ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2
KD025, a ROCK2 isoform-specific inhibitor, has an anti-adipogenic activity which is not mediated by ROCK2 inhibition. To identify the target, we searched binding targets of KD025 by using the KINOMEscan<sup>TM</sup> screening platform, and we identified casein kinase 2 (CK2) as a novel t...
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MDPI AG
2021-08-01
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| Online Access: | https://www.mdpi.com/1420-3049/26/16/4747 |
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| author | Nhu Nguyen Quynh Tran Kwang-Hoon Chun |
| author_facet | Nhu Nguyen Quynh Tran Kwang-Hoon Chun |
| author_sort | Nhu Nguyen Quynh Tran |
| collection | DOAJ |
| description | KD025, a ROCK2 isoform-specific inhibitor, has an anti-adipogenic activity which is not mediated by ROCK2 inhibition. To identify the target, we searched binding targets of KD025 by using the KINOMEscan<sup>TM</sup> screening platform, and we identified casein kinase 2 (CK2) as a novel target. KD025 showed comparable binding affinity to CK2α (<i>K</i><sub>d</sub> = 128 nM). By contrast, CK2 inhibitor CX-4945 and ROCK inhibitor fasudil did not show such cross-reactivity. In addition, KD025 effectively inhibited CK2 at a nanomolar concentration (IC<sub>50</sub> = 50 nM). We examined if the inhibitory effect of KD025 on adipocyte differentiation is through the inhibition of CK2. Both CX-4945 and KD025 suppressed the generation of lipid droplets and the expression of proadipogenic genes <i>Pparg</i> and <i>Cebpa</i> in 3T3-L1 cells during adipocyte differentiation. Fasudil exerted no significant effect on the quantity of lipid droplets, but another ROCK inhibitor Y-27632 increased the expression of <i>Pparg</i> and <i>Cebpa</i>. Both CX-4945 and KD025 acted specifically in the middle stage (days 1–3) but were ineffective when treated at days 0–1 or the late stages, indicating that CX-4945 and KD025 may regulate the same target, CK2. The mRNA and protein levels of CK2α and CK2β generally decreased in 3T3-L1 cells at day 2 but recovered thereafter. Other well-known CK2 inhibitors DMAT and quinalizarin inhibited effectively the differentiation of 3T3-L1 cells. Taken together, the results of this study confirmed that KD025 inhibits ROCK2 and CK2, and that the inhibitory effect on adipocyte differentiation is through the inhibition of CK2. |
| first_indexed | 2024-03-10T08:33:00Z |
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| language | English |
| last_indexed | 2024-03-10T08:33:00Z |
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| spelling | doaj.art-3ca41d4db52349b181cb1ee0d0111c412023-11-22T08:51:20ZengMDPI AGMolecules1420-30492021-08-012616474710.3390/molecules26164747ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2Nhu Nguyen Quynh Tran0Kwang-Hoon Chun1Gachon Institute of Pharmaceutical Sciences, College of Pharmacy, Gachon University, Incheon 21936, KoreaGachon Institute of Pharmaceutical Sciences, College of Pharmacy, Gachon University, Incheon 21936, KoreaKD025, a ROCK2 isoform-specific inhibitor, has an anti-adipogenic activity which is not mediated by ROCK2 inhibition. To identify the target, we searched binding targets of KD025 by using the KINOMEscan<sup>TM</sup> screening platform, and we identified casein kinase 2 (CK2) as a novel target. KD025 showed comparable binding affinity to CK2α (<i>K</i><sub>d</sub> = 128 nM). By contrast, CK2 inhibitor CX-4945 and ROCK inhibitor fasudil did not show such cross-reactivity. In addition, KD025 effectively inhibited CK2 at a nanomolar concentration (IC<sub>50</sub> = 50 nM). We examined if the inhibitory effect of KD025 on adipocyte differentiation is through the inhibition of CK2. Both CX-4945 and KD025 suppressed the generation of lipid droplets and the expression of proadipogenic genes <i>Pparg</i> and <i>Cebpa</i> in 3T3-L1 cells during adipocyte differentiation. Fasudil exerted no significant effect on the quantity of lipid droplets, but another ROCK inhibitor Y-27632 increased the expression of <i>Pparg</i> and <i>Cebpa</i>. Both CX-4945 and KD025 acted specifically in the middle stage (days 1–3) but were ineffective when treated at days 0–1 or the late stages, indicating that CX-4945 and KD025 may regulate the same target, CK2. The mRNA and protein levels of CK2α and CK2β generally decreased in 3T3-L1 cells at day 2 but recovered thereafter. Other well-known CK2 inhibitors DMAT and quinalizarin inhibited effectively the differentiation of 3T3-L1 cells. Taken together, the results of this study confirmed that KD025 inhibits ROCK2 and CK2, and that the inhibitory effect on adipocyte differentiation is through the inhibition of CK2.https://www.mdpi.com/1420-3049/26/16/4747KD025belumosudilROCK2casein kinaseCK2adipocyte differentiation |
| spellingShingle | Nhu Nguyen Quynh Tran Kwang-Hoon Chun ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2 Molecules KD025 belumosudil ROCK2 casein kinase CK2 adipocyte differentiation |
| title | ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2 |
| title_full | ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2 |
| title_fullStr | ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2 |
| title_full_unstemmed | ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2 |
| title_short | ROCK2-Specific Inhibitor KD025 Suppresses Adipocyte Differentiation by Inhibiting Casein Kinase 2 |
| title_sort | rock2 specific inhibitor kd025 suppresses adipocyte differentiation by inhibiting casein kinase 2 |
| topic | KD025 belumosudil ROCK2 casein kinase CK2 adipocyte differentiation |
| url | https://www.mdpi.com/1420-3049/26/16/4747 |
| work_keys_str_mv | AT nhunguyenquynhtran rock2specificinhibitorkd025suppressesadipocytedifferentiationbyinhibitingcaseinkinase2 AT kwanghoonchun rock2specificinhibitorkd025suppressesadipocytedifferentiationbyinhibitingcaseinkinase2 |