Presented by Haiping Yang The Clinical Research of Chimeric Antigen Receptor T-cell Immunotherapy

Adaptive Cell Transfer Therapy Adoptive cell therapy (ACT) is a treatment that uses a cancer patient’s own T lymphocytes with anti-tumour activity, expanded in vitroand reinfused into the patient with cancer.

 TIL( Tumor infiltration T-lymphocytes therapy)  TCR ( T-cell receptor therapy)  CAR-T (Chimeric antigen receptor T-cell therapy) Adaptive Cell Transfer Therapy

Cellular therapy has several pathways to the patient. Normal donor cells can be modified to inactivate their alloreactivity while being armed with antitumor CARs or TCRs, or a patient’s own cells can be modified with antitumor molecules. In the case of solid tumors,biopsy specimens can be used to isolate TILs for expansion. In most cases the patient will require some amount of conditioning before receiving antitumor lymphocyte infusions, and careful management of toxicities emerging from these therapies is also required. David M, Chimeric Antigen Receptor– and TCR-Modified T Cells Enter Main Street and Wall Street. The journal of immunology,2015

The first paper to demonstrate the regression of cancer using TIL for the immunotherapy of patients with metastatic melanoma. Rosenberg, S. A.et al.Use of tumor infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. Preliminary report. N. Engl. J. Med. 319, 1676–1680 (1988). TIL

The first paper demonstrating the adoptive cell transfer of lymphocytes transduced with a retrovirus encoding TCRs that recognize a cancer antigen can mediate anti-tumour responses in patients with metastatic melanoma. Morgan, R. A.et al.Cancer regression in patients after transfer of genetically engineered lymphocytes. Science 314, 126–129 (2006). TCR

CAR-T cells recognize tumor cells independent of their expression of human leukocyte antigen (HLA) molecules, tumors that escape conventional T cells by downregulating HLA and/or mutating components of the antigen processing machinery can be eliminated. CAR-T

T-cell differentiation

 Cytotoxic T-cell (CD8)  Helper T-cell (CD4)  Regulatory/suppressor T-cell  Memory T-cell Classfication of T-cell

ITAM: immunoreceptor tyrosine-based activation motif TCR complex :TCR, CD3, ζ TCR

CARs consist of fusion molecules and are typically comprised of an extracellular single chain variable fragment (scFv) of a monoclonal antibody (mAb) specific for a surface molecule on the tumor cell, a spacer domain that provides flexibility and optimizes T cell and target cell engagement, a transmembrane domain, and signaling modules that trigger T cell effector functions. Michael,Designing chimeric antigen receptors to effectively and safely target tumors. Current Opinion in Immunology 2015 CAR-T

Design of CAR T cells. First-generation CARs incorporated the CD3z-chain or similar signaling domains. Ab-based redirection of T cells was first described by Kuwana and refined by Eshhar. Roberts and Finney first described second- generation CARs incorporating CD28 or CD137 signaling domains. David M, Chimeric Antigen Receptor– and TCR-Modified T Cells Enter Main Street and Wall Street. The journal of immunology,2015

The clinical research of CAR-T  Porter DL, Levine BL, Kalos M, Bagg A, June CH. Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N Engl J Med. Aug  Grupp SA, Kalos M, Barrett D, et al. Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N Engl J Med. Apr  Ahmed N, Brawley VS, Hegde M, et al. Human Epidermal Growth Factor Receptor 2 (HER2) -Specific Chimeric Antigen Receptor-Modified T Cells for the Immunotherapy of HER2-Positive Sarcoma. J Clin Oncol. May  Maude SL, Frey N, Shaw PA, et al. Chimeric antigen receptor T cells for sustained remissions in leukemia. N Engl J Med. Oct  Kochenderfer JN, Dudley ME, Kassim SH, et al. Chemotherapy-refractory diffuse large B-cell lymphoma and indolent B-cell malignancies can be effectively treated with autologous T cells expressing an anti-CD19 chimeric antigen receptor. J Clin Oncol. Feb

 Porter DL, Levine BL, Kalos M, Bagg A, June CH. Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N Engl J Med. Aug ;365:725-33

Figure 1 Clinical Response in the Patient.

