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（由应用提供）：嵌合抗原受体（CAR）T细胞产物免疫疗法在临床试验中对癌症的治疗（包括急性淋巴细胞和慢性淋巴细胞性白血病）显示出了很大的希望。 CAR T细胞疗法已完成，该患者自己的T细胞（自体T细胞）经过基因设计以表达汽车。该受体是通过激活和信号传导功能融合到细胞内结构域的细胞外肿瘤结合部分完成的。不幸的是，购物车治疗具有复杂的多步骤制造过程，并且在运行临床试验的各个机构中没有实施标准化方法。标准化的制造过程有效，具有成本效益对于购物车治疗的广泛应用和最终成功至关重要。该项目的目的是开发刺激性反应性试剂系统（SRR），用于细胞分离和激活，这将有助于简化和改善自适应细胞疗法（例如CAR T细胞疗法）的制造过程。磁珠用于分离和激活CAR T细胞生产中的T细胞。这些珠具有表面结合的抗体，它们是微型（Dynal（r））或纳米（Miltenyi Biotec）大小的。由于每种方法都有缺点，因此在汽车T细胞制造过程中都普遍采用了技术。 Nexgenia的磁性纳米颗粒（MNP）技术旨在结合Microbead（快速磁分离）和纳米颗粒（快速目标结合）的有利属性，并在细胞操作后简单纳米颗粒去除的额外好处。实现这种理想组合的关键是Nexgenia MNP的独特刺激性响应性。这些MNP熊刺激反应性聚合物，它们从亲水性，单分离〜20 nm的直径颗粒变为微米大小的聚集体，以响应环境刺激，例如温度变化。 Nexgenia的SRR包括MNP和与类似的刺激性反应性聚合物结合的抗体。在低温下，聚合物 - 抗体结合迅速结合细胞表面抗原。在较高的温度下，聚合物 - 抗体结合与MNP聚集，从而支持快速磁性。靶细胞或细胞激活的分离。为了实现该项目的目标，Nexgenia将开发〜15°C的SRSS。 SRR将用于证明从人外周血单核细胞中分离出T细胞的可行性，其性能将以针对Dynal（R）CD3/28珠子为基准。然后，SRR将用于激活T细胞，从而在体外介导其膨胀。分离然后激活T细胞的SRR可以容易地整合到细胞疗法制造过程中，并导致更一致，更具特征的治疗。改进的汽车T细胞制造过程将有助于实现适应性细胞疗法的临床和商业成功，并最终扩大患者接受这些承诺疗法的机会。 Nexgenia的SRR可以解决这一巨大需求。