PEGylated megahemoglobin for use as a red blood cell substitute

聚乙二醇化巨血红蛋白用作红细胞替代品

• 批准号: 9975883
• 负责人: Pedro Cabrales
• 金额: $ 69.56万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-09 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975883
• 关键词: Address; Adverse reactions; Antibodies; Blood; Blood Banks; Blood Circulation; Blood Preservation; Blood Vessels; Blood flow; Bolus Infusion; Characteristics; Clinical Trials; Data; Emerging Communicable Diseases; Endothelium; Endothelium-Dependent Relaxing Factors; Environment; Erythrocyte Transfusion; Erythrocytes; Extravasation; Generations; Globin; Half-Life; Health care facility; Hemoglobin; Hemorrhagic Shock; Homeostasis; Hour; Human; Hydrogen Peroxide; Hypertension; Immune response; Immune system; Immunosuppression; Individual; Infusion procedures; Injections; Kinetics; Lesion; Ligands; Myocardial Infarction; Nitric Oxide; Organ; Oryctolagus cuniculus; Oxidative Stress; Oxides; Oxygen; Patients; Pheretima sieboldi; Physiological; Plant Roots; Polyethylene Glycols; Population; Proteins; Reaction; Religion and Spirituality; Research; Resistance; Resuscitation; Risk; Signal Transduction; Smooth Muscle; Surface; Systemic hypertension; Testing; Tissues; Toxic effect; Treatment Efficacy; United States; Vascular blood supply; Vasodilation; base; biophysical properties; clinical effect; conventional therapy; cost; disease transmission; disorder risk; hemodynamics; immunogenicity; improved; in vivo; molecular size; mortality; oxidation; prevent; side effect; therapeutic protein; transfusion medicine; vasoconstriction

Project Abstract In the United States (U.S.), demand for red blood cells (RBCs) for use in transfusion medicine is steadily growing and will exceed the supply with a projected shortage of 4 million units of RBCs by the year 2030. In addition, frequent seasonal blood shortages, decreasing donation rates, risks of disease transmission, the RBC storage lesion and other side-effects threaten the existing supply of RBCs. Therefore, a viable short-term alternative to donated human RBCs must be developed which is safe, available in large quantities at low cost, and free of the risks of disease transmission and immune suppression, as well as address concerns over religious objections to receiving transfused blood. RBC substitutes should be used to treat conditions in which the use of banked blood is unreasonable or for which there is no therapy. Despite decades of research and clinical trials, mammalian hemoglobin (Hb)-based oxygen (O2) carriers (HBOCs) are still plagued by severe side-effects, such as vasoconstriction at the microcirculatory level, systemic hypertension, myocardial infarction and increased mortality rates. It has been hypothesized that the side-effects associated with previous generations of HBOCs were caused by HBOC extravasation through the blood vessel wall into the tissue space, which led to scavenging of nitric oxide (NO). NO is also known as endothelium-derived relaxing factor (EDRF), and the endothelium (inner lining) of blood vessels uses NO to signal the surrounding smooth muscle to relax, resulting in vasodilation and increased blood flow. Previous generations of HBOCs were also easily oxidized in vivo, creating a significant amount of oxidative stress and tissue damage. All of these problems are a direct consequence of removing Hb from the protective environment of the RBC. We propose that many of these side-effects may be prevented or reduced by using naturally occurring acellular Earthworm Hb (erythrocruorin, LtEc), which has evolved to exist outside of RBCs. Unlike other Hbs (64 kDa), LtEc is naturally large (3.6 MDa) and should not extravasate through the pores lining the blood vessel wall. This feature should prevent/limit its vasoactivity and the extent of systemic hypertension. LtEc is also extremely stable and resistant to oxidation. Therefore, it should elicit limited oxidative stress and tissue toxicity compared to acellular HBOCs synthesized from mammalian Hbs. In addition, LtEc appears to have a much slower or non-existent NO scavenging rate compared to mammalian Hbs, which is the hypothesized root cause of vasoconstriction and hypertension observed in previous generations of acellular HBOCs. Interestingly, the O2 transport characteristics of LtEc are similar to human RBCs, which suggests it will effectively transport O2 to tissues and organs in vivo. Polyethylene glycol (PEG) conjugation to the surface of purified LtEc camouflages the molecule against recognition by the immune system and increases LtEc circulation half-life. Therefore, we hypothesize that PEG-LtEc will serve as an effective alternative to RBCs and will not cause the severe adverse reactions associated with previous generations of HBOCs formulated using acellular mammalian Hbs. To test this hypothesis, we propose 3 specific aims: Specific Aim 1: Purification, synthesis and biophysical characterization of LtEc/PEG-LtEc and individual globin subunits. Specific Aim 2: Assess and characterize the potential immunogenicity of LtEc, PEG-LtEc and denatured LtEc in rabbits. Specific Aim 3: Assess the ability of LtEc and PEG-LtEc to reestablish homeostasis after hemorrhagic shock.

