Skip to main content

Advertisement

SpringerLink
Log in

IL13Rα2-targeted third-generation CAR-T cells with CD28 transmembrane domain mediate the best anti-glioblastoma efficacy

  • Research
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Chimeric antigen receptor (CAR)-modified T (CAR-T) cell therapy has been proven to be a powerful tool for the treatment of cancer, however, the limits are obvious, especially for solid tumors. Therefore, constantly optimizing the structure of CAR to improve its therapeutic effect is necessary. In this study, we generated three different third-generation CARs targeting IL13Rα2, with the same scFv, but different transmembrane domains (TMDs) from CD4, CD8 or CD28 (IL13-CD4TM-28.BB.ζ, IL13-CD8TM-28.BB.ζ and IL13-CD28TM-28.BB.ζ). CARs were transduced into primary T cells using retroviruses. The anti-GBM efficacy of CAR-T cells was monitored by flow cytometry and real-time cell analysis (RTCA) in vitro and examined in two xenograft mouse models. The differentially expressed genes related to different anti-GBM activity were screened by high throughput RNA sequencing. We observed that T cells transduced with these three CARs have similar anti-tumor activity when co-cultured with U373 cells which expressed higher IL13Rα2 but exhibited different anti-tumor activity when co-cultured with U251 cells that expressed lower IL13Rα2. All the three groups of CAR-T cells can be activated by U373 cells, but only IL13-CD28TM-28.BB.ζ CAR-T cells could be activated and expressed increased IFN-γ after co-culturing with U251 cells. IL13-CD28TM-28.BB.ζ CAR-T cells exhibited the best anti-tumor activity in xenograft mouse models which can infiltrate into the tumors. The superior anti-tumor efficacy of IL13-CD28TM-28.BB.ζ CAR-T cells was partially owing to differentially expressed extracellular assembly, extracellular matrix, cell migration and adhesion-related genes which contribute to the lower activation threshold, increased cell proliferation, and elevated migration capacity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Availability of data and materials

Data available within the article or its supplementary materials.

References

  1. Stupp R, Taillibert S, Kanner A et al (2017) Effect of tumor-treating fields plus maintenance temozolomide versus maintenance temozolomide alone on survival in patients with glioblastoma: a randomized clinical trial. JAMA, J Am Med Assoc 318:2306–2316

    Article  CAS  Google Scholar 

  2. Bielamowicz K, Khawja S, Ahmed N (2013) Adoptive cell therapies for glioblastoma. Front Oncol 3:275

    Article  PubMed  PubMed Central  Google Scholar 

  3. Ahmed N, Salsman VS, Kew Y et al (2010) HER2-specific T cells target primary glioblastoma stem cells and induce regression of autologous experimental tumors. Clin Cancer Res Off J Am Assoc Cancer Res 16:474–485

    Article  CAS  Google Scholar 

  4. Sampson JH, Choi BD, Sanchez-Perez L et al (2014) EGFRvIII mCAR-modified T-cell therapy cures mice with established intracerebral glioma and generates host immunity against tumor-antigen loss. Clin Cancer Res Off J Am Assoc Cancer Res 20:972–984

    Article  CAS  Google Scholar 

  5. Hong JJ, Rosenberg SA, Dudley ME et al (2010) Successful treatment of melanoma brain metastases with adoptive cell therapy. Clin Cancer Res Off J Am Assoc Cancer Res 16:4892–4898

    Article  CAS  Google Scholar 

  6. Yaghoubi SS, Jensen MC, Satyamurthy N et al (2009) Noninvasive detection of therapeutic cytolytic T cells with 18F-FHBG PET in a patient with glioma. Nat Clin Pract Oncol 6:53–58

    Article  CAS  PubMed  Google Scholar 

  7. Sadelain M, Brentjens R, Riviere I, Park J (2015) CD19 CAR therapy for acute lymphoblastic Leukemia. Am Soc Clin Oncol Educ Book Am Soc Clin Oncol Annu Meet 35:e360-363

    Article  Google Scholar 

  8. Maus MV, Grupp SA, Porter DL, June CH (2014) Antibody-modified T cells: CARs take the front seat for hematologic malignancies. Blood 123:2625–2635

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Maude SL, Frey N, Shaw PA et al (2014) Chimeric antigen receptor T cells for sustained remissions in Leukemia. N Engl J Med 371:1507–1517

