Biological Treatment of Bacterial Keratitis

项目概要/摘要 细菌性角膜炎（BK）是一种角膜感染，如果不及时治疗，可导致失明。金黄色葡萄球菌（Sa）， 肺炎链球菌（Sp）和铜绿假单胞菌（Pa）是北美BK的三个主要原因。 BK的症状包括疼痛、发红、炎症和受影响角膜的混浊以及视力丧失（1）。 AmebaGone Inc.，(AG)与UW-Madison眼科部门合作开发了一种新的生物控制方法 AG拥有已颁发的专利，并拥有与使用网柱细胞（DC）相关的专业知识， 通过吞噬作用破坏致病菌的生物体。已颁发的专利广泛涵盖使用 DC吃生物膜缠结和自由生活（反硝化）细菌（“治疗阿米巴及其用途”; USPTO授予 专利8，551，471和后续专利US 8，551，671）。一项覆盖欧盟国家的专利已经提交 该公司拥有来自3个世界各地的收藏品的3,000多个DC菌株的许可权，这些菌株占据了不同的自然资源， 栖息地良性DC在10亿多年前从水生阿米巴中分化出来，其中一些是致病的。一期 AG鉴定了36种能够有效杀灭（细菌滴度降低> 4 log10）眼内Sa的Dicty菌株 温度在这些菌株中，AG鉴定出4种DC菌株，其可重复地萌发至高水平并形成凝胶 最大恢复时间从1小时增加到16小时。此外，AG测试了这些DC的安全性 菌株，并发现与载体对照相比，角膜病理学或不适没有增加。在二期工程中， AG建议：1）将DC的目标从针对Sa的单一作用扩展到BK-Pa的其他病原体， sp，2）。开发有效的配方并评估其中的DC稳定性。3)进行体内安全性和基础生物学试验， 眼内DC治疗，以及4）。量化DC鸡尾酒在治疗感染Sa、Pa或Sp的小鼠角膜中的功效， vivo.这些研究中产生的数据将使我们能够准备新药研究申请（IND）， 美国食品药品监督管理局（FDA）。

Project Summary/Abstract Bacterial keratitis (BK) is an infection of the cornea that, if left untreated, can cause blindness. Staphylococcus aureus (Sa), Streptococcus pneumoniae (Sp), and Pseudomonas aeruginosa (Pa) are three major causes of BK in North America. Symptoms of BK include pain, redness, inflammation, and opacity of the affected cornea and loss of visual acuity (1). AmebaGone Inc., (AG) partnered with UW-Madison Department of Ophthalmology to develop a novel biocontrol method to treat BK. AG holds issued patents and has expert knowledge related to use of Dictyostelid cells (DC), a benign, soil- dwelling organism known to destroy pathogenic bacteria through the phagocytosis. Issued patents broadly cover the use of DC to eat biofilm-enmeshed and free-living (planktonic) bacteria (“Therapeutic Ameba and Uses Thereof”; USPTO granted patent 8,551,471 and follow up patent US 8,551,671). A patent covering countries of the European Union has been filed and the firm has licensed rights to more than 3,000 DC strains from 3 worldwide collections occupying diverse natural habitats. Benign DC diverged from aquatic amoeba, some of which are pathogenic, over 1 billion years ago. In Phase I work, AG identified 36 Dicty strains capable of efficiently killing (> 4 log10 reduction in bacterial titers) Sa at eye temperature. Of these strains, AG identified 4 DC strains that reproducibly germinated to high levels and developed a gel formulation that increased the maximum recovery time from 1 to 16 hours. Additionally, AG tested safety of these DC strains and found no increase in corneal pathology or discomfort compared to the vehicle control. In the Phase II project AG proposes to: 1) Expand the target of DC from singular action against Sa to additional causative agents of BK- Pa and Sp, 2). Develop effective formulation and assess DC stability therein. 3) Conduct in vivo safety and basic biology testing of DC treatment in the eye, and 4). Quantify efficacy of DC cocktails in treating murine cornea infected with Sa, Pa, or Sp in vivo. The data generated in these studies will position us to prepare an Investigational New Drug (IND) application for the Food and Drug Administration (FDA).