Figure 2 Serum and Bone Marrow Cytokines before and after Chimeric Antigen Receptor T-Cell Infusion.

Figure 3.Expansion and Persistence of Chimeric Antigen Receptor T Cells In Vivo.

Failure  Morgan RA, et al. Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy. J Immunother 2013;36:133–151.  Morgan RA, Yang JC, Kitano M, Dudley ME,Laurencot CM, Rosenberg SA. Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2.Mol Ther 2010;18:843–851.  Parkhurst MR, et al. T cells targeting carcinoembryonic antigen can mediate regression of metastatic colorectal cancer but induce severe transient colitis. Mol Ther 2011;19:620–626.  Brentjens R, Yeh R, Bernal Y, Riviere I, SadelainM. Treatment of chronic lymphocytic leukemia with genetically targeted autologous T cells: case report of an unforeseen adverse event in a phase I clinical trial. Mol Ther 2010;18:666–668.

Challenges of CAR-T Target selection Optimize costimulatory signaling of T cell effector functions Toxicities (on-target but off-tumor toxicity) (The on-target toxicities result from the inability of engineered T cells to distinguish between normal cells and cancer cells that express the targeted Ag.)

Cytokine release syndrome Tumor lysis syndrome Neurologic toxicities Toxicities

合作双方事件日期价值简介备注 杨森制药 /Transposagen 2014 年 11 月 24 日 杨森制药支付 Transposagen 每个疗法 2.92 亿美元， 其中包括头款和其它收 益 异体 CAR-T 细胞疗法 Janssen 拥有双方合作的异体 CAR-T 疗法独家代理权。 诺华 / Oxford BioMedica2014 年 10 月 诺华支付 Oxford BioMedica 9000 万美 元其中包括 1400 万美 元头款 诺华获得 Oxford 慢病毒载体 LentiVector 应用于 CAR-T 免疫 疗法 CTL019 的非独家全球开发 和商业化权利 Oxford 已授予诺华此次合作所开发 的全部 CAR-T 产品的全球开发 和商业化权利。 Juno 制药 2014 年 8 月 B 轮募资 1.34 亿美元开发 CAR-T 细胞和 TCR T 细胞疗法 12 个月共融资超过 3 亿美元。 Juno 的技术来自 3 个过继 T 细胞疗法 最牛的研究机构： Fred Hutchinson Cancer Research Center 、 Memorial Sloan Kettering Cancer Center 、和 Seattle Children ’ s Research Institute Kite 制药 2014 年 7 月 1.28 亿美元 IPO 开发 CAR-T 细胞和 TCR 工程自体 T 细胞 疗法 建于 2009 年 Juno 制药 2014 年 4 月 A 轮募资 1.76 亿美元 开发 CAR-T 细胞和 TCR 工程自体 T 细胞 疗法 由 Fred Hutchinson Cancer Research 、 Memorial Sloan-Kettering Cancer Center 、和西雅图儿童医院 的科学家组建于 2013 年 Servier/Cellectis 2014 年 2 月 Servier 支付后者 1000 万头款 和每个产品最高 1.4 亿 美元其它收益 开发靶向 CD19 和 5 个固体肿瘤的 CAR- T 细胞疗法 Bluebird Bio 2013 年 6 月 1.16 亿美元 IPO 开发 CAR-T 细胞和其它癌症基因疗法 初建于 1992 年（ Genetix Pharmaceuticals ）后改名 Bluebird Bio 赛尔基因 /Bluebird Bio2013 年 3 月 赛尔基因支付未披露的头款和 每个产品最高 2.25 亿美 元其它收益 开发抗肿瘤 CAR-T 细胞疗法 诺华 / 宾夕法尼亚大学 2012 年 8 月未披露开发抗肿瘤 CAR-T 细胞疗法 Future perspectives of CAR-T