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

基本信息

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
项目类别：
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

项目摘要

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将非病毒基因转移至唾液腺的方法。该方法利用低频超声波脉冲破坏微泡，引起局部空化并在细胞膜上打开瞬时孔，使非病毒载体能够进入唾液腺细胞。这种提议的方法源自现有的超声辅助基因转移（UAGT）技术，我们小组最近证明该技术能够将稳健、稳定和持久的基因转移到唾液腺。与其他基因转移方法相比，UAGT 具有几个重要的优点，特别是假定缺乏免疫原性，因此理论上可以选择重新给药。基因转移到唾液腺提供了实现肽治疗药物的内源性生产和唾液分泌的机会。内源性生产克服了迄今为止限制新兴抗生物膜肽分子针对牙周菌群中个体不良行为者的效用的许多挑战。这些限制中最主要的是治疗剂本身的制造、稳定性和递送，以及在唾液中实现治疗剂的稳态、持续浓度的挑战。因此，这项研究的长期目标是为唾液腺UAGT在牙科医学中的应用奠定临床前基础。为了实现这一目标，我们将首先将已知的 UAGT 应用于唾液腺，并对该技术对唾液腺结构和活力的影响进行仔细的组织病理学评估。其次，我们将努力进行初步原理验证研究，其中我们将模型微生物（变形链球菌和铜绿假单胞菌）中具有已知抗生物膜活性的基因引入唾液腺，并在体外测定其生物膜抑制特性。接受治疗的动物产生的唾液。一旦这项技术在啮齿动物模型中被证明是可靠、安全和有效的，我们推测它可能是一种有吸引力的佐剂，甚至是处理牙周组织中破坏性微生物病原体的主要方法，特别是对于身体上无法进行日常活动的脆弱患者口腔卫生。公共健康相关性：在本申请中，我们建议使用非病毒基因治疗方法来解决口腔菌群的慢性牙周炎症和组织破坏的公共健康问题。具体来说，我们提出了一种抑制生物膜形成的方法，生物膜形成是一种自然过程，对于无法保持常规口腔卫生（即刷牙）的牙科患者来说，这种自然过程会加速。这项技术在公共卫生领域的直接应用是提供一种基于基因疗法的治疗方法，可以在间歇性牙周治疗之间架起脆弱患者的桥梁。