Targeting novel biotherapeutics to endothelium

针对内皮的新型生物治疗药物

• 批准号: 9096887
• 负责人: Vladimir R Muzykantov
• 金额: $ 54.96万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9096887
• 关键词: Acute Lung Injury; Adverse effects; Affect; Alveolar; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Apoptotic; Binding; Biological Products; Biological Response Modifier Therapy; Biology; Blood; Blood Vessels; CD31 Antigens; Cell Line; Cell membrane; Clinical; Disease Progression; Drug Delivery Systems; Edema; Effectiveness; Elements; Endothelial Cells; Endothelium; Endotoxins; Engineering; Epitopes; Extravasation; Genetic; Glycoproteins; Goals; Grant; Health; Hemorrhage; Homeostasis; Human; Inflammation; Inflammatory Response; Infusion procedures; Injury; Intervention; Lead; Lung; Measures; Mediating; Mediator of activation protein; Medical; Modeling; Molecular; Monoclonal Antibodies; Mus; Outcome; Oxidants; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Physiological; Pilot Projects; Plasma; Problem Solving; Production; Prophylactic treatment; Protein C; Pulmonary Inflammation; Recombinants; Regulation; Resistance; Role; Safety; Sepsis; Syndrome; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Thrombin; Thrombomodulin; Translations; Trauma; Vascular Endothelial Cell; activated Protein C; activated protein C receptor; base; clinical efficacy; cytokine; design; drug testing; effective therapy; improved; improved outcome; mortality; mouse model; mutant; novel; pre-clinical; prevent; prophylactic; prospective; protective effect; risk benefit ratio; sound; targeted treatment; vascular inflammation

 DESCRIPTION: The patho-physiological hallmark of acute lung injury (ALI) is severe derangement the interface of blood with the endothelial vascular lining. Endothelial cells (ECs) exert protective mechanisms including those mediated by thrombomodulin (TM), which inhibits thrombin's pathological activities in part through the production of activated protein C (APC) from its plasma precursor, via interactions between TM, thrombin, protein C, APC and the endothelial protein C receptor (EPCR) which is located in close proximity to TM in the endothelial plasmalemma. In ALI, TM and EPCR are functionally suppressed. Replenishment of this endothelial mechanism is an important therapeutic goal, but infusion of recombinant APC and TM did not improve the outcome of ALI, because problematic delivery and inability to partner with natural regulatory molecules limited effectiveness and lead to adverse effects. To solve these problems we fused TM to a single-chain variable fragment (scFv) of antibodies to PECAM, obtaining scFv/TM that provides protection superior to untargeted TM and APC in animal models of ALI. scFv/TM anchoring on endothelial cells in proximity to EPCR boosts protection. To circumvent the loss of endogenous EPCR typical of ALI, we use dual delivery of scFv/TM and scFv/EPCR, based on our recent serendipitous discovery of "collaborative enhancement" of endothelial binding of paired monoclonal antibodies directed to distinct PECAM epitopes. Here we will study the delivery, mechanisms and benefit/risk ratio of these fusions and embark on their clinical translation: Aim 1: Dual scFv/TM and scFv/EPCR targeting to endothelium. We will employ a combination of genetic and pharmacologic approaches to selectively modulate molecular regulation of the fusions in order to quantify their effects on ALI. Aim 2: Pre-clinical efficacy and safety in animals. Beneficial and adverse effects of anti-PECAM scFv/TM and scFv/EPCR will be characterized (vascular leakage, alveolar inflammatory response, morphologic and physiologic outcomes). Aim 3: Endothelial targeting scFv/TM in human lungs. To enable the clinical translation, we shall: A) Re-engineer the fusions using molecular elements applicable in humans; and, B) Study their targeting in isolated human lungs.

 描述：急性肺损伤（ALI）的病情 - 生理标志是严重进化的血液与内皮血管内膜的界面。 Endothelial cells (ECs) exercise protected mechanisms including those mediated by thrombomodulin (TM), which inhibits thrombin's pathological activities in part through the production of activated protein C (APC) from its plasma precursor, via interactions between TM, thrombin, protein C, APC and The endothelial protein C receptor (EPCR) which is located in close proximity to TM in the endothelial Plainsmalemma。在ALI中，TM和EPCR在功能上被抑制。补充这种内皮机制是一个重要的治疗目标，但是注入重组APC和TM并不能改善ALI的结果，因为有问题的分娩和无法与自然调节分子合作有限的有限效率并导致不良影响。为了解决这些问题，我们将TM与PECAM的抗体的单链可变片段（SCFV）融合在一起，获得了SCFV/TM，在ALI动物模型中，可提供优于未靶向的TM和APC的保护。 SCFV/TM锚定在靠近EPCR助推器保护的内皮细胞上。为了绕过ALI典型的内源性EPCR的损失，我们基于我们最近的偶然发现，使用SCFV/TM和SCFV/EPCR的双重递送，我们最近偶然发现了针对独特的PECAM占据的成对单克隆抗体的内皮结合的“协作增强”。在这里，我们将研究这些融合的输送，机制和益处/风险比，并开始进行临床翻译：AIM 1：双SCFV/TM和SCFV/EPCR靶向内皮。我们将采用遗传和药物方法的组合来选择性调节融合的分子调节，以量化其对ALI的影响。 目标2：动物的临床前效率和安全性。抗PECAM SCFV/TM和SCFV/EPCR的有益和不利影响将被表征（血管泄漏，肺泡炎症反应，形态和生理结果）。 AIM 3：人肺中的内皮靶向SCFV/TM。为了实现临床翻译，我们将：a）使用适用于人类的分子元素重新设计融合；并且，b）在孤立的人肺中研究其靶向。