Title

A Non-Natural Nucleoside with Combined Therapeutic and Diagnostic Activities Against Leukemia

Document Type

Article

Publication Date

3-5-2012

Publication Title

ACS Chemical Biology

Abstract

Acute lymphoblastic leukemia (ALL) is the most common type of childhood cancer, presenting with approximately 5,000 new cases each year in the United States. An interesting enzyme implicated in this disease is terminal deoxynucleotidyl transferase (TdT), a specialized DNA polymerase involved in V(D)J recombination. TdT is an excellent biomarker for ALL as it is overexpressed in ∼90% of ALL patients, and these higher levels correlate with a poor prognosis. These collective features make TdT an attractive target to design new selective anti-cancer agents against ALL. In this report, we evaluate the anti-leukemia activities of two non-natural nucleotides designated 5-nitroindolyl-2′-deoxynucleoside triphosphate (5-NITP) and 3-ethynyl-5-nitroindolyl-2′-deoxynucleoside triphosphate (3-Eth-5-NITP). Using purified TdT, we demonstrate that both non-natural nucleotides are efficiently utilized as TdT substrates. However, 3-Eth-5-NITP is poorly elongated, and this observation validates its activity as a chain-terminator for blunt-end DNA synthesis. Cell-based experiments validate that the corresponding non-natural nucleoside produces robust cytostatic and cytotoxic effects against leukemia cells that overexpress TdT. The strategic placement of the ethynyl moiety allows the incorporated nucleoside triphosphate to be selectively tagged with an azide-containing fluorophore via “click” chemistry. This reaction allows the extent of nucleotide incorporation to be quantified such that the anti-cancer effects of the corresponding non-natural nucleoside can be self-assessed. The applications of this novel nucleoside are discussed, focusing on its use as a “theranostic” agent that can improve the accuracy of dosing regimens and accelerate clinical decisions regarding therapeutic intervention.

Comments

This research was supported by funds from the National Institutes of Health (CA118408 to A.J.B.). E.A.M. was supported by the National Cancer Institute Training Programs in Cancer Pharmacology (CA148052). We would like to thank the Cytometry & Imaging Microscopy Core Facility of the Case Comprehensive Cancer Center supported by the NCI (P30 CA43703).

DOI

10.1021/cb300038f

Volume

7

Issue

6