WIN 2015 Symposium Radiation and immunology: a new therapeutic partnership

Transcription

1 WIN 215 Symposium Radiation and immunology: a new therapeutic partnership

2 Dr. Ralph Weichselbaum Conflict of Interest Nothing to Disclose

3 Limitations of Radiotherapy Radiotherapy cannot be given in high doses to large normal tissue volumes for very large primary tumors or widespread metastasis Normal tissue complications are common following high curative doses in some sites Need newer therapies for effective control of radioresistant histology e.g. Glioblastoma, large (T4), and micro and gross metastasis. Even with local control metastasis will defeat the purpose of treatment Immunotherapy?

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5 Courtesy of Joseph K. Salama, M.D. and Karl Farrey

6 Immunodeficiency Abrogates the Anti-tumor Effect of RT. (Blood, Vol. 114, Issue 3, , July 16, 29)

7 How Can RT Induce Antigen-specific T cells Priming? 5 d Harvest Draining Lymph Nodes, Spleen DLN SP Day 5x1 5 B16-SIY No RT Day 16 2Gy CFSE 2C transfer RT Increase antigen availability Reduced suppression Increase positive cytokines or other molecules Tolerance may be reversed? 2C CFSE

8 T cell Proliferation (cpm) Can RT increase the Cross-priming Capacity of Intratumoral DCs? Functional capacity of tumor infiltrating DCs directly ex vivo B16-SIY 14 days 2 Gy 25 2 *** days 5 CD11c from tumor 2C T cells Proliferation Assay

9 Total # CD8 Total # CD4 FoxP3- Total # CD9 Lymphocytes ] No RT ] 2 Gy Local RT Increases CD8 T cell Infiltration CD9 Lympocytes WT No RT WT RT Day 7 Post-RT Analysis of Tumor Infiltrating Lymphocytes (TIL) Local Chemokine Expression CD8 T cell CD4 (Non-Treg) WT No RT WT RT WT No RT WT RT 24hrs

10 Tumor Volume (mm 3 ) Tumor Volume (mm 3 ) Tumor Volume (mm 3 ) Type I Interferon is Essential for RT 25 WT WT RT 2 WT WT no RT IFNAR KO WT RT 15 IFNAR KO WT no RT 1 Interpretation: Type I interferon signaling on hematopoietic cells is required for tumor reduction following RT Days post tumor RT WT WT Control WT WT RT *** Days post RT WT KO Control WT KO RT *** Days post RT

11 Radiation & Interferon Induction Kim et. al. Blood 212

12 What are we actually modeling? No T cell response Generate Systemic cell Response Anti-PD-1 Anti-PD-L1 Anti-CTLA4 IR Qualitatively Enhance Existing T cell Response Anti-PD-1 Anti-PD-L1 Anti-CTLA4 Existing T cell Response Quantitativly Enhance Existing T cell Response

13 How to Improve the Radiation Response? Strategies that induce host Interferon production Immune Modulating Antibodies Anti-CTLA-4 (Ipilimumab) Anti-PD-1 / Anti-PD-L1 OX4

14 Blockade of PD-1 or CTLA-4 Signaling in Tumor Immunotherapy N Engl J Med. 212 Jun 28;366(26):

15 Immunologic Correlates of the Abscopal Effect in a Patient with Melanoma Michael A. Postow, M.D., Margaret K. Callahan, M.D., Ph.D., Christopher A. Barker, M.D., Yoshiya Yamada, M.D., Jianda Yuan, M.D., Ph.D., Shigehisa Kitano, M.D., Ph.D., Zhenyu Mu, M.D., Teresa Rasalan, B.S., Matthew Adamow, B.S., Erika Ritter, B.S., Christine Sedrak, B.S., Achim A. Jungbluth, M.D., Ramon Chua, B.S., Arvin S. Yang, M.D., Ph.D., Ruth-Ann Roman, R.N., Samuel Rosner, Brenna Benson, James P. Allison, Ph.D., Alexander M. Lesokhin, M.D., Sacha Gnjatic, Ph.D., and Jedd D. Wolchok, M.D., Ph.D. N Engl J Med 212; 366: March 8, 212

16 Case Report #1: Postow, NEJM, Gy x 3 16

17 Clinical Observations Radiation & anti-ctla-4

18 Radiation-Induced Equilibrium Is a Balance between Tumor Cell Proliferation and T Cell Mediated Killing

19 Three major outcomes after SBRT as monitored by MRI A: Early responses but relapse Hurthle cell thyroid 2 weeks before SBRT B: Cure or not 4months 9 months 1 months after SBRT (2Gyx1) Melanoma day 2 months 22 months after SBRT (2Gyx1)

