Employing biologics to probe GPCR signaling in maternal and fetal health

利用生物制剂探索孕产妇和胎儿健康中的 GPCR 信号传导

• 批准号: 10738487
• 负责人: Meredith Anne Skiba
• 金额: $ 12.43万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738487
• 关键词: Advisory Committees; Agonist; Antibodies; Antihypertensive Agents; Autoantibodies; B-Lymphocytes; Binding; Biological; Biological Assay; Biological Products; Biology; Birth; Cell Separation; Cells; Circulation; Committee Members; Complex; Cryoelectron Microscopy; Development; Discipline of obstetrics; Disease; Drug or chemical Tissue Distribution; Epitopes; Exhibits; Fetal Death; Fetal health; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Hypertension; Immunoglobulin Fragments; In Vitro; Institution; Interdisciplinary Study; Libraries; Ligand Binding; Ligands; Link; Maternal Health; Maternal Mortality; Mentorship; Methods; Molecular; Molecular Conformation; Mus; Oxytocin Receptor; Pathogenicity; Peptides; Pharmacology; Phase; Physiological; Physiological Processes; Pre-Eclampsia; Pregnancy; Premature Birth; Property; Protein Engineering; Receptor Activation; Receptor Signaling; Receptor, Angiotensin, Type 1; Recombinants; Regulation; Renal function; Reproductive Biology; Research; Sampling; Signal Pathway; Signal Transduction; Specificity; Structure; Therapeutic; Therapeutic Intervention; Training; Variant; Visualization; Work; antibody engineering; assisted reproduction; beta-arrestin; blood pressure reduction; clinically relevant; desensitization; fetal; heart function; improved outcome; in vivo; insight; intravenous administration; mouse model; nanobodies; pathogenic autoantibodies; pathophysiology of preeclampsia; peptide drug; pharmacokinetics and pharmacodynamics; pharmacologic; pregnancy disorder; pregnancy related death; programs; receptor; receptor function; recruit; reduce symptoms; response; skills; small molecule; structural biology; synthetic antibodies; therapeutic target; tool

Project Summary and Abstract G protein-coupled receptors (GPCRs) regulate many physiological processes during pregnancy. Oxytocin receptor (OXTR) signaling initiates labor and aberrant signaling through the angiotensin II type I receptor (AT1R) is associated with preeclampsia, a hypertensive disorder of pregnancy and leading cause of pregnancy-related deaths and premature births. GPCRs are therapeutic targets for complications arising during pregnancy and in assisted reproduction, but current small-molecule and peptide drugs exhibit off-target binding, require constant intravenous administration due to short half-lives in vivo, and in some cases are fetotoxic limiting their use. Differing pharmacokinetic and pharmacodynamic profiles and the enhanced specificity make antibodies attractive therapeutic molecules for targeting GPCRs during pregnancy, but the mechanisms antibodies use to modulate GPCR signaling is poorly understood. Previous studies with synthetic antibody fragments and endogenous pathogenic autoantibodies indicate that antibodies can regulate GPCR signaling in a way that does not mirror the action of prototypical small-molecule and peptide ligands. This proposal combines receptor pharmacology, structural biology, and antibody engineering to 1) characterize the mechanisms antibody fragments use to suppress AT1R signaling and demonstrate their ability to selectively target maternal AT1R to safely treat hypertension during pregnancy, 2) develop antibody selection strategies to mechanistically probe how synthetic antibody fragments stabilize distinct states of AT1R and OXTR to regulate the recruitment of signaling effectors and direct biological responses, 3) investigate the mechanisms by which pathogenic autoantibodies dysregulate AT1R signaling in preeclampsia. Collectively, the proposed research will determine how antibodies can be leveraged as tools to dissect the finer details of GPCR signaling, provide insight into the complex pathophysiology of preeclampsia, and identify new avenues to therapeutically target GPCRs to treat preeclampsia, regulate labor, and improve outcomes in assisted reproduction. The aims described in this proposal will offer the PI ample opportunities to expand her scientific skillset in structural pharmacology and antibody engineering, gain training in reproductive biology, and strengthen her expertise in GPCR biology. Mentorship from Dr. Andrew Kruse, advisory committee members, and collaborators will provide the PI with the skills needed to complete the proposed work and transition to independence. The extensive training plan for both scientific and professional development, combined with strong institutional support, will aid the PI in establishing an interdisciplinary research program studying the molecular function of pregnancy-related GPCRs.

项目摘要和摘要 G蛋白偶联受体(GPCRs)调节着妊娠过程中的许多生理过程。催产素 受体(OXTR)信号通过血管紧张素II I型受体(AT1R)启动分娩和异常信号 与先兆子痫有关，这是一种妊娠高血压疾病，也是与妊娠相关的主要原因 死亡和早产。GPCRs是治疗孕期和晚期并发症的靶点 辅助生殖，但目前的小分子和多肽药物表现出脱靶结合，需要恒定的 静脉给药由于体内半衰期短，在某些情况下是胎儿毒性的，限制了它们的使用。 不同的药代动力学和药效学特征以及增强的特异性使抗体 在怀孕期间靶向GPCRs的有吸引力的治疗分子，但抗体用于 对GPCRs信号的调控知之甚少。先前对合成抗体片段和 内源性致病自身抗体表明，抗体可以通过一种方式调节gpr信号。 不能反映典型的小分子和多肽配体的作用。这一建议结合了受体 药理学、结构生物学和抗体工程学，以1)表征抗体的机制 用于抑制AT1R信号的片段，并展示了它们选择性靶向母体AT1R的能力 安全治疗妊娠期高血压，2)开发抗体选择策略，从机制上探索 合成抗体片段如何稳定AT1R和OXTR的不同状态来调节 信号效应器和直接生物反应，3)研究致病机制 自身抗体在先兆子痫中失调AT1R信号转导。总的来说，拟议的研究将确定 抗体如何被用作工具来剖析GPCR信号的更细微细节，提供对 先兆子痫的复杂病理生理机制，并找出治疗靶向GPCRs的新途径 先兆子痫，调节分娩，并改善辅助生殖的结果。本文件中描述的目标 提案将为PI提供充分的机会来扩大她在结构药理学和 抗体工程，获得生殖生物学方面的培训，并加强她在GPCR生物学方面的专业知识。 安德鲁·克鲁斯博士、咨询委员会成员和合作者的指导将为PI提供 完成拟议工作并过渡到独立所需的技能。两人的广泛培训计划 科学和专业的发展，加上强有力的体制支持，将有助于国际和平研究所建立 一项研究妊娠相关GPCRs分子功能的跨学科研究计划。