The efficacy of third generation anti‑HER2 chimeric antigen receptor T cells in combination with PD1 blockade against malignant glioblastoma cells

  • Authors:
    • Luxi Shen
    • Hongzhi Li
    • Shufang Bin
    • Panyuan Li
    • Jie Chen
    • Haihua Gu
    • Weihua Yuan
  • View Affiliations

  • Published online on: August 5, 2019     https://doi.org/10.3892/or.2019.7263
  • Pages: 1549-1557
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Abstract

Without effective treatment, glioblastoma is one of the deadliest cancers worldwide. The aim of the present study was to explore whether combinational immunotherapy is effective for treating malignant glioblastoma in vitro. The therapeutic efficacy of third generation anti‑human epidermal growth factor receptor 2 (HER2) chimeric antigen receptor (CAR)‑T cells alone and in combination with PD1 blockade was investigated for the treatment of malignant glioblastoma cells in vitro. Anti‑HER2 CAR‑T cells were prepared by transducing activated primary human T cells with lentiviruses which expressed third generation anti‑HER2 CAR. The CAR‑positive cell ratio was detected using flow cytometry. The expression level of CAR was detected by western blot analysis. The binding of anti‑HER2 CAR‑T cells to HER2+ U251 glioblastoma cells was examined under a fluorescence microscope. The cytokine secretion of CAR‑T cells induced by target cells was analyzed via ELISA. The cytotoxicity of anti‑HER2 CAR‑T cells alone or in combination with anti‑programmed death‑1 (PD1) antibody against HER2+/PDL1+ U251 cells was examined using an LDH assay. The CAR‑positive cell ratio and expression level of CAR in prepared CAR‑T cells were both high enough. Anti‑HER2 CAR‑T cells could specifically bind to U251 cells. The IL‑2 and IFN‑γ secretion of CAR‑T cells increased after being co‑cultured with U251 cells, and further increased in the presence of anti‑PD1 antibody. Anti‑HER2 CAR‑T cells displayed a potent cytotoxicity against U251 cells. In addition, the presence of anti‑PD1 antibody further enhanced the efficacy of anti‑HER2 CAR‑T cells against U251 cells. The present results indicated that blocking PD1 immuno‑suppression can increase the activation of CAR‑T cells after they are activated by a targeting antigen. Third generation anti‑HER2 CAR‑T cells along with PD1 blockade have a great therapeutic potential for combatting malignant glioblastoma.

References

1 

Reardon DA and Mitchell DA: The development of dendritic cell vaccine-based immunotherapies for glioblastoma. Semin Immunopathol. 39:225–239. 2017. View Article : Google Scholar : PubMed/NCBI

2 

Ohba S and Hirose Y: Current and Future Drug Treatments for Glioblastomas. Curr Med Chem. 23:4309–4316. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Batash R, Asna N, Schaffer P, Francis N and Schaffer M: Glioblastoma multiforme, diagnosis and treatment; recent literature review. Curr Med Chem. 24:3002–3009. 2017. View Article : Google Scholar : PubMed/NCBI

4 

Hao L, Li T, Chang LJ and Chen X: Adoptive immunotherapy for B-cell malignancies using CD19-targeted chimeric antigen receptor T-cells: A systematic review of efficacy and safety. Curr Med Chem. Aug 1–2017.(Epub ahead of print). doi: 10.2174/0929867324666170801101842. View Article : Google Scholar : PubMed/NCBI

5 

Jacoby E, Bielorai B, Avigdor A, Itzhaki O, Hutt D, Nussboim V, Meir A, Kubi A, Levy M, Zikich D, et al: Locally produced CD19 CAR T cells leading to clinical remissions in medullary and extramedullary relapsed acute lymphoblastic leukemia. Am J Hematol. 93:1485–1492. 2018. View Article : Google Scholar : PubMed/NCBI

