Presented by Haiping Yang

1. The Clinical Research of Chimeric Antigen Receptor T-cell Immunotherapy Presented by Haiping Yang 2015.8.16

2. 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.

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

4. 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

5. TIL 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).

6. TCR 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).

7. CAR-T 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.

8. T-cell differentiation

9. Classfication of T-cell • Cytotoxic T-cell (CD8) • Helper T-cell (CD4) • Regulatory/suppressor T-cell • Memory T-cell

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

12. CAR-T 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

13. 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

14. 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 25 2011 • Grupp SA, Kalos M, Barrett D, et al. Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N Engl J Med. Apr 18 2013 • 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 20 2015 • Maude SL, Frey N, Shaw PA, et al. Chimeric antigen receptor T cells for sustained remissions inleukemia. N Engl J Med. Oct 16 2014 • 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 20 2015

15. 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 25 2011;365:725-33

16. Figure 1 Clinical Response in the Patient.

17. Figure 1 Clinical Response in the Patient.

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

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

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

21. 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.

22. 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.) * * * Challenges of CAR-T

23. Cytokine release syndrome Tumor lysis syndrome Neurologic toxicities Toxicities

24. Future perspectives of CAR-T

25. Thanks!