Stimuli-responsive reagents for improved cell isolations and activations

用于改善细胞分离和激活的刺激响应试剂

• 批准号: 9045281
• 负责人: Barrett J Nehilla
• 金额: $ 23.21万
• 依托单位: NEXGENIA, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9045281
• 关键词: Acute; Address; Adopted; Adoptive Immunotherapy; Affinity; Amines; Antibodies; Architecture; Area; Autologous; Behavior; Benchmarking; Binding; Blood; CD28 gene; CD3 Antigens; Caliber; Cancer Patient; Cell Separation; Cell Survival; Cell Therapy; Cell surface; Cells; Chronic Lymphocytic Leukemia; Clinical; Clinical Treatment; Clinical Trials; Complex; Development; Engineering; Enzyme-Linked Immunosorbent Assay; Excision; Flow Cytometry; Foundations; Generations; Genetic Engineering; Goals; High temperature of physical object; Human; In Vitro; Institution; Lead; Magnetic nanoparticles; Magnetism; Malignant Neoplasms; Measurement; Measures; Mediating; Methods; Microspheres; Nature; Particle Size; Patients; Performance; Peripheral Blood Mononuclear Cell; Phase; Polymers; Process; Production; Property; Reagent; Receptor Aggregation; Response to stimulus physiology; Running; Signal Transduction; Small Business Innovation Research Grant; Specificity; Stimulus; Structure; Surface; Surface Antigens; System; T cell therapy; T-Cell Activation; T-Lymphocyte; Technology; Temperature; Therapeutic; Ursidae Family; antibody conjugate; base; cancer clinical trial; chimeric antigen receptor; cold temperature; commercialization; cost; cost effective; crosslink; cytokine; design; extracellular; improved; magnetic beads; manufacturing process; nano; nanoparticle; novel; novel therapeutics; particle; public health relevance; receptor; response; success; tumor

 DESCRIPTION (provided by applicant): Chimeric antigen receptor (CAR) T cell adoptive immunotherapy has shown great promise in clinical trials for the treatment of cancers, including acute lymphoblastic and chronic lymphocytic leukemia. CAR T cell therapies are comprised of the patients own T cells (autologous T cells) that have been genetically engineered to express a CAR. This receptor is comprised of an extracellular tumor-targeted binding moiety fused to intracellular domains with activation and signaling functionalities. Unfortunately, CART therapy has a complex, multi-step manufacturing process, and no standardized approach has been implemented among various institutions running the clinical trials. An efficient, cost-effective an standardized manufacturing process is critical to the broad application and ultimate success of CART therapy. The goal of this project is to develop stimuli-responsive reagent systems (SRRS) for cell isolations and activations that will help to streamline and improve the manufacturing process of adoptive cellular therapies like CAR T cell therapy. Magnetic beads are used to isolate and activate T cells in CAR T cell manufacturing. These beads have surface-bound antibodies, and they are either micro- (Dynal(r)) or nano- (Miltenyi Biotec) sized. Neither technology has been adopted universally in CAR T cell manufacturing processes because each approach has shortcomings. Nexgenia's magnetic nanoparticle (mNP) technology is designed to combine the favorable attributes of microbeads (rapid magnetic separation) and nanoparticles (rapid target binding), with the additional benefit of simple nanoparticle removal after cell manipulations. The key to achieving this ideal combination is the unique stimuli-responsive nature of the Nexgenia mNP. These mNP bear stimuli-responsive polymers, and they change from hydrophilic, monodispersed ~20 nm diameter particles to micron-sized aggregates in response to an environmental stimulus like a temperature change. Nexgenia's SRRS comprise mNP and antibodies conjugated to similar stimuli-responsive polymers. At low temperatures, polymer-antibody conjugates rapidly bind cell surface antigens. At higher temperatures, the polymer-antibody conjugates aggregate with the mNP, facilitating rapid magnetic isolation of target cells or cell activation. To achieve the project's goal, Nexgenia will develop a SRSS with a temperature response of ~15°C. The SRRS will be used to demonstrate the feasibility of isolating T cells from human peripheral blood mononuclear cells and its performance will be benchmarked against Dynal(r) CD3/28 beads. The SRRS will then be used to activate T cells, thereby mediating their expansion in vitro. A SRRS that isolates and then activates T cells can be readily integrated into cell therapy manufacturing processes and lead to more consistent and better- characterized therapeutics. An improved CAR T cell manufacturing process would help to realize the clinical and commercial success of adoptive cell therapies and ultimately, expand patient access to these promising therapeutics. Nexgenia's SRRS can address this area of great need.

 描述（由申请人提供）：嵌合抗原受体（CAR）T细胞过继免疫疗法在治疗癌症的临床试验中显示出巨大的前景，包括由患者自身T细胞疗法组成的急性淋巴细胞和慢性淋巴细胞白血病。经过基因工程改造以表达 CAR 的细胞（自体 T 细胞）。该受体由与具有激活和信号传导功能的细胞内结构域融合的细胞外肿瘤靶向结合部分组成。不幸的是，CART疗法具有复杂的、多步骤的制造过程，并且在运行临床试验的各个机构之间尚未实施标准化方法，高效、具有成本效益的标准化制造过程对于CART的广泛应用和最终成功至关重要。该项目的目标是开发用于细胞分离和激活的刺激响应试剂系统（SRRS），这将有助于简化和改进过继性细胞疗法（如用于分离和激活 CAR T 细胞疗法）的制造过程。激活T细胞CAR T 细胞制造。这些珠子具有表面结合的抗体，并且它们是微米 (Dynal(r)) 或纳米 (Miltenyi Biotec) 尺寸的，这两种技术都没有在 CAR T 细胞制造过程中得到普遍采用，因为每种方法都有各自的特点。 Nexgenia 的磁性纳米颗粒 (mNP) 技术旨在结合微珠（快速磁分离）和纳米颗粒（快速目标结合）的有利属性，并具有简单的额外优点。实现这种理想组合的关键是 Nexgenia mNP 具有刺激响应聚合物，它们从亲水性、单分散的约 20 nm 直径颗粒转变为微米级聚集体。 Nexgenia 的 SRRS 包含与类似刺激响应聚合物缀合的 mNP 和抗体。在较高温度下，聚合物-抗体偶联物会与细胞表面抗原快速结合，从而促进靶细胞的快速磁分离或细胞激活。为了实现该项目的目标，Nexgenia 将开发具有SRRS 的温度响应约为 15°C，将用于证明从人外周血单核细胞中分离 T 细胞的可行性，其性能将以 Dynal(r) CD3/28 为基准。然后，SRRS 将用于激活 T 细胞，从而在体外介导其扩增。分离并激活 T 细胞的 SRRS 可以轻松整合到细胞治疗制造过程中，从而产生更一致、更具有特征的治疗方法。改进的 CAR T 细胞制造工艺将有助于实现过继细胞疗法的临床和商业成功，并最终扩大患者获得这些有前途的疗法的机会，可以解决这一领域的巨大需求。