北京大學(xué)第一醫(yī)院血液研究室朱平教授、博士后陳艷與華大基因公司侯勇等合作,，在骨髓增殖性腫瘤的研究方面取得突破性進(jìn)展,。該研究首次通過對(duì)血液腫瘤患者骨髓的單細(xì)胞全基因組外顯子測(cè)序,，探索骨髓增殖性腫瘤的細(xì)胞演化和惡性進(jìn)展規(guī)律,，相關(guān)論文3月2日在線發(fā)表于國(guó)際著名雜志《細(xì)胞》。
朱平介紹,，骨髓增殖性腫瘤是一種臨床表現(xiàn)相當(dāng)隱匿的疾病,，主要包括真性紅細(xì)胞增多癥、原發(fā)性血小板增多癥和骨髓纖維化,，分別由于紅細(xì)胞,、血小板或者纖維細(xì)胞過度增生造成。大量生長(zhǎng)的惡性細(xì)胞往往會(huì)堵塞小血管,，造成腦血管栓塞,、心肌梗死等致命疾病。過去由于缺乏有效的診斷手段,，臨床往往容易誤診為腦血管病和心血管病。
該研究課題組創(chuàng)建了單細(xì)胞全基因組外顯子測(cè)序法,，第一次通過基因分析,，清楚地展現(xiàn)了骨髓增殖性腫瘤（原發(fā)性血小板增多癥）患者體內(nèi)腫瘤發(fā)生時(shí)的細(xì)胞演化過程。JAK2基因突變?cè)诠撬柙鲋承阅[瘤很常見,，但研究人員發(fā)現(xiàn)這名患者卻未出現(xiàn)突變,，說明可能存在其他導(dǎo)致疾病發(fā)生的基因突變。研究人員利用顯微操作技術(shù),，從骨髓樣本中隨機(jī)選取82個(gè)骨髓細(xì)胞以及8個(gè)正?？谇火つど掀ぜ?xì)胞，進(jìn)行單個(gè)細(xì)胞測(cè)序和生物信息學(xué)分析,。結(jié)果發(fā)現(xiàn),，骨髓增殖性腫瘤患者的腫瘤細(xì)胞與正常細(xì)胞相比，基因圖譜完全不同，不同腫瘤細(xì)胞之間也有差異,，呈現(xiàn)遺傳多樣性,。這名患者的腫瘤曾經(jīng)有不同細(xì)胞同時(shí)存在多種基因突變的短暫時(shí)期，最終其中一個(gè)細(xì)胞群有明顯的生長(zhǎng)優(yōu)勢(shì),，發(fā)展成為腫瘤,。
朱平指出，這種新的測(cè)序方法開啟了一條嶄新的研究途徑,，除了可以闡明骨髓增殖性腫瘤的發(fā)生過程,，還可以用于探索其他類型腫瘤的起源過程。如果能充分了解腫瘤發(fā)生過程中多種基因突變出現(xiàn)的時(shí)間和相互作用,，就可以不斷發(fā)現(xiàn)腫瘤不同發(fā)展階段的新的診斷標(biāo)志,，也可以研發(fā)影響相關(guān)基因的藥物，尋找預(yù)防疾病發(fā)生的有效措施,。（生物谷 bioon.com）
doi:10.1016/j.cell.2012.02.028
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Single-CellExomeSequencing and MonoclonalEvolution of aJAK2-Negative Myeloproliferative Neoplasm
Yong Hou,，Luting Song，Ping Zhu,，Bo Zhang,，Ye Tao，Xun Xu,，F(xiàn)uqiang Li,，Kui Wu，Jie Liang,，Di Shao,，Hanjie Wu，Xiaofei Ye,，Chen Ye,，Renhua Wu，Min Jian,，Yan Chen,，Wei Xie，Ruren Zhang,，Lei Chen,，Xin Liu，Xiaotian Yao,，Hancheng Zheng,，Chang Yu，Qibin Li,，Zhuolin Gong,，Mao Mao，Xu Yang，Lin Yang,，Jingxiang Li,，Wen Wang，Zuhong Lu,，Ning Gu,，Goodman Laurie，Lars Bolund,，Karsten Kristiansen,，Jian Wang，Huanming Yang,，Yingrui Li,，Xiuqing Zhang，Jun Wang
Tumor heterogeneity presents a challenge for inferring clonal evolution and driver gene identification. Here, we describe a method for analyzing the cancer genome at asingle-cell nucleotide level. To perform our analyses, we first devised and validated a high-throughput whole-genome single-cellsequencing method using two lymphoblastoid cell line singlecells. We then carried out whole-exomesingle-cellsequencing of 90 cells from aJAK2-negative myeloproliferative neoplasm patient. The sequencing data from 58 cells passed our quality control criteria, and these data indicated that this neoplasm represented amonoclonalevolution. We further identified essential thrombocythemia (ET)-related candidate mutations such as SESN2 and NTRK1, which may be involved in neoplasm progression. This pilot study allowed the initial characterization of the disease-related genetic architecture at the single-cell nucleotide level. Further, we established asingle-cellsequencing method that opens the way for detailed analyses of a variety of tumor types, including those with high genetic complex between patients.
