Exploiting Endogenous Enzymes for Cancer-Cell Selective Metabolic Labeling of RNA in Vivo

Abstract

Tissues and organs are composed of many diverse cell types, making cell-specific gene expression profiling a major challenge. Herein we report that endogenous enzymes, unique to a cell of interest, can be utilized to enable cell-specific metabolic labeling of RNA. We demonstrate that appropriately designed "caged" nucleosides can be rendered active by serving as a substrate for cancer-cell specific enzymes to enable RNA metabolic labeling, only in cancer cells. We envision that the ease and high stringency of our approach will enable expression analysis of tumor cells in complex environments.

Synthesis and anticancer evaluation of acetylated-lysine conjugated gemcitabine prodrugs

“…HEK-293T (human embryonic kidney cells). 23 As shown in Fig. 2, when treated with 0.032 μM concentration of gemcitabine for 72 h, it induced 80% cell death with an IC 50 value of 0.01 μM.…”

“…In an attempt to characterize the specific mechanism of bioactivation of these prodrugs, human cysteine cathepsin L protease was used as it is overexpressed in various cancers and is associated with a number of prodrug activation processes. [20][21][22][23] Prodrug 1 was incubated with CTSL at 37 °C for 24 h and then analyzed by analytical HPLC chromatography. As shown in Fig.…”

“…18,19 The enhanced activity of CTSL in tumor cells could provide a specific environment for targeted cancer therapy, which has been successfully used for the development of anticancer prodrug strategies as well as selective metabolic labeling of RNA in cancer cells. [20][21][22][23] Herein, we present the synthesis of gemcitabine prodrugs with modifications on the 4-N-amino group where an L-or D-Lys(Ac) moiety is masked by different substitutions. The amide bonds in these prodrugs are theorized to be specifically cleaved by enriched CTSL inside cancer cells to finally release the parent compound gemcitabine.…”

“…18,19 The enhanced activity of CTSL in tumor cells could provide a specific environment for targeted cancer therapy, which has been successfully used for the development of anticancer prodrug strategies as well as selective metabolic labeling of RNA in cancer cells. 20–23…”

Molecular Engineering of Self-Immolative Bioresponsive MR Probes

“…Enzymes are associated with numerous physiological and pathological processes used as biomarkers to diagnose diseases and monitor therapeutic responses. − For example, β-galactosidase (β-gal) is a lysosomal hydrolase enzyme that converts lactose derivatives into monosaccharides. − An abnormally high activity of β-gal is closely associated with primary ovarian cancer. In contrast, a deficiency of β-gal is the cause of Morquio B syndrome and GM1 gangliosidosis diseases. ,− …”