Engineered Protein Oxygen Carriers: a novel blood substitute for trauma care

工程蛋白载氧体：一种用于创伤护理的新型血液替代品

• 批准号: 9899822
• 负责人: Bohdana Discher
• 金额: $ 20.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9899822
• 关键词: Affinity; Afghanistan; Ambulances; American; Amino Acid Substitution; Amino Acids; Anaerobic Bacteria; Anaphylaxis; Animals; Antibodies; Behavioral; Binding; Binding Sites; Biological Assay; Biological Markers; Biomedical Engineering; Biophysics; Blood; Blood Circulation; Blood Gas Analysis; Blood Pressure Monitors; Blood Substitutes; Blood Transfusion; Blood gas; Body Temperature; Caring; Carrier Proteins; Cells; Cessation of life; Charge; Chemicals; Clinical; Conflict (Psychology); Coupled; Crystallization; Custom; Data; Disease; Drug Kinetics; Emergency Care; Emergency Situation; Engineering; Ensure; Environment; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Escherichia coli; Failure; Flow Cytometry; Functional disorder; Gases; Gift Giving; Goals; Half-Life; Harvest; Health; Heart; Heme; Hemeproteins; Hemoglobin; Hemorrhage; Hemorrhagic Shock; Histology; Hour; Hypertension; IV Fluid; Innovative Therapy; Iraq; Kidney; Kinetics; Laceration; Lactated Ringer' s Solution; Lead; Libraries; Life; Ligands; Link; Liver; Lung; Mass Spectrum Analysis; Measures; Medical; Metabolic Clearance Rate; Metabolism; Military Personnel; Mitochondria; Modeling; Modernization; Molecular Weight; Myoglobin; Nitric Oxide; Organ; Outcome; Oxygen; Patients; Performance; Pharmacology; Physiological; Population; Powder dose form; Property; Protein Engineering; Proteins; Rattus; Refrigeration; Resolution; Resuscitation; Risk; Rodent; Rodent Model; Rural; Safety; Series; Serum; Shock; Site; Structure; Testing; Time; Tissues; Toxic effect; Transfusion; Translating; Traumatic Hemorrhage; Variant; Vascular blood supply; alpha helix; base; blood product; density; design; effective therapy; histological stains; immune activation; immunogenic; immunogenicity; immunoreaction; improved; novel; organ injury; pathogen; physical property; preclinical efficacy; preclinical safety; pressure; prevent; protein transport; screening; skills; success; time use; tissue oxygenation; transfusion medicine; trauma care; vasoconstriction

There is a pressing need for artificial oxygen (O2) carriers as an alternative to donated blood/blood products. For victims of life-threatening hemorrhage, transfusion is the most effective treatment, yet nearly 25% of the nation's blood centers are perennially in short supply. Additionally, the perishability of blood products coupled with blood's potential for immunologic reactions, makes the transfusion of blood products highly problematic in non-hospital settings. The issue of timely access to blood is particularly challenging in austere environments (i.e. battle field, rural site) where lengthy transports to definitive care may be required. Unfortunately, failure to transfuse blood within the “golden hour” increases the risk of poor clinical outcomes including death. The proposed studies aim to use modern advances in de novo designed artificial heme proteins to over come the design limitations of earlier emergency blood substitutes based on natural hemoglobin.. Engineered Protein O2 Carriers (EPOCs) are inherently more adaptable than hemoglobins, allowing direct control of gaseous ligand affinity. EPOCs also offer control over thermal stability (autoclavability), molecular weight and net charge (blood clearance rates), density of O2 binding sites per protein (transport capacity) as well as other desirable physical properties. Importantly, our preliminary results in a rodent hemorrhagic shock model suggest that EPOCs enhance tissue oxygen deliver compared to standard lactated Ringers resuscitation. This proposal will use diverse amino acid substitutions at three specific sites in contact with the O2 binding hemes in the EPOC frame to generate a large library of EPOC variants. These variants will be rapidly screened to select three EPOCs with high, medium, and low O2 affinity, while maintaining tight heme binding. They will also be selected for low nitric oxide binding to avoid potential counterproductive vascular constriction. An initial screening trial in rats using EPOCs to replace blood lost to hemorrhage will identify if any of the three classes of O2 affinities performs better than our current EPOC in circulation. The leading EPOC candidate will then be tested in larger rat trials for pharmacological safety, blood stability and clearance, and potential immunogenicity or toxicity. Another series of rodent trials will quantitate EPOC efficiency at treating severe hemorrhagic shock, in both a shock model with controlled bleeding and a freely hemorrhaging liver laceration shock model. By establishing the best properties for EPOCs in circulation and establishing their safety and efficacy, we will provide the means necessary to build a safe, effective and durable O2 carrier with the ultimate goal of expanding the nation's access to emergency “blood” products and revolutionizing transfusion medicine.

迫切需要人工氧气 (O2) 载体作为捐献血液的替代品 对于危及生命的出血患者来说，输血是最有效的治疗方法。 全国近 25% 的血液中心常年供不应求。 血液制品加上血液潜在的免疫反应，使得输血 产品在非医院环境中存在很大问题，及时获取血液的问题尤其严重。 在严峻的环境（即战场、农村地区）中具有挑战性，因为需要漫长的运输才能到达最终地点 不幸的是，未能在“黄金时间”内输血会增加输血风险。 不良临床结果（包括死亡）的风险。 拟议的研究旨在利用从头设计的人造血红素蛋白的现代进展 克服早期基于天然血红蛋白的紧急血液替代品的设计局限性。 工程蛋白 O2 载体 (EPOC) 本质上比血红蛋白更具适应性，因此可以 直接控制气态配体亲和力还可以控制热稳定性（耐高压灭菌）， 分子量和净电荷（血液清除率）、每个蛋白质的 O2 结合位点密度 （传输能力）以及其他理想的物理特性，我们的初步结果。 啮齿动物失血性休克模型表明，与其他药物相比，EPOC 可以增强组织氧输送 标准乳酸林格氏复苏。 该提案将在与蛋白质接触的三个特定位点使用不同的氨基酸取代 O2 结合 EPOC 框架中的血红素，生成大量 EPOC 变体库。 快速筛选以选择三种具有高、中、低 O2 亲和力的 EPOC，同时保持紧密 它们还将被选择用于低一氧化氮结合，以避免潜在的适得其反的效果。 使用 EPOC 替代因出血而流失的血液的大鼠的初步筛选试验。 将确定三类 O2 亲和力中的任何一类是否比我们当前流通的 EPOC 表现更好。 然后，领先的 EPOC 候选药物将在更大型的大鼠试验中进行药理学安全性、血液 稳定性和清除率，以及潜在的免疫原性或毒性。 定量 EPOC 在治疗严重失血性休克时的效率，在受控休克模型中 出血和自由出血肝裂伤休克模型。 通过确定流通中 EPOC 的最佳性能并确定其安全性和 功效，我们将提供必要的手段来建立一个安全、有效和持久的氧气载体 最终目标是扩大国家获得紧急“血液”产品的机会并进行革命性的变革 输血医学。