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A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
Published on: March 10, 2017
High-throughput screening identifies novel agents eliciting hypersensitivity in Fanconi pathway-deficient cancer
Eike Gallmeier1, Tomas Hucl, Jonathan R Brody
1The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins University, 1650 Orleans Street, Baltimore, MD 21231, USA.
Abstract:
Inactivation of the Fanconi anemia (FA) pathway occurs in diverse human tumors among the general population and renders those tumors hypersensitive to DNA interstrand-cross-linking (ICL) agents. The identification of novel agents to which FA pathway-deficient cells were hypersensitive could provide new therapeutic opportunities and improve our molecular understanding of the FA genes. Using high-throughput screening, we assessed the growth of isogenic human cancer cells that differed only in the presence or absence of single FA genes upon treatment with 880 active drugs and 40,000 diverse compounds. We identified several compounds to which FA pathway-deficient cells were more sensitive than FA pathway-proficient cells, including two groups of structurally related compounds. We further investigated the compound eliciting the strongest effect, termed 80136342. Its mechanism of action was distinct from that of ICL agents; 80136342 did not cause increased chromosomal aberrations, enhanced FANCD2 monoubiquitination, H2AX phosphorylation, p53 activation, or ICL induction. Similar to ICL agents, however, 80136342 caused a pronounced G(2) arrest in FA pathway-deficient cells. When applied in combination with ICL agents, 80136342 had at least additive toxic effects, excluding interferences on ICL-induced toxicity and facilitating a combinational application. Finally, we identified one particular methyl group necessary for the effects of 80136342 on FA-deficient cells. In conclusion, using high-throughput screening in an isogenic human FA cancer model, we explored a novel approach to identify agents eliciting hypersensitivity in FA pathway-deficient cells. We discovered several attractive candidates to serve as lead compounds for evaluating structure-activity relationships and developing therapeutics selectively targeting FA pathway-deficient tumors.
Insights
Researchers identified new drugs that kill cancer cells with a faulty Fanconi anemia (FA) pathway. This discovery offers potential new treatments for FA-deficient tumors and improves understanding of FA genes.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Fanconi anemia (FA) pathway inactivation is common in human tumors.
- FA-deficient tumors show hypersensitivity to DNA interstrand-cross-linking (ICL) agents.
- Identifying novel agents for FA-deficient cells offers therapeutic and research opportunities.
Purpose of the Study:
- To discover novel therapeutic agents targeting FA pathway-deficient cancer cells.
- To understand the molecular mechanisms of FA gene function and drug sensitivity.
- To explore new treatment strategies for FA-deficient tumors.
Main Methods:
- High-throughput screening of 880 drugs and 40,000 compounds.
- Utilized isogenic human cancer cell lines differing in FA gene status.
- Investigated the mechanism of action of a lead compound (80136342).
Main Results:
- Identified several compounds hypersensitive in FA-deficient cells.
- Compound 80136342 showed potent, distinct mechanism from ICL agents, causing G(2) arrest.
- 80136342 demonstrated additive toxicity with ICL agents and identified a key methyl group for its activity.
Conclusions:
- High-throughput screening in an isogenic FA cancer model is effective for identifying hypersensitive agents.
- Discovered promising lead compounds for developing targeted therapeutics for FA-deficient tumors.
- Novel agents like 80136342 offer new avenues for cancer therapy, particularly in combination treatments.
