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Interleukin-7-loaded oncolytic adenovirus improves CAR-T cell therapy for glioblastoma

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Abstract

Background

T cell with chimeric antigen receptors (CAR-T) has presented remarkable efficacy for blood cancer as an emerging immunotherapy. However, for solid tumors, the therapeutic efficacy is much impaired due to the lack of infiltration and persistence of CAR-T in tumor tissue. Thus, we constructed an interleukin-7-loaded oncolytic adenovirus and combined the use of oncolytic virus and CAR-T to improve the therapeutic outcome.

Methods

We constructed an interleukin-7-loaded oncolytic adenovirus (oAD-IL7) and a B7H3-targeted CAR-T and explored the efficacy of the single use of oAD-IL7, B7H3-CAR-T, or the combined therapy for glioblastoma in vitro and in vivo. The improved CAR-T anti-tumor efficacy was evaluated according to the proliferation, survival, persistence, exhaustion of T cells, and tumor regression.

Results

Constructed oAD-IL7 and B7H3-CAR-T presented moderate cytotoxicity during in vitro study, but failed to induce a thorough and persistent anti-tumor therapeutic efficacy in vivo. The combination of oAD-IL7 and B7H3-CAR-T in vitro resulted in enhanced T cell proliferation and reduced T cell apoptosis. The joint efficacy was further confirmed using tumor-bearing xenograft mice. During in vivo study, the mice treated with both oAD-IL7 and B7H3-CAR-T showed prolonged survival and reduced tumor burden. According to the ex vivo study, oAD-IL7 improved the proliferation and persistence of tumor-infiltrating B7H3-CAR-T, but failed to reverse the exhaustion.

Conclusions

Our results indicated that oAD-IL7 is a promising auxiliary therapy to improve the therapeutic efficacy of B7H3-CAR-T in glioblastoma by providing the activating signals for tumor-infiltrating T cells. Our results also lay the basis for the future clinical trials for the combination of IL7-loaded oncolytic adenovirus and CAR-T therapy for glioblastoma.

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Availability of data and material

All of the data relevant to this research are included in the article and supplementary files. Materials are available from corresponding author under reasonable request.

Abbreviations

CAR-T:

Chimeric antigen receptor T cells

DAMPs:

Damage-associated molecular patterns

GBM:

Glioblastoma

IL7:

Interleukin-7

MDSC:

Myeloid-derived suppressor cells

oAD:

Oncolytic adenovirus

PAMPs:

Pathogen-associated molecular patterns

scFv:

Single-chain variable fragment

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Acknowledgements

We thank Sheng Zenghua for the gift of H4 cell type. We also thank Cao Yi for the gift of glioblastoma tissue acquired from patients.

Funding

This work was supported by the National Natural Science Foundation of China (31471286 and 81772693), the National Major Scientific and Technological Special Project for Significant New Drugs Development (2015ZX09102010), the Postdoctoral Research Fund of Sichuan University (2018SCU12035), and Technology Innovation Project of Chengdu Science and Technology Bureau (No. 2019-YF05-00459-SN).

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Author notes

  1. Jianhan Huang and Meijun Zheng have contributed equally to this work.

Authors and Affiliations

  1. Department of Neurosurgery, West China Hospital, West China Medical School, Sichuan University, No. 37 Guo Xue Xiang, Chengdu, 610041, Sichuan Province, People’s Republic of China

    Jianhan Huang, Xin Tang, Yaxing Chen, Aijun Peng & Liangxue Zhou

  2. Department of Otolaryngology, Head and Neck Surgery, West China Hospital, West China Medical School, Sichuan University, No. 37 Guo Xue Xiang, Chengdu, 610041, Sichuan Province, People’s Republic of China

    Meijun Zheng

  3. State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, No. 37 Guo Xue Xiang, Chengdu, 610041, Sichuan Province, People’s Republic of China

    Zongliang Zhang, Xingchen Peng & Aiping Tong

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  1. Jianhan Huang
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Contributions

LZ raised the idea. JH designed the experiment and completed it with MZ. ZZ contributed to the xenograft model construction. XT and YC drafted the manuscript. AP completed the statistics analysis, and AT and XP revised the manuscript.

Corresponding authors

Correspondence to Aiping Tong or Liangxue Zhou.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

All animal experiments followed a protocol approved by the Institutional Animal Care and Use Committee of Sichuan University.

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Not required.

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Supplementary information

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Supplementary figure 1. Replication of oAD and oAD-IL7 in different GBM cell lines titrated by plaque assays

. The copies of both oAD and oAD-IL7 in U251 (A), U87 (B), A172 (C), T98G (D), and H4 (E) cell lines was titrated 24, 48, 72 hours after viral infection using plaque assay. Viral infection was performed at the MOI of 10. Supplementary figure 2. Cytotoxicity of oAD and oAD-IL7 in different GBM cell lines at different MOI. The cytotoxicity of oAD and oAD-IL7 for different GBM cell lines was measured at day 5 after infection at different MOI based on MTT analysis. (A-E) The 5 types of GBM cell lines. Supplementary figure 3. Construction of B7H3-CART based on multiple donors. The efficacy of CAR-T transduction (A) and activation (B) was monitored under T cells from different donators. Supplementary figure 4. The expression of B7-H3 and CXAR among glioblastoma according to The Cancer Genome Atlas (TCGA) Database. The prevalence (A) of B7H3 and CXAR and their relationship with patient survival (B) among GBM patients were analyzed based on the online data collected via TCGA database. Supplementary figure 5. Representative image of the cytotoxicity effect after serial coculturing B7H3-CART with oAD or oAD-IL7 infected U87 cell line. (A), (B), (C), (D) 2×105 U87 cells were infected by oAD or oAD-IL7 at the MOI of 10, and then cocultured with 1×106 untransduced T cells (UTD) or B7H3-CART for 3 days. The T cells were collected at day 4 for a repeated coculture assay, and the cytotoxicity of the second round of coculturing was measured at day 7 using MTT. Scale bar, 500μm. Supplementary figure 6. Plaque assay for the titration of oAD-IL7 in vivo. The infectious oAD-IL7 in the xenograft models was extracted by 3 consecutive freeze-thaw cycle, and quantified using plaque assay. The representative image is shown. Supplementary table 1. Nucleotide and amino acid sequence of B7H3-scfv. The B7H3-CART was constructed based on the B7H3-scfv. The amino and acid sequence of B7H3-scfv is shown in table. Red, heavy chain; Green, light chain; Yellow, linker.

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Huang, J., Zheng, M., Zhang, Z. et al. Interleukin-7-loaded oncolytic adenovirus improves CAR-T cell therapy for glioblastoma. Cancer Immunol Immunother 70, 2453–2465 (2021). https://doi.org/10.1007/s00262-021-02856-0

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