Developing aptamer probes for acute myelogenous leukemia detection and surface protein biomarker discovery

Journal of Hematology & Oncology

Ying Li¹  Samer Z Al-Quran¹  Christopher M Carter¹  Min Zhang¹  Kai Qiu¹  Wenjing Li¹  Guohua Jiang¹  Mingli Yang¹ 

[1] UF/Shands Medical Laboratory at Rocky Point, 4800 35th Drive, Gainesville, FL 32608, USA

关键词: Siglec-5;    Cell-SELEX;    Biomarker;    Aptamer;    Acute myeloid leukemia;   

Others  :  802159
DOI  :  10.1186/1756-8722-7-5

 received in 2013-11-02, accepted in 2013-12-24,  发布年份 2014

【 摘 要 】

Background

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.

Methods

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.

Results

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.

Conclusions

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.

【 授权许可】

   
2014 Yang et al.; licensee BioMed Central Ltd.

【 图 表 】

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