Core 2: Microbiome and Therapeutic Probiotics

核心2摘要/摘要 在人类身上将临床前的发现转化为可行的治疗方案的能力是一个重要的 挑战。最近对人类微生物群的研究表明，微生物 存在于我们身体内部和身体上的社区在健康和疾病方面发挥着重要作用。这个 肠道中蕴藏的微生物区系，很大程度上是由于细菌的数量和多样性 肠道中的微生物，对人类健康起着最关键的作用。因此，最近出现了一个 推动开发治疗肠道疾病的微生物疗法。主要功能 治疗益生菌优化的核心是为项目1和2提供基于微生物的 将改善肠道移植物抗宿主病的治疗方法。核心的目标1是分离新的肠道 能够产生高水平丁酸盐并具有抗炎作用的细菌 属性。我们已经开发了人类排泄物生物反应器，允许产生复合体 微生物群落和产丁酸酯细菌的中等吞吐量测试。核心2将 向项目1提供生产高水平丁酸盐以供测试的复杂社区。目标2 核心之一是将益生菌reuri开发成治疗载体，提供IL- 22、REG3A和REG3γ直接进入肠道。初步数据表明，路氏乳杆菌可以 分泌活性IL-22和REG3A。 我们对微生物组的基本了解的进展已经为更详细地了解奠定了基础 健康人和健康人之间的功能特性和元基因组差异的特征 疾病。在骨髓移植和移植物抗宿主病的背景下，几项研究具有广泛的差异 骨髓移植患者在GVHD前和GVHD期间肠道微生物区系之间的关系。这些研究表明， GVHD期间微生物多样性丧失，肠球菌增多。为了进一步了解 GVHD的发病机制，关键是破译特定毒株在GVHD中 微生物区系介导移植物抗宿主病。 在GVHD研究中缺乏一致的、特征良好的微生物群是一个主要的混淆 可以区分有严重GVHD风险的患者的因素。在过去的五年里，实验室 新上任的Core 2主管费思博士开发了高通量微生物分离管道， 高通量灵知生物群落筛选技术和高效计算算法 鉴定调节宿主生理的特定微生物菌株。通过这些微生物培养分离 方法，Core可以访问包含600多个菌株的微生物培养生物库 从14个个体中分离出来。因此，我们处于一个独特的位置，可以使用工具和试剂来 了解不同微生物群落对肠道发病机制的影响 GVHD模型。识别驱动或预防GVHD的特定微生物菌株将使 疾病病因的机制研究和促进GVHD创新疗法的开发 病人。 GVHD新疗法的一种高度创新的方法利用了 微生物。由结肠中的厌氧菌发酵产生的短链脂肪酸可以 对宿主的深刻影响，以及高水平的丁酸刺激肠道内Treg细胞的扩张。完毕 在过去的五年里，布里顿博士的实验室已经从 并对它们产生丁酸和其他短链脂肪酸的能力进行了筛选。 利用体外生物反应器，Core 2分离并鉴定了具有潜力的细菌菌株 移植物抗宿主病的治疗益处。此外，肠道细菌精密基因组工程的研究进展 而将合成生物技术应用于菌株培育的能力已经使 人类通过细菌产生的治疗性蛋白质成为现实。鲁氏乳杆菌与许多 哺乳动物的肠道，并在整个进化过程中与宿主物种共同进化。我们有 开发了一种人类勒伊氏菌株，作为一种治疗载体，可以通过 在没有抗生素选择的情况下，将人类基因插入到基因组中。

CORE 2 SUMMARY/ABSTRACT The ability to translate pre-clinical findings into viable therapeutic options in humans is a significant challenge. Recent work investigating the human microbiome has demonstrated that microbial communities that exist within and upon our bodies play a large role in health and disease. The microbiota that is harbored in the intestinal tract, due in large part to the vast number and diversity of microbes in the gut, plays the most crucial role in human health. Therefore, there has been a recent push to develop microbial-based therapeutics for the treatment of intestinal disease. The main function of the Therapeutic Probiotic Optimization Core is to provide Projects 1 and 2 with microbial-based therapeutics that will ameliorate intestinal GVHD. Objective 1 of the core is to isolate novel intestinal bacteria capable of producing high levels of butyrate and that also possess anti-inflammatory properties. We have developed human fecal bioreactors that allow for the generation of complex microbial communities and moderate throughput testing of bacteria that produce butyrate. Core 2 will provide Project 1 with complex communities that produce high levels of butyrate for testing. Objective 2 of the core is to develop probiotic Lactobacillus reuteri into a therapeutic delivery vehicle to provide IL- 22, REG3A and Reg3γ directly to the intestinal tract. Preliminary data demonstrates that L.reuteri can secrete active IL-22 and REG3A. Advances in our basic understanding of the microbiome have set the stage for more detailed characterizations of the functional properties and metagenomic differences between health and disease. In the context of BMT and GVHD, several studies have characterized broad differences between the gut microbiota of BMT patients before and during GVHD. These studies demonstrate a loss of microbial diversity and an increase Enterococci in during GVHD. To further understand the pathogenesis of GVHD, it is critical to decipher the mechanisms by which specific strains in a microbiota mediate GVHD. The lack of consistent, well-characterized microbiotas in GVHD studies represents a major confounding factor that could differentiate patients at risk for severe GVHD. Over the past five years, the laboratory of Dr. Faith, the new director of Core 2, has developed high-throughput microbial isolation pipelines, high-throughput gnotobiotic community screening technologies, and efficient computational algorithms to identify specific microbial strains that modulate host physiology. With these microbial culture isolation methods, the Core has access to a biobank of microbial culture collections with over 600 strains isolated from 14 individuals. We are therefore in a unique position to use the tools and reagents to understand the influence of diverse microbial communities on intestinal pathogenesis in experimental GVHD models. Identification of the specific microbial strains that drive or prevent GVHD will enable mechanistic studies of disease etiology and facilitate development of innovative therapeutics for GVHD patients. One highly innovative approach to new therapeutics for GVHD leverages the therapeutic potential of microbes. The production of short chain fatty acids via fermentation by anaerobes in the colon can have profound effects on the host, and high levels of butyrate stimulate Treg cell expansion in the gut. Over the past five years the laboratory of Dr. Britton has isolated several hundred microorganisms from the human gut and screened them for their ability to produce butyrate and other short chain fatty acids. Using in vitro bioreactors Core 2 has isolated and characterized bacterial strains with potential therapeutic benefit in GVHD. In addition, advances in the precision genome engineering of gut bacteria and the ability to apply synthetic biology technology to strain development has made the delivery of human therapeutic proteins via bacteria a reality. Lactobacillus reuteri is associated with many mammalian intestinal tracts and has co-evolved with host species throughout evolution. We have developed a human strain of L.reuteri as a therapeutic delivery vehicle that can survive transit through the human intestinal tract and inserted human genes into the genome without antibiotic selection.