PTPRZ1靶向RNA CAR T细胞在胶质母细胞瘤中发挥抗原特异性和旁观者抗肿瘤活性
PTPRZ1-Targeting RNA CAR T Cells Exert Antigen-Specific and Bystander Antitumor Activity in Glioblastoma
影响因子:8.20000
分区:医学1区 Top / 免疫学2区 肿瘤学2区
发表日期:2024 Dec 03
作者:
Darel Martinez Bedoya, Eliana Marinari, Suzel Davanture, Luis Castillo Cantero, Sarah Erraiss, Millicent Dockerill, Sofia Barluenga, Nicolas Winssinger, Karl Schaller, Philippe Bijlenga, Shahan Momjian, Christel Voize, Stéphanie R Tissot, Lana E Kandalaft, Philippe Hammel, Pierre Cosson, Paul R Walker, Valérie Dutoit, Denis Migliorini
摘要
嵌合抗原受体（CAR）T细胞疗法在B细胞恶性肿瘤患者中的巨大成功促使其转化为实体瘤。在胶质母细胞瘤（GBM）的情况下，临床试验显示出适度的功效，但是开发更有效的抗GBM CAR T细胞的努力正在进行中。在这项研究中，我们选择了蛋白酪氨酸磷酸酶受体型Z型（PTPRZ1）作为GBM治疗的靶标。我们从人类噬菌体显示库中分离了六个抗人类PTPRZ1单链可变片段，并以RNA格式产生了第二代汽车T细胞。使用患者来源的GBM PTPRZ1-KNOCKIN细胞系来选择显示高细胞毒性的汽车构建体，同时始终显示出高CAR表达（471_28Z）。掺入471_28Z的CAR T细胞能够释放IFNγ，IL2，TNFα，Granzyme B，IL17A，IL6和可溶性FASL，并显示出低强调信号传导。此外，他们在体外杀死后保持了效应子记忆表型。此外，在PTPRZ1阳性肿瘤细胞中染色后，有471_28Z CAR T细胞在对PTPRZ1阴性细胞系中杀死强烈的旁观者，但未杀死抗原阴性的非肿瘤细胞。在使用NOD/SCIDγ小鼠的原位异种移植肿瘤模型中，单剂量的抗PTPRZ1 CAR T细胞显着延迟了肿瘤的生长。综上所述，这些结果将PTPRZ1验证为GBM靶标，并促使抗PTPRZ1 CAR T细胞的临床翻译。
Abstract
The great success of chimeric antigen receptor (CAR) T-cell therapy in the treatment of patients with B-cell malignancies has prompted its translation to solid tumors. In the case of glioblastoma (GBM), clinical trials have shown modest efficacy, but efforts to develop more effective anti-GBM CAR T cells are ongoing. In this study, we selected protein tyrosine phosphatase receptor type Z (PTPRZ1) as a target for GBM treatment. We isolated six anti-human PTPRZ1 single-chain variable fragments from a human phage display library and produced second-generation CAR T cells in an RNA format. Patient-derived GBM PTPRZ1-knockin cell lines were used to select the CAR construct that showed high cytotoxicity while consistently displaying high CAR expression (471_28z). CAR T cells incorporating 471_28z were able to release IFNγ, IL2, TNFα, granzyme B, IL17A, IL6, and soluble FasL and displayed low tonic signaling. Additionally, they maintained an effector memory phenotype after in vitro killing. In addition, 471_28z CAR T cells displayed strong bystander killing against PTPRZ1-negative cell lines after preactivation by PTPRZ1-positive tumor cells but did not kill antigen-negative nontumor cells. In an orthotopic xenograft tumor model using NOD/SCIDγ mice, a single dose of anti-PTPRZ1 CAR T cells significantly delayed tumor growth. Taken together, these results validate PTPRZ1 as a GBM target and prompt the clinical translation of anti-PTPRZ1 CAR T cells.