6 

Wang D, Shi R, Wang Q and Li J: Extramedullary relapse of acute lymphoblastic leukemia after allogeneic hematopoietic stem cell transplantation treated by CAR T-cell therapy: A case report. Onco Targets Ther. 11:6327–6332. 2018. View Article : Google Scholar : PubMed/NCBI

7 

Jena B, Dotti G and Cooper LJ: Redirecting T-cell specificity by introducing a tumor-specific chimeric antigen receptor. Blood. 116:1035–1044. 2010. View Article : Google Scholar : PubMed/NCBI

8 

Potti A, Forseen SE, Koka VK, Pervez H, Koch M, Fraiman G, Mehdi SA and Levitt R: Determination of HER-2/neu overexpression and clinical predictors of survival in a cohort of 347 patients with primary malignant brain tumors. Cancer Invest. 22:537–544. 2004. View Article : Google Scholar : PubMed/NCBI

9 

Koka V, Potti A, Forseen SE, Pervez H, Fraiman GN, Koch M and Levitt R: Role of Her-2/neu overexpression and clinical determinants of early mortality in glioblastoma multiforme. Am J Clin Oncol. 26:332–335. 2003. View Article : Google Scholar : PubMed/NCBI

10 

Seliger B, Rongcun Y, Atkins D, Hammers S, Huber C, Storkel S and Kiessling R: HER-2/neu is expressed in human renal cell carcinoma at heterogeneous levels independently of tumor grading and staging and can be recognized by HLA-A2.1-restricted cytotoxic T lymphocytes. Int J Cancer. 87:349–359. 2000. View Article : Google Scholar : PubMed/NCBI

11 

Wang S, Saboorian MH, Frenkel E, Hynan L, Gokaslan ST and Ashfaq R: Laboratory assessment of the status of Her-2/neu protein and oncogene in breast cancer specimens: Comparison of immunohistochemistry assay with fluorescence in situ hybridisation assays. J Clin Pathol. 53:374–381. 2000. View Article : Google Scholar : PubMed/NCBI

12 

Li BT, Ross DS, Aisner DL, Chaft JE, Hsu M, Kako SL, Kris MG, Varella-Garcia M and Arcila ME: HER2 amplification and HER2 mutation are distinct molecular targets in lung cancers. J Thorac Oncol. 11:414–419. 2016. View Article : Google Scholar : PubMed/NCBI

13 

Press MF, Cordon-Cardo C and Slamon DJ: Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues. Oncogene. 5:953–962. 1990.PubMed/NCBI

14 

Whilding LM and Maher J: ErbB-targeted CAR T-cell immunotherapy of cancer. Immunotherapy. 7:229–241. 2015. View Article : Google Scholar : PubMed/NCBI

15 

Nowakowska P, Romanski A, Miller N, Odendahl M, Bonig H, Zhang C, Seifried E, Wels WS and Tonn T: Clinical grade manufacturing of genetically modified, CAR-expressing NK-92 cells for the treatment of ErbB2-positive malignancies. Cancer Immunol Immunother. 67:25–38. 2018. View Article : Google Scholar : PubMed/NCBI

16 

Liu G, Ying H, Zeng G, Wheeler CJ, Black KL and Yu JS: HER-2, gp100, and MAGE-1 are expressed in human glioblastoma and recognized by cytotoxic T cells. Cancer Res. 64:4980–4986. 2004. View Article : Google Scholar : PubMed/NCBI

17 

Ahmed N, Salsman VS, Kew Y, Shaffer D, Powell S, Zhang YJ, Grossman RG, Heslop HE and Gottschalk S: HER2-specific T cells target primary glioblastoma stem cells and induce regression of autologous experimental tumors. Clin Cancer Res. 16:474–485. 2010. View Article : Google Scholar : PubMed/NCBI

18 

Thomas CY, Chouinard M, Cox M, Parsons S, Stallings-Mann M, Garcia R, Jove R and Wharen R: Spontaneous activation and signaling by overexpressed epidermal growth factor receptors in glioblastoma cells. Int J Cancer. 104:19–27. 2003. View Article : Google Scholar : PubMed/NCBI

