Ultrasound-Assisted Gene Transfer of Anti-Biofilm Peptides to the Salivary Gland

超声辅助将抗生物膜肽基因转移至唾液腺

• 批准号: 8225139
• 负责人: Michael J. Passineau
• 金额: $ 11.03万
• 依托单位: ALLEGHENY-SINGER RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8225139
• 关键词: Address; Adjuvant; Alternative Therapies; Animals; Architecture; Area; Bacteria; Biological Factors; Bone Tissue; Cell membrane; Cells; Chronic; Debridement; Dental; Dental Hygiene; Diet; Dose; Equilibrium; Evaluation; Excision; Frequencies; Gender; Gene Delivery; Gene Transfer; Gene Transfer Techniques; Genes; Goals; Habits; Healed; Health; Host Defense; Human; In Vitro; Individual; Inflammation; Medical Device; Medicine; Methods; Microbial Biofilms; Microbubbles; Modeling; Mus; Non-Human Gene; Non-Viral Vector; Oral cavity; Oral health; Pathology; Patients; Peptides; Periodontal Diseases; Periodontium; Physiologic pulse; Physiological; Process; Production; Property; Proteins; Pseudomonas aeruginosa; Public Health; Reaction; Relative (related person); Reporting; Research; Rodent Model; Saliva; Salivary; Salivary Glands; Streptococcus mutans; Structure; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Tooth Loss; Toothbrushing; Ultrasonography; Viral Vector; antimicrobial; base; conventional therapy; design; gene therapy; healing; immunogenicity; microbial; microbicide; microorganism; non-viral gene therapy; oral bacteria; oral biofilm; pathogen; pre-clinical; public health relevance; research study; saliva secretion; salivary assay; soft tissue; therapeutic target; therapeutic transgene; toothbrush; transgene expression; Address; Adjuvant; Alternative Therapies; Animals; Architecture; Area; Bacteria; Biological Factors; Bone Tissue; Cell membrane; Cells; Chronic; Debridement; Dental; Dental Hygiene; Diet; Dose; Equilibrium; Evaluation; Excision; Frequencies; Gender; Gene Delivery; Gene Transfer; Gene Transfer Techniques; Genes; Goals; Habits; Healed; Health; Host Defense; Human; In Vitro; Individual; Inflammation; Medical Device; Medicine; Methods; Microbial Biofilms; Microbubbles; Modeling; Mus; Non-Human Gene; Non-Viral Vector; Oral cavity; Oral health; Pathology; Patients; Peptides; Periodontal Diseases; Periodontium; Physiologic pulse; Physiological; Process; Production; Property; Proteins; Pseudomonas aeruginosa; Public Health; Reaction; Relative (related person); Reporting; Research; Rodent Model; Saliva; Salivary; Salivary Glands; Streptococcus mutans; Structure; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Tooth Loss; Toothbrushing; Ultrasonography; Viral Vector; antimicrobial; base; conventional therapy; design; gene therapy; healing; immunogenicity; microbial; microbicide; microorganism; non-viral gene therapy; oral bacteria; oral biofilm; pathogen; pre-clinical; public health relevance; research study; saliva secretion; salivary assay; soft tissue; therapeutic target; therapeutic transgene; toothbrush; transgene expression

