骨转移和肺转移的骨肉瘤基因组学表现差异  

Osteosarcoma with bone metastasis or pulmonary metastasis show distinct genomic manifestations

作　　者：蔡震宇[1] 佘艳春 谢璐[1] 王涵 杜志业[1] 李原[2] 任婷婷[1] 许婕[1] 孙馨[1] 孙昆昆[3] 沈丹华[3] 汤小东[1] 郭卫[1] Cai Zhenyu;She Yanchun;Xie Lu;Wang Han;Du Zhiye;Li Yuan;Ren Tingting;Xu Jie;Sun Xin;Sun Kunkun;Shen Danhua;Tang Xiaodong;Guo Wei(Musculoskeletal Tumor Center,Peking University People's Hospital,Beijing 100044,China;Department of Nuclear Medicine,Peking Uiversity People's Hospital,Beijing 100044,China;Department of Tathology,Peking University People's Hospital,Beijing 100044,China)

机构地区：[1]北京大学人民医院骨肿瘤科,北京100044 [2]北京大学人民医院核医学科,北京100044 [3]北京大学人民医院病理科,北京100044

出　　处：《中华骨科杂志》2023年第9期581-590,共10页Chinese Journal of Orthopaedics

基　　金：国家自然科学基金(81972509,82072970)。

摘　　要：目的探讨初始骨转移与肺转移的骨肉瘤基因组学表现差异及发病机制。方法收集2021年5月1日至10月1日于北京大学人民医院诊治的38例高级别骨肉瘤患者肿瘤新鲜标本及部分石蜡包埋标本,男29例、女9例,年龄(19.6±2.2)岁(范围6~61岁)。38例中12例发生初始骨转移、26例发生初始肺转移,其中15例(40%,15/38)具有原发灶和转移灶的配对标本。应用Illumina NovaSeq 6000平台进行全外显子测序及转录组测序,并随治疗进程获取肿瘤演进过程中的新鲜配对标本,分析不同进展模式的骨肉瘤配对标本的全外显子组测序数据,进一步根据其基因型表现进行疾病亚分类,将基因学表现与临床过程进行关联。结果骨转移组以单核苷酸变异为主(83%,10/12),肺转移组以结构变异为主(58%,15/26)。骨转移组的肿瘤突变负荷为4.9(2.8,12.0)、新抗原负荷为743.0(316.5,1034.5),较肺转移组的2.4(1.4,4.5)和128.5(49.0,200.5)更高,差异均有统计学意义(P=0.010,P=0.003)。突变谱示骨转移组[0.33(0.13,0.60)]和肺转移组[0.10(0.00,0.13)]在与年龄有关的基因特征1上的差异有统计学意义(P=0.005)。以随机表法每组各选择3例患者进行多色免疫荧光染色,骨转移组中1例发现了三级淋巴结构。通过对配对标本进行进化图谱分析发现骨肉瘤配对标本的已知基因呈高度保守。结论以单核苷酸变异为主而非结构变异的骨肉瘤可能容易发生初始骨转移,此类患者年龄更大,肿瘤微环境的免疫源性可能更高。Objective To investigate the genomic manifestation and pathogenesis of osteosarcoma with different relapse pattens,which were respectively initially presented with bone metastasis or pulmonary metastasis.Methods From May 1,2021 to October 1,2021,38 fresh tumor specimens and some paraffin-embedded specimens of high-grade osteosarcoma were collected in Peking University People's Hospital,including 29 males and 9 females,aged 19.6±2.2 years(range,6-61 years).Among the 38 cases,12 cases had initial bone metastasis(group A)and 26 cases had initial lung metastasis(group B),of which 15 cases(40%,15/38)had paired specimens of primary and metastatic lesions.Based on Illumina NovaSeq 6000,we analyzed whole-exome sequencing(WES)as well as transcriptome for osteosarcoma with paired samples in different relapse patterns.During all their treatment courses,we also collected their paired samples to reveal these tumors'evolution.We sought to redefine disease subclassifications for osteosarcoma based on genetic alterations and correlate these genetic profiles with clinical treatment courses to elucidate potential evolving cladograms.Results We found that osteosarcoma in group A mainly carried single-nucleotide variations(83%,10/12),displaying higher tumor mutation burden[4.9(2.8,12.0)&2.4(1.4,4.5),P=0.010]and neoantigen load[743.0(316.5,1,034.5)&128.5(49.0,200.5),P=0.003],while those in group B mainly exhibit structural variants(58%,15/26).The mutation spectrum showed that there was a significant difference in age-related gene imprinting 1 between the bone metastasis group and the lung metastasis group(P=0.005).Samples were randomly selected from group A(3 patients)to investigate immunologic landscape by multiplex immunohistochemistry,from which we noticed tertiary lymphatic structure from one patient from group A.High conservation of reported genetic sequencing over time was found in their evolving cladograms.Conclusion Osteosarcoma with mainly single-nucleotide variations other than structural variants might exhibit biological

关 键 词：骨肉瘤 肿瘤转移 基因组学 染色体结构变异 单核苷酸变异 

分 类 号：R738.1[医药卫生—肿瘤]