Development of an Antimicrobial Cellular Therapeutic for Inhibition of Biofilm Fo

描述（由申请人提供）：慢性和感染伤口显著影响数百万美国公民的生活质量，并与并发症风险增加相关，包括截肢的破坏性后果。截至2006年，糖尿病足溃疡感染每年导致82，000例截肢，据估计，这些数字每年将增加10%。此外，在对几种类型的慢性伤口（包括糖尿病足溃疡、静脉性腿部溃疡和压力性溃疡）的广泛研究中，发现60%含有细菌生物膜。美国国立卫生研究院估计，80%的人类传染病与细菌生物膜的形成直接相关。此外，广泛认识到细菌生物膜对抗生素治疗具有高度抗性。有越来越多的证据表明，生物膜形成是皮肤伤口愈合的主要障碍。因此，迫切需要新的策略来破坏这些伤口中的生物膜形成。 该快速通道SBIR转化研究提案的具体目标旨在推进Stratatech Corporation创建的抗微生物活皮肤替代品- 2009年3月;17（3）：562-9）。Stratatech已与工业界、学术界和美国军队建立了一系列跨学科的合作伙伴关系，以测试其在预防或破坏多重耐药细菌形成生物膜方面的有效性。第一阶段的里程碑将1）证明hCAP-18/LL-37分泌的hCAP-18/LL-37是有效的生物膜形成在体外，2）开发效力测定，以监测这种生物活性和3）优化理想的生产和储存条件，为这种活的皮肤替代组织。II期研究将：1）确定CRAFFETHENENCE组织在体外预防或破坏细菌生物膜形成的功效，2）证明CRAFFETHENCE组织在生物膜感染的鼠和猪体内伤口模型中对抗生物膜形成的功效，3）实施并验证cGMP生产工艺和最终临床使用的效价测定，以及4）证明了用于产生Tumor GraftEnhance的晚期传代细胞是核型稳定的、非致瘤性的、不含病毒外源因子，并保持转基因稳定性和完整性。Stratatech的最终目标是商业化的皮肤替代品，用于治疗由于细菌定植和伤口病原体形成生物膜而难以愈合的皮肤伤口。本提案的研究旨在生成临床前数据，这些数据是支持将植入物GraftEnhance技术转化为人体临床试验所需的。 细菌生物膜最近已被医学界认为是伤口愈合的重要障碍，并且美国国立卫生研究院估计，80%的人类感染性疾病与细菌生物膜形成直接相关，但尚未开发和销售特异性靶向生物膜感染的生物治疗选择。这一转化型快速通道SBIR提案的目标是建立一系列合作伙伴关系，以加快创新抗菌治疗剂的开发，旨在通过预防或破坏由多药耐药细菌形成的伤口生物膜来加速伤口愈合。专门设计用于破坏生物膜的培养人类皮肤替代品的开发将针对数百万患有难以愈合的皮肤伤口的患者的需求，通过消除愈合障碍，同时促进愈合，减少疼痛，并减少其他主要并发症的可能性。

DESCRIPTION (provided by applicant): Chronic and infected wounds significantly affect the quality of life for millions of US citizens and are associated with increased risk of complications including the devastating consequence of amputation. As of 2006, infections of diabetic foot ulcers resulted in 82,000 amputations each year and it has been estimated that these numbers will increase by 10% annually. Moreover, in an extensive study of several types of chronic wounds including diabetic foot ulcers, venous leg ulcers, and pressure ulcers, 60% were found to contain bacterial biofilms. The National Institutes of Health estimates that 80% of human infectious diseases are directly related to bacterial biofilm formation. Furthermore, it is widely recognized that bacterial biofilms are highly resistant to antibiotic treatment. There is a growing body of evidence indicating that biofilm formation acts as a major impediment to the healing of cutaneous wounds. Accordingly, there is an urgent need for novel strategies to disrupt biofilm formation in these wounds. The specific aims for this Fast-Track SBIR translational research proposal are designed to advance the development of ExpressGraftEnhance tissue, an antimicrobial living skin substitute created by Stratatech Corporation that has been shown to inhibit the growth of multidrug-resistant bacteria in an in vivo burn infection model (Mol Ther. 2009 Mar;17(3):562-9). Stratatech has established an interdisciplinary series of collaborative partnerships with industry, academia, and the U.S. armed forces to test ExpressGraftEnhance its effectiveness in preventing or disrupting biofilm formation by multidrug-resistant bacteria. Phase I milestones will 1) demonstrate that hCAP-18/LL-37 secreted by ExpressGraftEnhance is efficacious against biofilm formation in vitro, 2) develop potency assays to monitor this bioactivity and 3) optimize ideal production and storage conditions for this living skin substitute tissue. The Phase II study will: 1) determine the efficacy ExpressGraftEnhance tissue in preventing or disrupting bacterial biofilm formation in vitro, 2) demonstrate the efficacy of ExpressGraftEnhance tissue against biofilm formation in both murine and porcine in vivo wound models of biofilm infection, 3) implement and qualify cGMP manufacturing processes and potency assays for ultimate clinical use and 4) demonstrate that late passage cells used to create ExpressGraftEnhance are karyotypically stable, non-tumorigenic, free of viral adventitious agents, and maintain transgene stability and integrity. Stratatech's ultimate goal is to commercialize the ExpressGraftEnhance skin substitute for use in treating cutaneous wounds that are refractory to healing due to bacterial colonization and biofilm formation by wound pathogens. The studies of this proposal are designed to generate preclinical data that are required to support translation of the ExpressGraftEnhance technology into human clinical trials. Bacterial biofilms have become recently become recognized by the medical community as significant barriers to wound healing, and the National Institutes of Health estimates that 80% of human infectious diseases are directly related to bacterial biofilm formation yet no biological treatment options that specifically target biofilm infection have been developed and marketed. The goal of this translational Fast-Track SBIR proposal is to develop a series of collaborative partnerships to expedite the development of an innovative antimicrobial therapeutic aimed at accelerating the healing of wounds by preventing or disrupting wound biofilms formed by multidrug-resistant bacteria. The development of a cultured human skin substitute specifically engineered to disrupt biofilms would target the needs of the millions of patients afflicted with hard-to-heal skin wounds by both eliminating the impediment to healing while also promoting healing, reducing pain, and reducing the likelihood of other major complications.