Clinicopathologic and Molecular Characterization of Adult 1p/19q Non-codeleted Oligodendrogliomas
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摘要:
目的 探讨成人1p/19q未共缺失的“少突胶质细胞瘤”中的临床病理特征与及其他分子标记物相关性。 方法 收集326例手术切除后组织病理诊断为少突胶质细胞瘤、间变少突胶质细胞瘤、少突-星形细胞瘤及间变少突-星形细胞瘤的病例。采用荧光原位杂交（FISH）检测1p/19q的共缺失状态，采用直接测序法检测IDH1/2、TP53、TERT 启动子突变状态，采用免疫组化染色检测ATRX、PDGFRA、EGFR、 CIC、FUBP1、INA、PTEN表达水平，采用甲基化特异性PCR（MSP）方法检测（MGMT）甲基化水平。 结果 326例成人少突胶质细胞瘤的1p/19q状态检测结果为37.6%的肿瘤1p/19q未共缺失，超过一半的1p/19q未共缺失的“少突胶质细胞瘤”缺乏星形细胞特征性标记物p53和ATRX表达。1p/19q未共缺失的“少突胶质细胞瘤”队列中，年龄较小（<45岁）、WHO 2级和典型少突胶质细胞瘤组织形态学的患者，预后较好（P < 0.01），典型少突胶质细胞瘤组织形态学与TERTp突变状态、p53表达、EFGR表达、PDGFRA表达相关（P < 0.05）。FUBP1、TERT、MGMT、PDGFRA、EGFR、PTEN、INA和CIC的表达未检测到显著的预后价值（P > 0.05）。 结论 依据2021年第5版WHO中枢神经系统肿瘤分类，星形细胞瘤不能解释所有的1p/19q未共缺失的“少突胶质肿瘤”，1p/19q未共缺失的“少突胶质细胞瘤”可能形成弥漫性胶质瘤的一个不同亚群。 -
关键词:
- 少突胶质细胞肿瘤 /
- 1p/19q未共缺失 /
- 分子标记物
Abstract:Objective To investigate the clinicopathologic and other molecular markers features of oligodendrogliomas in adults with non-codeletion of 1p/19q. Methods 326 cases of oligodendrogliomas, anaplastic oligodendrogliomas, oligoastrocytomas, and anaplastic oligoastrocytomas diagnosed by histopathology after surgical resection were collected. The polymorphism of chromosome 1p and 19q were analyzed by fluorescence in situ hybridization （FISH）. Isocitrate dehydrogenase （IDH） 1/2 mutation, TP53 and promoter of telomerase reverse transcriptase （TERTp） were detected by Sanger sequencing. The expression levels of ATRX, PDGFRA, EGFR, CIC, FUBP1, INA, and PTEN were detected by immunohistochemical staining, and methylation specific PCR （MSP） was used to detect the methylation of MGMT promotor. Results The 1p/19q status of 326 adult oligodendrogliomas showed that 37.6% of the tumors were 1p/19q non-codeletion, and more than half of the 1p/19q non-codeleted oligodendrogliomas showed lack of expression of p53 and ATRX, the defining astrocytic markers. In the cohort of 1p/19q non-codeleted "oligodendrogliomas", patients with younger age （<45 years）, WHO grade 2, and typical oligodendroglioma histomorphology had a better prognosis（P < 0.01）, and the typical histomorphology of oligodendrogliomas was associated with TERTp mutation status, p53 expression, EFGR expression, PDGFRA expression（P < 0.05）. The expression of FUBP1, TERT, MGMT, PDGFRA, EGFR, PTEN, INA, and CIC was not found to have significant prognostic value（P > 0.05）. Conclusion According to the 5th edition of the 2021 WHO Classification of Tumors of the Central Nervous System, astrocytomas do not account for all 1p/19q non-codeleted oligodendrogliomas, and 1p/19q non-codeleted oligodendrogliomas may represent a distinct subgroup of diffuse gliomas. -