项目摘要 在美国，用于输血医学的红细胞（RBC）的需求稳步增长， 红细胞的供应量将不断增加，预计到2030年将超过供应量，短缺400万单位。在 此外，频繁的季节性血液短缺，献血率下降，疾病传播的风险， 红细胞贮存损伤和其他副作用威胁着红细胞的现有供应。一个可行的短期 必须开发捐献的人红细胞的替代品，其是安全的，可以以低成本大量获得， 没有疾病传播和免疫抑制的风险，并解决 宗教反对接受输血。红细胞替代品应用于治疗以下疾病： 库存血液的使用不合理或没有治疗方法。尽管经过数十年的研究和 在临床试验中，哺乳动物血红蛋白（Hb）基氧（O2）载体（HBOCs）仍然受到严重的 副作用，如微循环水平的血管收缩、全身性高血压、心肌收缩、 梗死和死亡率增加。 据推测，与前几代HBOC相关的副作用是由 通过HBOC通过血管壁外渗到组织间隙中，这导致清除一氧化氮 氧化物（NO）。NO也被称为内皮源性舒张因子（EDRF），并且内皮（内 血管内膜）使用NO向周围平滑肌发出放松信号，导致血管舒张， 血流量增加。前几代的HBOCs也容易在体内氧化，产生显著的 氧化应激和组织损伤的量。所有这些问题都是去除Hb的直接后果 从红细胞的保护环境中分离出来。 我们认为，许多这些副作用可以预防或减少使用天然存在的 无细胞蚯蚓血红蛋白（erythrocuorin，LtEc），它已经进化到存在于红细胞之外。与其他HBS不同 (64 kDa），LtEc天然较大（3.6MDa），不应通过衬于血液的孔外渗 血管壁该特征应防止/限制其血管活性和全身性高血压的程度。LtEc是 也非常稳定和抗氧化。因此，它应该引起有限的氧化应激和组织 与由哺乳动物Hbs合成的非细胞HBOC相比的毒性。此外，LtEc似乎有一个 与哺乳动物Hbs相比，NO清除速率慢得多或不存在，这是假设的根源 在前几代无细胞HBOC中观察到的血管收缩和高血压的原因。 有趣的是，LtEc的O2转运特性与人类RBC相似，这表明它将 有效地将O2输送到体内组织和器官。聚乙二醇（PEG）缀合至 纯化的LtEc使该分子抵抗免疫系统的识别并增加LtEc 循环半衰期因此，我们假设PEG-LtEc将作为RBC的有效替代品， 将不会引起与前几代HBOC相关的严重不良反应， 非细胞哺乳动物血红蛋白为了验证这一假设，我们提出了三个具体目标： 具体目标1：LtEc/PEG-LtEc和单个球蛋白的纯化、合成和生物物理表征 亚单位。 具体目的2：评估和表征LtEc、PEG-LtEc和变性LtEc的潜在免疫原性 家兔 具体目标3：评估LtEc和PEG-LtEc在失血性休克后重建体内平衡的能力。

项目成果

Application of negative tissue interstitial pressure improves functional capillary density after hemorrhagic shock in the absence of volume resuscitation.

• DOI: 10.14814/phy2.14783
• 发表时间: 2021-03
• 期刊: Physiological reports
• 影响因子: 2.5
• 作者: Jani VP;Jani VP;Munoz CJ;Govender K;Williams AT;Cabrales P
• 通讯作者: Cabrales P

Control of systemic inflammation through early nitric oxide supplementation with nitric oxide releasing nanoparticles.

• DOI: 10.1016/j.freeradbiomed.2020.09.025
• 发表时间: 2020-12
• 期刊: Free radical biology & medicine
• 影响因子: 7.4
• 作者: Williams AT;Muller CR;Govender K;Navati MS;Friedman AJ;Friedman JM;Cabrales P
• 通讯作者: Cabrales P

Do Skeletal Dynamics Mediate Sugar Uptake and Transport in Human Erythrocytes?

骨骼动力学是否介导人类红细胞的糖摄取和运输？

• DOI: 10.1016/j.bpj.2018.01.041
• 发表时间: 2018
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Asaro,RobertJ;Zhu,Qiang;Cabrales,Pedro;Carruthers,Anthony
• 通讯作者: Carruthers,Anthony

What determines blood viscosity at the highest city in the world?

世界最高城市的血液粘度由什么决定？

• DOI: 10.1113/jp280206
• 发表时间: 2020
• 期刊: The Journal of physiology
• 作者: Cabrales,Pedro;Govender,Krianthan;Williams,AlexanderT
• 通讯作者: Williams,AlexanderT

Apohemoglobin-haptoglobin complex attenuates the pathobiology of circulating acellular hemoglobin and heme.

脱辅基血红蛋白-触珠蛋白复合物减弱循环无细胞血红蛋白和血红素的病理学。

• DOI: 10.1152/ajpheart.00136.2020
• 发表时间: 2020
• 期刊: American journal of physiology. Heart and circulatory physiology
• 作者: Munoz,CarlosJ;Pires,IvanS;Baek,JinHyen;Buehler,PaulW;Palmer,AndreF;Cabrales,Pedro
• 通讯作者: Cabrales,Pedro