Figure 3. High-throughput genetic screens and compound screens identify CDK4 and XPO1 as potential targets.
(a) Summary of RNAi, CRISPR-Cas9 and small-molecule screens. (b) Comparison of CLF-PED-015-T sensitivity of pan-CDK and XPO1 inhibitors with CCLE cell lines. Values are based on a robust z score. (c) shRNA (n=2) validation of dependency on CDK4. Error bars represent mean±s.d. for at least three independent experiments. (d) Effects of palbociclib on cell viability. Error bars represent mean±s.d. for four independent experiments. (e) shRNA (n=2) targeting XPO1. Error bars represent mean±s.d. for at least three independent experiments. (f) Effects of KPT-330 on cell viability. Error bars represent mean±s.d. for three independent experiments. (g) Schema depicts implantable microdevice used for in vivo assessment of drugs. (h) Sample images of tumour regions in which drugs released from the microdevices cause apoptosis as measured by cleaved caspase-3 expression (brown cells) for the indicated agents. Scale bars, 200 μm. (i) Quantitative analysis of apoptotic index for each of the tested agents. Error bars represent mean±s.d. *P value by the student's t-test=0.05. (j) Waterfall plot indicating the tumour response following treatment for mice harbouring CLF-PED-015-T subcutaneous xenografts following 25 days of treatment. Once tumours grew to 100–200 mm3, mice were treated with doxorubicin, palbociclib, KPT-330 or the combination of palbociclib and KPT-330 (see Methods section) and monitored over 25 days. *P values by the student's t-test <0.05, **P<0.005.