Drug resistance is a major obstacle in the conquest of bacterial infections. This proposal targets a critical issue in the treatment of bacterial infectious diseases: the need to develop strategies aimed at preserving the effectiveness of currently available aminoglycoside antibiotics. The investigators focus on resistance to the aminoglycoside amikacin mediated by the aminoglycoside 6'-N-acetyltransferase AAC(6')-Ib, which can also modify several other aminoglycoside antibiotics. The long term goal of this research project is to develop the use of antisense oligonucleotides as tools to selectively inhibit the expression of aac(6')-Ib and other related genes.
The specific aims for this grant proposal are: 1a) Identification of synthetic oligonucleotides and analogs that promote cleavage of aac(6')-Ib mRNA by RNase H or RNase P. 1b) Bioavailability studies of synthetic oligonucleotides and oligonucleotide analogues. RNase H or RNase P can mediate inhibition of gene expression by degradation of mRNA in the presence of appropriate antisense molecules. The general strategy of this specific aim will consist of the utilization of two conceptually different approaches to identify regions in the mRNA available for interaction with oligonucleotides. With this information, oligodeoxynucleotides and analogues will be designed to inhibit the expression of aac(6')-Ib by inducing RNase H degradation of the mRNA. The information will also be utilized for the development of antisense oligoribonucleotides that inhibit the expression of aac(6')-Ib by inducing RNase P degradation of the mRNA. Following, the investigators will initiate studies on the cell uptake of oligonucleotides and analogues. 2a) Development of peptide nucleic acid (PNA) molecules that inhibit expression of AAC(6')-Ib. 2b) Bioavailability studies of PNA molecules. A novel strategy to inhibit gene expression using antisense technology is now available with the synthesis of peptide nucleic acids (PNAs). PNAs may be developed as antimicrobial agents in prokaryotic systems. The investigators will test the in vitro and in vivo activity of several PNA molecules to interfere with the expression of aac(6')-Ib. The investigators will then study the bioavailability of naked and liposome encapsulated PNA molecules.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
1R15AI047115-01
Application #
6083937
Study Section
Special Emphasis Panel (ZRG1-SSS-K (01))
Project Start
2000-06-01
Project End
2005-05-31
Budget Start
2000-06-01
Budget End
2005-05-31
Support Year
1
Fiscal Year
2000
Total Cost
$127,516
Indirect Cost
Name
California State University Fullerton
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
106670755
City
Fullerton
State
CA
Country
United States
Zip Code
92831
Davies-Sala, Carol; Jani, Saumya; Zorreguieta, Angeles et al. (2018) Identification of the Acinetobacter baumannii Ribonuclease P Catalytic Subunit: Cleavage of a Target mRNA in the Presence of an External Guide Sequence. Front Microbiol 9:2408
Tran, Tung; Chiem, Kevin; Jani, Saumya et al. (2018) Identification of a small molecule inhibitor of the aminoglycoside 6'-N-acetyltransferase type Ib [AAC(6')-Ib] using mixture-based combinatorial libraries. Int J Antimicrob Agents 51:752-761
Chiem, Kevin; Hue, Fong; Magallon, Jesus et al. (2018) Inhibition of aminoglycoside 6'-N-acetyltransferase type Ib-mediated amikacin resistance by zinc complexed with clioquinol, an ionophore active against tumors and neurodegenerative diseases. Int J Antimicrob Agents 51:271-273
Jani, Saumya; Jackson, Alexis; Davies-Sala, Carol et al. (2018) Assessment of External Guide Sequences' (EGS) Efficiency as Inducers of RNase P-Mediated Cleavage of mRNA Target Molecules. Methods Mol Biol 1737:89-98
Stietz, Maria S; Lopez, Christina; Osifo, Osasumwen et al. (2017) Evaluation of the electron transfer flavoprotein as an antibacterial target in Burkholderia cenocepacia. Can J Microbiol 63:857-863
Ramirez, Maria S; Tolmasky, Marcelo E (2017) Amikacin: Uses, Resistance, and Prospects for Inhibition. Molecules 22:
Jackson, Alexis; Jani, Saumya; Sala, Carol Davies et al. (2016) Assessment of configurations and chemistries of bridged nucleic acids-containing oligomers as external guide sequences: a methodology for inhibition of expression of antibiotic resistance genes. Biol Methods Protoc 1:
Chiem, Kevin; Jani, Saumya; Fuentes, Brooke et al. (2016) Identification of an Inhibitor of the Aminoglycoside 6'-N-Acetyltransferase type Ib [AAC(6')-Ib] by Glide Molecular Docking. Medchemcomm 7:184-189
Traglia, German Matías; Dixon, Chelsea; Chiem, Kevin et al. (2015) Draft Genome Sequence of Empedobacter (Formerly Wautersiella) falsenii comb. nov. Wf282, a Strain Isolated from a Cervical Neck Abscess. Genome Announc 3:
Davies-Sala, Carol; Soler-Bistué, Alfonso; Bonomo, Robert A et al. (2015) External guide sequence technology: a path to development of novel antimicrobial therapeutics. Ann N Y Acad Sci 1354:98-110

Showing the most recent 10 out of 44 publications