Development of chimeric antigen receptor-expressing macrophages for enhanced phagocytosis of systemic amyloid

开发表达嵌合抗原受体的巨噬细胞以增强系统性淀粉样蛋白的吞噬作用

• 批准号: 10263880
• 负责人: JONATHAN S WALL
• 金额: $ 17.5万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10263880
• 关键词: Alzheimer' s Disease; Amyloid; Amyloid Fibrils; Amyloidosis; Animals; Antibody Response; B-Lymphocytes; Binding; CD19 gene; CD47 gene; Cardiac; Cell Therapy; Cell secretion; Cells; Clinic; Complement; Cytoplasmic Tail; Data; Deposition; Development; Disease; Dyes; Eating; Elements; Engineering; Evaluation; Exhibits; Extracellular Domain; Fluorescence; Functional disorder; Goals; Heart; Human; Immune System Diseases; Immune system; Immunotherapy; In Vitro; Kidney; Label; Laboratories; Light; Light-Chain Immunoglobulins; Liver; MHC Class I Genes; Mediating; Modeling; Monitor; Monoclonal Antibodies; Mus; Organ; Pathogenesis; Pathology; Patients; Peptide antibodies; Peptides; Peripheral Nerves; Phagocytes; Phagocytosis; Phagolysosome; Plasma Cells; Prognosis; Proteins; Quality of life; Research; Signal Transduction; Spleen; Structure; Survival Rate; Systemic disease; Technology; Therapeutic; Tissues; Translating; Transmembrane Domain; base; chemotherapy; chimeric antigen receptor; exhaustion; extracellular; fluorophore; improved; macrophage; neoplastic cell; novel; novel strategies; novel therapeutics; optical imaging; phagocytosis receptor; receptor binding; research clinical testing; uptake

Amyloidosis is a devastating pathology that is associated not only with the development of Alzheimer's disease, but also with the lesser known, but similarly devastating, disorder immunoglobulin light chain- associated (AL) amyloidosis. Patients with AL develop amyloid in abdominothoracic organs and peripheral nerves leading to organ dysfunction which is often fatal. The amyloid deposits in systemic diseases are immunologically inert – they are not recognized or cleared by phagocytic cells of the immune system (macrophages [Mφ]) and do not illicit an antibody response. In patients presenting with significant cardiac amyloidosis, the prognosis is poor with a median survival of ~9 mos. Treatment of AL amyloidosis generally involves anti-plasma cell chemotherapy and immunotherapy to suppress plasma cell secretion of the amyloid-forming light chain protein. However, clearance of tissue amyloid has now become a major goal of many of the novel therapeutics being developed for these patients. Consequently, amyloid-reactive monoclonal antibodies have been developed that can be used to opsonize deposits and induce clearance by Mφ. Despite disappointing results from late stage clinical evaluation of certain of these mAbs, these trials demonstrated that clearance of amyloid and improvement in organ function can be achieved. The goal of this exploratory proposal is to generate two chimeric antigen receptor (CAR) constructs, expressed in macrophages, to induce specific recognition and enhanced phagocytosis of AL amyloid. To facilitate amyloid binding, the CARs will incorporate either the scFv domain of the amyloid reactive 11-1F4 mAb or the multi-amyloid binding peptide p5+14. The cytoplasmic domain of the CAR, from the murine FcR, will be incorporated to signal phagocytosis of the amyloid following receptor binding. Phagocytosis of synthetic AL amyloid fibrils and patient-derived AL amyloid extracts (labeled with the pH-sensitive dye pHrodo red) by CAR-phagocytic (P) Mφ will be quantified in vitro by monitoring the increased fluorescence emission of the fluorophore as the labeled amyloid enters the acidified phagolysosome. Additionally we will use small animal optical imaging to study co-localization with, and phagocytosis of, fluorophore-labeled human amyloid by CAR- P Mφ in mice. Quantitatively significant improvement in phagocytosis of AL amyloid due to the presence of the CAR will be assessed by comparison with CAR-negative Mφ and direct opsonization of amyloid using 11-1F4. The amyloid-directed CAR-P Mφ is a novel approach to the treatment of AL and related systemic amyloidoses. The long term goal of this study is to provide support for an engineered, cell-based amyloid clearance paradigm for the treatment of amyloidosis. In light of positive data from these studies, we hope that the amyloid-directed CAR-P technology can be optimized for efficacy and utility, and translated to the clinic to provide meaningful benefit to patients by enhancing quality of life and prolonging patient survival.

淀粉样变性是一种毁灭性的病理学，不仅与阿尔茨海默氏症的发展有关 疾病，但还具有鲜为人知但具有灾难性的，无序的免疫球蛋白轻链 相关（Al）淀粉样变性。在腹部胸部器官和周围内发育淀粉样蛋白的患者 导致器官功能障碍的神经通常是致命的。全身性疾病中的淀粉样蛋白沉积物是 免疫学惰性 - 没有被免疫系统的吞噬细胞识别或清除 （巨噬细胞[Mφ]），并且不会非法抗体反应。在出现明显心脏的患者中 淀粉样变性，预后较差，中位存活率约为9个月。 Al淀粉样变性的治疗通常涉及抗血浆细胞化疗和免疫疗法 抑制淀粉样蛋白的光链蛋白的血浆细胞分泌。但是，组织清除率 现在，淀粉样蛋白已成为为这些患者开发的许多新型疗法的主要目标。 因此，已经开发了可用于调理的淀粉样蛋白反应性单克隆抗体 沉积并通过mφ诱导清除。尽管对后期临床评估的结果令人失望 这些mab中的某些mab，这些试验表明淀粉样蛋白的清除和器官功能的改善 可以实现。 该探索性建议的目的是生成两个嵌合抗原受体（CAR）构建体，即 在巨噬细胞中表达，以诱导特异性识别和增强的Al淀粉样蛋白吞噬作用。到 促进淀粉样蛋白结合，这些汽车将结合淀粉样蛋白反应性11-1f4的SCFV结构域 mAb或多淀粉样蛋白结合肽P5+14。汽车的细胞质结构域，来自鼠FCR， 将纳入受体结合后淀粉样蛋白的吞噬作用。合成的吞噬作用 Al淀粉样蛋白原纤维和患者来源的AL淀粉样蛋白提取物（用pH敏感的染料Phrodo Red标记） 通过监测增加 标记淀粉样蛋白的荧光团进入认证的吞噬体。另外，我们将使用小动物 光学成像研究与荧光团标记的人淀粉样蛋白的共定位和吞噬作用 小鼠的pmφ。由于存在，由于存在，Al淀粉样蛋白的吞噬作用有所改善 使用11-1F4的CAR-CORNGATICMφ和淀粉样蛋白的直接调整将评估CAR。 淀粉样蛋白导向的CAR-PMφ是一种治疗Al和相关全身治疗的新方法 淀粉样蛋白。这项研究的长期目标是为基于细胞的淀粉样蛋白提供支持 清除范式治疗淀粉样变性。鉴于这些研究的积极数据，我们希望 淀粉样蛋白导向的CAR-P技术可以优化以提高效率和实用性，并将其转化为诊所 通过提高生活质量并延长患者的生存，为患者提供有意义的好处。