Biotechnological Research Support Division, FASMAC Co., Ltd, Atsugi, Kanagawa, 243-0021, Japan.
Department of Rare Diseases Research, Institute of Medical Science, St. Marianna University School of Medicine, Kawasaki, Kanagawa, 216-8512, Japan.
Commun Biol. 2022 Jun 2;5(1):535. doi: 10.1038/s42003-022-03467-w.
Both natural viral infections and therapeutic interventions using viral vectors pose significant risks of malignant transformation. Monitoring for clonal expansion of infected cells is important for detecting cancer. Here we developed a novel method of tracking clonality via the detection of transgene integration sites. RAISING (Rapid Amplification of Integration Sites without Interference by Genomic DNA contamination) is a sensitive, inexpensive alternative to established methods. Its compatibility with Sanger sequencing combined with our CLOVA (Clonality Value) software is critical for those without access to expensive high throughput sequencing. We analyzed samples from 688 individuals infected with the retrovirus HTLV-1, which causes adult T-cell leukemia/lymphoma (ATL) to model our method. We defined a clonality value identifying ATL patients with 100% sensitivity and 94.8% specificity, and our longitudinal analysis also demonstrates the usefulness of ATL risk assessment. Future studies will confirm the broad applicability of our technology, especially in the emerging gene therapy sector.
无论是自然的病毒感染还是使用病毒载体的治疗干预，都存在恶性转化的重大风险。监测感染细胞的克隆扩增对于检测癌症很重要。在这里，我们开发了一种通过检测转基因整合位点来跟踪克隆性的新方法。RAISING（无基因组 DNA 污染干扰的整合位点快速扩增）是一种比现有方法更敏感、更廉价的替代方法。它与 Sanger 测序的兼容性，结合我们的 CLOVA（克隆性值）软件，对于那些无法使用昂贵的高通量测序的人来说至关重要。我们分析了 688 名感染逆转录病毒 HTLV-1 的个体的样本，该病毒会导致成人 T 细胞白血病/淋巴瘤（ATL），以此来模拟我们的方法。我们定义了一个克隆性值，可以识别出 100%敏感和 94.8%特异性的 ATL 患者，我们的纵向分析也证明了这种方法在评估 ATL 风险方面的有用性。未来的研究将证实我们的技术具有广泛的适用性，特别是在新兴的基因治疗领域。
