Some very potent chemotherapeutics used in the clinic function by stabilizing covalent protein-DNA adducts formed as obligatory intermediates in normal cellular function. Despite the demonstrated effectiveness of these drugs, there has been no effort to systematically identify all targets of drugs in this class. Our proposal describes a novel approach that combines stringent enrichment of covalent protein adducts upon drug treatment with unbiased proteomics to identify and discover drug targets. Using this approach, we have confirmed known targets of topotecan and etoposide, and we have identified novel targets of these drugs, thereby establishing the power of this approach as a discovery tool. We propose to apply this approach to identify the targets of six chemotherapeutic drugs that function by this mechanism: etoposide, doxorubicin, topotecan, decitabine (5-aza-dC), azacitidine (5-aza-C) and olaparib, and to validate these targets. Success in these experiments will identify new targets of drugs in current use, and establish a general approach for determining mechanisms of drug action to enable future identification of novel drug-target pairs.

Public Health Relevance

Some very potent chemotherapeutics trap complexes made between proteins and DNA. However, approaches to screen for new drugs and targets in this class are slow and focus on single protein targets. We have developed an approach that can be used to discover nucleic acid-protein adducts and we will use this approach to identify and validate novel targets of six chemotherapeutic drugs used in the clinic.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21CA194876-02
Application #
9031087
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Arya, Suresh
Project Start
2015-03-10
Project End
2017-02-28
Budget Start
2016-03-01
Budget End
2017-02-28
Support Year
2
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Washington
Department
Pharmacology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195