Key words:
- Oligodendrogliomas /
- 1p/19q non-codeletion /
- Molecular maker
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图 1 不同的1p/19q编码的病理分型
Figure 1. Pathological classification of different 1p/19q frequency
图 4 成人“少突胶质细胞瘤”EGFR、PDGFRA、INA、CIC、FUBP1和PTEN的免疫组化结果（×20）
A：EGFR；B：PDGFRA；C：INA；D：CIC；E：FUBP1；F：PTEN。
Figure 4. Immunohistochemical results of EGFR，PDGFRA，INA，CIC，FUBP1，and PTENin adult oligodendroglioma （×20）
图 5 不同临床病理特征及分子标记物的相关性分析
AGE45年龄45岁；Gender性别；Histological type（O vs OA）组织学类型（少突胶质细胞瘤vs少突星形细胞瘤）；Histological grade组织学分级；No frontal lobe vs frontal lobe 非额叶vs额叶；Operation（no total vs total） 非全切vs全切；IDH异柠檬酸脱氢酶；TERT端粒酶逆转录酶；TP53肿瘤蛋白P53；MGMTp methylation O-6-甲基鸟嘌呤-DNA甲基转移酶启动器甲基化；ATRX expression α-地中海贫血/精神发育迟滞综合征X染色体相关基因表达；EGFR expression表皮生长因子受体表达；PDGFRA expression血小板衍生生长因子受体α表达；PTEN expression磷酸酶与张力蛋白同源物表达；INA expression INA表达；FUBP1expression 远端上游元件结合蛋白1；CIC expression CIC表达。*在 0.05 级别（双尾），相关性显著；**在 0.01 级别（双尾），相关性显著。
Figure 5. Correlation analysis between various clinicopathological and molecular factors
图 6 1p/19q非共缺失的“少突胶质细胞瘤”的Kaplan-Merier分析
A～B:年龄与1p/19q非共缺失的“少突胶质细胞瘤”患者预后的关系；C～D:组织病理分级与1p/19q非共缺失的“少突胶质细胞瘤”患者预后的关系；E～F:组织病理分类与1p/19q非共缺失的“少突胶质细胞瘤”患者预后的关系。
Figure 6. Kaplan-Meier survival analysis in 1p/19q non-codeleted oligodendrogliomas
表 1 IDH1/2、TERT 、TP53基因检测引物序列
Table 1. IDH1/2，TERT、TP53 gene detection primer sequence
基因 引物 序列（5' to 3'） 扩增产物范围（bp） 退火温度（ ℃） IDH 1 F CGGTCTTCAGAGAAGCCATT 122 60 R CACATTATTGCCAACATGAC IDH 2 F AGCCCATCATCTGCAAAAAC 150 60 R CTAGGCGAGGAGCTCCAGT TERT
PromoterF GTCCTGCCCCTTCACCTTC 274 68 R AGCACCTCGCGGTAGTGG TP53 Exon2 2F1 AGGTGACCCAGGGTTGGA 231 64 2R1 TCCCACAGGTCTCTGCTAGG TP53 Exon3 3F1 CCCCCTAGCAGACCTGT 190 64 3R1 TGGGTGAAAAGAGCAGTCAG TP53 Exon4 4F5 CCTGGTCCTCTGACTGCTCT 242 64 4R5 TTCTGGGAAGGGACAGAAGA TP53 Exon5 5F3 GTTCTTTCGTCGGCTCTTC 357 60 5R3 GGGCCAGACCTAAGAGCAAT TP53 Exon 6 6F3 GCTGGGGCTGGAGAGACGACAG 260 64 6R3 TACTGCTCACCTGGAGGGCCACTG TP53 Exon 7 7F7 CAAGGCGCACTGGCCTCAT 216 64 7R1 GTCAGAGGCAAGCAGAGGCT TP53 Exon 8 8F3 CAAGGGTGGTTGGGAGTAGA 327 60 8R3 AGGAAAGAGGCAAGGAAAGG TP53 Exon 9 9F2 GACCAAGGGTGCAGTTATGC 196 60 9R2 CGGCATTTTGAGTGTTAGACTG TP53 Exon 10 10F4 TGCATGTTGCTTTTGTACCG 235 60 10R1 AAGGGGCTGAGGTCACTCAC TP53 Exon 11 11F1 GGGCACAGACCCTCTCACT 223 60 11R1 CAAAGACCCAAAACCCAAAA 
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表 2 MGMT 启动子甲基化引物序列
Table 2. MGMT promoter methylation primer sequence
