Depletion of creatine phosphagen energetics with a covalent creatine kinase inhibitor
Narek Darabedian1,2, Wenzhi Ji3, Mengyang Fan3
1Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA.
Abstract:
Creatine kinases (CKs) provide local ATP production in periods of elevated energetic demand, such as during rapid anabolism and growth. Thus, creatine energetics has emerged as a major metabolic liability in many rapidly proliferating cancers. Whether CKs can be targeted therapeutically is unknown because no potent or selective CK inhibitors have been developed. Here we leverage an active site cysteine present in all CK isoforms to develop a selective covalent inhibitor of creatine phosphagen energetics, CKi. Using deep chemoproteomics, we discover that CKi selectively engages the active site cysteine of CKs in cells. A co-crystal structure of CKi with creatine kinase B indicates active site inhibition that prevents bidirectional phosphotransfer. In cells, CKi and its analogs rapidly and selectively deplete creatine phosphate, and drive toxicity selectively in CK-dependent acute myeloid leukemia. Finally, we use CKi to uncover an essential role for CKs in the regulation of proinflammatory cytokine production in macrophages.
Insights
Researchers developed a novel inhibitor targeting creatine kinases (CKs), crucial for cancer cell energy. This inhibitor selectively targets CKs, showing promise for treating certain cancers and understanding immune responses.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Biology
Background:
- Creatine kinases (CKs) are vital for ATP production during high energy demands.
- Dysregulated creatine energetics represent a metabolic vulnerability in proliferating cancers.
- Lack of potent and selective CK inhibitors has hindered therapeutic targeting.
Purpose of the Study:
- To develop a selective covalent inhibitor targeting CKs.
- To investigate the therapeutic potential of CK inhibition in cancer.
- To explore the role of CKs in cellular processes beyond energy metabolism.
Main Methods:
- Leveraging an active site cysteine unique to CK isoforms for inhibitor design.
- Utilizing deep chemoproteomics to identify target engagement.
- Co-crystallography to elucidate the inhibition mechanism.
- Cellular assays to assess inhibitor efficacy and selectivity.
Main Results:
- Developed CKi, a selective covalent inhibitor of creatine phosphagen energetics.
- CKi selectively engages the active site cysteine of CKs in cellular environments.
- Co-crystal structure confirmed CKi binding and inhibition of creatine kinase B.
- CKi treatment depleted creatine phosphate and induced selective toxicity in CK-dependent acute myeloid leukemia cells.
- CKi revealed an essential role for CKs in regulating proinflammatory cytokine production in macrophages.
Conclusions:
- CKi is a potent and selective inhibitor of CKs, validating CKs as a therapeutic target.
- CK inhibition demonstrates selective anti-cancer activity in specific leukemia contexts.
- CKs play a significant role in regulating inflammatory responses in macrophages.
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