Gymnotic Delivery of LNA/DNA Oligomers into Bacterial

GERMAN M. TRAGLIA; CAROL G DAVIES SALA; BROOKE FUENTES; ALFONSO J. C. SOLER BISTUÉ; ANGELES ZORREGUIETA; MARÍA SOLEDAD RAMÍREZ; MARCELO E TOLMASKY

New Orleans

Congreso; ASM 2015 - 115th General Meeting American Society for Microbiology; 2015

ASM

Background: The need for new therapeutic agents due to the spread of antibiotic resistant bacteria led to development ofdifferent antisense technology strategies for drug design. However, the successful utilization of antisense compounds asantibacterials presents numerous challenges, among them their penetration into the cytosol. While delivery vehicles havebeen researched, relatively recent breakthroughs showed gene silencing in a variety of cells by delivery of nakedoligonucleotides (gymnosis). Here we show gymnotic delivery of DNA oligomers containing locked nucleic acids (LNAs)(LNA/DNA oligomers) into bacterial pathogens.Results: Uptake of labeled LNA/DNA oligomer by cells in stationary phase was most efficient in A. baumannii with anaverage of 24.23% ± 8.9 of the cells internalizing the compound. Other bacteria like A. xyloxosidans, E. cloacae, and P.aeruginosa showed internalization levels between 9 and 16%. The rest of the bacteria assayed did not showed significantlevels of uptake in the conditions tested. CLSM analysis confirmed that the oligomers were internalized rather associated tothe cells.Conclusions: Gymnotic delivery of LNA/DNA oligomers inside bacterial cells was successful for some bacteria. Although itoccurred at a relatively low frequency, it is a starting point to look for strategies to achieve high frequency of gymnotic uptake.The results suggest that utilization of LNA/DNA oligomers may contribute to overcoming the problem of uptake, a stumblingblock in development of antisense compounds as antibacterials.