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Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer

  • Authors:
    • Zengqin Zhan
    • Chuncha Zhan
    • Lianqiao Li
    • Dan Lan
    • Guya Meng
    • Fan Chen
  • View Affiliations / Copyright

    Affiliations: Department of Oncology, Guangzhou Development District Hospital, Guangzhou, Guangdong 510700, P.R. China, Department of General Practice, Tianyuan Community Health Service Center, Tianhe, Guangzhou, Guangdong 510000, P.R. China, Department of Oncology, People's Hospital of Sandu County, Guizhou 558100, P.R. China
    Copyright: © Zhan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 132
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    Published online on: February 10, 2026
       https://doi.org/10.3892/ol.2026.15485
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Abstract

The clinical efficacy of immune checkpoint inhibitors (ICIs) in colorectal cancer (CRC) remains limited. Modulation of the glucagon‑like peptide‑1 receptor (GLP‑1R) may enhance T‑cell‑mediated antitumor responses. The present study aimed to evaluate the antitumor effects of the GLP‑1R antagonist Exendin 9‑39 (Exe‑9) combined with anti‑programmed cell death protein‑1 (PD‑1) treatment in preclinical CRC models. Using in vitro co‑culture assays, ELISA and in vivo murine models, alongside immunohistochemical and molecular analyses of clinical samples, HT‑29 and MC38‑OVA colon cancer cell lines were co‑cultured in vitro with activated T cells in the presence of Exe‑9. In vivo, male BALB/c mice were injected with MC38 to establish a CRC model and nude mice were used to assess T‑cell dependency. To evaluate this synergistic effect, BALB/c mice with CRC were treated with Exe‑9, anti‑PD‑1 or a combination. Additionally, clinical CRC samples were analyzed to assess the association of GLP‑1R expression with the immunotherapy response. Exe‑9 significantly enhanced T‑cell‑mediated cytotoxicity in CRC cell lines and reduced tumor growth in immunocompetent CRC mice; however, this effect was not observed in nude mice. Furthermore, combination therapy with the GLP‑1R antagonist and anti‑PD‑1 yielded an improved antitumor effect compared with either treatment alone, and high GLP‑1R expression in clinical samples correlated with poor ICI response. These findings suggest that GLP‑1R antagonism potentiates T‑cell‑mediated antitumor immunity and may provide a promising adjunctive therapeutic strategy for patients with CRC when combined with ICIs in the future.
View Figures

Figure 1

Glucagon-like peptide-1 receptor
antagonist enhances T-cell-mediated cytotoxicity against colon
cancer cells. Colon cancer cell lines HT-29 and MC38-OVA were
cultured either individually or co-cultured with activated T cells
for 48 h in the presence of Exe-9 at concentrations of 10 and 50
nM. (A) Viability of HT-29 colon cancer cells cultured alone or in
co-culture with activated T cells for 48 h, treated with Exe-9 (10
or 50 nM) or control. Viability was assessed by crystal violet
staining; quantitative spectrophotometric data are shown (right
panel); (B) viability of MC38-OVA colon cancer cells under the same
culture and treatment conditions as in, (A) quantified by crystal
violet staining (right panel, spectrophotometric data); (C) IFN-γ
levels in culture supernatants from the 48-h co-cultures described
in (A) and (B), measured by ELISA; (D) TNF-α levels in the same
supernatants as in (C), determined by ELISA.*P<0.05, **P<0.01
and ***P<0.001. Exe-9, Exendin 9–39; IFN-γ, interferon-γ; TNF-α,
tumor necrosis factor-α.

Figure 2

Glucagon-like peptide-1 receptor
antagonist enhances T-cell-mediated antitumor efficacy in mice. (A)
A schematic representation of the experimental timeline. Mice were
subcutaneously injected with MC38 tumor cells to generate a
colorectal cancer model, after which Exe9 was administered
intratumorally on days 10, 12, 14 and 16. (B) Representative images
of excised tumor tissues collected on day 20. (C) Tumor volumes
were measured at regular intervals during the treatment period. (D)
Endpoint tumor volumes and (E) tumor weights are depicted to
illustrate the difference between groups. (F) Immunohistochemical
analysis was conducted to assess the infiltration of
CD8+ T cells in tumor sections (scale bar, 100 µm). (G)
In a parallel study using nude mice, tumor volumes of (H)
individual mice and (I) their average value and (J) tumor weights,
were similarly measured to determine the dependence of the
antitumor effects on T-cell-mediated immunity. Data are presented
as mean ± SEM. ***P<0.001. SC, subcutaneous; Exe-9, Exendin
9–39; i.t., intratumoral injections.

