Nanoparticle mediated delivery of 2'-O-methyl-RNA leads to efficient telomerase inhibition and telomere shortening in human lung cancer cells.
A promising approach for treatment of non-small cell lung cancer (NSCLC) is based on the inhibition of telomerase in cancer cells. The antisense oligonucleotide 2'-O-methyl-RNA binding to the RNA component of telomerase acts as a selective telomerase inhibitor. We developed chitosan-coated polylactide-coglycolide (PLGA) nanoparticles to mediate efficient delivery of 2'-O-methyl-RNA into human lung cancer cells. Cellular uptake of the inhibitor mediated by chitosan-coated PLGA nanoparticles was greatly enhanced compared to the uptake of antisense oligonucleotide alone as shown by flow cytometry analysis. Confocal laser scanning microscopy clearly demonstrated internalization of 2'-O-methyl-RNA. 2'-O-methyl-RNA-nanoparticle complexes exhibited nearly no acute cytotoxicity in human lung cancer cells and did not influence the viability of primary tumor lung fibroblasts. Human NSCLC A549 cells treated with 2'-O-methyl-RNA-nanoparticle complexes showed 87% viability compared to untreated control cells. 2'-O-methyl-RNA delivered by nanoparticle complexes inhibited telomerase activity in a sequence-specific manner. During long-term treatment (15 weeks) telomerase activity was continuously reduced by approximately 80%. Furthermore, nanoparticle mediated delivery of 2'-O-methyl-RNA resulted in significant telomere shortening from 5.9kb to 4kb (p=0.008) in A549 cells. In summary, our data demonstrate that nanoparticle mediated delivery of 2'-O-methyl-RNA induces effective telomerase inhibition and telomere shortening in human lung cancer cells and therefore represents a novel and promising strategy for the treatment of lung cancer.
Beisner
Julia
J
Dr. Margarete Fischer - Bosch-Institute of Clinical Pharmacology, University of Tuebingen, Stuttgart, Germany. julia.beisner@ikp-stuttgart.de
Dong
Meng
M
Taetz
Sebastian
S
Nafee
Noha
N
Griese
Ernst-Ulrich
EU
Schaefer
Ulrich
U
Lehr
Claus-Michael
CM
Klotz
Ulrich
U
Mürdter
Thomas E
TE
D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D018014:Gene Transfer Techniques; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000628:therapy*; D018613:Microscopy, Confocal; D053758:Nanoparticles; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage* / Q000235:genetics; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000235:genetics; D016615:Telomere / Q000235:genetics / Q000473:pathology
C467846:2'-O-(2-(methylthio)ethyl) RNA; D009838:Oligodeoxyribonucleotides; D019098:Telomerase