Selection of a Nuclease-Resistant RNA Aptamer Targeting CD19

Cancers

Katrien Van Roosbroeck¹  George A. Calin²  William G. Wierda²  Alessandra Ferrajoli²  Deborah Rotoli³  Carla L. Esposito³  Silvia Catuogno³  Vittorio de Franciscis³  Annamaria Sandomenico⁴  Menotti Ruvo⁴  Gianluca Santamaria⁵ 

[1] Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA;Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA;IEOS—Istituto per l’endocrinologia e l’oncologia “Gaetano Salvatore”, CNR, 80131 Naples, Italy;Istituto di Biostrutture e Bioimmagini, CNR, 80134 Naples, Italy;Klinikum Rechts der Isar, Department of Regenerative Medicine in Cardiovascular Disease, Technical University of Munich, 81675 Munich, Germany;

关键词: CD19;    aptamer;    B-cells;    cell-SELEX;   

DOI  :  10.3390/cancers13205220

来源: DOAJ

【 摘 要 】

The transmembrane glycoprotein cluster of differentiation 19 (CD19) is a B cell–specific surface marker, expressed on the majority of neoplastic B cells, and has recently emerged as a very attractive biomarker and therapeutic target for B-cell malignancies. The development of safe and effective ligands for CD19 has become an important need for the development of targeted conventional and immunotherapies. In this regard, aptamers represent a very interesting class of molecules. Additionally referred to as ‘chemical antibodies’, they show many advantages as therapeutics, including low toxicity and immunogenicity. Here, we isolated a nuclease-resistant RNA aptamer binding to the human CD19 glycoprotein. In order to develop an aptamer also useful as a carrier for secondary reagents, we adopted a cell-based SELEX (Systematic Evolution of Ligands by EXponential Enrichment) protocol adapted to isolate aptamers able to internalise upon binding to their cell surface target. We describe a 2′-fluoro pyrimidine modified aptamer, named B85.T2, which specifically binds to CD19 and shows an exquisite stability in human serum. The aptamer showed an estimated dissociation constant (K_D) of 49.9 ± 13 nM on purified human recombinant CD19 (rhCD19) glycoprotein, a good binding activity on human B-cell chronic lymphocytic leukaemia cells expressing CD19, and also an effective and rapid cell internalisation, thus representing a promising molecule for CD19 targeting, as well as for the development of new B-cell malignancy-targeted therapies.

【 授权许可】

Unknown