Personalisierte Krebsimmuntherapie mit CAR-modifizierten T-Zellen

Transcription

1 Personalisierte Krebsimmuntherapie mit CAR-modifizierten T-Zellen Neue Hightech Medizin gegen Krebs Fortschritte und Herausforderungen Thomas Nerreter, Michael Hudecek Max Eder Research Group T Cell Engineering, Department of Medicine II, University of Würzburg 11. Mai 2016

2 Cancer immunotherapy on the fast track Milestones accomplished: - anti-pd1 / anti-ctla4 check-point inhibitors mediate unspecific dis-inhibition of immune cells; clinical trials demonstrate efficacy in solid tumors (melanoma) (Topalian et al. NEJM 2012; Wolchok et al. NEJM 2013) - tumor-reactive CAR T cells induce durable complete remissions in patients with advanced chemoradiotherapy-refractory leukemia and lymphoma (Kalos et al. Science Transl. Med. 2011; Grupp et al. NEJM 2013)

3 Genetic modification of T cells with chimeric antigen receptors (CARs) CARs are synthetic receptors that bind surface molecules on tumor cells independent from HLA CAR Targeting element (scfv) Spacer Transmembrane Domain Co-stimulatory Domain (e.g. CD28 or 4-1BB) Signaling Domain CD3z

4 Genetic modification of T cells with chimeric antigen receptors (CARs) CARs are synthetic receptors that bind surface molecules on tumor cells independent from HLA Personalized immunotherapy with single infusion of patient-derived CAR T cells

5 Cancer immunotherapy on the fast track Ambition: Make CAR T cells a universal, broadly available and affordable treatment CAR clinical trials world map. Source: clinical trials.gov, May 06, 2016

6 Cancer immunotherapy on the fast track Ambition: Make CAR T cells a universal, broadly available and affordable treatment CAR clinical trials world map. Source: clinical trials.gov, May 06, 2016

7 CAR T-cell therapy emerging opportunities and challenges Ambition: Make CAR T cells a universal, broadly available and affordable treatment Current status: Variable cell products Viral gene transfer B-cell malignancies Mixed bag of random CD8 + & CD4 + T cells Expensive, small batches Novel targets needed Novel technologies and applications: Defined & consistent T-cell composition Non-viral gene transfer CARs for solid tumors

8 Physiologic course of a cellular immune response Naїve T cells (CD45RA +, CD62L + ) Effector T cells (CD45RO +, CD62L - CCR7 -, perforin hi, granzyme B hi ) Effector Memory T cells (CD45RA/RO +, CD62L - ) Central Memory T cells (CD45RO +, CD62L + ) I. Antigen challenge expansion phase 2. Antigen clearance contraction phase Apoptosis (1) After TCR engagement, naive T-cells (TN) undergo proliferation and programmed differentiation, resulting in large numbers of effector T-cells (TE) (2) After antigen clearance most TE die, leaving a small number of central and effector memory T-cells (TCM/TEM) that persist long-term and respond to antigen re-exposure

9 Do T-cells differ in their ability to persist after adoptive transfer? Persistence of TEM-derived T-cells Ancestry Fate after adoptive transfer TEM (CD45RO +, CD62L + ) Effector T-cells (CD45RO +, CD62L - ) Apoptosis Persistence of TCM-derived T-cells TCM (CD45RO +, CD62L + ) Apoptosis TCM TEM TN (CD45RA +, CD62L + )? Apoptosis Berger et al Effector T-cells derived from central memory T-cells possess an intrinsic programming that allows them to persist for extended periodes after transfer Fate of effector T-cells derived from naive T-cells not definitively established

10 What are optimal killer & helper T-cell subsets for CAR transfer? Sorting of naive, central and effector memory T cells modification with CD19-CAR Sommermeyer & Hudecek et al., Leukemia 2016 CD4 + naive + CD8 + central memory CAR T cells: max. performance in vitro and in vivo

