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. 2020 Jun:86:21-27.e2.
doi: 10.1016/j.exphem.2020.05.001. Epub 2020 May 8.

Cyclosporine enhances the sensitivity to lenalidomide in MDS/AML in vitro

Xiaofei He  1 , Aixia Dou  1 , Saran Feng  1 , Ashley Roman-Rivera  1 , Caleb Hawkins  1 , Lauren Lawley  1 , Jiajia Zhang  2 , Mark Wunderlich  3 , Benjamin Mizukawa  4 , Stephanie Halene  5 , Amisha Patel  5 , Jing Fang  6
Affiliations

Cyclosporine enhances the sensitivity to lenalidomide in MDS/AML in vitro

Xiaofei He et al. Exp Hematol. 2020 Jun.

Abstract

Our previous study revealed that expression of G protein-coupled receptor 68 (GPR68) was upregulated in MDSL cells, a cell line representing myelodysplastic syndromes (MDS), in response to lenalidomide (LEN), and mediated a calcium/calpain proapoptotic pathway. Isx, a GPR68 agonist, enhanced the sensitivity to LEN in MDSL cells. The fact that Isx is not a U.S. Food and Drug Administration-approved drug prompts us to look for alternative candidates that could enhance the sensitivity of LEN in MDS as well as other hematologic malignancies, such as acute myeloid leukemia (AML). In the study described here, we found that regulator of calcineurin 1 (RCAN1), an endogenous inhibitor of calcineurin (CaN), was upregulated in MDSL cells in response to LEN, possibly through degradation of IKZF1. Consistently, cyclosporin (Cys), a pharmacological inhibitor of CaN, inhibited the activity of CaN and induced apoptosis in MDSL cells, indicating that CaN provided a prosurvival signal in MDSL cells. In addition, Cys enhanced the cytotoxic effect of LEN in MDS/AML cell lines as well as primary bone marrow cells from MDS patients and AML patient-derived xenograft models. Intriguingly, pretreatment with LEN reversed the suppressive effect of Cys on T-cell activation. Our study suggests a novel mechanism of action of LEN in mediating cytotoxicity in MDS/AML via upregulation of RCAN1, thus inhibiting the CaN prosurvival pathway. Our study also suggests that Cys enhances the sensitivity to LEN in MDS/AML cells without compromising T-cell activation.

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Conflict of interest statement

Conflict of interest disclosure The authors declare no competing financial interests.

Figures

Figure 1.
Figure 1.. Depression of RCAN1 in MDSL cells in response to LEN.
(A-B) Expression of RCAN1 mRNA (A) and protein (B) in MDSL cells after treated with DMSO or 10μM LEN for 4 days (n=3). (C) Expression of RCAN1 mRNA in MDSL cells expressing shCTL or shIKZF1 (n=3). (D-E) Expression of RCAN1 mRNA (D) and protein (E) in MDSL cells expressing shCTL or shRCAN1 (n=4). (F) CaN activity in lysates of MDSL cells expressing shCTL or shRCAN1 (n=2).
Figure 2.
Figure 2.. The combined effect of LEN and Cys on MDS cells.
(A) CaN activity in lysates of MDSL cells after treated with DMSO or 10μM Cys for 4 days (n=2). (B-C) Representative flow cytometric analysis (B) and frequency (C) of Annexin V+ cells in MDSL cells after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 20μM Cys for additional 3 days (n=2). Annexin V+ cells were analyzed within CD34− cells. (D) Colonies produced by MDSL cells in methylcellulose in the presence of DMSO, 10μM LEN, or 10μM LEN plus 10μM Cys (n=3). (E-F) Representative flow cytometric analysis (E) and frequency (F) of Annexin V+ cells in BM cells from patient MDS_1 after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 10 μM Cys for additional 2 days (n=3). (G) Frequency of Annexin V+ cells in BM cells from patient MDS_2 after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 10 μM Cys for additional 2 days (n=3). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001
Figure 3.
Figure 3.. The combined effect of LEN and Cys on AML and T cells.
(A-B) Representative flow cytometric analysis (A) and frequency (B) of Annexin V+ cells in TF-1 cells after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 20μM Cys for additional 3 days (n=2). Annexin V+ cells were gated on CD34− cells. (C-D) Representative flow cytometric analysis (C) and frequency (D) of Annexin V+ cells in PDX model (PDX_1) after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 10μM Cys for additional 3 days (n=3). (E-F) Representative flow cytometric analysis (E) and frequency (F) of Annexin V+ cells in PDX model (PDX_2) after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 10μM Cys for additional 3 days (n=3). Annexin V+ cells were gated on total cells. (G-H) Representative flow cytometric analysis (G) and frequency (H) of Annexin V+ cells in PDX model (PDX_3) after treatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with control (Ctl) or 10μM Cys for additional 3 days (n=3). Annexin V+ cells were gated on total cells. (I) Measurement of splenocyte expansion via MTS after treatment with DMSO, 10μM LEN, or 10μM LEN plus 10μM Cys for 2 days (n=3~4, left panel). Measurement of splenocyte expansion via MTS after pretreatment with DMSO or 10μM LEN for 2 days, followed by concurrent treatment with 10μM Cys for additional 2 days (n=3~4, right panel). *, P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001
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References

    1. Pan B and Lentzsch S, The application and biology of immunomodulatory drugs (IMiDs) in cancer. Pharmacol Ther, 2012. 136(1): p. 56–68. - PubMed
    1. Fang M, et al., Outcome of patients with acute myeloid leukemia with monosomal karyotype who undergo hematopoietic cell transplantation. Blood, 2011. 118(6): p. 1490–4. - PMC - PubMed
    1. Cancer, et al., Pretreatment cytogenetics add to other prognostic factors predicting complete remission and long-term outcome in patients 60 years of age or older with acute myeloid leukemia: results from Cancer and Leukemia Group B 8461. Blood, 2006. 108(1): p. 63–73. - PMC - PubMed
    1. Kronke J, et al., Lenalidomide causes selective degradation of IKZF1 and IKZF3 in multiple myeloma cells. Science, 2014. 343(6168): p. 301–5. - PMC - PubMed
    1. Lu G, et al., The myeloma drug lenalidomide promotes the cereblon-dependent destruction of Ikaros proteins. Science, 2014. 343(6168): p. 305–9. - PMC - PubMed

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