引物 序列 MSP-MGMT-F 5'-GGATATGTTGGGATAGTT-3' MSP-MGMT-R 5'-CCAAAAACCCCAAACCC-3' MGMT-MF 5'-TTTCGACGTTCGTAGGTTTTCGC-3' MGMT-MR 5'-GCACTCTTCCGAAAACGAAACG-3' MGMT-UF 5'-TTTGTGTTTTGATGTTTGTAGGTTTTTGT-3' MGMT-UR 5'-AACTCCACACTCTTCCAAAAACAAAACA-3'
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表 3 患者临床基本资料[n（%）]
Table 3. Clinical characteristics [n（%）]
类别 n 占比 年龄 （岁） 44.08±11.08 性别 327 男 181（55.4） 女 146（44.6） 组织类型 327 少突胶质细胞瘤 123（37.6） 间变性少突胶质瘤 71（21.7） 少突星形细胞瘤 100（30.6） 间变性少突星形细胞瘤 33（10.1） 组织分级 327 WHO2 223（68.2） WHO3 104（31.8） 肿瘤位置 324 额叶 197（60.8） 颞叶 41（12.7） 顶叶 14（4.3） 枕叶 8（2.5） 超过一侧脑叶 50（15.4） 其他部位 14（4.3） 手术方式 263 全切 188（71.5） 非全切 75（28.5） 术后治疗 246 联合放化疗 129（52.4） 单纯放疗 64（26） 单纯化疗 12（4.9） 未治疗 41（16.7）
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表 4 不同分子标记物的Kaplan-Meier生存分析
Table 4. Kaplan-Meier survival analysis of various molecular markers
分子标记物 n 中位OS HR（95%CI） χ2 P 中位PFS HR（95%CI） χ2 P IDH 92 野生型 22 42（23.3～103.4） 2.151 0.143 27.0（2.0～52.0） 1.739 0.187 突变型 70 102（84.6～119.6） 70.0（40.3～99.7） TERT启动子 67 野生型 37 93.6（44.7～142.5） 0.012 0.914 48.0（0～106.7） 0.018 0.894 突变型 30 109.1（24.1～194.1） 62.0（8.7～115.3） PDGFRA 63 阳性 37 93.6（73.2～114.0） 2.954 0.086 57 （20.3～93.7） 0.911 0.340 阴性 26 70.0（20.3～119.7） 36（28.2～43.8） EGFR 68 Negative/weak 47 48（26.3～69.7） 0.014 0.100 48（26.3～69.7） 2.654 0.103 Moderate/strong 21 96.8（44.7～148.9） 96.8（44.7～148.9） PTEN 54 阳性 32 93.6 （46.8～136.4） 0.612 0.434 54.1（26.0～82.2） 0.12 0.750 阴性 22 91.6 （46.8～136.4） 53.6（17.0～90.3） INA 58 阳性 27 93.6（78.2～109.0） 0.263 0.608 62（45.2～78.8） 0.701 0.401 阴性 31 94.0（45.7～142.3） 54.1（0～78.7） CIC 58 阳性 44 86.4（54.5～118.2] 0.036 0.800 62 （38.2～85.8） 1.073 0.300 阴性 14 97.3（71.6～123.1） 34.9（0～75.2） FUBP1 47 阳性 42 94.0（57.9～130.1） 2.073 0.150 62（37.0～87.0） 3.212 0.073 阴性 5 44.2（40.7～47.7） 31.7（15.2～48.2） N：病例数；OS：总生存期；PFS：无进展生存期；NA：不可用。
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[1] Louis D N,Perry A,Wesseling P,et al. The 2021 WHO classification of tumors of the central nervous system: A summary[J]. Neuro Oncol,2021,23(8):1231-1251. doi: 10.1093/neuonc/noab106 [2] 王樑,潘亚文,屈延,等. 2021年世界卫生组织中枢神经系统肿瘤分类(第五版)成人型弥漫性胶质瘤分类解读[J]. 中国现代神经疾病杂志,2021,21(9):783-790. doi: 10.3969/j.issn.1672-6731.2021.09.008 [3] Li Y X, Shi Z, Aibaidula A, et al. Not all 1p/19q non-codeleted oligodendroglial tumors are astrocytic [J]. Oncotarget, 2016, 7（40）: 64615-64630.Li Y X,Shi Z,Aibaidula A,et al. Not all 1p/19q non-codeleted oligodendroglial tumors are astrocytic [J]. Oncotarget,2016,7(40): 64615-64630. [4] Bettegowda C,Agrawa