Obtaining a leukemic cell line with ectopic expression of human protein CD73 as a tool for validation of anti-CD73 antibody

Abstract

Acute lymphoblastic leukemia (ALL) is a type of cancer that is characterized for chromosomal abnormalities and genetic changes during differentiation and proliferation of lymphoid precursor cells and occurs mainly in children. A subtype of ALL, the Philadelphia-like ("Ph like") is related to high rates of cancer recidive. One of the methods used to identificate this type of cancer is qRT-PCR, which uses 15 marker genes such as NT5E, which encodes the CD73 enzyme. This enzyme is an ecto-5'-nucleotidase that has a significant role in the development of tumors. This protein is responsible for hydrolyzing extracellular AMP (adenosine monophosphate) in adenosine, which is an important anti-inflamatory and immunosupressive agent. The expression of CD73 is positively associated with tumor growth, metastasis, angiogenesis and with poor prognosis for several types of cancers. Our research group obtained an therapeutic anti-CD73 antibody using the hybridoma methodology. The next step is the validation of its therapeutic potential in vitro and in vivo. Thereby, this project intends to obtain a leukemic human cell line with ectopic expression of CD73, using the RS4;11 cell line, which is a model of LLA-B. This cell will be transduced with human NT5E gene and with reporter gene (GFP/luciferase) to serve as a tool for validation of de anti-CD73 antibody in vitro.