11 CAR T cells from defined subsets: superior outcome?! CAR T cells from PBMC - random composition - total dose: 8x10 5 CAR T cells from defined subsets - 1:1 ratio of CD8T CM :CD4T N - same total dose: 8x10 5 Clinical trial at Fred Hutch, Seattle; PIs: Turtle & Maloney Turtle et al. CD 9 CA T cells of defined CD4 + :CD8 + composition in adult B cell ALL patients. J Clin Invest (Epub, doi: /jci85309) Hudecek & Sommermeyer, Leukemia 2015 Superior anti-tumor effect when defined T-cell subsets are used for CAR modification Achieve more consistent outcome, enhance the efficacy and safety of immunotherapy

12 CAR T-cell therapy emerging opportunities and challenges Ambition: Make CAR T cells a universal, broadly available and affordable treatment Current status: Variable cell products Viral gene transfer B-cell malignancies Mixed bag of random CD8 + & CD4 + T cells Expensive, small batches Novel targets needed Novel technologies and applications: Defined & consistent T-cell composition Non-viral gene transfer CARs for solid tumors

13 Non-viral CAR gene transfer: sleeping beauty transposition - cut and paste gene translocation: transposon and transposase, e.g. B X Field et al. PLoS Huang et al. Mol Ther 2008 Low gene transfer rate & high toxicity with conventional DNA plasmids

14 SB gene transfer into human T cells: minicircles vs. plasmids Gene Transfer Viability Minicircle: CD19-CAR Mock PBS MC LV Mock untreated Monjezi, Hudecek et al. submitted Superior gene transfer rate & cell viability; fully GMP-compliant & cost-effective

15 CAR T-cell therapy emerging opportunities and challenges Ambition: Make CAR T cells a universal, broadly available and affordable treatment Current status: Variable cell products Viral gene transfer B-cell malignancies Mixed bag of random CD8 + & CD4 + T cells Expensive, small batches Novel targets needed Novel technologies and applications: Defined & consistent T-cell composition Non-viral gene transfer CARs for solid tumors

16 CAR T-cell therapy is the next big thing CD19-CAR T-cells vs. acute leukemia (ALL) Toxicities include tumor lysis syndrome, cytokine storm, depletion of normal B cells

17 Targeting B-cell tumors with CD19-CARs clinical expericence UPenn (PI Grupp): 30 pediatric & adult pts r/r ALL (NEJM 2014) - 90% CR; 6-month overall survival 78%, event-free 67% - 2 pt had failed CD3x19 BiTE treatment; 15 pts had undergone prior HCT: no GvHD MSKCC (PI Davila): 16 pts r/r ALL (Science Transl. Med 2014) - 14 pts CR, incl. Ph + ALL and s.p. allo-hct - monitoring of cytokine levels/crp; short CAR T cell persistence in some pts (bridge to allo-hct) NIH (PI Kochenderfer): 15 pts DLBCL/CLL 8 CR up to 22 months follow-up, 4 PR (JCO 2014) - toxicities: fever, hypotension, reversible neurological Baylor (PI Savoldo): 6 pts r/r NHL no objective response (JCI 2011) FHCRC (PI Turtle/Maloney): >40 pts w ALL, CLL, FL, DLBCL: >90% CR (ASH 2014) - defined composition of cell product: CD8 T CM : CD4 bulk (1:1) - 2x10 5 CAR T cells/kg bw; fewer toxicities Group/Ref. scfv Spacer domain Signaling Outcome UPenn / Kalos-Grupp FMC63 CD8a Hinge+TM (48 AA) 4-1BB_CD3ζ ++ MSKCC / Davila SC25J1 CD28 extracellular (39 AA) CD28_CD3ζ ++ NIH / Kochenderfer FMC63 CD28 extracellular (39 AA) CD28_CD3ζ +/++ Baylor / Savoldo FMC63 IgG1-Fc (CH2-CH3) (217 AA) CD28_CD3ζ --- FHCRC / Turtle FMC63 IgG4-Fc (Hinge) (12 AA) 4-1BB_CD3ζ ++