19 

Andersson U, Guo D, Malmer B, Bergenheim AT, Brannstrom T, Hedman H and Henriksson R: Epidermal growth factor receptor family (EGFR, ErbB2-4) in gliomas and meningiomas. Acta Neuropathol. 108:135–142. 2004. View Article : Google Scholar : PubMed/NCBI

20 

Kuramitsu S, Yamamichi A, Ohka F, Motomura K, Hara M and Natsume A: Adoptive immunotherapy for the treatment of glioblastoma: Progress and possibilities. Immunotherapy. 8:1393–1404. 2016. View Article : Google Scholar : PubMed/NCBI

21 

Tlsty TD and Coussens LM: Tumor stroma and regulation of cancer development. Annu Rev Pathol. 1:119–150. 2006. View Article : Google Scholar : PubMed/NCBI

22 

Denko NC: Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer. 8:705–713. 2008. View Article : Google Scholar : PubMed/NCBI

23 

Zheng Y, Zha Y and Gajewski TF: Molecular regulation of T-cell anergy. EMBO Rep. 9:50–55. 2008. View Article : Google Scholar : PubMed/NCBI

24 

Gajewski TF, Schreiber H and Fu YX: Innate and adaptive immune cells in the tumor microenvironment. Nat Immunol. 14:1014–1022. 2013. View Article : Google Scholar : PubMed/NCBI

25 

Al-Zoughbi W, Huang J, Paramasivan GS, Till H, Pichler M, Guertl-Lackner B and Hoefler G: Tumor macroenvironment and metabolism. Semin Oncol. 41:281–295. 2014. View Article : Google Scholar : PubMed/NCBI

26 

Zou W and Chen L: Inhibitory B7-family molecules in the tumour microenvironment. Nat Rev Immunol. 8:467–477. 2008. View Article : Google Scholar : PubMed/NCBI

27 

Keir ME, Butte MJ, Freeman GJ and Sharpe AH: PD-1 and its ligands in tolerance and immunity. Annu Rev Immunol. 26:677–704. 2008. View Article : Google Scholar : PubMed/NCBI

28 

Yuan W, Chen J, Cao Y, Yang L, Shen L, Bian Q, Bin S, Li P, Cao J, Fang H, et al: Comparative analysis and optimization of protocols for producing recombinant lentivirus carrying the anti-Her2 chimeric antigen receptor gene. J Gene Med. 20:e30272018. View Article : Google Scholar : PubMed/NCBI

29 

Wang C, Hu W, Shen L, Dou R, Zhao S, Shan D, Yu K, Huang R and Li H: Adoptive antitumor immunotherapy in vitro and in vivo using genetically activated erbB2-specific T cells. J Immunother. 37:351–359. 2014. View Article : Google Scholar : PubMed/NCBI

30 

Zhong XS, Matsushita M, Plotkin J, Riviere I and Sadelain M: 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. 2010. View Article : Google Scholar : PubMed/NCBI

31 

Brentjens RJ, Santos E, Nikhamin Y, Yeh R, Matsushita M, La Perle K, Quintas-Cardama A, Larson SM and Sadelain M: Genetically targeted T cells eradicate systemic acute lymphoblastic leukemia xenografts. Clin Cancer Res. 13:5426–5435. 2007. View Article : Google Scholar : PubMed/NCBI

32 

Finney HM, Akbar AN and Lawson AD: Activation of resting human primary T cells with chimeric receptors: Costimulation from CD28, inducible costimulator, CD134, and CD137 in series with signals from the TCR zeta chain. J Immunol. 172:104–113. 2004. View Article : Google Scholar : PubMed/NCBI

