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 医疗辅助设备后，CNS，肺，心脏和肾脏并发症的成本估算值 需要血液泵，例如，在泵的心肺旁路（CPB）上，估计为8000万美元 十年中，美国的个别州。 复杂的手术，溶血以及随后的血红蛋白（HB）释放到循环中 - 已知是多器官有毒和血管活性 - 是对更密集管理的相关贡献者 患者 仅通过肾脏伤害，每年约为90亿美元，每年30万人死亡和平均 住院时间增加3.5天/患者该项目的重点是了解 溶血，HB和血红素与次级末端器官损伤相关 心肺旁路（通常持续4小时）。 在这种情况下的裂解，重点是基础理解 HB和血红素以及这些毒素的最终器官病理分析的治疗策略 前瞻性确定HB的水平 心脏手术患者的血浆和尿液。 HB，血红素和铁结合和清除蛋白，触觉球蛋白，血红素和转移蛋白分别。 此外，我们将使用此数据来构建最终器官损伤的等离子体和泌尿率标记。 一个确定Hb毒素浓度的限制的生物动力学模型，该模型与最终器官损伤相关 标记 最后，我们根据我们的丰富经验，为生物工程设计了一种新颖的HB，血红素和蛋白质清道夫 通过研究HB毒素。 虽然同时隔离haptoglobin与Cohn分级糊剂IV。 复合并进一步纯化以产生Apo-Hb-Haptoglobin复合物。 在高血红素暴露状态下结合血红素，血红素配体中的安全铁，并安全地清除复合物 单核细胞和巨噬细胞。 将HB二聚体清除至单核细胞和巨噬细胞，而释放的APO-HB二聚体降解 通过蛋白质和无害的循环清除。 计划在泵CPB的动物模型中进行一系列概念验证研究，以定义apo-的能力 HB-热蛋白的剂量以防止最终器官损伤。