PTPRZ1靶向RNA CAR T细胞在胶质母细胞瘤中表现出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

Cancer Immunology Research

影响因子:8.2

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

DOI: 10.1158/2326-6066.CIR-23-1094

摘要

嵌合抗原受体（CAR）T细胞疗法在治疗B细胞恶性肿瘤方面取得巨大成功，推动其向实体瘤的转化。在胶质母细胞瘤（GBM）中，临床试验显示效果有限，但开发更有效的抗GBM CAR T细胞的努力仍在进行中。本研究选择蛋白酪氨酸磷酸酶受体Z型（PTPRZ1）作为GBM的治疗靶点。我们从人源噬菌体展示文库中分离出六个抗人PTPRZ1的单链可变片段（scFv），并制备了第二代RNA型CAR T细胞。利用患者来源的PTPRZ1敲入的GBM细胞系筛选出具有高细胞毒性且持续表达高水平CAR的构建（471_28z）。含有471_28z的CAR T细胞能够释放IFNγ、IL2、TNFα、颗粒酶B、IL17A、IL6及可溶性FasL，表现出低的张力信号。它们在体外杀伤后仍保持效应记忆表型。此外，471_28z CAR T细胞在预激活的肿瘤细胞中对PTPRZ1阴性细胞系表现出强大的旁观者杀伤作用，但不会杀伤抗原阴性非肿瘤细胞。在NOD/SCIDγ小鼠的正统异种移植模型中，单次注射抗PTPRZ1 CAR T细胞显著延缓肿瘤生长。综上所述，这些结果验证了PTPRZ1作为GBM治疗靶点的有效性，并推动抗PTPRZ1 CAR T细胞的临床转化。

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.