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.

迫切需要人造氧（O2）载体作为捐献血液/血液的替代品 产品.对于危及生命的出血患者，输血是最有效的治疗方法，但 全国近25%的血液中心常年供应不足。此外， 血液制品加上血液的潜在免疫反应，使得输血 产品在非医院环境中存在很大问题。及时获得血液的问题尤其重要， 在严峻的环境中（即战场，农村地区）具有挑战性， 可能需要小心。不幸的是，未能在“黄金时间”内输血， 不良临床结局的风险，包括死亡。 拟议的研究旨在利用现代先进的从头设计的人工血红素蛋白 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， 直接控制气体配体亲和力。EPOC还提供对热稳定性（高压灭菌性）的控制， 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 （输送能力）以及其它所需的物理性质。重要的是，我们的初步结果 啮齿类动物失血性休克模型表明，与对照组相比， 标准乳酸林格氏复苏。 该提议将在与蛋白质结合的三个特定位点处使用不同的氨基酸取代。 O2结合血红素的EPOC框架，以产生一个大的EPOC变体库。这些变体将 快速筛选，以选择具有高，中，低O2亲和力的三种EPOC，同时保持紧密的 血红素结合他们也将被选择为低一氧化氮结合，以避免潜在的适得其反 血管收缩在大鼠中使用EPOC替代失血的初步筛选试验 将确定三种O2亲和力中的任何一种是否比我们目前流通中的EPOC表现更好。 然后，将在更大的大鼠试验中测试领先的EPOC候选药物的药理学安全性，血液 稳定性和清除率以及潜在的免疫原性或毒性。另一系列啮齿动物试验将 定量EPOC治疗严重失血性休克的效率，在休克模型和对照模型中， 出血和自由落体式肝裂伤休克模型。 通过确定EPOC在流通中的最佳特性，并确定其安全性， 有效性，我们将提供必要的手段，建立一个安全，有效和持久的O2载体， 最终目标是扩大国家获得紧急“血液”产品的机会， 输血医学