in Beijing Chaoyang Hospital, Capital Medical University. According to the disease status, the patients were divided into active MM (aMM, with CRAB symptoms) group and responding MM (responding MM, achieving partial response or above PR) group. The levels of LILRB1, CD36 and LEF-1 in plasma and bone marrow plasma were measured by Enzyme linked immunosorbent assay (ELISA), and optical genome mapping (OGM) was used to detect cytogenetic abnormalities in 6 MM patients. The correlation between the above results and the main clinical features such as hypercalcemia, renal damage, anemia symptoms, M protein and high-risk genes of the patients was comprehensively analyzed. Results The levels of LILRB1, CD36 and LEF-1 in the plasma of aMM patients were significantly higher than those of HC (P < 0.010), and also significantly higher than those of rMM patients (P < 0.050). The levels of LILRB1 and LEF-1 in the bone marrow plasma of aMM patients were significantly higher than those of HC (P < 0.010) and rMM patients (P < 0.050), while the level of CD36 in aMM patients was not significantly different from that in rMM patients (P > 0.050), but was significantly higher than that in HC patients (P < 0.050). Correlation analysis showed that in the plasma, LEF-1 was significantly positively correlated with LILRB1 (r = 0.403, P < 0.010), LEF-1 was significantly positively correlated with CD36 (r = 0.340, P < 0.050), and LILRB1 was significantly positively correlated with CD36 (r = 0.406, P < 0.010). In the bone marrow plasma, LEF-1 was significantly positively correlated with LILRB1 (r = 0.528, P < 0.010), while the correlation between LEF-1 and CD36 (r = 0.080, P = 0.612) was not significant, and LILRB1 was significantly positively correlated with CD36 (r = 0.343, P < 0.050). In the plasma of MM patients, LILRB1 was significantly positively correlated with IgG-type M protein (r = 0.457, P < 0.050) and IgA-type M protein (r = 0.512, P < 0.050), CD36 was significantly positively correlated with IgA-type M protein (r = 0.740, P < 0.010), and LEF-1 was significantly positively correlated with IgG-type M protein (r = 0.410, P < 0.050) and IgA-type M protein (r = 0.685, P < 0.010). In the bone marrow plasma of MM patients, LILRB1 was significantly positively correlated with the ratio of involved to uninvolved free light chains in plasma (r = 0.387, P < 0.050), and LEF-1 was significantly positively correlated with the ratio of involved to uninvolved free light chains in plasma (r = 0.330, P < 0.050) and IgA-type M protein (r = 0.664, P < 0.010). In the plasma, LILRB1 was significantly positively correlated with LDH (r = 0.657, P < 0.010), TG (r = 0.724, P < 0.010) and extramedullary lesions (r = 0.477, P < 0.050); CD36 was significantly positively correlated with TG (r = 0.489, P < 0.050), MLR (r = 0.396, P < 0.050) and extramedullary lesions (r = 0.417, P < 0.050); LEF-1 was significantly positively correlated with t(4;14) (r = 0.585, P < 0.050) and double-hit (r=0.585, P<0.050). In the bone marrow plasma, LILRB1 was significantly negatively correlated with albumin (r = -0.509, P < 0.050); CD36 was significantly negatively correlated with hemoglobin (r = -0.425, P < 0.050) and HDL-C (r = -0.498, P < 0.050), but was significantly positively correlated with TP53 (r = 0.561, P < 0.050); LEF-1 was significantly positively correlated with TP53 (r = 0.579, P < 0.050) and extramedullary lesions (r = 0.400, P < 0.050).

