Sleeping Beauty Transposon Mutagenesis Identifies Genes Driving the Initiation and Metastasis of Uterine Leiomyosarcoma
Sleeping Beauty Transposon Mutagenesis Identifies Genes Driving the Initiation and Metastasis of Uterine Leiomyosarcoma
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DOI：
10.1158/0008-5472.can-21-0356
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发表时间：
2021-11-01
期刊：
影响因子：
11.2
通讯作者：
Copeland, Neal G.
中科院分区：
文献类型：
--
作者：
Kodama, Michiko;Shimura, Hiroko;Copeland, Neal G.
子宫平滑肌肉瘤是一种发生于子宫平滑肌的恶性肿瘤。由于其罕见性、侵袭性和极差的预后，驱动ULMS的分子机制仍然难以捉摸。为了鉴定驱动ULMS的候选癌症基因（CCG），我们在子宫肌层特异性、PTEN敲除、KRAS突变（PTEN KO/KRAS）小鼠中进行了体内睡美人（SB）转座子诱变筛选。ULMS在SB PTEN KO/KRAS小鼠中迅速发展，但在PTEN KO/KRAS小鼠中没有，这证明SB诱变对于在该模型中驱动ULMS至关重要。随后对这些肿瘤中的SB插入位点进行测序，鉴定出19个ULMS CCG，这些CCG在已知的癌症基因中显著富集。其中，Zfp 217和Sfmbt 2在肿瘤发生的早期阶段起作用，似乎是癌基因。与配对的正常子宫平滑肌相比，ZFP 217的人类同源物ZNF 217的表达在人类ULMS中升高，其中其与患者预后呈负相关。抑制ZNF 217抑制，而过表达诱导，增殖，存活，迁移，和人ULMS的干性。在第二次离体ULMS SB转移筛选中，鉴定了可驱动ULMS转移至肺的三种CCG。这些CCG之一，Nrd 1（人类NRDC），与原发性ULMS相比，在人类转移性肿瘤中表现出更强的表达，并与患者生存率呈负相关。NRDC敲低损害迁移和粘附而不影响细胞增殖，而过表达则具有相反的效果。总之，这些结果揭示了新的机制驱动ULMS肿瘤发生和转移，并确定ZNF 217和NRDC作为潜在的目标ULMS therapy.Significance：在体内睡美人转座子诱变筛选确定候选癌症基因，驱动子宫平滑肌肉瘤的起始和进展，并可能作为治疗靶点。
Uterine leiomyosarcoma (ULMS) is a malignancy, which arises from the uterine smooth muscle. Because of its rarity, aggressive nature, and extremely poor prognosis, the molecular mechanisms driving ULMS remain elusive. To identify candidate cancer genes (CCG) driving ULMS, we conducted an in vivo Sleeping Beauty (SB) transposon mutagenesis screen in uterine myometrium-specific, PTEN knockout, KRAS mutant (PTEN KO/KRAS) mice. ULMS quickly developed in SB PTEN KO/KRAS mice, but not in PTEN KO/KRAS mice, demonstrating the critical importance of SB mutagenesis for driving ULMS in this model. Subsequent sequencing of SB insertion sites in these tumors identified 19 ULMS CCGs that were significantly enriched in known cancer genes. Among them, Zfp217 and Sfmbt2 functioned at early stages of tumor initiation and appeared to be oncogenes. Expression of ZNF217, the human homolog of ZFP217, was shown to be elevated in human ULMS compared with paired normal uterine smooth muscle, where it negatively correlated with patient prognosis. Inhibition of ZNF217 suppressed, whereas overexpression induced, proliferation, survival, migration, and stemness of human ULMS. In a second ex vivo ULMS SB metastasis screen, three CCGs were identified that may drive ULMS metastasis to the lung. One of these CCGs, Nrd1 (NRDC in humans), showed stronger expression in human metastatic tumors compared with primary ULMS and negatively associated with patient survival. NRDC knockdown impaired migration and adhesion without affecting cell proliferation, whereas overexpression had the opposite effect. Together, these results reveal novel mechanism driving ULMS tumorigenesis and metastasis and identify ZNF217 and NRDC as potential targets for ULMS therapy.Significance: An in vivo Sleeping Beauty transposon mutagenesis screen identifies candidate cancer genes that drive initiation and progression of uterine leiomyosarcoma and may serve as therapeutic targets.