Application of antisense PNA as antibiotics against periodontal pathogens

Research Project

Project/Area Number	24659925
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Periodontal dentistry
Research Institution	Osaka Dental University (2014) Okayama University (2012-2013)
Principal Investigator	MAEDA Hiroshi 大阪歯科大学, 歯学部, 教授 (00274001)
Co-Investigator(Kenkyū-buntansha)	TAKASHIBA Shogo 岡山大学, 医歯薬学総合研究科, 教授 (50226768) KOKEGUCHI Susumu 岡山大学, 医歯薬学総合研究科, 准教授 (10144776) KITAMATSU Mizuki 近畿大学, 理工学部, 講師 (60379716) YAMASHIRO Keisuke 岡山大学, 医歯薬学総合研究科, 助教 (30581087)
Research Collaborator	MINESHIBA Fumi 岡山大学, 医歯薬学総合研究科, 研究員 (90346461) ISOSHIMA Daichi 岡山大学, 医歯薬学総合研究科
Project Period (FY)	2012-04-01 – 2015-03-31
Project Status	Completed (Fiscal Year 2014)
Budget Amount *help	¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000) Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000) Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000) Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Keywords	歯周病 / 歯周病原細菌 / ペプチド核酸 / アンチセンス / キャリアーペプチド / アンチセンス医療
Outline of Final Research Achievements	The final goal of this research project is to design peptide nucleic acids (PNA) with a carrier peptide and apply them as species-specific antisense drugs against periodontal pathogens. Prior to the application of antisense PNA, effective carrier peptides were selected. Total of 64 peptides (10 amino acids) were designed by referring to previous reports and were synthesized. Fluorescent label (tmp-red) was added to each peptide, and bacterial uptake of the peptides was examined by measuring the strength of the fluorescence. The synthesized peptides were incubated with periodontal pathogen Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Escherichia coli, and the fluorescence inside the bacterial cells was measured. As a results, a peptide with the sequence of Tmr-KFFKFFKFFK-NH2 demonstrated the most effective uptake of P. gingivalis. This peptide will be an effective carrier for antisense PNA against P. gingivalis.