Detection of BRAF V600E mutation in radiological Langerhans cell histiocytosis-associated neurodegenerative disease using droplet digital PCR analysis
Detection of BRAF V600E mutation in radiological Langerhans cell histiocytosis-associated neurodegenerative disease using droplet digital PCR analysis
复制标题
使用液滴数字 PCR 分析检测放射性朗格汉斯细胞组织细胞增多症相关神经退行性疾病中的 BRAF V600E 突变
DOI：
10.1007/s12185-023-03588-w
复制
发表时间：
2023
影响因子：
2.1
通讯作者：
Shioda Yoko
中科院分区：
文献类型：
--
作者：
Shimizu Soichiro;Sakamoto Kenichi;Kudo Ko;Morimoto Akira;Shioda Yoko
朗格汉斯细胞组织细胞病相关神经退行性疾病（LCH-ND）是继发于LCH的最严重的晚期并发症，具有渐进性、破坏性和不可逆性。在外周血单个核细胞（PBMC）中检测到BRAFV 600 E突变，即使在没有活动性LCH病变的情况下，也被认为是临床LCH-ND的标志，表现为异常的影像学表现和神经系统症状。然而，在无症状放射学LCH-ND（rLCH-ND）而无活动性LCH病变且仅表现为异常影像学表现的患者的PBMC中检测到BRAFV 600 E突变尚不清楚。在这项研究中，我们使用液滴数字聚合酶链反应（ddPCR）分析了无活动性LCH病变的rLCH-ND患者（n= 5）PBMC和细胞游离DNA（cfDNA）中的BRAFV 600 E突变。PBMCs中BRAFV 600 E突变的检出率为60%。3例阳性病例的突变等位基因频率分别为0.049%、0.027%和0.015%。然而，cfDNABRAFV 600 E突变在所有患者中仍未检测到。检测PBMCs中BRAFV 600 E突变等位基因可能有助于在发生LCH-ND的高风险患者中识别无症状rLCH-ND，包括CNS风险部位复发或中枢性尿崩症患者。
Langerhans cell histiocytosis-associated neurodegenerative disease (LCH-ND) is the most serious late complication secondary to LCH and is gradually progressive, destructive, and irreversible. Detection of theBRAFV600E mutation in peripheral blood mononuclear cells (PBMCs), even in the absence of active LCH lesions, is considered a sign of clinical LCH-ND, presenting with both abnormal imaging findings and neurological symptoms. However, the detection of theBRAFV600E mutation in PBMCs of patients with asymptomatic radiological LCH-ND (rLCH-ND) without active LCH lesions who present only with abnormal imaging findings is unknown. In this study, we analyzed theBRAFV600E mutations in PBMCs and cell-free DNA (cfDNA) of patients with rLCH-ND without active LCH lesions (n= 5) using a droplet digital polymerase chain reaction (ddPCR) assay. TheBRAFV600E mutation in PBMCs was detected in three out of five (60%) cases. The mutant allele frequencies in the three positive cases were 0.049%, 0.027%, and 0.015%, respectively. However, the cfDNABRAFV600E mutation remained undetected in all patients. Detection of theBRAFV600E mutant allele in PBMCs may be helpful in identifying asymptomatic rLCH-ND in patients at high risk for developing LCH-ND, including those with relapses at CNS risk sites or central diabetes insipidus.