I-Corps: Developing polymerized hemoglobin using enzymatic ligation

这个i-Corps项目的更广泛的影响/商业潜力是开发用于兽医和人类应用的血液替代品。由于战争、自然灾害和流行病/地方病等全球性灾难，献血需求正在上升。此外，军方可能对野战准备好的氧气(O2)载体感兴趣，这种载体可以为野战中可能导致严重失血的失血性休克病例提供一种替代输血的方案。目前从创伤和无血的情况下复苏的临床标准是使用晶体，如乳酸林格氏溶液。晶体材料随处可得，安全且价格低廉。这些解决方案用于恢复血容量和血压，允许血液流动继续，但没有氧气携带能力，对血液流动和氧气输送有不同程度的负面影响。理想的复苏液需要能够维持血容量和血压，而不过度稀释携氧能力和血浆蛋白，或降低血液粘度。这项拟议的技术为血液替代品提供了一种替代方法，称为“O2桥”，或基于血红蛋白的氧气载体(HBOC)。HBOC有可能被改造成复制甚至优化循环血液的关键生物物理参数，从而能够生产维持生命的产品。这项拟议的技术旨在将氧气有效地输送到组织中，与全血或红细胞一样有效。此外，这种HBOC溶液可以执行这一功能，而不会有疾病传播、输血反应或其他与输血相关的并发症的风险。这个I-Corps项目是基于开发一种基于血红蛋白的氧气载体，该载体是从过期的捐赠人体血液中生产的。拟议的技术使用酶连接(转谷氨酰胺酶)来生产大尺寸聚合的人血红蛋白(PolyhHb)。在这个过程中，红血球被从膜上剥离，留下血红蛋白被提纯并加工成具有功能的生物相容溶液。大的、基于聚合血红蛋白的氧气载体(HBOC)溶液具有循环半衰期较长的优势，这表明全身和网状内皮铁转运的标志物存在差异。这项技术已经在小动物身上进行了测试，并在贫血条件下显示了有希望的结果。此外，它还与其他HBOC设计和其他血液产品进行了测试。建议的技术可用于创建在无法控制的出血环境中具有实际用途的功能性HBOC解决方案。该奖项反映了NSF的法定使命，并已通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this I-Corps project is the development of a blood substitute for use in veterinary and human applications. The demand for donated blood is rising due to global catastrophes such as war, natural disasters, and pandemics/endemics. Additionally, the military may have interest in field-ready oxygen (O2) carriers that can provide an alternative solution to impractical blood transfusions for cases of hemorrhagic shock in the field that may lead to severe blood loss. The current clinical standard for resuscitation from trauma and the absence of blood is the use of crystalloids, such as lactated Ringer’s solution. Crystalloids are widely available, safe, and inexpensive. These solutions are used to restore blood volume and blood pressure, allowing blood flow to continue, but do not have (O2)-carrying capacity and have varying negative effects on blood flow and O2 delivery. The ideal resuscitation fluid needs to be able to maintain blood volume and blood pressure, without excessively diluting O2-carrying capacity, and plasma proteins, or reducing blood viscosity. The proposed technology offers an alternative approach to blood substitutes, termed “O2 bridges”, or hemoglobin-based oxygen carrier (HBOC). HBOCs have the potential to be engineered to replicate, or even optimize the critical biophysical parameters of circulating blood, enabling the production of a life-sustaining product. The proposed technology is designed to transport O2 to tissues as effectively as whole blood or packed red blood cells. In addition, this HBOC solution may perform this function without the risk of disease transmission, transfusion reactions, or other transfusion-related complications.This I-Corps project is based on the development of a hemoglobin-based oxygen carrier that is produced from expired donated human blood. The proposed technology uses enzymatic ligation (transglutaminase) to produce large-size polymerized human hemoglobin (PolyhHb). In this process, red blood cells are stripped from their membrane leaving the hemoglobin protein to be purified and processed into a functional biocompatible solution. Large, polymerized hemoglobin-based oxygen carrier (HBOC) solutions have the advantage of longer circulatory half-life indicative of the difference in markers of systemic and reticuloendothelial iron transport. This technology has been tested in small animals and has shown promising results in anemic conditions. In addition, it has been tested against other HBOC designs and other blood products. The proposed technology may be used to create a functional HBOC solution that has practical use in uncontrollable hemorrhage environments.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.