DESCRIPTION (provided by applicant): The present proposal will optimize and apply a method for non-viral gene transfer to the salivary glands. This method utilizes low frequency ultrasound pulses to destroy microbubbles, causing local cavitation and opening transient pores in cell membranes enabling non-viral vectors to enter cells of the salivary gland. This proposed method is derived from existing ultrasound-assisted gene transfer (UAGT) techniques that our group has recently shown is capable of robust, stable, and long-lasting gene transfer to the salivary glands. UAGT has several important advantages over other gene transfer methods, in particular a presumed lack of immunogenicity and therefore the theoretical option to re-dose. Gene transfer to the salivary glands presents the opportunity to achieve endogenous production and salivary secretion of peptide therapeutics. Endogenous production overcomes many of the challenges that have thus far limited the utility of emerging anti-biofilm peptide molecules for targeting individual bad actors in the periodontal flora. Chief among these limitations are manufacturing, stability and delivery of the therapeutics itself, as well as the challenge of achieving a steady-state, sustained concentration of the therapeutic in the saliva. The long-term goal of this research therefore is to lay the pre-clinical groundwork for UAGT to the salivary glands for application in dental medicine. In order to achieve this goal, we will first apply what is already known regarding UAGT to the salivary gland and undertake a careful histopathological evaluation of the effect of this technique on salivary gland architecture and viability. Secondly, we will endeavor an initial proof-of-principle study wherein we will introduce genes with known anti-biofilm activity in model microorganisms (S. mutans and P. aeruginosa) to the salivary glands and assay in vitro the biofilm-inhibiting properties of saliva produced by treated animals. Once this technology has been proven to be reliable, safe and effective in the rodent model, we speculate that it may be an attractive adjuvant or even a primary approach to dealing with destructive microbial pathogens in the periodontium, particularly in vulnerable patients physically incapable of routine oral hygiene. PUBLIC HEALTH RELEVANCE: In this application, we propose to use a non-viral gene therapy approach to address the public health issue of chronic periodontal inflammation and tissue destruction by a subset of oral flora. Specifically, we propose a method of inhibiting biofilm formation, a natural process that is accelerated in the most vulnerable to of dental patients who are unable to maintain routine oral hygiene (i.e. tooth brushing). The immediate application of this technology to public health is to provide a gene therapy-based treatment that may bridge vulnerable patients between episodic periodontal treatments.

描述（由申请人提供）：本提案将优化并应用一种非病毒基因转移的方法，以转移到唾液腺。该方法利用低频超声脉冲来破坏微泡，导致局部气蚀并在细胞膜中打开瞬态孔，使非病毒载体能够进入唾液腺的细胞。该提出的方法来自现有的超声辅助基因转移（UAGT）技术，该技术最近显示，这些技术能够具有稳健，稳定和持久的基因转移到唾液腺。 UAGT比其他基因转移方法具有多个重要优势，特别是假定缺乏免疫原性，因此是重新剂量的理论选择。基因转移到唾液腺为实现肽疗法的内源性生产和唾液分泌提供了机会。内源性生产克服了迄今为止限制新兴抗双膜肽分子的效用，这些挑战限制了靶向牙周植物群中个体不良参与者的效用。这些局限性的主要是治疗剂本身的制造，稳定性和交付，以及在唾液中实现稳定，持续浓度的稳定，持续浓度的挑战。因此，这项研究的长期目标是将UAGT的临床前基础奠定到用于牙科医学中的唾液腺。为了实现这一目标，我们将首先将有关UAGT已知的内容应用于唾液腺，并对该技术对唾液腺结构和生存能力的影响进行仔细的组织病理学评估。其次，我们将努力一项初步的原则研究，其中我们将在模型微生物（S. mutans and P. eruginosa）中引入具有已知抗生物胶片活性的基因，以在体外在体外测定由治疗动物产生的唾液的生物膜抑制性质。一旦这项技术被证明在啮齿动物模型中是可靠，安全和有效的，我们推测它可能是一种有吸引力的佐剂，甚至是牙周上处理破坏性微生物病原体的主要方法，尤其是在脆弱的患者中，身体上无能为力。 公共卫生相关性：在此应用中，我们建议采用非病毒基因治疗方法来解决慢性牙周炎症和组织破坏的公共卫生问题。具体而言，我们提出了一种抑制生物膜形成的方法，这是一种自然过程，在无法维持常规口腔卫生的牙科患者最容易受到加速（即牙齿刷牙）。该技术在公共卫生中的立即应用是提供一种基于基因治疗的治疗方法，该治疗可能会弥合情节性牙周治疗之间的脆弱患者。