Lipid Bodies and Prostaglandin Production in Labor

描述（由申请人提供）：本提案描述了一个严格的五年培训计划，旨在促进生殖医学独立学术生涯的发展。主要研究者在俄亥俄州州立大学（OSU）的围产期研究实验室（LPR）完成了博士后研究。在此经验的基础上，该计划旨在通过在几位备受推崇的生物医学科学家的实验室中学徒来扩展特定的科学技能。该计划将侧重于炎症的生物学和生物化学方面的高级培训，特别强调其在人类分娩中的作用。主要研究者的科学发展将由咨询委员会的成员指导。申办者是一位备受尊敬的生殖生物学家，在足月和早产的分子方面具有专业知识，培训了许多临床研究员，博士后研究员和研究生。临床顾问是早产临床方面的权威。每一位共同导师都是根据所采用的研究方法的具体方面的广泛专业知识选择的。该研究将集中于确定脂质体（LB）在羊膜上皮细胞内花生四烯酸（AA）储存、动员和代谢中的作用。随着妊娠的进行，大量LB的积累是羊膜细胞的一个显著形态学特征，尽管这些结构的作用尚未明确。在白细胞和其他与炎症相关的细胞中，LB与酯化AA的储存有关。此外，与AA的动员和氧化代谢相关的酶可能被招募到这些结构中。AA是形成被认为对分娩过程至关重要的前列腺素（PC）的专性前体。导致PG形成的代谢途径已经建立;然而，协调这一过程的生理机制仍然不完全确定。在某种程度上，这是由于系统的复杂性：PG生物合成是一个高度动态和密切调控的过程，需要多种酶（包括磷脂酶，环加氧酶和PG合酶亚型）的瞬时偶联，否则分离到不同的细胞区室。待检验的具体假设是，羊水细胞LB通过组织代谢前体（AA）和合成酶之间的功能偶联，可能对分娩所需PG的产生起关键作用。具体目标包括：1）确定劳动对LB积累的影响; 2）确定LB与AA动员和代谢所必需的酶结合的程度; 3）确定LB扰动对PG产生的影响;和4）确定AA代谢受抑制对LB形成的影响。具体的实验将涉及补充生化，分子，免疫学和形态学评估，这将显着扩展实习生的实验室技能。在俄勒冈州立大学的LPR提供了一个理想的环境，为医生科学家的培训，无论是在资源和智力支持方面。通过妇产科医生和各个学科的基础科学家之间的长期合作，LPR成为转化研究的独特平台。培训将通过在另外两个实验室轮换来补充，每个实验室都配备了进行拟议研究所需的资源，此外还有研究生教学课程。

DESCRIPTION (provided by applicant): This proposal describes a rigorous five-year training program intended to facilitate the development of an independent academic career in reproductive medicine. The prinicipal investigator has completed a postdoctoral fellowship in the Laboratory of Perinatal Research (LPR) at The Ohio State University (OSU). Building upon this experience, this program has been designed to enable expansion of specific scientific skills through apprenticeship in the laboratories of several highly-regarded biomedical scientists. The program will focus on advanced training in the biological and biochemical aspects of inflammation, with specific emphasis on its role in human parturition. The principal investigator's scientific development will be mentored by members of an advisory committee. The sponsor, a well-respected reproductive biologist with expertise in the molecular aspects of term and preterm labor, has trained numerous clinical fellows, postdoctoral fellows, and graduate students. The clinical advisor is an authority on the clinical aspects of preterm labor. Each of the co-mentors was chosen based on extensive expertise in specific aspects of the research methodologies to be employed. The research will focus on defining the role of lipid bodies (LBs) in arachidonic acid (AA) storage, mobilization, and metabolism within cells of the amnion epithelium. The accumulation of numerous LBs with advancing gestation is a striking morphological feature of amniotic cells, although the role of these structures has not been clearly defined. In leukocytes and other cells associated with inflammation, LBs have been implicated in the storage of esterified AA. Furthermore, enzymes associated with the mobilization and oxidative metabolism of AA may be recruited to these structures. AA is the obligate precursor for the formation of the prostaglandins (PCs) considered critical to the process of parturition. The metabolic pathway leading to PG formation has been established; however, the physiological mechanisms that orchestrate this process remain incompletely defined. In part, this is due to the complexity of the system: PG biosynthesis is a highly dynamic and intimately regulated process, requiring the transient coupling of multiple enzymes (including phospholipases, cyclooxygenases, and PG synthase isoforms) that are otherwise segregated to different cellular compartments. The specific hypothesis to be tested is that amniotic cell LBs, by organizing the functional coupling between metabolic precursor (AA) and synthetic enzymes, may factor critically into the generation of PGs necessary for labor. The specific aims include: 1) determining the influence of labor on LB accumulation; 2) establishing the degree to which LBs associate with enzymes necessary for AA mobilization and metabolism; 3) determining the effects of LB perturbation on PG production; and 4) determining the effects of inhibited AA metabolism on LB formation. Specific experiments will involve complementary biochemical, molecular, immunological, and morphological assessments, which will significantly extend the laboratory skills of the trainee. The LPR at OSU provides an ideal environment for the training of physician-scientists, both in terms of resources and intellectual support. Through long-standing collaborative efforts between obstetrician-gynecologists and basic scientists from a variety of disciplines, the LPR serves as a unique platform for translational research. Training will be supplemented through rotations in two additional laboratories, each of which is fully equipped with the resources necessary to conduct the proposed studies, in addition to didactic graduate course work.