去甲基化治疗后癌基因的去甲基化和上调。
Demethylation and Up-Regulation of an Oncogene after Hypomethylating Therapy.
机构信息
From the Department of Pathology, Brigham and Women's Hospital (Y.-C.L., C. Gao, Jun Liu, J.Y., L. Chai), Harvard Stem Cell Institute, Harvard Medical School (A.I.J., M.A.B., D.G.T.), and the Department of Medical Oncology, Dana-Farber Cancer Institute (R.P.) - all in Boston; the Division of Hematology, Department of Medicine, Taipei Veterans General Hospital (Y.-C.L.), and the Faculty of Medicine and the Program in Molecular Medicine, Institute of Biopharmaceutical Sciences, School of Life Science, National Yang Ming Chiao Tung University (Y.-C.L., J.-H.L.) - both in Taipei, Taiwan; the Cancer Science Institute of Singapore, Singapore (J.K., Y.V.L., H.Y., M.A.B., D.G.T.); the Department of Biomedicine and Prevention, University of Rome Tor Vergata (E.F., G.F., L.V., C. Gurnari, M.T.V.), and UniCamillus-Saint Camillus International University of Health Sciences (E.F.), Rome, and Cellular Signaling Laboratory, Department of Biomedical and Neuromotor Sciences, University of Bologna (M.Y.F., L. Cocco), and IRCCS Azienda Ospedaliero-Universitaria di Bologna, Istituto di Ematologia "Seràgnoli" (C.F.), Bologna - all in Italy; the National Clinical Research Center for Blood Diseases and State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China (Z.X., Jinqin Liu, H.H.); and the School of Medical Sciences and Lowy Cancer Research Centre (J.A.I.T., J.E.P.) and Prince of Wales Clinical School and Lowy Cancer Research Centre (A.U., J.E.P.), Faculty of Medicine, University of New South Wales, Sydney, and the Department of Hematology, Prince of Wales Hospital, Randwick, NSW (J.E.P.) - both in Australia.
出版信息
N Engl J Med. 2022 May 26;386(21):1998-2010. doi: 10.1056/NEJMoa2119771.
BACKGROUND
Although hypomethylating agents are currently used to treat patients with cancer, whether they can also reactivate and up-regulate oncogenes is not well elucidated.
METHODS
We examined the effect of hypomethylating agents on , a known oncogene that plays an important role in myelodysplastic syndrome and other cancers. Paired bone marrow samples that were obtained from two cohorts of patients with myelodysplastic syndrome before and after treatment with a hypomethylating agent were used to explore the relationships among changes in expression, treatment response, and clinical outcome. Leukemic cell lines with low or undetectable expression were used to study the relationship between methylation and expression. A locus-specific demethylation technology, CRISPR-DNMT1-interacting RNA (CRISPR-DiR), was used to identify the CpG island that is critical for expression.
RESULTS
up-regulation after treatment with hypomethylating agents was observed in 10 of 25 patients (40%) in cohort 1 and in 13 of 43 patients (30%) in cohort 2 and was associated with a worse outcome. Using CRISPR-DiR, we discovered that demethylation of a CpG island within the 5' untranslated region was critical for expression. In cell lines and patients, we confirmed that treatment with a hypomethylating agent led to demethylation of the same CpG region and up-regulation of expression.
CONCLUSIONS
By combining analysis of patient samples with CRISPR-DiR technology, we found that demethylation and up-regulation of an oncogene after treatment with a hypomethylating agent can indeed occur and should be further studied. (Funded by Associazione Italiana per la Ricerca sul Cancro and others.).
背景
虽然低甲基化剂目前被用于治疗癌症患者，但它们是否也能重新激活和上调癌基因尚不清楚。
方法
我们研究了低甲基化剂对 的作用， 是一种已知的癌基因，在骨髓增生异常综合征和其他癌症中起重要作用。我们使用来自两个骨髓增生异常综合征患者队列的配对骨髓样本，在接受低甲基化剂治疗前后，探索 表达变化、治疗反应和临床结局之间的关系。使用低表达或未检测到 的白血病细胞系研究 甲基化与表达之间的关系。我们使用一种局灶性去甲基化技术，CRISPR-DNMT1 相互作用 RNA（CRISPR-DiR），来确定对 表达至关重要的 CpG 岛。
结果
在队列 1 的 25 名患者中有 10 名（40%）和队列 2 的 43 名患者中有 13 名（30%）在接受低甲基化剂治疗后观察到 上调，并且与更差的结局相关。使用 CRISPR-DiR，我们发现 5'非翻译区 CpG 岛内的去甲基化对 表达至关重要。在细胞系和患者中，我们证实低甲基化剂治疗导致同一 CpG 区域的去甲基化和 表达的上调。
结论
通过将患者样本分析与 CRISPR-DiR 技术相结合，我们发现低甲基化剂治疗后癌基因的去甲基化和上调确实会发生，应进一步研究。（由意大利癌症研究协会等资助）。
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