Analysis of gene expression profile of non-Hodgkin lymphoma in children based on next-generation sequencing technology
10.3760/cma.j.cn115356-20240630-00099
- VernacularTitle:基于二代测序技术检测儿童非霍奇金淋巴瘤基因表达谱分析
- Author:
Ping ZHU
1
;
Chunmei WANG
Author Information
1. 郑州大学第一附属医院儿童血液与肿瘤科，郑州 450052
- Keywords:
Lymphoma, non-Hodgkin;
Child;
High-throughput nucleotide sequencing;
Circulating tumor DNA;
Mutation;
Prognosis
- From:
Journal of Leukemia & Lymphoma
2025;34(6):349-356
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the gene expression profile of children with non-Hodgkin lymphoma (NHL) and to analyze the clinical significance of circulating tumor DNA (ctDNA) in children with NHL based on next-generation sequencing technology.Methods:A retrospective case series study was conducted. The clinical data of 46 children with NHL who underwent genetic analysis of 122 lymphoma related genes whole exome in pathological tissues and (or) peripheral blood before treatment by using NGS technology in the First Affiliated Hospital of Zhengzhou University from July 2019 to August 2023 were collected. Among the 46 children, 19 cases had the simple extraction of peripheral blood to isolate ctDNA and 19 cases had the simple acquisition of pathological tissue specimens genome DNA (gDNA) for genetic testing, and 8 cases had peripheral blood ctDNA and tissue samples at the same time for detection. The gene expression profile of the whole group and different pathologic types of children were summarized. Kaplan Meier method was used to analyze the overall survival of children with mutations and those without mutations, and the corresponding relationship between peripheral blood ctDNA detected by NGS and gene mutations detected by biopsy gDNA was analyzed.Results:Among the 46 children with NHL, 38 cases (82.6%) were male and 8 cases (17.4%) were female; the median age [ M ( Q1, Q3)] was 9 (6, 11) years. There were 5 cases of ALK-positive anaplastic large cell lymphoma, 16 cases of Burkitt lymphoma (BL), 9 cases of B-cell lymphoblastic lymphoma (B-LBL), 10 cases of T-cell lymphoblastic lymphoma (T-LBL), 4 cases of high-grade B-cell lymphoma-not otherwise specified (HGBL-NOS), 1 case of small intestine NK/T-cell lymphoma, and the remaining 1 case was classified as aggressive B-cell lymphoma which was not further classified due to insufficient tissue specimens. A total of 9 fusion genes were detected in 18 cases (39.1%). The median number of mutated genes was 1 (0, 4) (range from 0 to 6). Among 46 cases, 26 cases (56.5%) had at least 1 gene mutation, and 18 cases (39.1%) had ≥ 3 gene mutations. Among 36 mutant genes, the top 5 mutation rates were MYC (17.4%, 8/46), DDX3X (17.4%, 8/46), NRAS (13.0%, 6/46), NOTCH1 (10.8%,5/46), PHF6 (10.8%, 5/46), TP53 (8.7%, 4/46), ID3 (8.7%, 4/46), ARID1A (8.7%, 4/46), PTEN (6.5%,3/46), FOXO1 (6.5%,3/46), KMT2D (6.5%, 3/46). Among 8 pediatric cases with DDX3X gene mutation, 5 cases were BL and 8 cases were all male; among 5 BL pediatric cases with DDX3X gene mutation, 4 cases were accompanied with MYC gene mutation. There were great differences in gene expression profile among different pathological types of NHL in children. Ras/protein phosphatase/MARK signaling pathway related gene mutations occurred in 20 cases (43.5%), mainly in children with B-LBL. Mutations associated with transcription factor regulation and epigenetic modification occurred in 28.3% (13 cases) and 23.9% (11 cases), respectively. The mutation rates of PI3K-AKt and JAK-STAT signaling pathway were 15.2% (7 cases) and 13.0% (6 cases), respectively, in which PHF6, ID3 and ARID1A with high mutation rate mainly occurred in BL children. The median follow-up was 21.25 (13.55, 29.20) months, and the median overall survival time was 21.10 (11.72, 29.20) months. At the end of last follow-up time, 40 cases (87.0%) survived and 6 cases (13.0%) died. Among 16 cases with BL, the median overall survival time was 12.5 months (95% CI: 0.8-24.2 months) in the DDX3X mutation group (5 cases), 23.2 months (95% CI:19.0-27.4 months) in the DDX3X non-mutation group (11 cases), and the difference in the overall survival was not statistically significant between the 2 groups ( P = 0.214). A total of 16 mutated genes were identified in 8 cases of children who obtained peripheral blood ctDNA and tissue samples gDNA at the same time, 5 mutated genes found in ctDNA were detected in the corresponding tissues gDNA, while 16 mutated genes were found in tissue gDNA detection. ctDNA and gDNA test results were identical in 2 of the 8 pediatric cases. Relevant mutations were detected in the tissue samples gDNA in 5 cases, but ctDNA test was negative. Conclusions:Genetic mutation analysis based on NGS technology revealed genetic heterogeneity among different NHL subtypes and involved multiple signaling pathways, indicating that different pathological types of NHL have different molecular pathogenesis. ctDNA analysis based on NGS technology can assist tissue biopsy to dynamically monitor gene changes within tumors.