20 RT-induced dominancy can be relapse for years (1) (2) (3) before SBRT Post SBRT 1 month Post SBRT 11 months (4) (5) (6) Post SBRT 25 months Post SBRT 49 months relapse Post SBRT 57 months

21 T cell Tumor Cell Interactions The specificity and magnitude of the T cell response against tumor antigens determines the outcome between tumor suppression/elimination and tumor outgrowth. Immunotherapy, or modification of existing or induced (vaccination) T cells responses, can push this interaction in the direction of tumor elimination. R D Schreiber et al. Science 211;331:

22 Tumor Volume (mm 3 ) Tumor Volume (mm 3 ) Ablative RT Can Suppress Local Tumor Regrowth and Induce Stable Disease A Tubo B B16-SIY 6 Ctrl (n=5) RT(n=23) 6 Ctrl (n=5) RT (n=29) 4 15Gy 4 25Gy Days after tumor challenge Days after tumor challenge Liang H, Deng LF et al. The Journal of Immunology (11)

23 Tumor Volume (mm 3 ) Percentage of survival cells Fraction of survival cells Radio-sensitivity of Cancer Cells was not Correlated with the Efficacy of RT 7-8 days later Remove tumor NR R collagenase, 2, 5 or 1 Gy 7 days Clonogenic assay Non-RT control 21 days post IR Partial Response (PR) 7 days R Dormant (D) A B C 7 days post RT Non-RT control Non-responsive (NR) responsive (R) Days post RT Gy 1Gy Non-RT NR R days post RT Non-RT PR D p=.57 no-ir 2Gy 5Gy 1Gy

24 Percentage of survival cells Percentage of survival cells Relative apoptosis Host immune responses, not the radiosensitivity of cancer cells, correlate with efficacy of RT 4 3 Non-RT NR R 1 8 Non-RT PR S 2.5 ** Gy 1Gy 4 2 Gy 2Gy 5Gy 1Gy.5. area CD8 area Ki67 Ki-67 TUNEL CD8 Apoptosis co-localized with CD8 cells

25 IFN cells / 2x1 5 DLNs PD-L1 Blockade Breaks the Equilibrium of Stable Disease to Favor Tumor Regression Tumor Volume (mm 3 ) Tumor-bearing (%) A 2 anti-pd-l1 B 15 1 * Ctrl RT RT anti-pd-l1 5 anti-pd-l Days post RT Days after RT C 2 15 *** ** 3T3KB (Control) 3T3NKB 1 5 * Ctrl D D PDL-1 RT

26 Infiltration of T cells: A better predictor for colorectal cancer patients survival Galon et al, Science (5795):196-4

27 PD-1/PD-L1 Checkpoint 27

28 Clinical Activity of Anti PD-1 Antibody in the Efficacy Population Objective Response Objective- Response Rate Melanoma 26/94 28% Non small-cell lung Cancer Renal-Cell Cancer 14/76 18% 9/33 27% N Engl J Med 212; 366:

29 Association between Pretreatment Tumor PD-L1 Expression and Clinical Response N Engl J Med 212; 366:

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31 Irradiation and anti PD-L1 treatment synergistically promote antitumor immunity in mice

32 % of Max % of Max % of Max % of Max % of Max % of Max Expression of PD-L1 and PD-1 in the tumor microenvironment A CD11c CD11b Gr1 CD11 F4/8 CD Iso type Non-RT RT <PE-A> PE-A <PE-A> <PE-A> PD-L1 B CD8 T CD4 T Isotype Non-RT RT <PE-A> <PE-A> PD-1

33 Tumor Volume (mm 3 ) Volume(mm 3 ) Tumor Volume (mm 3 ) Tumor Volume (mm 3 ) Anti-PD-L1 blockade synergizes with Radiation A B C TUBO 8 Ctrl PD-L1 6 RT RT PD-L1 4 2 *** D Days after tumor challenge ** MC Days after tumor challenge * *** Ctrl PDL-1 RT RT PDL Naive (n=4) RT PD-L1 (n=4) Days after tumor rechallenge Days post RT control PD-L1 RT RT PD-L1

34 Tumor Volume (mm 3 ) IFN cells / 2x15 DLNs CD8 T Cells are Required for Combination Therapy with IR and PD-L1 Blockade A 8 anti-pd-l1 Ctrl PD-L1 6 RT RT PD-L1 RT PD-L1 CD8 4 B ** ** *** 3T3KB 3T3NKB 2 * Days after tumor challenge Ctrl PD-L1 RT PD-L1 9 days post RT

