Engineering a novel biomaterial for oxygen transport applications

设计用于氧传输应用的新型生物材料

• 批准号: 10322431
• 负责人: Paul Werner Buehler
• 金额: $ 63.66万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322431
• 关键词: Animal Model; Animals; Antioxidants; Arteriosclerosis; Biocompatible Materials; Biotechnology; Blood; Blood Circulation; Blood Preservation; Blood Volume; Caliber; Cardiac; Cattle; Cavia; Chemicals; Clinical; Clinical Trials; Complex; Complex Mixtures; Development; Diet; Dose; Drug Kinetics; Electron Spin Resonance Spectroscopy; Emerging Communicable Diseases; Endothelium; Engineering; Enzymes; Erythrocytes; Event; Exhibits; Extravasation; Fatty acid glycerol esters; Generations; Glutaral; Goals; Hemoglobin; Hemorrhage; Heterogeneity; High Fat Diet; Human; Image; Impairment; Individual; Injury; Laboratories; Lead; Light; Measures; Membrane; Methods; Modeling; Modification; Molecular; Molecular Weight; Myocardial Infarction; Nanostructures; Nitric Oxide; Oxidative Stress; Oxygen; Oxygen saturation measurement; Phase III Clinical Trials; Physiological; Plasma; Polymers; Population; Proteins; RNA Splicing; Recombinants; Regulation; Research; Reticuloendothelial System; Risk; Role; Sampling; Signal Transduction; Structure; Sucrose; Systemic hypertension; Testing; Therapeutic; Tissues; Toxic effect; Trans-Splicing; Transfusion; Translating; Ultrafiltration; Vascular Permeabilities; Vascular blood supply; Whole Blood Exchange Transfusion; Work; base; biophysical techniques; design; design and construction; dimer; endothelial dysfunction; femoral artery; hemoglobin polymer; hypertensive; innovation; intein; molecular size; monomer; mutant; nonalcoholic steatohepatitis; novel; oxidative damage; oxygen transport; physical property; pre-clinical; preclinical trial; primary endpoint; repaired; response; screening; side effect; survival outcome; tissue injury; tissue oxygenation; transfusion medicine; vascular injury; vasoconstriction

Abstract Hemoglobin (Hb)-based oxygen (O2) carriers (HBOCs) are currently being developed as red blood cell (RBC) substitutes for use in transfusion medicine. Despite significant commercial development, recent late stage clinical results of polymerized hemoglobin (PolyHb) solutions (i.e. Hemopure (OPK Biotech, Cambridge, MA), a glutaraldehyde polymerized bovine Hb; and PolyHeme (Northfield Laboratories Inc., Evanston, IL), a glutaraldehyde polymerized pyridoxylated human Hb) hamper further development. Both of these commercial products elicit vasoconstriction at the microcirculatory level, and lead to the development of systemic hypertension and oxidative tissue damage. These side-effects are hypothesized to occur either by a nitric oxide (NO) scavenging or oxygen (O2) oversupply mechanism, and are both exacerbated by PolyHb extravasation into the tissue space. In light of these 2 potential mechanisms, it is apparent that PolyHb size will have a profound impact on the extent of vasoconstriction, systemic hypertension and oxidative tissue toxicity. However, commercial PolyHb products are complex mixtures with broad size distributions defined only by the size cutoff of the ultrafiltration membranes used in their manufacture. Furthermore, these mixtures are known to contain up to 1% of individual tetrameric Hb molecules and a significantly higher proportion of lower molecular weight (MW) Hb oligomers (80% with MW < 500 kDa). Hence, the side-effects observed during clinical/pre-clinical trials are attributed to a mixture of low MW Hb polymers with different sizes and points of chemical modification, and not to any one, single PolyHb molecule. This precludes precise characterization of how individual components of these complex PolyHb mixtures interact with the vasculature. An important advance would therefore be the ability to produce molecularly uniform, monodisperse, and high MW PolyHb nanostructures. In this application, we hypothesize that the molecular diameter and topology of recombinant PolyHb (rPolyHb) will regulate vasoactivity and oxidative injury to tissues. To test our hypothesis we propose the following specific aims: Specific Aim 1: Use orthogonal split splicing inteins to produce well-defined, monodisperse, high MW rPolyHb nanostructures. Specific Aim 2a: Analyze the role of endothelial function on the development of vasoactivity and oxidative tissue injury to rPolyHbs of varying size. Specific Aim 2b: Evaluate the pharmacokinetics of rPolyHbs in normal guinea pigs and HFSD guinea pigs. Specific Aim 3: Evaluate the ability of rPolyHbs to restore tissue oxygenation and optimize survival in severe blood loss. The proposed work is both significant and innovative, since it seeks to develop safe and efficacious rPolyHbs for use in transfusion medicine. In addition, state-of-the-art biophysical techniques and two unique animal models will be used to understand rPolyHb physiological responses and determine the clinical potential of these novel materials.

摘要 基于血红蛋白(Hb)的氧(O2)载体(HBOC)目前正被开发为红细胞 用于输血医学的(RBC)替代品。尽管商业发展显著，但最近晚些时候 聚合血红蛋白(PolyHb)溶液(即血液净化(OPK Biotech， 马萨诸塞州坎布里奇)，一种戊二醛聚合的牛Hb；和Polyheme(诺斯菲尔德实验室公司， 伊利诺伊州埃文斯顿)，一种戊二醛聚合的吡哆醇基化的人Hb)阻碍了进一步的开发。两个都是 这些商业产品在微循环水平上引起血管收缩，并导致 全身性高血压和氧化组织损伤。这些副作用被认为是由一种 一氧化氮(NO)清除或氧(O2)供应过剩的机制，两者都被PolyHb加剧 渗入组织间隙。根据这两种可能的机制，显然PolyHb大小将 对血管收缩程度、全身性高血压和氧化组织毒性有深远影响。 然而，商用PolyHb产品是复杂的混合物，具有广泛的粒度分布，仅由 制造过程中使用的超滤膜的截止值。此外，这些混合物是 已知含有高达1%的单个四聚体Hb分子，且低Hb分子所占比例要高得多 相对分子质量(MW)Hb低聚物(80%，分子量为500 kDa)。因此，期间观察到的副作用 临床/临床前试验归因于低分子量Hb聚合物的混合物，这些聚合物具有不同的大小和点 化学修饰，而不是对任何一个，单个聚Hb分子。这就排除了对 这些复杂的聚Hb混合物的各个成分如何与血管系统相互作用。 因此，一个重要的进展将是能够产生分子均匀、单分散和 高相对分子质量的多Hb纳米结构。在这个应用中，我们假设分子直径和 重组多聚Hb(RPolyHb)的拓扑结构将调节血管活性和对组织的氧化损伤。至 为了验证我们的假设，我们提出了以下具体目标： 具体目标1：使用正交分割拼接整数来产生定义明确、单分散、高分子量的 RPolyHb纳米结构。 特定目标2a：分析内皮功能在血管活性和氧化发展中的作用 不同大小的rPolyHbs的组织损伤。 特异靶2b：评价rPolyHbs在正常豚鼠和HFSD豚鼠体内的药代动力学 猪。 具体目标3：评估rPolyHbs恢复组织氧合和优化存活的能力 严重失血。 拟议的工作既有意义又有创新，因为它寻求发展安全和有效的 用于输血医学的rPolyHbs。此外，最先进的生物物理技术和两种独特的 动物模型将被用来理解rPolyHb的生理反应并确定其临床潜力 这些新奇的材料。