Degnin Michelle, Agarwal Anupriya, Tarlock Katherine, Meshinchi Soheil, Druker Brian J, Tognon Cristina E
*Knight Cancer Institute, Oregon Health and Science University ‡Howard Hughes Medical Institute, Portland, OR †Fred Hutchinson Cancer Research Center, Seattle, WA.
J Pediatr Hematol Oncol. 2017 Oct;39(7):e359-e366. doi: 10.1097/MPH.0000000000000946.
The ability to assess antileukemic drug activity on primary patient samples is a powerful tool in determining potential drug targets and selection of therapeutic agents with biological and functional rationale. We previously established small molecule inhibitor screens for use on freshly isolated leukemia cells for this purpose. Here we describe a method that produces functional small molecule inhibitor screening results using cryopreserved primary acute myeloid leukemia cells. This method was established to take advantage of biorepositories containing archival material, such as those established by the Children's Oncology Group, and to enable validation of potential pathway dependencies uncovered by genomic analysis. Various conditions used to thaw and culture cryopreserved specimens were assessed for effect on viability, differentiation, and the ability to recapitulate sensitivity results obtained on fresh samples. The most reproducible results were obtained by quick-thawing and culturing samples in cytokine rich media before performing drug screens. Our data suggest that cytokine-enriched media aids in maintaining the viability and numbers required to perform functional analysis on cryopreserved leukemia cells. This method can aid in producing informative data on therapeutic targeting and precision medicine efforts in leukemia by making use of biorepositories and bio banks.
评估抗白血病药物对患者原代样本活性的能力,是确定潜在药物靶点以及基于生物学和功能原理选择治疗药物的有力工具。此前,我们为此目的建立了用于新鲜分离白血病细胞的小分子抑制剂筛选方法。在此,我们描述一种使用冷冻保存的原发性急性髓系白血病细胞产生功能性小分子抑制剂筛选结果的方法。建立该方法是为了利用包含存档材料的生物样本库,如儿童肿瘤组建立的样本库,并验证基因组分析发现潜在的通路依赖性。评估了用于解冻和培养冷冻保存标本的各种条件对活力、分化以及重现新鲜样本敏感性结果能力的影响。通过在进行药物筛选前快速解冻并在富含细胞因子的培养基中培养样本,获得了最可重复的结果。我们的数据表明,富含细胞因子的培养基有助于维持对冷冻保存的白血病细胞进行功能分析所需的活力和细胞数量。该方法通过利用生物样本库和生物银行,有助于生成有关白血病治疗靶点和精准医学研究的信息性数据。