Ultra-sensitive Sequencing Identifies High Prevalence of Clonal Hematopoiesis-Associated Mutations throughout Adult Life
Ultra-sensitive Sequencing Identifies High Prevalence of Clonal Hematopoiesis-Associated Mutations throughout Adult Life
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DOI：
10.1016/j.ajhg.2017.05.013
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发表时间：
2017-07-06
影响因子：
9.8
通讯作者：
Hoischen, Alexander
中科院分区：
文献类型：
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作者：
Acuna-Hidalgo, Rocio;Sengul, Hilal;Hoischen, Alexander
克隆造血是造血干细胞体细胞突变的结果，这为突变细胞提供了优势，推动其克隆扩张，并可能导致白血病。克隆造血驱动突变 (CHDM) 会随着正常衰老而发生，并且在超过 10% 的 65 岁以上个体中已检测到这些突变。我们的目的是研究成人一生中 CHDM 的患病率和特征。我们开发了一种基于单分子分子倒置探针 (smMIP) 的靶向重测序测定法，将高通量与超高灵敏度相结合。使用 smMIP，我们从 20 至 69 岁人群对照的 2,000 多个血液 DNA 样本中筛选了 100 多个 CHDM 基因座。筛选的基因座包括 40 个已知在突变时驱动克隆造血的区域和 64 个新的候选基因座。我们在整个队列中发现了 224 个体细胞突变，其中 216 个是已知驱动基因（DNMT3A、JAK2、GNAS、TET2 和 ASXL1）的编码突变，包括 196 个点突变和 20 个插入缺失。我们的检测方法灵敏度得到提高，使我们能够检测变异等位基因频率低至 0.001 的突变。超过 20% 的 60 至 69 岁个体和 3% 的 20 至 29 岁个体中发现 CHDM，尽管筛查了一组有限的位点，但该患病率大约是之前报道的患病率的两倍。我们的研究结果支持克隆造血相关突变的发生是与衰老相关的一种广泛机制，表明由于携带体细胞突变的细胞克隆进化而产生的嵌合现象是健康人类所有年龄段发生的普遍机制。
Clonal hematopoiesis results from somatic mutations in hematopoietic stem cells, which give an advantage to mutant cells, driving their clonal expansion and potentially leading to leukemia. The acquisition of clonal hematopoiesis-driver mutations (CHDMs) occurs with normal aging and these mutations have been detected in more than 10% of individuals >= 65 years. We aimed to examine the prevalence and characteristics of CHDMs throughout adult life. We developed a targeted re-sequencing assay combining high-throughput with ultra-high sensitivity based on single-molecule molecular inversion probes (smMIPs). Using smMIPs, we screened more than 100 loci for CHDMs in more than 2,000 blood DNA samples from population controls between 20 and 69 years of age. Loci screened included 40 regions known to drive clonal hematopoiesis when mutated and 64 novel candidate loci. We identified 224 somatic mutations throughout our cohort, of which 216 were coding mutations in known driver genes (DNMT3A, JAK2, GNAS, TET2, and ASXL1), including 196 point mutations and 20 indels. Our assay's improved sensitivity allowed us to detect mutations with variant allele frequencies as low as 0.001. CHDMs were identified in more than 20% of individuals 60 to 69 years of age and in 3% of individuals 20 to 29 years of age, approximately double the previously reported prevalence despite screening a limited set of loci. Our findings support the occurrence of clonal hematopoiesis-associated mutations as a widespread mechanism linked with aging, suggesting that mosaicism as a result of clonal evolution of cells harboring somatic mutations is a universal mechanism occurring at all ages in healthy humans.