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）载体作为捐赠血/血液的替代品 产品。对于威胁生命的出血的惊喜，输血是最有效的治疗方法 该国近25％的血液中心长期供应。另外，易腐性 血液产物以及血液的免疫反应潜力，使血液输血 在非医院环境中的产品极为问题。及时获得血液的问题尤其是 在严峻的环境（即战场，粗糙地点）中具有挑战性 可能需要注意。不幸的是，未能在“黄金时光”内传播血液会增加 包括死亡在内的临床结果不良的风险。 拟议的研究旨在利用从头设计的现代进步设计的人工血红素蛋白 超越基于天然血红蛋白的早期紧急血液替代品的设计限制。 工程蛋白O2载体（EPOC）本质上比血红蛋白更适应 EPOC还提供了对热稳定性（高灭菌性）的控制， 分子量和净电荷（血液清除率），每个蛋白质的O2结合位点的密度 （运输能力）以及其他理想的物理特性。重要的是，我们的初步结果 啮齿动物出血性休克模型表明，EPOC与 标准的乳酸铃声复苏。 该提案将在与与该地点接触的三个特定地点使用不同的氨基酸取代 EPOC框架中的O2结合下摆，以生成大型EPOC变体库。这些变体会 快速筛选以选择具有高，中和低O2亲和力的三个EPOC，同时保持紧密 血红素结合。还将选择低一氧化氮结合以避免潜在的适得其反 血管收缩。使用EPOC在大鼠中进行初步筛查试验，以取代流血流血的血液 将确定三类的O2亲和力中的任何一个都比我们目前的循环中的EPOC更好。 然后，领先的EPOC候选人将在较大的大鼠试验中进行药物安全，血液的测试 稳定性和清除率以及潜在的免疫原性或毒性。另一个系列的啮齿动物试验将 量化EPOC在处理严重出血性休克时的EPOC效率，这两种电击模型都具有受控 出血和自由出血肝间隙冲击模型。 通过在流通中建立最佳EPOC的特性，并确定其安全性 有效，我们将提供必要的手段，以建造安全，有效和耐用的O2载体 扩大国家获得紧急“血液”产品并彻底改变国家的最终目标 输血医学。