Developing aptamer probes for acute myelogenous leukemia detection and surface protein biomarker discovery.
Developing aptamer probes for acute myelogenous leukemia detection and surface protein biomarker discovery.
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DOI：
10.1186/1756-8722-7-5
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发表时间：
2014-01-09
影响因子：
28.5
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Yang M;Jiang G;Li W;Qiu K;Zhang M;Carter CM;Al-Quran SZ;Li Y
大多数急性髓性白血病（AML）患者仍然死于他们的疾病。为了提高AML患者的生存率，需要新的策略来发现用于AML检测和靶向治疗的生物标志物。基于适体的技术的优点之一是独特的基于细胞的选择过程，这使我们能够有效地选择细胞特异性适体，而无需知道细胞表面存在哪些靶分子。NB 4 AML细胞系用作选择单链DNA适体的靶细胞群。在确定所选适体对白细胞的亲和力后，将适体用于临床样本中的人骨髓白细胞和AML细胞的表型。然后用生物素标记的核酸适体富集并鉴定其靶蛋白。从所选适体池（JH 6、JH 19和K19）表征三种新适体。所有这些都可以选择性地识别Kd在低纳摩尔范围（2.77至12.37 nM）的骨髓细胞。生物素标记的K19适体探针的靶标被鉴定为Siglec-5，这是一种低丰度的表面膜蛋白，其表达可用作粒细胞成熟的生物标志物并用于表型AML。更重要的是，Siglec-5表达可用于检测人骨髓标本中低浓度的AML细胞，并可作为白血病治疗的潜在靶点。我们已经展示了一种流水线方法，用于开发单链DNA适体探针，对临床标本中的AML细胞进行表型分析，然后鉴定适体识别的靶蛋白。开发的适体探针和鉴定的Siglec-5蛋白可能在我们未来的临床实践中用于白血病细胞检测和治疗。
The majority of patients with acute myelogenous leukemia (AML) still die of their disease. In order to improve survival rates in AML patients, new strategies are necessary to discover biomarkers for the detection and targeted therapy of AML. One of the advantages of the aptamer-based technology is the unique cell-based selection process, which allows us to efficiently select for cell-specific aptamers without knowing which target molecules are present on the cell surface. The NB4 AML cell line was used as the target cell population for selecting single stranded DNA aptamers. After determining the affinity of selected aptamers to leukocytes, the aptamers were used to phenotype human bone marrow leukocytes and AML cells in clinical specimens. Then a biotin-labelled aptamer was used to enrich and identify its target surface protein. Three new aptamers were characterized from the selected aptamer pools (JH6, JH19, and K19). All of them can selectively recognize myeloid cells with Kd in the low nanomole range (2.77 to 12.37 nM). The target of the biotin-labelled K19 aptamer probe was identified as Siglec-5, a surface membrane protein in low abundance whose expression can serve as a biomarker of granulocytic maturation and be used to phenotype AML. More importantly, Siglec-5 expression can be used to detect low concentrations of AML cells in human bone marrow specimens, and functions as a potential target for leukemic therapy. We have demonstrated a pipeline approach for developing single stranded DNA aptamer probes, phenotyping AML cells in clinical specimens, and then identifying the aptamer-recognized target protein. The developed aptamer probes and identified Siglec-5 protein may potentially be used for leukemic cell detection and therapy in our future clinical practice.
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DOI：
10.1007/978-1-59745-557-2_5
发表时间：
2009
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Cerchia, Laura;Giangrande, Paloma H;McNamara, James O;de Franciscis, Vittorio
通讯作者：
de Franciscis, Vittorio
影响因子：
11.4
作者：
通讯作者：
--
影响因子：
7
作者：
Mallikaratchy, Prabodhika;Tang, Zhiwen;Tan, Weihong
通讯作者：
Tan, Weihong
影响因子：
2.6
作者：
Nguyen, Dzung H.;Ball, Edward D.;Varki, Ajit
通讯作者：
Varki, Ajit
影响因子：
7.2
作者：
Gold, Larry;Janjic, Nebojsa;Zichi, Dom
通讯作者：
Zichi, Dom