Cell-Free DNA Sequencing of Intraocular Fluid as Liquid Biopsy in the Diagnosis of Vitreoretinal Lymphoma.
Cell-Free DNA Sequencing of Intraocular Fluid as Liquid Biopsy in the Diagnosis of Vitreoretinal Lymphoma.
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眼内液游离 DNA 测序作为液体活检诊断玻璃体视网膜淋巴瘤
DOI：
10.3389/fonc.2022.932674
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发表时间：
2022
影响因子：
4.7
通讯作者：
Chang, Qing
中科院分区：
文献类型：
--
作者：
Gu, Junxiang;Jiang, Tingting;Liu, Shixue;Ping, Bo;Li, Ruiwen;Chen, Wenwen;Wang, Ling;Huang, Xin;Xu, Gezhi;Chang, Qing
为了寻求新的诊断方法，我们改进了无细胞DNA（cfDNA）测序的工作流程，并评估了其在玻璃体视网膜淋巴瘤（VRL）标本中的可行性;初步分析了突变谱的潜在诊断价值。这项研究是一项诊断性试验。入选VRL患者23例23眼和炎性眼病患者25例25眼。通过诊断性玻璃体切除术获得约500μl未稀释的玻璃体液和10 ml稀释的玻璃体液，并送去进行细胞病理学检查。将500μ 1稀释的玻璃体液备用用于cfDNA测序。对于cfDNA测序，将DNA片段化程序添加到工作流程中以提高提取效率;分析检测到的突变以获得潜在的诊断模型。比较细胞病理学和cfDNA测序的敏感性和特异性。初步分析其临床表现与基因型的相关性。对23只VRL眼和20只炎性眼病眼进行CfDNA测序。VRL相关突变基因包括MYD 88（18只眼，78%），ETV 6（11只眼，48%），PIM 1（11眼，48%），BTG 2（7只眼，30%），IRF 4（7只眼，30%），CD 79 B（6只眼，26%），LRP 1B以MYD 88和ETV 6基因突变为基础的Logistic回归分析对VRL的诊断有一定的价值（P<0.001，校正R2 = 0.789，敏感性0.913，特异性0.950），玻璃体细胞学检查的敏感性和特异性分别为0.826和1.000。突变谱的进一步分析显示，携带CD 79 B突变的患者倾向于具有更高的眼内白细胞介素-10水平（P=0.030），CARD 11突变与眼部发病时的年轻年龄相关（P=0.039），并且颅内受累的患者携带更多的BTG 2基因多位点突变（P=0.013）。CfDNA测序的改进工作流程在有限量的玻璃体液中具有良好的可行性。基于突变的Logistic模型可为VRL的诊断提供可靠的线索。
To seek novel diagnostic approaches, we improved the workflow of cell-free DNA (cfDNA) sequencing and evaluated its feasibility in vitreoretinal lymphoma (VRL) specimens; the profile of mutations was preliminarily analyzed for potential diagnostic value. The study was a diagnostic trial. 23 eyes of 23 patients with VRL and 25 eyes of 25 patients with inflammatory eye diseases were enrolled. Approximate 500μl undiluted vitreous humor and 10ml diluted vitreous fluid was obtained through diagnostic vitrectomy and sent for cytopathological examinations. 500μl of the diluted vitreous fluid was spared for cfDNA sequencing. For cfDNA sequencing, DNA fragmentation procedure was added to the workflow to improve the extraction efficiency; mutations detected were analyzed for potential diagnostic model. The sensitivity and specificity of the cytopathology and cfDNA sequencing were compared. The clinical manifestations were preliminarily analyzed for potential correlations with the genotypes. CfDNA sequencing was accomplished in 23 eyes with VRL and 20 eyes with inflammatory eye diseases. VRL-related mutated genes included MYD88 (18 eyes, 78%), ETV6 (11 eyes, 48%), PIM1 (11 eyes,48%), BTG2 (7 eyes, 30%), IRF4 (7 eyes, 30%), CD79B (6 eyes, 26%), LRP1B (6 eyes, 26%), etc. Logistic regression based on the mutations of MYD88 and ETV6 was of the potential for the diagnosis of VRL (P<0.001, adjusted R2 = 0.789, sensitivity 0.913, specificity 0.950); by comparison, the sensitivity and specificity of the vitreous cytopathology were 0.826 and 1.000, respectively. Further analysis of the mutation profile showed that patients carrying CD79B mutation tended to have higher intraocular interleukin-10 level (P=0.030), that CARD11 mutation was correlated with younger age at ocular onset (P=0.039), and that patients with intracranial involvement carried more multiple-site mutations in the BTG2 gene (P=0.013). The improved workflow of CfDNA sequencing is of sound feasibility in a limited amount of vitreous humor. The logistic model based on the mutations could help to provide reliable clues for the diagnosis of VRL.
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影响因子：
2.7
作者：
Soliman SE;Alhanafy AM;Habib MSE;Hagag M;Ibrahem RAL
通讯作者：
Ibrahem RAL
影响因子：
11.5
作者：
Kaseb, Ahmed O.;Sanchez, Nora S.;Sen, Shiraj;Kelley, Robin K.;Tan, Benjamin;Bocobo, Andrea G.;Lim, Kian H.;Abdel-Wahab, Reham;Uemura, Marc;Pestana, Roberto Carmagnani;Qiao, Wei;Xiao, Lianchun;Morris, Jeffrey;Amin, Hesham M.;Hassan, Manal M.;Rashid, Asif;Banks, Kimberly C.;Lanman, Richard B.;Talasaz, AmirAli;Mills-Shaw, Kenna R.;George, Bhawana;Haque, Abedul;Raghav, Kanwal P. S.;Wolff, Robert A.;Yao, James C.;Meric-Bernstam, Funda;Ikeda, Sadakatsu;Kurzrock, Razelle
通讯作者：
Kurzrock, Razelle
影响因子：
8.1
作者：
Hiemcke-Jiwa, Laura S.;Ten Darn-van Loon, Ninette H.;Kuiper, Jonas J. W.
通讯作者：
Kuiper, Jonas J. W.
影响因子：
20.3
作者：
Klapper, Wolfram;Kreuz, Markus;Siebert, Reiner
通讯作者：
Siebert, Reiner
影响因子：
3.3
作者：
Shi, Huimin;Zhou, Xian;Wang, Qingping
通讯作者：
Wang, Qingping