Distinct BTK inhibitors differentially induce apoptosis but similarly suppress chemotaxis and lipid accumulation in mantle cell lymphoma

  • BMC Cancer. 2021 Jun 26;21(1):732. doi: 10.1186/s12885-021-08475-3.
Zhuojun Liu   #  1  2 ,  Jia Liu   #  1  2 ,  Tianming Zhang  1  2 ,  Lin Li  1  2 ,  Shuo Zhang  2 ,  Hao Jia  2 ,  Yuanshi Xia  2 ,  Mingxia Shi  3 ,  Jing Zhang  1  2 ,  Shuhua Yue  2 ,  Xiaofang Chen  4 ,  Jian Yu  5  6
Affiliations
  • 1. Interdisciplinary Institute of Cancer Diagnosis and Treatment, Beijing Advanced Innovation Center for Biomedical Engineering, Beihang University, Beihang University, Beijing, 100083, China.
  • 2. School of Biological Science and Medical Engineering, Beihang University, Beijing, 100083, China.
  • 3. Department of Hematology, the First Affiliated Hospital of Kunming Medical University, Kunming, China.
  • 4. School of Biological Science and Medical Engineering, Beihang University, Beijing, 100083, China. [email protected].
  • 5. Interdisciplinary Institute of Cancer Diagnosis and Treatment, Beijing Advanced Innovation Center for Biomedical Engineering, Beihang University, Beihang University, Beijing, 100083, China. [email protected].
  • 6. School of Biological Science and Medical Engineering, Beihang University, Beijing, 100083, China. [email protected].
  • # Contributed equally.
PMID: 34174847 DOI: 10.1186/s12885-021-08475-3
Abstract

Background: The more selective second-generation Btk inhibitors (BTKi) Acalabrutinib and Zanubrutinib and the first-generation BTKi Ibrutinib are highlighted by their clinical effectiveness in mantle cell lymphoma (MCL), however, similarities and differences of their biological and molecular effects on anti-survival of MCL cells induced by these BTKi with distinct binding selectivity against Btk remain largely unknown.

Methods: AlamarBlue assays were performed to define cytotoxicity of BTKi against MCL cells, Jeko-1 and Mino. Cleaved PARP and Caspase-3 levels were examined by immunoblot analysis to study BTKi-induced apoptotic effects. Biological effects of BTKi on MCL-cell chemotaxis and lipid droplet (LD) accumulation were examined in Jeko-1, Mino and primary MCL cells via Transwell and Stimulated Raman scattering imaging analysis respectively. Enzyme-linked immunoassays were used to determine CCL3 and CCL4 levels in MCL-cell culture supernatants. RNA-seq analyses identified BTKi targets which were validated by quantitative RT-PCR (qRT-PCR) and immunoblot analysis.

Results: Acalabrutinib and Zanubrutinib induced moderate Apoptosis in Ibrutinib high-sensitive JeKo-1 cells and Ibrutinib low-sensitive Mino cells, which was accompanied by cleaved PARP and Caspase-3. Such effects might be caused by the stronger ability of Ibrutinib to upregulate the expression of pro-apoptotic genes, such as HRK, GADD45A, and ATM, in JeKo-1 cells than in Mino cells, and the expression of such apoptotic genes was slightly changed by Acalabrutinib and Zanubrutinib in both JeKo-1 and Mino cells. Further, Acalabrutinib, Zanubrutinib and Ibrutinib reduced MCL-cell chemotaxis with similar efficiency, due to their similar abilities to downmodulate chemokines, such as CCL3 and CCL4. Also, these three BTKi similarly suppressed MCL-cell LD accumulation via downregulating lipogenic factors, DGAT2, SCD, ENPP2 and ACACA without significant differences.

Conclusion: BTKi demonstrated differential capacities to induce MCL-cell Apoptosis due to their distinct capabilities to regulate the expression of apoptosis-related genes, and similar biological and molecular inhibitory effects on MCL-cell chemotaxis and LD accumulation.

Keywords
Acalabrutinib; Apoptosis; Chemotaxis; Ibrutinib; Lipid droplet accumulation; Mantle cell lymphoma; Zanubrutinib.
Products