18 CD19-CARs with long IgG-Fc spacer: inferior function in vivo? CD19-CAR (long IgG-Fc spacer) T cell Fc-receptor (e.g. CD64) Myeloid cell pre post T cells Hudecek et al., Cancer Immunol Res 2014 Function of CARs with long IgG-Fc spacer is restored by removal of Fc binding motifs

19 ROR1 is a novel CAR target on B-cell malignancies and epithelial cancers ROR1 is expressed on CLL, MCL, t(1;19)all but not mature normal B cells Extend applications of CAR T-cell therapy to prevalent solid tumors Candidate antigens on solid tumors: - CAIX - CEA - ErbB2/HER2 - GD2 - Mesothelin - MUC1 - PSMA -... ROR1 ROR1 ROR1 Construction of ROR1-specific CARs that confer anti-tumor reactivity in vitro/in vivo (Hudecek et al. Blood 2010, Clin Cancer Res 2013, Cancer Immunol Res. 2014)

20 Expression of ROR1 on solid tumors potential for clinical applications ROR1 is expressed by a variety of solid tumors, including triple-negative breast cancer MDA-MB-231: A) Bulk B) CD44 + /24 -/low C) GD2 + Zhang et al Hudecek et al and unpublished data ROR1 on subsets of breast, lung, renal, pancreatic cancers, sarcoma and neuroblastoma

21 Are ROR1 + epithelial cancers recognized and eliminated by ROR1-CAR T cells? R12 ROR1-CAR vs. MDA-MB-231 Triple-negative breast cancer NSG: 2x10 6 MDA-231 s.c. 5x10 6 T cells i.v. d14 Tumor engraftment R12 ROR1-CAR control T cells d42 Hudecek et al. 2013; Hudecek, unpublished data ROR1-CAR T cells confer potent reactivity against ROR1 + breast, renal and lung cancers

22 Interim conclusions Adoptive immunotherapy with CAR T cells is emerging as effective modality for cancer therapy We have established a CAR platform for applications in hematologic malignancies / solid tumors Leukemia/Solid: ROR1 Myeloma: SLAMF7, BCMA AML: nd Lymphoma: CD19 - We have developed algorithms to optimize CAR design and enhance tumor recognition - Deriving CAR T-cell products from defined subsets of CD8 + and CD4 + T cells to enhance therapeutic efficacy and safety Hudecek et al. Blood 2010, Clin Cancer Res 2013, Cancer Immunol Res 2014 and 2015, Leukemia 2015 and directions Launch clinical trials of CAR T-cell therapy in Germany Ongoing effort to establish GMP manufacturing and obtain regulatory approval Derive more detailed insight into CAR function Antigen engagement, immune synapse formation, signalling Advanced CAR T-cell engineering Non-viral delivery systems & targeted gene integration Maintain T-cell function in immunosuppressive tumor environment Smart or universal CAR T cells

23 University of Würzburg Hermann Einsele Team Medizinische Klinik II and CCC Hudecek Lab Tea Gogishvili Sabrina Prommersberger Andreas Mades Julia Wegner Sophia Danhof Tanja Stüber Julian Rydzek Razieh Monjezi Katrin Mestermann Lars Wallstabe Hardik Jetani Estefania G. Garcia Silke Frenz Elke Spirk Irene C. Cadenas Silvia Koch Tissue Engineering: Heike Walles & team Vielen Dank! Christoph Rader, Scripps Florida Johannes Huppa, Med. Uni Vienna Zoltán Ivics, Csaba Miskey, PEI Martin Schleef, Marco Schmeer, PF Halvard Bönig, BSD Frankfurt Ulf Grawunder, NBE Therapeutics Stanley R. Riddell, Fred Hutch Michael C. Jensen, eattle Children s Dirk Busch, TUM Max Eder Programm Junges Kolleg

24 Questions?!