Engineering of an Avidity-Optimized CD19-Specific Parallel Chimeric Antigen Receptor That Delivers Dual CD28 and 4-1BB Co-Stimulation.
Engineering of an Avidity-Optimized CD19-Specific Parallel Chimeric Antigen Receptor That Delivers Dual CD28 and 4-1BB Co-Stimulation.
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DOI：
10.3389/fimmu.2022.836549
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发表时间：
2022
影响因子：
7.3
通讯作者：
Maher J
中科院分区：
文献类型：
--
作者：
Halim L;Das KK;Larcombe-Young D;Ajina A;Candelli A;Benjamin R;Dillon R;Davies DM;Maher J
共刺激对嵌合抗原受体（CAR）T细胞的功能至关重要。先前，我们证明了双重共刺激可以通过平行（p）CAR架构有效地利用，其中含有CD 28的第二代CAR与含有4-1BB的嵌合共刺激受体（CCR）共表达。当与线性汽车相比时，pCAR工程化的T细胞在一系列临床前模型中引发上级抗肿瘤活性。由于CD 19是细胞免疫治疗的最佳验证的临床靶标，因此我们在本研究中评估了一组CD 19特异性CAR和pCAR T细胞。首先，我们通过CD 19特异性FMC 63 scFv（VH结构域）的丙氨酸扫描诱变产生了一组单链抗体片段（scFv），并将这些片段掺入第二代CD 28 + CD 3汽车中。所得的CAR T细胞组证明了广泛的CD 19结合能力和对表达CD 19的肿瘤细胞的亲合力。然后通过共表达靶向FMC 63 scFv的CCR与4-1BB胞内结构域将每个scFv修饰的CAR转化为pCAR。当与含有未修饰或突变的FMC 63 scFv的第二代汽车相比时，每种pCAR在具有已建立的Nalm-6白血病异种移植物的小鼠中显示出肿瘤再刺激潜力和IL-2释放的显著增强、耗竭标志物表达的降低和治疗功效的增强。这些数据加强了pCAR平台通过有效提供双重共刺激来提供增强的抗肿瘤活性的证据。对于中等亲合力CAR T细胞和衍生的pCAR，注意到最大的抗肿瘤活性，提高了效应子对靶细胞亲合力是功效的重要决定因素的可能性。
Co-stimulation is critical to the function of chimeric antigen receptor (CAR) T-cells. Previously, we demonstrated that dual co-stimulation can be effectively harnessed by a parallel (p)CAR architecture in which a CD28-containing second generation CAR is co-expressed with a 4-1BB containing chimeric co-stimulatory receptor (CCR). When compared to linear CARs, pCAR-engineered T-cells elicit superior anti-tumor activity in a range of pre-clinical models. Since CD19 is the best validated clinical target for cellular immunotherapy, we evaluated a panel of CD19-specific CAR and pCAR T-cells in this study. First, we generated a panel of single chain antibody fragments (scFvs) by alanine scanning mutagenesis of the CD19-specific FMC63 scFv (VH domain) and these were incorporated into second generation CD28+CD3ζ CARs. The resulting panel of CAR T-cells demonstrated a broad range of CD19 binding ability and avidity for CD19-expressing tumor cells. Each scFv-modified CAR was then converted into a pCAR by co-expression of an FMC63 scFv-targeted CCR with a 4-1BB endodomain. When compared to second generation CARs that contained an unmodified or mutated FMC63 scFv, each pCAR demonstrated a significant enhancement of tumor re-stimulation potential and IL-2 release, reduced exhaustion marker expression and enhanced therapeutic efficacy in mice with established Nalm-6 leukemic xenografts. These data reinforce the evidence that the pCAR platform delivers enhanced anti-tumor activity through effective provision of dual co-stimulation. Greatest anti-tumor activity was noted for intermediate avidity CAR T-cells and derived pCARs, raising the possibility that effector to target cell avidity is an important determinant of efficacy.
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DOI：
10.1097/cji.0b013e3181ac6138
发表时间：
2009-09
期刊：
Journal of immunotherapy (Hagerstown, Md. : 1997)
影响因子：
--
作者：
Kochenderfer JN;Feldman SA;Zhao Y;Xu H;Black MA;Morgan RA;Wilson WH;Rosenberg SA
通讯作者：
Rosenberg SA
影响因子：
17.1
作者：
Katsarou, Afroditi;Sjostrand, Maria;Naik, Jyoti;Mansilla-Soto, Jorge;Kefala, Dionysia;Kladis, Georgios;Nianias, Alexandros;Ruiter, Ruud;Poels, Renee;Sarkar, Irene;Patankar, Yash R.;Merino, Elena;Reijmers, Rogier M.;Frerichs, Kristine A.;Yuan, Huipin;de Bruijn, Joost;Stroopinsky, Dina;Avigan, David;van de Donk, Niels W. C. J.;Zweegman, Sonja;Mutis, Tuna;Sadelain, Michel;Groen, Richard W. J.;Themeli, Maria
通讯作者：
Themeli, Maria
影响因子：
5.1
作者：
Chervin, A. S.;Stone, J. D.;Kranz, D. M.
通讯作者：
Kranz, D. M.
影响因子：
46.9
作者：
Maher, J;Brentjens, RJ;Sadelain, M
通讯作者：
Sadelain, M
影响因子：
4.6
作者：
Park S;Shevlin E;Vedvyas Y;Zaman M;Park S;Hsu YS;Min IM;Jin MM
通讯作者：
Jin MM