IPSC-derived anti-KIT CAR-T cell generation: functional optimization and in vivo application in a transplantable mouse gastrointestinal stromal tumor (GIST) model

IPSC衍生的抗KIT CAR-T细胞生成：功能优化和在可移植小鼠胃肠道间质瘤（GIST）模型中的体内应用

• 批准号: 441925459
• 负责人: Dr. Michael Rassner
• 金额: --
• 依托单位: Center for iPS Cell Research and Application (CiRA)
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/441925459?language=en
• 关键词: IPSC; derived; anti; KIT; CAR

Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the gastrointestinal tract. 85-90% of GIST patients harbor activating mutations in cKIT or PDGFRA, allowing the application of tyrosine kinase inhibitors (TKI). In patients with primary or secondary TKI resistance, therapeutic options are limited. In vitro and in vivo mice data as well as preliminary patient observations imply that immunotherapeutic approaches might be successful in GIST patients, including adoptive T cell transfer using chimeric antigen receptors (CAR-T cells). However, CAR-T cell therapies are often restricted by low numbers of generated T cells, which frequently present with an unfavorable phenotype. This might be circumvented by generating CAR-T cells from inducible pluripotent stem cells (iPSCs). In previous studies, CAR-T cells originating from iPSCs possessed a phenotype resembling -T cells with diminished antitumoral activity. The aim of the present research proposal is to improve antitumoral functionality by generating CAR-T cells targeting cKIT from iPSC. The generated anti-KIT CAR-T-iPSCs will be investigated for their phenotype and metabolism, as it has been demonstrated that CAR-T cell activity strongly depends on these profiles. By variations of culture conditions, cytokines, drugs or genetic/epigenetic modifications, functionally optimized CAR-T-iPSCs will be generated. To prove their in vitro and in vivo antitumoral activity, these cells will be tested in GIST cell co-cultures and a mouse xenograft GIST model. Finally, the anti-KIT CAR-T iPSCs will be further improved by targeted knockout of HLA-antigens and the T cell receptor to reduce allorejection and graft-versus-host-disease.

胃肠道间质瘤是胃肠道最常见的间叶性肿瘤。85-90%的GIST患者在cKIT或PDGFRA中携带激活突变，允许应用酪氨酸激酶抑制剂（TKI）。在原发性或继发性TKI耐药患者中，治疗选择有限。体外和体内小鼠数据以及初步患者观察结果表明，免疫治疗方法可能在GIST患者中取得成功，包括使用嵌合抗原受体（CAR-T细胞）的过继性T细胞转移。然而，CAR-T细胞疗法通常受到生成的T细胞数量少的限制，这些T细胞通常具有不利的表型。这可以通过从诱导性多能干细胞（iPSC）中产生CAR-T细胞来规避。在先前的研究中，源自iPSC的CAR-T细胞具有类似于具有减弱的抗肿瘤活性的CAR-T细胞的表型。本研究提案的目的是通过从iPSC产生靶向cKIT的CAR-T细胞来改善抗肿瘤功能。将研究产生的抗KIT CAR-T-iPSC的表型和代谢，因为已经证明CAR-T细胞活性强烈依赖于这些特征。通过改变培养条件、细胞因子、药物或遗传/表观遗传修饰，将产生功能优化的CAR-T-iPSC。 为了证明其体外和体内抗肿瘤活性，将在GIST细胞共培养物和小鼠异种移植GIST模型中测试这些细胞。最后，抗KIT CAR-T iPSC将通过靶向敲除HLA抗原和T细胞受体来进一步改善，以减少同种异体排斥反应和移植物抗宿主病。