Skip to main content

Table 3 Combinations of immunotherapies including ICB in preclinical GBM models

From: Immunotherapy for glioblastoma: the promise of combination strategies

Combination treatment Protocol Cell line and model Outcome Reference
anti-PD-1
anti-LAG-3
• Tumor implantation: 1.3 × 105 cells, striatum
• anti-PD-1: 200 μg, IP
 - on d7, d10, d12, and d14 (early schedule)
 - on d10, d12, and d14 (late schedule)
• anti-LAG-3: 200 μg, IP
 - on d7 and d10 (early schedule)
 - on d10 and d12 (late schedule)
GL261-Luc orthotopic
syngeneic
• Early treatment with anti-LAG-3 and/or anti-PD-1 significantly improved survival
• Late treatment with anti-LAG-3 did not significantly improve survival, but the combination did
• The global immunological profiles were not different between the different treatment arms
• Immune memory was established in long-term survivors
Harris-Bookman S., et al. (2018) [137]
anti-PD-1
anti-TIGIT
• Tumor implantation: 1.3 × 105 cells, striatum
• anti-PD-1: 200 μg, on d10, d12, and d14, IP
• anti-TIGIT: 200 μg, IP, every other day for a total of 5 doses starting on d8, d10, d12 or d14
(4 ≠ schedules: A, B, C, D)
GL261-Luc orthotopic
syngeneic
• Combination therapy improved long-term survival following each schedule
• Combination therapy increased immune cell tumor infiltration and cytokine production
• Tumor-infiltrating DCs were reduced following anti-TIGIT and anti-PD-1 combination treatment
• Immune memory was established in long-term survivors
Hung AL., et al. (2018) [138]
anti-CTLA-4
anti-PD-L1
1-MT
• Tumor implantation: 4 × 105 cells
• anti-CTLA-4: 100 μg loading dose followed by 3 × 50 μg maintenance doses every 3 days, IP
• anti-PD-L1: 500 μg loading dose followed by 3 × 200 μg maintenance doses every 3 days, IP
• 1-MT: in the drinking water over 30 days, starting on d7 for early blockade or on d14 for late blockade
GL261 orthotopic
syngeneic
• Early blockade with the triple combination cured 100% of mice, reduced Treg infiltration and increased IFN-γ-secreting CD8 T cell infiltration
• Late blockade prolonged survival (78% long-term survival rate) and reduced Treg infiltration but also reduced brain-infiltrating T cells
Wainwright DA., et al. (2014) [139]
anti-GITR agonist
SRS
• Tumor implantation: 1.3 × 105 cells, striatum
• SRS: 10 Gy radiation (1.9 Gy/min), d10, focal
• anti-GITR: 10 mg/kg, 3x, on d10, d13, and d16, IP
GL261-Luc orthotopic
syngeneic
• Combination therapy improved survival
• The combination increased the CD8 effector T cell/Treg ratio
• Immune memory was established in long-term survivors
Patel MA., et al. (2016) [140]
anti-PD-1
anti-OX40 agonist
GVAX (whole tumor cell vaccine)
• Tumor implantation: 7.5 × 104 cells, right striatum
• anti-PD-1: 200 μg, on d3, d6, and d9, IP
• anti-OX40: 250 μg, on d3, d6, and d9, IP
• GVAX: 1 × 106 irradiated (35 Gy) GL261-GMCSF cells, on d3, d6, and d9, SC
GL261
orthotopic
syngeneic
• The anti-PD-1 + anti-OX40 dual combination improved survival and the CD8/Treg ratio
• The anti-PD-1 + GVAX dual combination improved survival and the CD8/Treg ratio and increased brain-infiltrating CD8 T cells
• The triple combination led to 100% long-term survival with an increase in IFN-γ- and IL-2-secreting splenocytes and the CD4/CD8 ratio
• Immune memory is established in long-term survivors
Jahan N., et al. (2019) [141]
anti-PD-L1
Neoantigen peptide vaccine
• Tumor implantation: 5 × 104 cells
• anti-PD-L1: on d7, d9, and d11, IP
• Vaccine: 50 μg of each peptide + 100 μg polyIC adjuvant, on d3, d6, and d9, SC
CT2A
orthotopic
syngeneic
The combination therapy significantly improved mouse survival (60% long-term survivors) Liu CJ., et al. (2020) [142]
anti-PD-1
anti-TIM3
SRS
• Tumor implantation: 1.3 × 105 cells, left striatum
• SRS: 10 Gy radiation (1.9 Gy/min), d10, focal, using the Small Animal Radiation Research Platform (SARRP)
• anti-PD-1: 200 μg, on d10, d12, and d14
• anti-TIM-3: 250 μg, on d7, d11, and d-15
GL261-Luc orthotopic
syngeneic
• The anti-PD-1 and dual therapies improved survival and led to long-term survival
• The triple combination led to 100% remission