35 % of CD45 cells % of CD45 cells Gr-1 <FITC-A>: Gr1 <FITC-A>: Gr1 <FITC-A>: Gr1 <FITC-A>: Gr1 IR and PD-L1 blockade induce the reduction of MDSCs A 1 5 Isotype Ctrl αpd-l1 IR IRαPD-L1 15.1% 5.71% 3.96%.27% <PerCP-Cy5-5-A>: CD11b <PerCP-Cy5-5-A>: CD11b CD11b <PerCP-Cy5-5-A>: CD11b <PerCP-Cy5-5-A>: CD11b B * ** Day 1 after IR 4 Day 3 after IR Isotype ctrl 3 PD-L1 IR IR PD-L1 2 ** 1 CD11b Gr1 CD11b *** F4/8 T CD8 T CD4 CD11b Gr1 CD11b F4/8 T CD8 T CD4

36 Gr-1 <PE-A>: Gr-1 <PE-A>: Gr-1 <PE-A>: Gr-1 % (MDSCs in CD45 TILs) IR αpd-l1 Isotype Ctrl Intercellular distance between Gr1 and CD8 T ( m) A Gr1 CD8 Merge B 8 6 Isotype Ctrl IR PD-L1 ** 4 2 Gr1 CD8α Caspase3 C Isotype Ctrl IRαPD-L1 IRαPD-L1αCD8 19.9% % % D ** ** <PE-Cy7-A>: CD11b <PE-Cy7-A>: CD11b CD11b <PE-Cy7-A>: CD11b Isotype Ctrl IR PD-L1 IR PD-L1 CD8 CD8 Figure 5

37 Tumor Volume(mm 3 ) Annexin V MDSC(%) Annexin V MDSC(%) Tumor Volume (mm 3 ) Ly-6C <FITC-A>: Ly6C <FITC-A>: Ly6C <FITC-A>: Ly6C <FITC-A>: Ly6C <FITC-A>: Ly6C <FITC-A>: Ly6C A Gr1 activated CD8 T Gr1 alone Gr1 Naïve CD8 T isotype αtnf-α αifn-γ αtnf-ααifn-γ % 12.6% 42.5% 22.8% 38.7% 2.5% <APC-A>: Annexin-V <APC-A>: Annexin-V <APC-A>: Annexin-V 1 2 Annexin-V <APC-A>: Annexin-V <APC-A>: Annexin-V <APC-A>: Annexin-V B C D Gr1 alone Gr1 naive CD8 T *** *** ***** rat IgG1 -TNF -IFNg -TNF -IFNg Gr1 activated CD8 T TNF- IFN Concentration (ng/ml) Isotype Ctrl TNFRhIgG IR PD-L1 IR PD-L1TNFRhIgG Days after tumor challenge * E 6 non-ir IR IR1A8 4 F 2 ** Days after tumor challenge Figure 6

38 % of total splenocytes % of total splenocytes 6 4 Day 1 after RT 6 4 Day 3 after RT Isotype ctrl PD-L1 IR IR PD-L1 2 2 * CD11bGr1 CD11bF4/8 CD11c B B22 T CD8 T CD4 CD11bGr1 CD11bF4/8 CD11c B B22 T CD8 T CD4 Supplementary Fig. 4

39 Schematic of proposed mechanism for tumor destruction induced by IR and PD-L1 blockade Myeloid cells B7-H1 PD-1 CD8 T cells IR TNFα Antigen B7-H1 Tumor cells Antigen MDSCs

40 Questions to be answered: What is mechanism behind the synergistic combination of PD-L1/ PD-1 blockade? Does radiation provide a window for anti-pd-l1/pd-1 axis inhibitors? What is the best sequence/timing for RT and Ab? Does RT induce PD-L1 directly? If so, what is the mechanism? Study the effect of RT and anti-pd-l1 in several different tumor models? (e.g. B16.SIY, MC38, Autochthonous Tumor Models) What changes in tumor microevironment after the combination of radiation and anti-pd-l1/pd-1? A) Immune infiltration B) cytokine/chemokine profile Is anti-pd-1 superior to anti-pd-l1 when combined with radiation?

41 The IRDS is expressed by primary breast cancer and a wide variety of other human tumors in a manner resembling Nu61/SCC61. Weichselbaum R R et al. PNAS 28;15:

42 IRDS-positive clones can be selected by tumor microenvironment due to their resistance to the host defense systems Khodarev, Roizman, Weichselbaum, Clin Cancer Res 212

43 Acknowledgements Weichselbaum Lab Radiation and Cellular Oncology Liufu Deng Hua Laura Liang Mike Beckett Helena Mauceri Byron Burnette Nikolai Khodarev Tom Darga Fu Lab. Department of Pathology/Immunology Yang-Xing Fu, M.D., Ph.D