Targeting genetic alterations of oncogenes by moleculartargeted agents (MTA) is an effective approach for treating cancer. However, there are still no clinical MTA options for many cancers, including esophageal cancer. We used a short hairpin RNA library to screen for a new oncogene in the esophageal cancer cell line KYSE70 and identified YES proto-oncogene 1 (YES1) as having a significant impact on tumor growth. An analysis of clinical samples showed that YES1 gene amplification existed not only in esophageal cancer but also in lung, head and neck, bladder, and other cancers, indicating that YES1 would be an attractive target for a cancer drug. Because there is no effective YES1 inhibitor so far, we generated a YES1 kinase inhibitor, CH6953755. YES1 kinase inhibition by CH6953755 led to antitumor activity against YES1-amplified cancers in vitro and in vivo. Yes-associated protein 1 (YAP1) played a role downstream of YES1 and contributed to the growth of YES1amplified cancers. YES1 regulated YAP1 transcription activity by controlling its nuclear translocation and serine phosphorylation. These findings indicate that the regulation of YAP1 by YES1 plays an important role in YES1-amplified cancers and that CH6953755 has therapeutic potential in such cancers. Significance: These findings identify the SRC family kinase YES1 as a targetable oncogene in esophageal cancer and describe a new inhibitor for YES1 that has potential for clinical utility. See related commentary by Rai, p. 5702

In the course of screening for a new type of androgen receptor (AR) antagonist, we isolated a novel compound, arabilin, with two structural isomers, spectinabilin and SNF4435C, produced by Streptomyces sp. MK756-CF1. Structure elucidation on the basis of the spectroscopic properties showed that arabilin is a novel polypropionate-derived metabolite with a p-nitrophenyl group and a substituted c-pyrone ring. Arabilin competitively blocked the binding of androgen to the ligand-binding domain of AR in vitro. In addition, arabilin inhibited androgen-induced prostate-specific antigen mRNA expression in prostate cancer LNCaP cells.

<p>Anti-proliferative activity of various MTAs against 66 cancer cell lines.</p>

<p>Supplementary Figure S1. YES1 gene is amplified and YES1 expression is related to poor prognosis in esophageal cancer and lung cancer in clinical. Supplementary Figure S2. YES1 gene amplification including the upregulation of YES1 expression. Supplementary Figure S3. Cell growth inhibition profile of various molecular targeted agents. Supplementary Figure S4. Antiproliferative activity of dasatinib against cancer cell lines expressing YES1-WT or YES1-GK. Supplementary Figure S5. The contribution of SFK other than YES1 in YES1-amplified cancer cell lines. Supplementary Figure S6. The antitumor activity of dasatinib against YES1-amplified xenograft tumor. Supplementary Figure S7. Dasatinib inhibited YAP1 downstream signal via YES1 kinase inhibition against YES1-amplified cancer cell. Supplementary Figure S8. YAP1 and DAPI staining in treatment with CH6953755. Supplementary Figure S9. YAP1 localization change in xenograft tumor tissue by CH6953755 treatment.</p>

<p>Supplementary materials and methods</p>

<div>Abstract<p>Targeting genetic alterations of oncogenes by molecular-targeted agents (MTA) is an effective approach for treating cancer. However, there are still no clinical MTA options for many cancers, including esophageal cancer. We used a short hairpin RNA library to screen for a new oncogene in the esophageal cancer cell line KYSE70 and identified YES proto-oncogene 1 (<i>YES1</i>) as having a significant impact on tumor growth. An analysis of clinical samples showed that <i>YES1</i> gene amplification existed not only in esophageal cancer but also in lung, head and neck, bladder, and other cancers, indicating that YES1 would be an attractive target for a cancer drug. Because there is no effective YES1 inhibitor so far, we generated a YES1 kinase inhibitor, CH6953755. YES1 kinase inhibition by CH6953755 led to antitumor activity against <i>YES1</i>-amplified cancers <i>in vitro</i> and <i>in vivo</i>. Yes-associated protein 1 (YAP1) played a role downstream of YES1 and contributed to the growth of <i>YES1</i>-amplified cancers. YES1 regulated YAP1 transcription activity by controlling its nuclear translocation and serine phosphorylation. These findings indicate that the regulation of YAP1 by YES1 plays an important role in <i>YES1</i>-amplified cancers and that CH6953755 has therapeutic potential in such cancers.</p>Significance:<p>These findings identify the SRC family kinase YES1 as a targetable oncogene in esophageal cancer and describe a new inhibitor for YES1 that has potential for clinical utility.</p><p><i>See related commentary by Rai, p. 5702</i></p></div>

<p>Anti-proliferative activity against KYSE70 expressing mock, YES1-WT and SRC-WT</p>

<p>Kinase selectivity profile of CH6953755. The values of % competition at 0.01 or 0.1 μmol/L CH6953755 in DiscoveRx's KINOMEscan for 456 kinases including mutated kinases.</p>