Bioengineering a novel therapeutic protein complex to minimize the effects of medical device induced hemolysis

生物工程新型治疗性蛋白质复合物可最大程度地减少医疗设备引起的溶血的影响

• 批准号: 10380296
• 负责人: Paul Werner Buehler
• 金额: $ 70.2万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10380296
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Affinity; Animal Model; Animals; Attenuated; Binding; Binding Proteins; Binding Sites; Biological Assay; Biological Markers; Biomedical Engineering; Blood Circulation; Brain; Cardiac; Cardiac Output; Cardiac Surgery procedures; Cardiopulmonary Bypass; Cells; Cessation of life; Chemicals; Clinical; Complex; Complication; Cytolysis; Data; Dimerization; Dose; Drug or chemical Tissue Distribution; Effectiveness; Engineering; Erythrocytes; Extracorporeal Membrane Oxygenation; Fractionation; Functional disorder; Goals; Hamsters; Haptoglobins; Health Care Costs; Heart; Heme; Heme Iron; Hemeproteins; Hemoglobin; Hemoglobin concentration result; Hemolysis; Hemopexin; Hospitalization; Hour; Human; In Vitro; Individual; Injury to Kidney; Intestines; Iron; Kidney; Kinetics; Lead; Length of Stay; Ligands; Lung; Medical; Medical Device; Methods; Microvascular Permeability; Modeling; Monitor; Morbidity - disease rate; Operative Surgical Procedures; Organ; Outcome; Paste substance; Patients; Plasma; Procedures; Process; Proteins; Proteolysis; Public Health; Pump; Rattus; Renal function; Risk; Sampling; Secure; Self-Help Devices; Severities; Supportive care; System; Testing; Therapeutic; Therapeutic Intervention; Therapeutic Uses; Toxic effect; Toxin; Transferrin; Urine; Ventricular; apohemoglobin; base; blood pump; cell injury; clinically relevant; cost; cost estimate; design; dimer; experience; haptoglobin-hemoglobin complex; heart function; heme a; hemodynamics; improved; in vivo; individual patient; instrument; large scale production; lung injury; macrophage; monocyte; mortality; mortality risk; novel; novel therapeutics; organ injury; prevent; prospective; protein complex; proteostasis; pulmonary function; response; urinary

PROJECT SUMMARY Cost estimates for CNS, pulmonary, cardiac, and renal complications following medical assist devices requiring blood pumps, for example on pump cardiopulmonary bypass (CPB) is estimated at $80 million per individual states in the US over a ten-year period. Because of the need for extended CPB (> 4hours) use in complex surgeries, hemolysis, and the subsequent release of hemoglobin (Hb) into the circulation – which is known to be multi-organ toxic and vasoactive – is a relevant contributor to more intensive management of patients. To date, no effective strategy to remove toxic by-products of Hb are available. The burden caused by kidney injury alone accounts for approximately $9 billion/year, 300,000 deaths/year and an average increase in hospital stay of 3.5 days/patient This project focuses on understanding the contribution of hemolysis, Hb and heme associated with secondary end organ injury following extended on pump cardiopulmonary bypass (typically lasting up to 4 hours). To understand medical device related red blood cell lysis in this setting, we propose a strategy that is focused on sequentially understanding the contributions of Hb and heme and a therapeutic strategy to attenuate the end organ pathophysiology of these toxins. We will prospectively determine the levels of Hb, heme and iron as well as the concentrations of each toxin in plasma and urine of cardiac surgery patients. Simultaneously we will determine the concentrations of Hb, heme and iron binding and clearance proteins, haptoglobin, hemopexin and transferrin, respectively. Further we will determine plasma and urinary markers of end organ injury. We will use this data to construct a biokinetic model that determines the limits of Hb toxin concentrations that associate with end organ injury markers. The goal of this effort will be to define the need and timing for therapeutic interventions. To this end we have bioengineered a novel Hb, heme and protein scavenger based on our extensive experience with studying Hb toxicity. The novel protein construct is prepared by generating apo-Hb in multi-step process, while simultaneously isolating haptoglobin from Cohn fractionation paste IV. Finally, the two proteins are complexed and further purified to generate the apo-Hb-haptoglobin complex. This novel protein construct binds heme in high heme exposure states, secures iron in the heme ligand and safely clears the complex to monocytes and macrophages. Alternatively, in high Hb exposures apo-Hb exchanges binding sites on haptoglobin clearing Hb dimers to monocytes and macrophages, while released apo-Hb dimers are degraded by proteolysis and harmlessly cleared from circulation. To test the effect of the novel complex we have planed a range of proof-of-concept studies in animal models of on pump CPB to define the ability of apo- Hb-haptoglobin dosing to prevent end organ injury.

项目摘要 使用医疗辅助设备后CNS、肺、心脏和肾脏并发症的成本估计 需要血液泵，例如泵式心肺转流术（CPB）， 在美国的各个州，在十年的时间里。由于需要延长CPB（> 4小时）， 复杂的手术，溶血，以及随后释放的血红蛋白（Hb）进入循环-这是 已知具有多器官毒性和血管活性-是更强化管理的相关因素， 患者迄今为止，没有有效的策略来去除Hb的有毒副产物。造成的负担 仅肾损伤就占约90亿美元/年，30万人死亡/年， 住院时间增加3.5天/患者本项目侧重于了解 持续泵上后溶血、Hb和血红素与继发性终末器官损伤相关 心肺转流（通常持续长达4小时）。了解医疗器械相关红细胞 在这种情况下，我们提出了一个战略，重点是顺序理解的贡献， 血红蛋白和血红素以及减轻这些毒素的终末器官病理生理学的治疗策略。我们将 前瞻性地测定Hb、血红素和铁的水平以及每种毒素的浓度， 心脏手术患者的血浆和尿液。同时，我们将确定 血红蛋白、血红素和铁结合和清除蛋白、触珠蛋白、血红素结合蛋白和转铁蛋白。 此外，我们将确定终末器官损伤的血浆和尿液标记物。我们将使用这些数据来构建 确定与终末器官损伤相关的Hb毒素浓度限值的生物动力学模型 标记。这项工作的目标是确定治疗干预的需要和时机。本 最后，根据我们丰富的经验，我们已经生物工程化了一种新的血红蛋白，血红素和蛋白质清除剂 研究Hb毒性。该新型蛋白质构建体通过在多步过程中产生apo-Hb来制备， 同时从科恩分级糊IV中分离触珠蛋白。最后，这两种蛋白质是 复合并进一步纯化以产生apo-Hb-触珠蛋白复合物。这种新的蛋白质结构 在高血红素暴露状态下结合血红素，将铁固定在血红素配体中，并安全地清除复合物， 单核细胞和巨噬细胞。或者，在高Hb暴露下，apo-Hb交换了 结合珠蛋白将Hb二聚体清除到单核细胞和巨噬细胞，而释放的apo-Hb二聚体被降解 通过蛋白水解无害地从循环中清除。为了测试我们的小说情结的效果， 计划在体外循环的动物模型中进行一系列概念验证研究，以确定apo的能力， Hb-触珠蛋白给药以防止终末器官损伤。