    Article  PubMed  PubMed Central  Google Scholar 

  10. Abramson JS, McGree B, Noyes S et al (2017) Anti-CD19 CAR T cells in CNS diffuse large-B-Cell lymphoma. N Engl J Med 377:783–784

    Article  PubMed  Google Scholar 

  11. Brown CE, Starr R, Aguilar B et al (2012) Stem-like tumor-initiating cells isolated from IL13Ralpha2 expressing gliomas are targeted and killed by IL13-zetakine-redirected T Cells. Clin Cancer Res Off J Am Assoc Cancer Res 18:2199–2209

    Article  CAS  Google Scholar 

  12. Fichtner-Feigl S, Terabe M, Kitani A et al (2008) Restoration of tumor immunosurveillance via targeting of interleukin-13 receptor-alpha 2. Can Res 68:3467–3475

    Article  CAS  Google Scholar 

  13. Jarboe JS, Johnson KR, Choi Y, Lonser RR, Park JK (2007) Expression of interleukin-13 receptor alpha2 in glioblastoma multiforme: implications for targeted therapies. Can Res 67:7983–7986

    Article  CAS  Google Scholar 

  14. Joshi BH, Puri RA, Leland P et al (2008) Identification of interleukin-13 receptor alpha2 chain overexpression in situ in high-grade diffusely infiltrative pediatric brainstem glioma. Neuro Oncol 10:265–274

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zeng J, Zhang J, Yang YZ et al (2020) IL13RA2 is overexpressed in malignant gliomas and related to clinical outcome of patients. Am J Transl Res 12:4702–4714

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Iwami K, Shimato S, Ohno M et al (2012) Peptide-pulsed dendritic cell vaccination targeting interleukin-13 receptor alpha2 chain in recurrent malignant glioma patients with HLA-A*24/A*02 allele. Cytotherapy 14:733–742

    Article  CAS  PubMed  Google Scholar 

  17. Kunwar S, Prados MD, Chang SM et al (2007) Direct intracerebral delivery of cintredekin besudotox (IL13-PE38QQR) in recurrent malignant glioma: a report by the Cintredekin Besudotox Intraparenchymal Study Group. J Clin Oncol Off J Am Soc Clin Oncol 25:837–844

    Article  CAS  Google Scholar 

  18. Brown CE, Badie B, Barish ME et al (2015) Bioactivity and safety of il13ralpha2-redirected chimeric antigen receptor CD8+ T cells in patients with recurrent glioblastoma. Clin Cancer Res Off J Am Assoc Cancer Res 21:4062–4072

    Article  CAS  Google Scholar 

  19. Brown CE, Alizadeh D, Starr R et al (2016) Regression of glioblastoma after chimeric antigen receptor T-cell therapy. N Engl J Med 375:2561–2569

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Brown CE, Aguilar B, Starr R et al (2018) Optimization of IL13Ralpha2-targeted chimeric antigen receptor T cells for improved anti-tumor efficacy against glioblastoma. Mol Ther J Am Soc Gene Therapy 26:31–44

    Article  CAS  Google Scholar 

  21. Kim K, Gwak HS, Han N et al (2021) Chimeric antigen receptor T cells with modified interleukin-13 preferentially recognize IL13Ralpha2 and suppress malignant glioma: a preclinical study. Front Immunol 12:715000

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Yin Y, Rodriguez JL, Li N et al (2022) Locally secreted BiTEs complement CAR T cells by enhancing killing of antigen heterogeneous solid tumors. Mol Ther 30:2537–2553

    Article  CAS  PubMed  Google Scholar 

  23. Zhong XS, Matsushita M, Plotkin J, Riviere I, Sadelain M (2010) Chimeric antigen receptors combining 4–1BB and CD28 signaling domains augment PI3kinase/AKT/Bcl-XL activation and CD8+ T cell-mediated tumor eradication. Mol Ther 18:413–420

    Article  CAS  PubMed  Google Scholar 

  24. Majzner RG, Rietberg SP, Sotillo E et al (2020) Tuning the antigen density requirement for CAR T-cell activity. Cancer Discov 10:702–723