Figure 3

Glucagon-like peptide-1 receptor
antagonist induces tumor-specific CD8+ T-cell
replication. (A) Number of IFN-γ-secreting cells quantified by
ELISpot after restimulation of splenocytes from Exe-9-treated mice
with MC38 tumor cells for 20 h; (B) number of IFN-γ-secreting cells
quantified by ELISpot after restimulation of splenocytes from
Exe-9-treated mice with the MHC-I-restricted AH1 peptide for 20 h;
(C) Pearson correlation between the number of IFN-γ-secreting cells
(from MC38-restimulated group) and tumor volume; (D) Pearson
correlation between the number of IFN-γ-secreting cells (from AH1
peptide-restimulated group) and tumor volume. Data are presented as
mean ± SEM. *P<0.05. Exe-9, Exendin 9–39; sec, secreting; IFN-γ,
interferon-γ.

Figure 4

A combination of glucagon-like
peptide-1 receptor antagonist and immune checkpoint inhibitors
synergistically suppresses colorectal cancer growth in vivo.
Mice bearing MC38 tumors were administered intratumoral injections
of the glucagon-like peptide-1 receptor antagonist Exe-9 on days
10, 12, 14 and 16 or intraperitoneal injections of the anti-PD-1
antibody on days 10, 13, 16 and 19 or both treatments as the (A)
schematic representation of experimental design demonstrates. (B)
Representative excised tumor images collected on day 20
post-treatments. (C) Tumor growth curves during the treatment
period generated by regular measurements of tumor dimensions using
digital calipers. (D) Tumor volumes and (E) weights quantified at
the endpoint. Data are presented as mean ± SEM. ***P<0.001.
Exe-9, Exendin 9–39; PD-1, programmed cell death protein-1; SC,
subcutaneous; i.t., intratumoral injections; i.p.,
intraperitoneal.

Figure 5

High GLP-1R expression correlates
with reduced efficacy of anti-programmed cell death protein 1
immunotherapy in patients with CRC. Tumor specimens were collected
from 11 patients with CRC treated with immune checkpoint inhibitors
and subsequently categorized into responder (n=5) and non-responder
(n=6) based on therapeutic outcomes. (A) Quantitative polymerase
chain reaction analysis was performed to measure the mRNA
expression levels of GLP-1R in tumor tissues. (B) Patients were
further stratified into high and low receptor expression groups
based on the median expression value. Pearson's correlation
analysis was conducted to evaluate the relationship between
receptor expression and treatment responsiveness. (C)
Representative immunohistochemical images (scale bar, 100 µm) and
(D) western blotting analysis illustrates the protein expression of
GLP-1R in responders and non-responders. Data are presented as mean
± SEM. *P<0.05, **P<0.01 and ***P<0.001. GLP-1R,
glucagon-like peptide-1 receptor; CRC, colorectal cancer.
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Copy and paste a formatted citation
Spandidos Publications style
Zhan Z, Zhan C, Li L, Lan D, Meng G and Chen F: Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer. Oncol Lett 31: 132, 2026.
APA
Zhan, Z., Zhan, C., Li, L., Lan, D., Meng, G., & Chen, F. (2026). Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer. Oncology Letters, 31, 132. https://doi.org/10.3892/ol.2026.15485
MLA
Zhan, Z., Zhan, C., Li, L., Lan, D., Meng, G., Chen, F."Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer". Oncology Letters 31.4 (2026): 132.
Chicago
Zhan, Z., Zhan, C., Li, L., Lan, D., Meng, G., Chen, F."Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer". Oncology Letters 31, no. 4 (2026): 132. https://doi.org/10.3892/ol.2026.15485
Copy and paste a formatted citation
x
Spandidos Publications style
Zhan Z, Zhan C, Li L, Lan D, Meng G and Chen F: Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer. Oncol Lett 31: 132, 2026.
APA
Zhan, Z., Zhan, C., Li, L., Lan, D., Meng, G., & Chen, F. (2026). Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer. Oncology Letters, 31, 132. https://doi.org/10.3892/ol.2026.15485
MLA
Zhan, Z., Zhan, C., Li, L., Lan, D., Meng, G., Chen, F."Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer". Oncology Letters 31.4 (2026): 132.
Chicago
Zhan, Z., Zhan, C., Li, L., Lan, D., Meng, G., Chen, F."Potential treatment benefits of a GLP‑1R antagonist in combination with immune checkpoint inhibitors in colorectal cancer". Oncology Letters 31, no. 4 (2026): 132. https://doi.org/10.3892/ol.2026.15485
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