Development of GPC2-directed chimeric antigen receptors using mRNA for pediatric brain tumors.
Development of GPC2-directed chimeric antigen receptors using mRNA for pediatric brain tumors.
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DOI：
10.1136/jitc-2021-004450
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发表时间：
2022-09
影响因子：
10.9
通讯作者：
Bosse, Kristopher R.
中科院分区：
文献类型：
--
作者：
Foster, Jessica B.;Griffin, Crystal;Rokita, Jo Lynne;Stern, Allison;Brimley, Cameron;Rathi, Komal;Lane, Maria, V;Buongervino, Samantha N.;Smith, Tiffany;Madsen, Peter J.;Martinez, Daniel;Delaidelli, Alberto;Sorensen, Poul H.;Wechsler-Reya, Robert J.;Kariko, Katalin;Storm, Phillip B.;Barrett, David M.;Resnick, Adam C.;Maris, John M.;Bosse, Kristopher R.
儿童脑瘤是儿童癌症死亡的主要原因，迫切需要创新的治疗方法。Glypican2(GPC2)是一种在神经母细胞瘤中表达的细胞表面癌蛋白，针对神经母细胞瘤已经开发了靶向免疫治疗。这项工作旨在表征GPC2在儿童脑肿瘤中的表达，并建立一种针对这一靶点的mRNA CAR T细胞方法。我们用RNA测序、免疫组织化学和流式细胞术研究了GPC2在一组儿童原发脑瘤样本和细胞系中的表达。为了通过过继细胞疗法靶向大脑中的GPC2并减轻潜在的炎性神经毒性，我们使用优化的mRNA来创建瞬时嵌合抗原受体(CAR)T细胞。我们使用高度GPC2特异性的全人D3单链可变片段开发了四个mRNACAR T细胞结构，用于临床前测试。我们发现GPC2在多种儿童脑肿瘤类型中都有高表达，包括髓母细胞瘤、多层玫瑰花环的胚胎性肿瘤、其他中枢神经系统胚胎性肿瘤，以及可明确的高度恶性胶质瘤亚型。接下来，我们对表达GPC2的神经母细胞瘤细胞进行了体外细胞毒性试验，对CAR配置进行了验证和优先排序，其中轻到重的单链可变片段配置被证明是优越的。我们将两个最有效的GPC2导向CAR构建物的测试扩展到表达GPC2的髓母细胞瘤和高级别胶质瘤细胞系，在多种模型中显示了显著的GPC2特异性细胞死亡。最后，在原位髓母细胞瘤模型中，每两周局部区域注射2-400万 百万GPC2导向的mRNACAR T细胞可显著诱导肿瘤消退，并显著延长侵袭性原位丘脑弥漫性中线胶质瘤异种移植模型的存活期。未观察到GPC2介导的CAR T细胞相关的神经或全身毒性。综上所述，这些数据表明，GPC2是一种在多种儿童恶性脑肿瘤中高度差异表达的细胞表面蛋白，可以通过局部传递mRNACAR T细胞来安全地靶向，为GPC2指导的儿童脑肿瘤免疫治疗的临床翻译奠定了基础。
Pediatric brain tumors are the leading cause of cancer death in children with an urgent need for innovative therapies. Glypican 2 (GPC2) is a cell surface oncoprotein expressed in neuroblastoma for which targeted immunotherapies have been developed. This work aimed to characterize GPC2 expression in pediatric brain tumors and develop an mRNA CAR T cell approach against this target. We investigated GPC2 expression across a cohort of primary pediatric brain tumor samples and cell lines using RNA sequencing, immunohistochemistry, and flow cytometry. To target GPC2 in the brain with adoptive cellular therapies and mitigate potential inflammatory neurotoxicity, we used optimized mRNA to create transient chimeric antigen receptor (CAR) T cells. We developed four mRNA CAR T cell constructs using the highly GPC2-specific fully human D3 single chain variable fragment for preclinical testing. We identified high GPC2 expression across multiple pediatric brain tumor types including medulloblastomas, embryonal tumors with multilayered rosettes, other central nervous system embryonal tumors, as well as definable subsets of highly malignant gliomas. We next validated and prioritized CAR configurations using in vitro cytotoxicity assays with GPC2-expressing neuroblastoma cells, where the light-to-heavy single chain variable fragment configurations proved to be superior. We expanded the testing of the two most potent GPC2-directed CAR constructs to GPC2-expressing medulloblastoma and high-grade glioma cell lines, showing significant GPC2-specific cell death in multiple models. Finally, biweekly locoregional delivery of 2–4 million GPC2-directed mRNA CAR T cells induced significant tumor regression in an orthotopic medulloblastoma model and significantly prolonged survival in an aggressive orthotopic thalamic diffuse midline glioma xenograft model. No GPC2-directed CAR T cell related neurologic or systemic toxicity was observed. Taken together, these data show that GPC2 is a highly differentially expressed cell surface protein on multiple malignant pediatric brain tumors that can be targeted safely with local delivery of mRNA CAR T cells, laying the framework for the clinical translation of GPC2-directed immunotherapies for pediatric brain tumors.
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DOI：
10.1073/pnas.1706055114
发表时间：
2017-08-08
影响因子：
11.1
作者：
Li, Nan;Fu, Haiying;Ho, Mitchell
通讯作者：
Ho, Mitchell
影响因子：
28.2
作者：
通讯作者：
--
影响因子：
29.4
作者：
Beatty GL;O'Hara MH;Lacey SF;Torigian DA;Nazimuddin F;Chen F;Kulikovskaya IM;Soulen MC;McGarvey M;Nelson AM;Gladney WL;Levine BL;Melenhorst JJ;Plesa G;June CH
通讯作者：
June CH
影响因子：
4.1
作者：
Almåsbak H;Aarvak T;Vemuri MC
通讯作者：
Vemuri MC
DOI：
10.1158/1078-0432.ccr-15-0428
发表时间：
2015-09-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Brown CE;Badie B;Barish ME;Weng L;Ostberg JR;Chang WC;Naranjo A;Starr R;Wagner J;Wright C;Zhai Y;Bading JR;Ressler JA;Portnow J;D'Apuzzo M;Forman SJ;Jensen MC
通讯作者：
Jensen MC