Nardi V, Raz T, Cao X, Wu C J, Stone R M, Cortes J, Deininger M W N, Church G, Zhu J, Daley G Q
Division of Hematology/Oncology, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Children's Hospital Boston, Boston, MA 02115, USA.
Oncogene. 2008 Jan 31;27(6):775-82. doi: 10.1038/sj.onc.1210698. Epub 2007 Aug 6.
Resistance to molecularly targeted chemotherapy, and the development of novel agents that are active against resistant forms of target proteins create the need for a sensitive and quantitative assay to monitor drug-resistant mutations in patients to guide treatment and assess response. Here, we describe an application of the polymerase colony (polony) method to identify and quantify known point mutations in the BCR-ABL oncogene in patients with chronic myelogenous leukemia who evolve resistance to ABL kinase inhibitors. The assay can detect mutations with a sensitivity of 10(-4), quantify the burden of drug-resistant cells, and simultaneously monitor the dynamics of several coexisting mutations. As a proof of concept, we analysed blood samples from three patients undergoing therapy with ABL kinase inhibitors and found that the patients' response to therapy correlated with our molecular monitoring. We were also able to detect mutations emerging in patients long before clinical relapse. Therefore, the polony assay could be applied to a larger patient sample to assess the utility of early mutation detection in patient-specific treatment decisions. Finally, this methodology could be a valuable research tool to shed light on the natural behavior of mutations pre-existing kinase inhibitors therapy and either disappearing over time or slowly taking over.
对分子靶向化疗产生耐药性，以及研发对耐药形式的靶蛋白具有活性的新型药物，使得需要一种灵敏且定量的检测方法来监测患者体内的耐药突变，以指导治疗并评估疗效。在此，我们描述了一种聚合酶集落（polony）方法的应用，用于鉴定和定量慢性髓性白血病患者中对ABL激酶抑制剂产生耐药性的BCR-ABL癌基因中的已知点突变。该检测方法能够以10^(-4)的灵敏度检测突变，定量耐药细胞的负荷，并同时监测几种共存突变的动态变化。作为概念验证，我们分析了三名接受ABL激酶抑制剂治疗患者的血样，发现患者对治疗的反应与我们的分子监测结果相关。我们还能够在临床复发之前很久就检测到患者体内出现的突变。因此，聚合酶集落检测方法可应用于更大规模的患者样本，以评估早期突变检测在针对患者的治疗决策中的效用。最后，这种方法可能是一种有价值的研究工具，有助于阐明在激酶抑制剂治疗之前就已存在、随着时间推移要么消失要么逐渐占据主导的突变的自然行为。