33 

Roger A, Finet A, Boru B, Beauchet A, Mazeron JJ, Otzmeguine Y, Blom A, Longvert C, de Maleissye MF, Funck-Brentano E and Saiag P: Efficacy of combined hypo-fractionated radiotherapy and anti-PD-1 monotherapy in difficult-to-treat advanced melanoma patients. Oncoimmunology. 7:e14421662018. View Article : Google Scholar : PubMed/NCBI

34 

Taquin H, Fontas E, Massol O, Chevallier P, Balloti R, Beranger G, Lacour JP, Passeron T and Montaudie H: Efficacy and safety data for checkpoint inhibitors in advanced melanoma under real-life conditions: A monocentric study conducted in Nice from 2010 to 2016. Ann Dermatol Venereol. 145:649–658. 2018. View Article : Google Scholar : PubMed/NCBI

35 

Wang Y, Zhang X, Yang L, Xue J and Hu G: Blockade of CCL2 enhances immunotherapeutic effect of anti-PD1 in lung cancer. J Bone Oncol. 11:27–32. 2018. View Article : Google Scholar : PubMed/NCBI

36 

Zibetti Dal Molin G, Abrahao CM, Coleman RL and Maluf FC: Response to pembrolizumab in a heavily treated patient with metastatic ovarian carcinosarcoma. Gynecol Oncol Res Pract. 5:62018. View Article : Google Scholar : PubMed/NCBI

37 

Li S, Siriwon N, Zhang X, Yang S, Jin T, He F, Kim YJ, Mac J, Lu Z, Wang S, et al: Enhanced cancer immunotherapy by chimeric antigen receptor-modified t cells engineered to secrete checkpoint inhibitors. Clin Cancer Res. 23:6982–6992. 2017. View Article : Google Scholar : PubMed/NCBI

38 

Serganova I, Moroz E, Cohen I, Moroz M, Mane M, Zurita J, Shenker L, Ponomarev V and Blasberg R: Enhancement of PSMA-directed CAR adoptive immunotherapy by PD-1/PD-L1 blockade. Mol Ther Oncolytics. 4:41–54. 2017. View Article : Google Scholar : PubMed/NCBI

39 

Liu X, Ranganathan R, Jiang S, Fang C, Sun J, Kim S, Newick K, Lo A, June CH, Zhao Y and Moon EK: A chimeric switch-receptor targeting PD1 augments the efficacy of second-generation car t cells in advanced solid tumors. Cancer Res. 76:1578–1590. 2016. View Article : Google Scholar : PubMed/NCBI

40 

Cherkassky L, Morello A, Villena-Vargas J, Feng Y, Dimitrov DS, Jones DR, Sadelain M and Adusumilli PS: Human CAR T cells with cell-intrinsic PD-1 checkpoint blockade resist tumor-mediated inhibition. J Clin Invest. 126:3130–3144. 2016. View Article : Google Scholar : PubMed/NCBI

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October 2019
Volume 42 Issue 4

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Copy and paste a formatted citation
APA
Shen, L., Li, H., Bin, S., Li, P., Chen, J., Gu, H., & Yuan, W. (2019). The efficacy of third generation anti‑HER2 chimeric antigen receptor T cells in combination with PD1 blockade against malignant glioblastoma cells. Oncology Reports, 42, 1549-1557. https://doi.org/10.3892/or.2019.7263
MLA
Shen, L., Li, H., Bin, S., Li, P., Chen, J., Gu, H., Yuan, W."The efficacy of third generation anti‑HER2 chimeric antigen receptor T cells in combination with PD1 blockade against malignant glioblastoma cells". Oncology Reports 42.4 (2019): 1549-1557.
Chicago
Shen, L., Li, H., Bin, S., Li, P., Chen, J., Gu, H., Yuan, W."The efficacy of third generation anti‑HER2 chimeric antigen receptor T cells in combination with PD1 blockade against malignant glioblastoma cells". Oncology Reports 42, no. 4 (2019): 1549-1557. https://doi.org/10.3892/or.2019.7263