N,Jiao Y,et al. Mutations in CIC and FUBP1 contribute to human oligodendroglioma[J]. Science,2011,333(6048):1453-1455. doi: 10.1126/science.1210557 [5] Ducray F,Mokhtari K,Criniere E,et al. Diagnostic and prognostic value of alpha internexin expression in a series of 409 gliomas[J]. Eur J Cancer,2011,47(5):802-808. doi: 10.1016/j.ejca.2010.11.031 [6] Eigenbrod S,Roeber S,Thon N,et al. α-Internexin in the diagnosis of oligodendroglial tumors and association with 1p/19q status[J]. J Neuropathol Exp Neurol,2011,70(11):970-978. doi: 10.1097/NEN.0b013e3182333ef5 [7] Brito C,Azebedo A,Esreves S,et al. Clinical insights gained by refining the 2016 WHO classification of diffuse gliomas with: EGFR amplification,TERT mutations,PTEN deletion and MGMT methylation[J]. BMC Cancer,2019,19(1):968. doi: 10.1186/s12885-019-6177-0 [8] Martinho O,Longatto-filho A,Lambros M B,et al. Expression,mutation and copy number analysis of platelet-derived growth factor receptor A (PDGFRA) and its ligand PDGFA in gliomas[J]. Br J Cancer,2009,101(6):973-982. doi: 10.1038/sj.bjc.6605225 [9] Louis D N,Ohgaki H,Wiestler O D,et al. The 2007 WHO classification of tumours of the central nervous system[J]. Acta Neuropathol,2007,114(2):97-109. doi: 10.1007/s00401-007-0243-4 [10] Reuss D E,Sahm F,Schrimpf D,et al. ATRX and IDH1-R132H immunohistochemistry with subsequent copy number analysis and IDH sequencing as a basis for an“integrated”diagnostic approach for adult astrocytoma,oligodendroglioma and glioblastoma[J]. Acta Neuropathol,2015,129(1):133-146. doi: 10.1007/s00401-014-1370-3 [11] Takami H,Yoshida A,Fukushima S,et al. Revisiting TP53 mutations and Immunohistochemistry—a comparative study in 157 diffuse gliomas[J]. Brain Pathol,2015,25(3):256-265. doi: 10.1111/bpa.12173 [12] Canale M,Petracci E,Delmonte A,et al. Impact of TP53 mutations on outcome in EGFR-mutated patients treated with firstline tyrosine kinase inhibitors[J]. Clin Cancer Res,2017,23(9):2195-2202. doi: 10.1158/1078-0432.CCR-16-0966 [13] Chan A K,Pang J C,Chung N Y,et al. Loss of CIC and FUBP1 expressions are potential markers of shorter time to recurrence in oligodendroglial tumors[J]. Mod Pathol,2014,27(3):332-342. doi: 10.1038/modpathol.2013.165 [14] Gusyatiner O,Hegi M E. Glioma epigenetics: From subclassification to novel treatment options[J]. Semin Cancer Biol,2018,51:50-58. doi: 10.1016/j.semcancer.2017.11.010 [15] 苏玉金,杜江,王军梅,等. 568 例少突胶质细胞肿瘤中染色体 1p/19q 缺失和多体特点分析[J]. 