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Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors
Published on: September 20, 2016
Cancer-associated loss-of-function mutations implicate DAPK3 as a tumor-suppressing kinase
John Brognard1, You-Wei Zhang, Lorena A Puto
1Signalling Networks in Cancer Group, Cancer Research UK, Paterson Institute for Cancer Research, The University of Manchester, Manchester, United Kingdom. jbrognard@picr.man.ac.uk
Abstract:
Cancer kinome sequencing studies have identified several protein kinases predicted to possess driver (i.e., causal) mutations. Using bioinformatic applications, we have pinpointed DAPK3 (ZIPK) as a novel cancer-associated kinase with functional mutations. Evaluation of nonsynonymous point mutations, discovered in DAPK3 in various tumors (T112M, D161N, and P216S), reveals that all three mutations decrease or abolish kinase activity. Furthermore, phenotypic assays indicate that the three mutations observed in cancer abrogate the function of the kinase to regulate both the cell cycle and cell survival. Coexpression of wild-type (WT) and cancer mutant kinases shows that the cancer mutants dominantly inhibit the function of the WT kinase. Reconstitution of a non-small cell lung cancer cell line that harbors an endogenous mutation in DAPK3 (P216S) with WT DAPK3 resulted in decreased cellular aggregation and increased sensitivity to chemotherapy. Our results suggest that DAPK3 is a tumor suppressor in which loss-of-function mutations promote increased cell survival, proliferation, cellular aggregation, and increased resistance to chemotherapy.
Insights
Death-associated protein kinase 3 (DAPK3) mutations are linked to cancer. Loss-of-function mutations in this kinase impair cell cycle regulation and promote tumor growth and chemotherapy resistance.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Cancer kinome sequencing identifies potential driver mutations in protein kinases.
- DAPK3 (ZIPK) is identified as a novel cancer-associated kinase with functional mutations.
Purpose of the Study:
- To investigate the functional impact of DAPK3 mutations found in various tumors.
- To determine the role of DAPK3 in cancer development and its potential as a tumor suppressor.
Main Methods:
- Bioinformatic analysis to identify DAPK3 mutations.
- Evaluation of kinase activity for nonsynonymous point mutations (T112M, D161N, P216S).
- Phenotypic assays to assess DAPK3 function in cell cycle and survival.
- Coexpression studies of wild-type (WT) and mutant DAPK3.
- Reconstitution experiments in non-small cell lung cancer cells.
Main Results:
- All three identified DAPK3 mutations (T112M, D161N, P216S) decrease or abolish kinase activity.
- Cancer-associated DAPK3 mutations abrogate its function in regulating cell cycle and cell survival.
- Mutant DAPK3 dominantly inhibits the function of WT DAPK3.
- Restoring WT DAPK3 in lung cancer cells with a P216S mutation reduced cellular aggregation and increased chemotherapy sensitivity.
Conclusions:
- DAPK3 functions as a tumor suppressor.
- Loss-of-function mutations in DAPK3 promote cancer progression by increasing cell survival, proliferation, and cellular aggregation.
- These mutations also contribute to increased resistance to chemotherapy, highlighting DAPK3 as a potential therapeutic target.
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