Abstract 1444: The new therapy strategy for treatment of peripheral T cell lymphomas: CD30-targeted CAR-modified T cell therapy
Authors: Yang Wu, Dan Chen, Rong Ma, Jun-ying Zhang, Yuan Zhang, Hai-xia Cao, Zhuo Wang, Chang-wen Jing, Si-wen Liu, Ji-feng Feng, Jian-Zhong Wu
Journal: Cancer Research (2019)

Abstract Objective: Peripheral T-cell lymphoma (PTCL) is a type of non-hodgkin lymphoma with high metastasis and poor prognosis. The guidelines (NCCN guidelines, 2017) recommended CHOEP or CHOP chemotherapy regimen for treatment of PTCL. Unfortunately, the 5-year survival rate of PTCL is lower than 30% according to the present clinical data. Establishing new effective treatment methods and strategies for PTCL has been an urgent problem. Car-T cell therapy is a promising therapy for individuals with lymphoma. In USA, CD19 Car-T cell therapy has been proved to be effective for acute and chronic B-cell lymphoma treatment in more than 1000 clinic trials. However, there is currently no known Car-T cell therapy for PTCL, one of the most prevalent malignancies in Asia. In this study, to provide a safe and effective treatment for the recurrence and malignant CD30 positive PTCL, we establish the world-first CD30 Car-T cell therapy strategy and test its effect in PTCL. Method: The extracellular fragment gene sequences of CD30 were obtained from CD30 positive tumor tissues of PTCL patients and cloned into a recombinant plasmid to obtained CD30 protein. Then, the CD30 targeting single-chain antibody fragment (scFv) gene sequences were obtained from CD30 positive monoclonal hybridoma cell. The T cells were stimulated by using OKT3, CD28 and IL-2 from peripheral blood mononuclear cell (PBMC), then transfected with CD30 Lenti-CAR. The numbers of CD30 positive T cells were counted by flow cytometry after 14 days culture. The cytotoxic function of Car-T cells to CD30+ PTCL cells were detected using chromium release assay (CRA) and real-time cell electronic sensing assay (xCelligence station, ACEA biosciences). Results: The SDS-PAGE data shows that the molecular weight of CD30 protein was 45KDa, and the mass spectrometry data indicate that amino acid sequences were consistent with the prediction. The flow cytometry data shows 35% CD30 positive Car-T cells after transfection with CD30 Lenti-Car at day of 14. The Car-T cells can significantly inhibit the 299 cells proliferation at 20:1 effector-to-target ratios after 12 hours. The real-time cell electronic sensing data shows the Car-T cells can significantly inhibit the 299 cells proliferation at effector-to-target ratios of 5:1, 10:1, 20:1 and 40:1 after 6 hours co-culture. Conclusions: In summary, we have shown a strategy for treating CD30 positive PTCL with Car-T cell therapy. We obtained a CD30 antibody with high specificity and affinity though immunized of mouse by CD30 antigen which sequences comes from the CD30 positive PTCL tissue. We also established real-time cell electronic sensing method to assessing cytotoxic function of Car-T cells and Car-T cells show high efficiency to cytotoxic of Karpas 299 cells. Therefore, this new CD30 Car-T cells therapy strategy may provide an effective and safe treatment of PTCL. Note: This abstract was not presented at the meeting. Citation Format: Yang Wu, Dan Chen, Rong Ma, Jun-ying Zhang, Yuan Zhang, Hai-xia Cao, Zhuo Wang, Chang-wen Jing, Si-wen Liu, Ji-feng Feng, Jian-Zhong Wu. The new therapy strategy for treatment of peripheral T cell lymphomas: CD30-targeted CAR-modified T cell therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1444.