中国医刊,2017,52(3):57-60. doi: 10.3969/j.issn.1008-1070.2017.03.018 [16] Cairncross G,Wang M,Shaw E,et al. Phase III trial of chemoradiotherapy for anaplastic oligodendroglioma: long-term results of RTOG 9402[J]. J Clin Oncol,2013,31(3):337-343. doi: 10.1200/JCO.2012.43.2674 [17] Giannini C,Burger P C,Berkey B A,et al. Anaplastic oligodendroglial tumors: refining the correlation among histopathology,1p 19q deletion and clinical outcome in Intergroup Radiation Therapy Oncology Group Trial 9402[J]. Brain Pathol,2008,18(3):360-369. doi: 10.1111/j.1750-3639.2008.00129.x [18] Ohba S,Kuwahara K,Yamada S,et al. Correlation between IDH,ATRX,and TERT promoter mutations in glioma[J]. Brain Tumor Pathol,2020,37(2):33-40. doi: 10.1007/s10014-020-00360-4 [19] 张成娟,魏冰,郭永军,等. 胶质瘤中TERT启动子突变的研究进展[J]. 河南医学研究,2017,26(6):1018-1019. doi: 10.3969/j.issn.1004-437X.2017.06.026 [20] Liu H,Xu C,Diplas B H,et al. Cancer-associated SMARCAL1 loss-of-function mutations promote alternative lengthening of telomeres and tumorigenesis in telomerase-negative glioblastoma cells[J]. Neuro Oncol,2023,25(9):1563-1575. [21] Nandakumar P,Mansouri A,Das S. The role of ATRX in glioma biology[J]. Front Oncol,2017,7:236. doi: 10.3389/fonc.2017.00236 [22] Killela P J,Reitman Z J,Jiao Y,et al. TERT promoter mutations occur frequently in gliomas and a subset of tumors derived from cells with low rates of self-renewal[J]. Proc Natl Acad Sci U S A,2013,110(15):6021-6026. doi: 10.1073/pnas.1303607110 [23] Bedics G,Szőke P,Bátai B,et al. Novel,clinically relevant genomic patterns identified by comprehensive genomic profiling in ATRX-deficient IDH-wildtype adult high-grade gliomas[J]. Sci Rep,2023,13(1):18436. [24] Eckel-Passow J E,Lachance D H,Molinaro A M,et al. Glioma groups based on 1p/19q,IDH,and TERT promoter mutations in tumors[J]. Sci RepN Engl J Med,2015,372(26):2499-2508. doi: 10.1056/NEJMoa1407279 [25] 樊小龙,杨子辰. 成人型弥漫性胶质瘤的分子分型研究进展[J]. 北京师范大学学报(自然科学版),2023,59(1):38-42. doi: 10.12202/j.0476-0301.2022153 [26] Sun Y,Zhang W,Chen D,et al. A glioma classification scheme based on coexpression modules of EGFR and PDGFRA[J]. Proc Natl Acad Sci U S A,2014,111(9):3538-3543. doi: 10.1073/pnas.1313814111 [27] 孙晓玲,张晓娟,张清泉,等. TP53、EGFR、PDGF在星形胶质细胞瘤进展中的表达及意义[J]. 广东医学,2019,40(12):1750-1753. [28] Chen C C L,Deshmukh S,Jessa S,et al. Histone H3.3G34-mutant interneuron progenitors co-opt PDGFRA for gliomagenesis [J]. Cell,2020,183(6): 1617-1633 e22. [29] Horbinski C,Hobbs J,Cieply K,et al. EGFR expression stratifies oligodendroglioma behavior [J]. Am J Pathol,2011,179(4): 1638-1644. -
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