Key words: Multiple myeloma, LILRB1, CD36, LEF-1, HNF1β gene, prognosis

摘要： 多发性骨髓瘤（Multiple Myeloma，MM）是一种恶性浆细胞肿瘤，其特征是骨髓中浆细胞的克隆性增殖，伴血钙升高（Hypercalcemia，C）、肾损害（Renal Insufficiency，R）、贫血（Anemia，A）和骨损害（Lytic Bone Lesions，B）等CRAB症状。尽管蛋白酶体抑制剂、免疫调节剂、抗CD38单克隆抗体、CAR-T细胞疗法等新型治疗策略已经显示出一定的疗效，但患者仍不可避免地复发或病情进展。目的 近年来，随着对MM分子机制的深入研究，发现涉及遗传学异常、代谢重编程和免疫逃逸等多方面肿瘤微环境异常，但其具体机制尚未完全阐明。人白细胞免疫球蛋白样受体B1（Leukocyte Immunoglobulin-like Receptor B1，LILRB1）和分化簇36（Cluster of Differentiation 36，CD36）与人体代谢异常和免疫逃逸相关，淋巴增强因子1（Lymphoid Enhancer-binding Factor-1，LEF-1）表达于T细胞和前B细胞，并参与早期淋巴细胞的发育，作为Wnt/β-catenin信号通路的重要转录因子，其水平升高与肿瘤发生、病情进展和耐药性密切相关。肝细胞核因子1β（Hepatocyte Nuclear Factor 1 Beta，HNF1β）基因位于人类染色体17q12区域，其缺失可导致β-catenin活化和下游效应因子LEF-1表达增加。在皮肤肿瘤中，LEF-1的突变可以导致P53活性显著降低，且在人肿瘤细胞和LEF-1突变小鼠模型中可观察到LEF-1和P53之间的密切关系。方法 MM患者的诊断、分期和治疗反应评估根据2014年国际骨髓瘤工作组（International Myeloma Working Group，IMWG）的标准进行。收集2023年12月至2025年1月在首都医科大学附属北京朝阳医院石景山院区收住院的MM患者76例，健康对照组（HC）33例。根据疾病状态分为活动性MM（active MM或aMM，具备CRAB症状）组、反应MM（responding MM，取得部分治疗反应PR以上疗效）组。采用酶联免疫吸附试验（Enzyme linked immunosorbent assay, ELISA）方法检测血浆和骨髓浆中LILRB1、CD36和LEF-1水平，同时使用光学基因组图谱技术（Optical Genome Mapping，OGM）对其中6例MM患者的细胞遗传学异常进行检测。将上述结果与患者的高钙血症、肾损害、贫血症状、M蛋白等主要临床特征以及高危基因进行相关性综合分析。结果 aMM组血浆和骨髓浆中LILRB1、CD36和LEF-1水平均显著高于HC（均为P<0.010）和rMM患者（均为P<0.050）；aMM组骨髓浆中LILRB1和LEF-1水平显著高于HC组（P<0.010）和rMM组（P<0.050），CD36水平在aMM组与rMM组间差异无统计学意义（P>0.050），但显著高于HC组（P<0.050）。相关性分析显示，血浆中LILRB1、CD36和LEF-1相互间均呈显著正相关；骨髓浆中，LILRB1与CD36和LEF-1呈显著正相关。在血浆中，MM组患者LILRB1和LEF-1 均与IgG型M蛋白（分别为r = 0.457，P<0.050和r = 0.410，P<0.050）和IgA型M蛋白（分别为r = 0.512，P<0.050和r = 0.685，P<0.010）呈显著正相关；CD36与IgA型M蛋白（r = 0.740，P<0.010）呈显著正相关。在骨髓浆中，MM组患者LILRB1和LEF-1 与血浆受累与非受累游离轻链比值（分别为r = 0.387，P<0.050和r = 0.330，P<0.050）呈正相关。在骨髓浆中，LILRB1与白蛋白（r = -0.509，P<0.050）呈显著负相关；CD36与血红蛋白（r = -0.425，P<0.050）、HDL-C（r = -0.498，P<0.050）呈显著负相关，而与TP53（r = 0.561，P<0.050）呈显著正相关；骨髓浆中LEF-1与IgA型M蛋白（r = 0.664，P<0.010）、TP53（r = 0.579，P<0.050）和髓外病变（r = 0.400，P<0.050）呈显著正相关。在血浆中，LILRB1与LDH（r = 0.657，P<0.010）、TG（r = 0.724，P<0.010）和髓外病变（r = 0.477，P<0.050）呈显著正相关；CD36与TG（r = 0.489，P<0.050）、MLR（r = 0.396，P<0.050）和髓外病变（r = 0.417，P<0.050）呈显著正相关；LEF-1与t(4;14)（r = 0.585，P<0.050）和双打击（r = 0.585，P<0.050）均呈显著正相关。

关键词: 多发性骨髓瘤, LILRB1, CD36, LEF-1, HNF1β基因, 预后