title: Combination Therapy with Anti-PD-1, Anti-TIM-3, and Focal Radiation Results in Regression of Murine Gliomas.
abstract: OBJECTIVE
  Checkpoint molecules like programmed death-1 (PD-1) and T-cell immunoglobulin mucin-3 (TIM-3) are negative immune regulators that may be upregulated in the setting of glioblastoma multiforme. Combined PD-1 blockade and stereotactic radiosurgery (SRS) have been shown to improve antitumor immunity and produce long-term survivors in a murine glioma model. However, tumor-infiltrating lymphocytes (TIL) can express multiple checkpoints, and expression of ≥2 checkpoints corresponds to a more exhausted T-cell phenotype. We investigate TIM-3 expression in a glioma model and the antitumor efficacy of TIM-3 blockade alone and in combination with anti-PD-1 and SRS.
  
  
  METHODS
  C57BL/6 mice were implanted with murine glioma cell line GL261-luc2 and randomized into 8 treatment arms: (i) control, (ii) SRS, (iii) anti-PD-1 antibody, (iv) anti-TIM-3 antibody, (v) anti-PD-1 + SRS, (vi) anti-TIM-3 + SRS, (vii) anti-PD-1 + anti-TIM-3, and (viii) anti-PD-1 + anti-TIM-3 + SRS. Survival and immune activation were assessed.
  
  
  RESULTS
  Dual therapy with anti-TIM-3 antibody + SRS or anti-TIM-3 + anti-PD-1 improved survival compared with anti-TIM-3 antibody alone. Triple therapy resulted in 100% overall survival (P < 0.05), a significant improvement compared with other arms. Long-term survivors demonstrated increased immune cell infiltration and activity and immune memory. Finally, positive staining for TIM-3 was detected in 7 of 8 human GBM samples.
  
  
  CONCLUSIONS
  This is the first preclinical investigation on the effects of dual PD-1 and TIM-3 blockade with radiation. We also demonstrate the presence of TIM-3 in human glioblastoma multiforme and provide preclinical evidence for a novel treatment combination that can potentially result in long-term glioma survival and constitutes a novel immunotherapeutic strategy for the treatment of glioblastoma multiforme. Clin Cancer Res; 23(1); 124-36. ©2016 AACR.
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Kim
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Mira A
authors.initials: MA
authors.lastname: Patel
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Antonella
authors.initials: A
authors.lastname: Mangraviti
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Eileen S
authors.initials: ES
authors.lastname: Kim
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Debebe
authors.initials: D
authors.lastname: Theodros
authors.affiliation: Department of Radiation Oncology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Esteban
authors.initials: E
authors.lastname: Velarde
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Ann
authors.initials: A
authors.lastname: Liu
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Eric W
authors.initials: EW
authors.lastname: Sankey
authors.affiliation: Flow Cytometry Core, Sidney Kimmel Comprehensive Cancer Center, Baltimore, Maryland.
authors.forename: Ada
authors.initials: A
authors.lastname: Tam
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Haiying
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Mathios
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Jackson
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Sarah
authors.initials: S
authors.lastname: Harris-Bookman
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Tomas
authors.initials: T
authors.lastname: Garzon-Muvdi
authors.affiliation: Department of Dermatology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Mary
authors.initials: M
authors.lastname: Sheu
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Allison M
authors.initials: AM
authors.lastname: Martin
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Betty M
authors.initials: BM
authors.lastname: Tyler
authors.affiliation: Department of Radiation Oncology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Phuoc T
authors.initials: PT
authors.lastname: Tran
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Xiaobu
authors.initials: X
authors.lastname: Ye
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Olivi
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Janis M
authors.initials: JM
authors.lastname: Taube
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.|Department of Pathology, Johns Hopkins University, Baltimore, Maryland.|Department of Oncology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Burger
authors.affiliation: Department of Oncology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Charles G
authors.initials: CG
authors.lastname: Drake
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Henry
authors.initials: H
authors.lastname: Brem
authors.affiliation: Department of Oncology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Drew M
authors.initials: DM
authors.lastname: Pardoll
authors.affiliation: Department of Neurosurgery, Johns Hopkins University, Baltimore, Maryland. mlim3@jhmi.edu.
authors.forename: Michael
authors.initials: M
authors.lastname: Lim
mesh_terms: D000818:Animals; D000074322:Antineoplastic Agents, Immunological / Q000494:pharmacology*; D015415:Biomarkers; D003131:Combined Modality Therapy; D004195:Disease Models, Animal; D005260:Female; D005910:Glioma / Q000276:immunology / Q000378:metabolism* / Q000473:pathology* / Q000628:therapy; D000072597:Hepatitis A Virus Cellular Receptor 2 / Q000037:antagonists & inhibitors*; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D016246:Lymphocytes, Tumor-Infiltrating / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D051379:Mice; D058990:Molecular Targeted Therapy; D061026:Programmed Cell Death 1 Receptor / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016634:Radiosurgery / Q000379:methods; D016019:Survival Analysis; D016896:Treatment Outcome; D059016:Tumor Microenvironment / Q000187:drug effects / Q000276:immunology
chemical_list: D000074322:Antineoplastic Agents, Immunological; D015415:Biomarkers; C442241:Havcr2 protein, mouse; D000072597:Hepatitis A Virus Cellular Receptor 2; D061026:Programmed Cell Death 1 Receptor