Middle East Journal of Cancer

Modulation of CTLA-4, STAT3 and LAG-3 Expression by Ibrutinib and Idelalisib in Leukemic Cells of Patients with Chronic Lymphocytic Leukemia

Document Type : Original Article(s)

Authors

Fatemeh Mousavi-Mirkalaei 1
Saeid Taghiloo 1
Ali-Asghar Ghasemi 1
Ehsan Zaboli 2, 3
Mohammad Eslami-Jouybari 2, 3
Ramin Shekarriz 2, 3
Leila Mirzakhani 2, 3
Zohreh Ehsani 2, 3
Maryam Alizadeh-Foroutan 2, 3
Hossein Asgarian-Omran 1, 2

1 Department of Immunology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

2 Gastrointestinal Cancer Research Center, Non-Communicable Diseases Institute, Mazandaran University of Medical Sciences, Sari, Iran

3 Department of Hematology and Oncology, Imam Khomeini Hospital, Mazandaran University of Medical Science, Sari, Iran

https://doi.org/10.30476/mejc.2026.105538.2238
Abstract
Background: Small molecule inhibitors are the new therapeutic approaches for many cancers, but their exact mechanisms in tumor evasion and expansion remains unknown. The present study aimed to evaluate the expression of cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), lymphocyte-activation gene 3 (LAG-3) and signal transducers and activators of transcription 3 (STAT3) in isolated leukemic cells from patients with chronic lymphocytic leukemia (CLL) following treatment with ibrutinib and idelalisib.
Method: In this in-vitro experimental study, leukemic B cells were isolated from CLL patients, cultured and treated with ibrutinib and idelalisib for 72 hours. The optimal IC50 values of 5 and 15 µM were determined for ibrutinib and idelalisib, respectively. Following treatment, the mRNA expression of these genes was measured by real-time polymerase chain reaction assay using β-actin as a housekeeping control. One-way analysis of variance (ANOVA) test was employed for multiple comparisons. P-values of <0.05 were considered to be statistically significant.
Results: Isolation of CLL cells showed the purity of >97% as confirmed by flow cytometry. Leukemic cells indicated a significance reduction in cell viability following treatment with applied drugs compared with the untreated group. Treatment with ibrutinib showed a relative decrease in CTLA-4 (P = 0.09) and a relative increase in the LAG-3 and the STAT3 expression. Idelalisib indicated a significant increase in the STAT3 expression (P = 0.04) and also modulation in the CTLA-4 and LAG-3 expression (P > 0.05).
Conclusion: These data show that ibrutinib and idelalisib not only serve as cytotoxic drugs, but also influence the immune escape mechanisms of CLL cells by disrupting the signaling pathways which should be considered for further treatment approaches, especially for combinational strategies.

Highlights

Hossein Asgarian-Omran (google scholar)

Saeid Taghiloo (google scholar)

 

Keywords

Chronic lymphocytic leukemia
Ibrutinib
Idelalisib
CTLA-4
LAG-3
Subjects

How to cite this article:

Mousavi-Mirkalaei F, Taghiloo S, Ghasemi AA, Zaboli E, Eslami-Jouybari M, Shekarriz R, et al. Modulation of CTLA-4, STAT3 and LAG-3 expression by ibrutinib and idelalisib in leukemic cells of patients with chronic lymphocytic leukemia. Middle East J Cancer. 2026; 17(3): 197-209. doi: 10.30476/ mejc.2026.105538.2238.

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