• The triple combination improved the immune profile of the TME and the cytokine profile of both CD4 and CD8 T cells (increased the CD8/Treg ratio, decreased the frequency of FoxP3+ Tregs, and increased the production of the inflammatory cytokines IFN-γ, TNF-α, and IL-17a)
• Immune memory was established in long-term survivors
Kim JE., et al. (2017) [143]
anti-CTLA-4
anti-4-1BB agonist
SRS
• Tumor implantation: 1.3 × 105 cells, left striatum
• SRS: 10 Gy radiation (1.9 Gy/min), d10, focal, using the SARRP
• anti-4-1BB: 200 μg 3x, on d11, d14, and d17, IP
• anti-CTLA-4: 800 μg 3x, on d11, d17, and d23, IP
GL261-Luc orthotopic
syngeneic
• The SRS + anti-CTLA-4 dual therapy prolonged survival, but only the triple combination led to long-term survival
• The triple therapy and double immunotherapy led to higher TILs (CD4 and CD8 T cells)
• Immune memory was established in long-term survivors and was glioma-specific
Belcaid Z., et al. (2014) [144]
anti-PD-1
anti-CXCR4
• Tumor implantation: 1.3 × 105 cells, left striatum
• anti-PD-1: 200 μg, on d10, d12, and d14, IP
• anti-CXCR4: 200 μg, on d10, d12, and d14, IP
GL261-Luc orthotopic
syngeneic
• The combination improved survival
• Combination therapy decreased populations of suppressive myeloid cells in the brain
• Combination therapy decreased the CD4/CD8 and Treg/CD8 ratios in the brain to a higher extent than did the monotherapies
• The combination and monotherapies increased the levels of circulating inflammatory antitumor cytokines
• Immune memory was established in long-term survivors
Wu A., et al. (2019) [145]
anti-PD-1
Antiangiogenic therapy (anti-VEGF + anti-Ang-2)
• Tumor implantation: 1 × 105 cells, striatum
• anti-PD-1: 10 mg/kg, 2x/week starting on d10 for a total of 8 doses, IP
• Antiangiogenic therapy: 25 mg/kg (anti-VEGF) and 5.6 mg/kg (anti-Ang-2), 2x/week starting on d5 until symptom apparition, SC
GL261
orthotopic
syngeneic
• The triple therapy significantly improved survival compared to antiangiogenic therapy alone
• The triple therapy increased CD8 TIL numbers and decreased MDSCs and Tregs in the brain
The antiangiogenic therapy efficacy was improved by the addition of anti-PD-1 therapy
Di Tacchio M., et al. (2019) [146]
anti-PD-1
anti-CTLA-4
G47Δ-mIL12 (= Oncolytic herpes simplex virus expressing IL-12)
• Tumor implantation: 2 × 104 cells (005 GSCs) or 1 × 104 cells (CT-2A), striatum
• anti-PD-1: 10 mg/kg, on d8, 11 and d14 (005 GSCs) or on d10, d13 and d16 (CT-2A), IP
• anti-CTLA4: 5 mg/kg, on d8, d11 and d14 (005 GSCs) or on d10, d13 and d16 (CT-2A), IP
• G47Δ-mIL12: 5 × 105 PFU, on d8 (005 GSCs) or on d10 (CT-2A), intratumoral
mouse 005 GSCs or
CT-2A
orthotopic
syngeneic
• Individual ICB only minimally extended survival in 005 GBM; combination of G47Δ-mIL12 with individual ICB modestly improved efficacy
• G47Δ-mIL12 decreased the % of 005 cells and Tregs and induced M1-like polarization in TAMs; these effects are further increased with the triple combination
• Dual ICBs significantly increased CD8 T cells in the brain, triple combination increased CD8 T cells and decreased Tregs
• Triple combination resulted in 89% or 50% long term survival (005 GSCs or CT-2A, respectively)
Immune memory was established in long-term survivors
Saha D., et al. (2017) [147]
anti-PD-1
anti-CTLA-4
anti-IL-6
anti-CD40 agonist
• Tumor implantation: 3 × 105 genetically engineered mouse tumor cells or 2 × 105 GL261 cells
• anti-PD-1: 200 μg on d9, d12, d15 and d18, IP
• anti-CTLA-4: 200 μg on d9, d12, d15 and d18, IP
• anti-IL-6: 200 μg on d9, d12, d15 and d18, IP
• anti-CD40: 100 μg on d12, IP
RCAS-genetically engineered model or GL261 orthotopic syngeneic • Dual targeting of IL-6 and CD40 sensitized GBM to ICBs; survival was significantly improved following triple combination
• Dual targeting of IL-6 and CD40 reduced tumor growth, triple combination blocked it
• All treatments induced a decrease in immunosuppressive TAM activity
• All treatments, except anti-CD40 alone, decreased the expression of immunosuppressive cytokines (IL-10, TGFβ)
• Only triple combination induced an increase in TILs and in IFN-γ-secreting CD8 T cells
Yang F. et al. (2021) [148]