Immunotherapy: Efficiently Targeting Pediatric Neuroblastoma and Glioblastoma by the Combinatorial Therapy of Il-21 Secretion Oncolytic Virus and Anti-Ror1 Car Nk Cells
Author Type(s)
Faculty
Document Type
Abstract
Publication Date
2022
DOI
10.1016/S1465-3249(22)00323-1
Journal Title
Cytotherapy
Department
Pediatrics
Abstract
Background & Aim: Background: Children with recurrent and/or metastatic neuroblastoma (NB) and glioblastoma multiforme (GBM) have a dismal event-free survival (<25%) (Chu/Cairo, JITC, 2021). Novel therapies are desperately needed for these poor risk patients. ROR1 is highly expressed on the majority of NB and GBM. Our group has successfully expanded functional and active peripheral blood NK cells (exPBNK) with irradiated feeder cells and electroporated CAR mRNA to exPBNK (Chu/Cairo, Cancer Immunol Res, 2015). Oncolytic herpes simplex viruses (oHSVs) are a promising experimental therapy and have been safely used in clinical trials for a wide range of cancers (Cassady, et al, JITC, 2021). Objectives: To determine the anti-tumor efficacy of the combinatorial therapy of C134-based human IL21 expression with anti-ROR1 CAR engineered exPBNK cells against ROR1+ NB and GBM. Methods, Results & Conclusion: Methods: ExPBNK cells were expanded and electroporated with anti-ROR1-CAR mRNA as we previously described (Chu/Cairo, Cancer Immunol Res, 2015). C021 was generated by modifying C134 to express human IL-21 gene. The supernatants of C134 and C021 (C134+hIL21) were generated as previously (Figure Presented) described (Cassady, et al, JITC, 2021). In vitro cytotoxicity of anti-ROR1 CAR NK against NB and GBM cell lines were examined at different E:T ratios. IFN-g, granzyme and perforin levels were evaluated by ELISA assays. In vivo anti- tumor effect was examined utilizing human NB tumor xenografted NSG mice (Chu/Cairo, JITC, 2021). Results: C021 infected NB cells (MOI 0.025) generated hIL-21 in cell supernatants at 24 hours post infection (hpi) and peaked at 72hpi. The combination of C021 and anti-ROR1 CAR exPBNK cells significantly enhanced the killing of NB and GBM cells than controls including C134 + anti-ROR1 CAR exPBNK cells (p<0.05) (Fig.1A). The enhanced killing function of anti-ROR1 CAR exPBNK by C021 was associated with significantly enhanced secretion of IFN-g (p<0.05) (Fig.1B), granzyme B (p<0.05) and perforin (p<0.05) and significantly enhanced expression of NK activating marker CD25 (p<0.05). Our in vivo animal study showed that the combination of C021 and anti-ROR1 CAR exPBNK cells reduced tumor burden in human NB xenografted NSG mice compared to untreated group (p<0.05) and anti-ROR1 CAR exPBNK treated group (P=0.056) (Fig.2). Conclusion: Our data demonstrated (Figure Presented) the anti-tumor efficacy of the combination of C021 with anti-ROR1 CAR exPBNK cells targeting NB and GBM in vitro and in vivo.
Recommended Citation
Chu, Y., Tian, M., Cassady, K., Saini, U., Cripe, T., Lee, D. A., Riddell, S., & Cairo, M. S. (2022). Immunotherapy: Efficiently Targeting Pediatric Neuroblastoma and Glioblastoma by the Combinatorial Therapy of Il-21 Secretion Oncolytic Virus and Anti-Ror1 Car Nk Cells. Cytotherapy, 24 (5), S119-S120. https://doi.org/10.1016/S1465-3249(22)00323-1
