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

密苏里大学突变小鼠资源和研究中心 - 应用研究第 2 部分

• 批准号: 9923244
• 负责人: Aaron Ericsson
• 金额: $ 3.47万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9923244
• 关键词: Animal Housing; Animals; Antibiotics; Applied Research; Area; Award; Biocompatible Materials; Biomedical Research; Cecum; Cells; Characteristics; Collaborations; Collection; Communities; Complex; Data; Development; Discipline of Nursing; Disease; Embryo Transfer; Engineering; Ensure; Enterohepatic Circulation; Etiology; Experimental Designs; Feces; Filament; Fostering; Funding; Gastrointestinal Contents; Gastrointestinal tract structure; Germ-Free; Housing; Human; Individual; Laboratory mice; Large Intestine; Lead; Longitudinal Studies; Lower Gastrointestinal Tract; Medicine; Methods; Mind; Missouri; Molecular; Molecular Analysis; Monoclonal Antibody R24; Mus; Mutant Strains Mice; Neonatal; Oral; Pathway interactions; Phenotype; Physiological Processes; Physiology; Polymers; Predisposition; Productivity; Prospective Studies; Reporting; Reproducibility; Research; Research Personnel; Resistance; Resources; Role; Sampling; Severity of illness; Small Business Innovation Research Grant; Small Intestines; Stomach; Surveys; System; Testing; Therapeutic; Time; Translations; Universities; Upper digestive tract structure; Xenobiotics; base; cohort; colonization resistance; comparative; fecal microbiota; gut microbiota; high reward; high risk; innovation; longitudinal animal study; member; metallicity; microbial; microbiota; microbiota profiles; mouse model; non-invasive system; nutrition; particle; prospective test; success; tool; virtual

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.

应用研究第二部分--项目总结 在确定复杂的肠道微生物区系(GM)的特征和 对疾病的抵抗力或易感性，研究人员经常试图评估它们之间的因果关系 通过在抗性动物和敏感动物之间转移转基因基因而产生的关联。有几种方法可以做到这一点 因此，包括将转基因受者的胚胎移植到假孕转基因供者，交叉培养新生儿转基因 接受者给转基因捐赠者喂奶，简单地与动物合住，或者试验性地使用准备好的泥浆 从捐赠者的粪便到无菌或经抗生素治疗的转基因接受者(粪便微生物区系转移，FMT)。是否 无论是否成功转移表型，很少有研究仔细检查或报告GM的有效性 从捐赠者转移到接受者，很少有研究直接比较一种以上的转基因转移方法。 此外，已知捐赠者和接受者转基因体内微生物多样性的差异会影响 转基因生物转移成功。因此，应用研究第2部分(具体目标1)的长期目标是 比较使用上述方法和控制良好和彻底的转基因转移的效率 以转基因接受者和捐赠者为特征，他们拥有低多样性和高多样性的转基因特征。具体目标1是测试 转基因胚胎移植、交叉培养、共居和FMT相互作用的效率 低多样性转基因捐助者和高多样性接受者之间的转移，反之亦然，并确定 转基因转移的程度是否与表型也转移的程度有关。 此外，虽然粪便是一种非侵入性样本，允许对个体进行纵向采样 对于小鼠，人们普遍认为粪便只反射下胃肠道(GIT)。许多生理上的 过程和疾病机制依赖于存在于上胃肠道的GM，需要多次 用于纵向研究的小鼠的末端队列，因为上消化道的非侵入性死亡前采样不是 目前是可能的。考虑到这一点，应用研究的长期目标第2节(具体目标 2)开发和验证用于下游测序的非侵入性上Git采样系统 方法：研究方法。与生物材料工程师团队合作，具体目标2开发聚合物/金属 基于颗粒的系统，能够在口腔灌胃后选择性地从上消化道收集细菌细胞， 然后从粪便中提取和分离，用于下游分子分析。的发展。 这样的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 用于检查上腹部的纵向研究。此外，这些方法将彻底改变人类 药物，并将很快获得专利，并可能导致后续资金通过 机制。总体而言，应用研究第二部分的目标将显著增强 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，并 以前无法访问的GIT样本区域。