Development of non-viral, ligand-dependent, EPHB4-specific chimeric antigen receptor T cells for treatment of rhabdomyosarcoma.

Published on Mar 5, 2021in Molecular Therapy - Oncolytics4.115
· DOI :10.1016/J.OMTO.2021.03.001
Hiroshi Kubo1
Estimated H-index: 1
(Kyoto Prefectural University of Medicine),
Shigeki Yagyu14
Estimated H-index: 14
(Kyoto Prefectural University of Medicine)
+ 6 AuthorsHajime Hosoi33
Estimated H-index: 33
(Kyoto Prefectural University of Medicine)
Abstract Ephrin type-B receptor 4 (EPHB4), expressed in tumors including rhabdomyosarcoma, is a suitable target for chimeric antigen receptor (CAR)-T cells. Ligand independent activation of EPHB4 causes cell proliferation and malignant transformation in rhabdomyosarcoma, whereas ligand-dependent stimulation of EPHB4 induces apoptosis in rhabdomyosarcoma. Therefore, we hypothesized that ligand-based EPHB4-specific CAR-T cells may kill rhabdomyosarcoma cells without stimulating downstream cell proliferation mechanisms. We developed novel CAR-T cells by targeting EPHB4 via EPHRIN B2, a natural ligand of EPHB4. Generating EPHB4-CAR-T cells via piggyBac transposon (PB)-based gene transfer resulted in sufficient T cell expansion and CAR positivity (78.5 ± 5.9 %). PB-EPHB4-CAR-T cells displayed a dominant stem cell memory fraction (59.4 ± 7.2 %) as well as low PD-1 expression (0.60 ± 0.21 %) after 14 d of expansion. The PB-EPHB4-CAR-T cells inhibited EPHB4-positive tumor cells without activating cell proliferation downstream of EPHB4, even after multiple tumor re-challenges, and suppressed tumor growth in xenograft-bearing mice. Therefore, PB-EPHB4-CAR-T cells possess a memory-rich fraction without early T cell exhaustion, and show potential as promising therapeutic agents for treating rhabdomyosarcoma and other EPHB4-positive tumors.
📖 Papers frequently viewed together
93 Citations
5 Authors (Avery D. Posey, ..., Carl H. June)
310 Citations
#1Sheng-an Su (ZJU: Zhejiang University)H-Index: 10
#2Yao Xie (ZJU: Zhejiang University)H-Index: 4
Last. Meixiang Xiang (ZJU: Zhejiang University)H-Index: 26
view all 6 authors...
EphrinB2, a membrane-tethered ligand preferentially binding to its receptor EphB4, is ubiquitously expressed in all mammals. Through the particular bidirectional signaling, EphrinB2 plays a critical role during the development of cardiovascular system, postnatal angiogenesis physiologically and pathologically, and cardiac remodeling after injuries as an emerging role. This review highlights the pivotal involvement of EphrinB2 in heart, from developmental cardiogenesis to pathological cardiac rem...
4 CitationsSource
#1Robbie G. Majzner (Stanford University)H-Index: 17
#2Crystal L. Mackall (Stanford University)H-Index: 89
Emerging data from chimeric antigen receptor (CAR) T-cell trials in B-cell malignancies demonstrate that a common mechanism of resistance to this novel class of therapeutics is the emergence of tumors with loss or downregulation of the target antigen. Antigen loss or antigen-low escape is likely to emerge as an even greater barrier to success in solid tumors, which manifest greater heterogeneity in target antigen expression. Potential approaches to overcome this challenge include engineering CAR...
191 CitationsSource
#1Joseph A. Fraietta (UPenn: University of Pennsylvania)H-Index: 22
#2Simon F. Lacey (UPenn: University of Pennsylvania)H-Index: 63
Last. J. Joseph Melenhorst (UPenn: University of Pennsylvania)H-Index: 56
view all 37 authors...
Tolerance to self-antigens prevents the elimination of cancer by the immune system1,2. We used synthetic chimeric antigen receptors (CARs) to overcome immunological tolerance and mediate tumor rejection in patients with chronic lymphocytic leukemia (CLL). Remission was induced in a subset of subjects, but most did not respond. Comprehensive assessment of patient-derived CAR T cells to identify mechanisms of therapeutic success and failure has not been explored. We performed genomic, phenotypic a...
426 CitationsSource
#1Erhao Zhang (CPU: China Pharmaceutical University)H-Index: 5
#1Erhao Zhang (CPU: China Pharmaceutical University)H-Index: 1
Last. Hanmei Xu (CPU: China Pharmaceutical University)H-Index: 16
view all 3 authors...
The potential for adoptive cell immunotherapy as a treatment against cancers has been demonstrated by the remarkable response in some patients with hematological malignancies using autologous T cells endowed with chimeric antigen receptors (CARs) specific for CD19. Clinical efficacy of CAR-T cell therapy for the treatment of solid tumors, however, is rare due to physical and biochemical factors. This review focuses on different aspects of multiple mechanisms of immunosuppression in solid tumors....
