The Mutant Mouse Resource and Research Center at the University of Missouri - Applied Research Section 2

应用研究第2组-计划摘要 在确定复杂肠道微生物群（GM）的特征与 研究人员经常试图评估这些疾病的因果关系， 通过在抗性动物和易感动物之间转移转基因，有几种方法可以做到 因此，包括转基因受体胚胎移植到假孕转基因供体，新生转基因交叉培养， 接受者与护理转基因捐赠者，简单地共同饲养动物，或实验性地施用制备的浆液 从供体粪便到无菌或经驱虫处理的转基因受体（粪便微生物群转移，FMT）。是否 无论转基因是否成功，很少有研究仔细检查或报道转基因的效率。 从捐赠者到接受者的转移，很少有研究直接比较一种以上的转基因转移方法。 此外，已知供体和受体GM内微生物多样性的差异会影响 转基因成功。因此，应用研究第2部（具体目标1）的长远目标如下： 比较转基因转移的效率，使用上述方法和良好的控制和彻底 特征的转基因接受者和捐助者窝藏低和高多样性的转基因概况。具体目标1是测试 利用胚胎移植、交叉培养、共同饲养和FMT进行转基因移植的效率， 多样化程度低的全球机制捐助方和多样化程度高的接受方之间的资金转移，反之亦然，并确定 转基因转移的程度是否与表型转移的程度相关。 此外，虽然粪便代表允许个体纵向取样的非侵入性样品，但是， 对于小鼠，公认的是粪便仅反映下胃肠道（GIT）。许多生理 过程和疾病机制依赖于存在于上GIT的GM，需要多个 用于纵向研究的小鼠终末队列，因为上GIT的非侵入性生前取样不是 目前可能。有鉴于此，应用研究第2节（具体目标）的长期目标 2)是开发和验证一个非侵入性的上GIT采样系统，用于下游测序 方法.与生物材料工程师团队合作，Specific Aim 2开发聚合物/金属 基于颗粒的系统，其能够在口服管饲后从上GIT选择性地收集细菌细胞， 然后从粪便中回收并分离用于下游分子分析。的发展 这样一个系统将通过减少所需的小鼠数量来推进比较医学的三个R 用于检查上消化道的纵向研究此外，这些方法将彻底改变人类 药物，并将立即获得专利，并可能导致随后的资金，通过多个 机制等总的来说，应用研究的目标第2节将大大提高能力， 来自广泛领域的研究人员以知情的方式操纵实验室小鼠的转基因， 以前无法访问的GIT样本区域。

Applied Research section 2 – Project Summary Following identification of associations between characteristics of a complex gut microbiota (GM) and resistance or susceptibility to disease, investigators frequently attempt to evaluate the causality of those associations by transferring the GM between resistant and susceptible animals. There are several ways to do so including embryo transfer of GM recipients into pseudopregnant GM donors, cross-fostering of neonatal GM recipients to nursing GM donors, simply co-housing animals, or experimentally administering a slurry prepared from donor feces to germ-free or antibiotic-treated GM recipients (fecal microbiota transfer, FMT). Whether successful or not in transferring the phenotype, few studies closely examine or report the efficiency of GM transfer from donor to recipient, and very few studies directly compare more than one method of GM transfer. Moreover, differences in the microbial diversity within donor and recipient GMs are known to influence the success of GM transfer. Thus the long-term objectives of Applied Research section 2 (Specific Aim 1) are to compare the efficiency of GM transfer using the aforementioned methods and well-controlled and thoroughly characterized GM recipients and donors harboring low and high-diversity GM profiles. Specific Aim 1 is to test the efficiency of GM transfer using embryo transfer, cross-fostering, co-housing, and FMT, in reciprocal transfers between low-diversity GM donors and high-diversity recipients and vice versa, and to determine whether the degree of GM transfer is associated with the degree to which the phenotype is also transferred. Furthermore, while feces represents a non-invasive sample allowing for longitudinal sampling of individual mice, it is well-recognized that feces reflects only the lower gastrointestinal tract (GIT). Many physiological processes and disease mechanisms are dependent on the GM present in the upper GIT, necessitating multiple terminal cohorts of mice for longitudinal studies as non-invasive ante mortem sampling of the upper GIT is not currently possible. With that in mind, the long-term objective of Applied Research section 2 (Specific Aim 2) is to develop and validate a system of non-invasively sampling the upper GIT for downstream sequencing methods. Working with a team of biomaterials engineers, Specific Aim 2 to develop a polymeric/metallic particle-based system capable of collecting bacterial cells selectively from the upper GIT following oral gavage, and then being retrieved and isolated from feces for downstream molecular analyses. The development of such a system would advance the three ‘R’s of comparative medicine by reducing the required number of mice for longitudinal studies examining the upper GIT. Moreover, these methods would revolutionize human medicine and would be imminently patentable and likely to lead to subsequent funding via multiple mechanisms. Collectively, the Aims of Applied Research section 2 will significantly enhance the ability of researchers from a wide range of fields to manipulate the GM of laboratory mice in an informed manner, and to sample regions of the GIT that were previously inaccessible.