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Kochenderfer JN, Wilson WH, Janik JE et al (2010) Eradication of B-lineage cells and regression of lymphoma in a patient treated with autologous T cells genetically engineered to recognize CD19. Blood 116:4099–4102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Brentjens RJ, Riviere I, Park JH et al (2011) Safety and persistence of adoptively transferred autologous CD19-targeted T cells in patients with relapsed or chemotherapy refractory B-cell leukemias. Blood 118:4817–4828

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Bridgeman JS, Hawkins RE, Bagley S, Blaylock M, Holland M, Gilham DE (2010) The optimal antigen response of chimeric antigen receptors harboring the CD3zeta transmembrane domain is dependent upon incorporation of the receptor into the endogenous TCR/CD3 complex. J Immunol 184:6938–6949

    Article  CAS  PubMed  Google Scholar 

  28. Wang D, Aguilar B, Starr R, Alizadeh D, Brito A, Sarkissian A, Ostberg JR, Forman SJ, Brown CE (2018) Glioblastoma-targeted CD4+ CAR T cells mediate superior antitumor activity. JCI Insight. https://doi.org/10.1172/jci.insight.99048

    Article  PubMed  PubMed Central  Google Scholar 

  29. Ramello MC, Benzaid I, Kuenzi BM et al (2019) An immunoproteomic approach to characterize the CAR interactome and signalosome. Sci Signal 12:568

    Article  Google Scholar 

  30. Muller YD, Nguyen DP, Ferreira LMR et al (2021) The CD28-transmembrane domain mediates chimeric antigen receptor heterodimerization with CD28. Front Immunol 12:639818

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Watanabe K, Terakura S, Martens AC et al (2015) Target antigen density governs the efficacy of anti-CD20-CD28-CD3 zeta chimeric antigen receptor-modified effector CD8+ T cells. J Immunol 194:911–920

    Article  CAS  PubMed  Google Scholar 

  32. Stone JD, Aggen DH, Schietinger A, Schreiber H, Kranz DM (2012) A sensitivity scale for targeting T cells with chimeric antigen receptors (CARs) and bispecific T-cell Engagers (BiTEs). Oncoimmunology 1:863–873

    Article  PubMed  PubMed Central  Google Scholar 

  33. Nerreter T, Letschert S, Gotz R et al (2019) Super-resolution microscopy reveals ultra-low CD19 expression on myeloma cells that triggers elimination by CD19 CAR-T. Nat Commun 10:3137

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

Not applicable.

Funding

Beijing Municipal Science and Technology Commission, Brain Science Research Fund (Z16110000021636).

Author information

Author notes

  1. Aiqin Gu, Yue Bai and Can Zhang have contributed equally to this work.

Authors and Affiliations

  1. The Clinical Center of Gene and Cell Engineering, Beijing Shijitan Hospital, Capital Medical University, No. 10, Iron Medicine Road, Yang Fang Dian, Haidian District, Beijing, 100038, China

    Aiqin Gu, Yue Bai, Can Zhang, Chang Xu, Zhijing An, Ying Zhang, Yi Hu & Xiaosong Zhong

  2. Carriage Therapeutics for Affiliation, Beijing, China

    Xiaosong Zhong

  3. River Hill High School, Clarksville, USA

    Steven H. Zhong

Authors
  1. Aiqin Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yue Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Can Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chang Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhijing An
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ying Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Steven H. Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yi Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xiaosong Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

AG designed experiments, collected and analyzed data, processed the pictures and wrote manuscripts; YB participated in flow cytometry analysis; FW participated in cytokine detection; CZ participated in preparation of CAR-T cells; CX participated in mouse experiments; ZA and YZ participated in cell culture; SZ participated in statistical analysis; YH and XZ supervised the project and designed the study. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yi Hu or Xiaosong Zhong.

Ethics declarations

Conflict of interest

No competing interest.

Ethical approval and consent to participate

This research was approved by the Beijing Shijitan Hospital Institutional Review Board.

Consent for publication

Written informed consent was obtained from all the participants.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gu, A., Bai, Y., Zhang, C. et al. IL13Rα2-targeted third-generation CAR-T cells with CD28 transmembrane domain mediate the best anti-glioblastoma efficacy. Cancer Immunol Immunother 72, 2393–2403 (2023). https://doi.org/10.1007/s00262-023-03423-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-023-03423-5

Keywords

Search

Navigation