33 CitationsSource
#1Daisuke Morita (Shinshu University)H-Index: 5
#2Nobuhiro Nishio (Nagoya University)H-Index: 21
Last. Yozo Nakazawa (Shinshu University)H-Index: 25
view all 14 authors...
Adoptive T cell therapy using chimeric antigen receptor (CAR)-modified T cells is a promising cancer immunotherapy. We previously developed a non-viral method of gene transfer into T cells using a piggyBac transposon system to improve the cost-effectiveness of CAR-T cell therapy. Here, we have further improved our technology by a novel culture strategy to increase the transfection efficiency and to reduce the time of T cell manufacturing. Using a CH2CH3-free CD19-specific CAR transposon vector a...
27 CitationsSource
#1Yinnan Chen (ASU: Arizona State University)H-Index: 7
#2Hongmei Zhang (SJTU: Shanghai Jiao Tong University)H-Index: 1
Last. Yanmin Zhang (Xi'an Jiaotong University)H-Index: 34
view all 3 authors...
Abstract Eph receptors and their Eph receptor-interacting (ephrin) ligands together form an important cell communication system with diverse roles. Experimental evidence demonstrated Eph receptor bidirectional signaling with both tumor-suppressing and tumor-promoting activities in cancer cells. The tyrosine kinase EphB4, a member of the Eph receptor family, has been associated with tumor angiogenesis, growth and metastasis, thus making it a valuable and attractive target for drug design for ther...
26 CitationsSource
#1Biliang Hu (UPenn: University of Pennsylvania)H-Index: 14
#2Jiangtao Ren (UPenn: University of Pennsylvania)H-Index: 12
Last. Carl H. June (UPenn: University of Pennsylvania)H-Index: 169
view all 8 authors...
Summary The effects of transgenically encoded human and mouse IL-18 on T cell proliferation and its application in boosting chimeric antigen receptor (CAR) T cells are presented. Robust enhancement of proliferation of IL-18-secreting human T cells occurred in a xenograft model, and this was dependent on TCR and IL-18R signaling. IL-18 augmented IFN-γ secretion and proliferation of T cells activated by the endogenous TCR. TCR-deficient, human IL-18-expressing CD19 CAR T cells exhibited enhanced p...
142 CitationsSource
#2Megan M. Cleary (OHSU: Oregon Health & Science University)H-Index: 5
Last. Charles Keller (OHSU: Oregon Health & Science University)H-Index: 37
view all 15 authors...
Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma affecting children and is often diagnosed with concurrent metastases. Unfortunately, few effective therapies have been discovered that improve the long-term survival rate for children with metastatic disease. Here we determined effectiveness of targeting the receptor tyrosine kinase, EphB4, in both alveolar and embryonal RMS either directly through the inhibitory antibody, VasG3, or indirectly by blocking both forward and reverse sign...
3 CitationsSource
#1Nabil Ahmed (BCM: Baylor College of Medicine)H-Index: 26
#2Vita S. Brawley (BCM: Baylor College of Medicine)H-Index: 13
Last. Stephen GottschalkH-Index: 55
view all 33 authors...
Importance Glioblastoma is an incurable tumor, and the therapeutic options for patients are limited. Objective To determine whether the systemic administration of HER2-specific chimeric antigen receptor (CAR)–modified virus-specific T cells (VSTs) is safe and whether these cells have antiglioblastoma activity. Design, Setting, and Participants In this open-label phase 1 dose-escalation study conducted at Baylor College of Medicine, Houston Methodist Hospital, and Texas Children’s Hospital, patie...
240 CitationsSource
#1Miyuki Tanaka (Center for Cell and Gene Therapy)H-Index: 18
#2Haruko Tashiro (Center for Cell and Gene Therapy)H-Index: 10
Last. Cliona M. RooneyH-Index: 110
view all 12 authors...
Purpose: The multiple mechanisms used by solid tumors to suppress tumor-specific immune responses are a major barrier to the success of adoptively transferred tumor-specific T cells. As viruses induce potent innate and adaptive immune responses, we hypothesized that the immunogenicity of viruses could be harnessed for the treatment of solid tumors if virus-specific T cells (VST) were modified with tumor-specific chimeric antigen receptors (CAR). We tested this hypothesis using VZV-specific T cel...
37 CitationsSource
Cited By0
Objectives null Chimeric antigen receptor (CAR)-T cell therapy possesses the potential to cause unexpected on-target toxicities that may be life-threatening. Non-human primates (NHPs) share considerable structural homology and expression profiles of most proteins with humans and are therefore utilised as an animal model for non-clinical safety studies. We have developed a lymphodepleted NHP model by conditioning the animals with immunosuppressive chemotherapy designed to simulate clinical practi...