Abstract 2448: Meiosis activating sterols counteract KRas-driven epithelial carcinogenesis via an LXR-dependent mechanism

Abstract

Abstract Sterols are structural components of lipid membranes which can exert potent biological activities by regulating cell surface receptor trafficking and stability. Arrest of the sterol pathway at the level of SC4MOL (sterol C4-methyl oxydase-like) or NSDHL (NADPH-dependent steroid dehydrogenase-like) specifically antagonizes signaling and trafficking of the epidermal growth factor receptor (EGFR) (Sukhanova, 2013). SC4MOL and NSDHL catalyze two sequential steps of oxidative decarboxylation of the C4-methyl groups of meiosis activating sterols (MAS). The protein binding MAS has not been identified. We investigated whether the anti-EGFR activities of MAS metabolites are mediated via their interaction with the Liver X Receptor (LXR) so that LXR and its transcriptional targets induce subsequent deregulation of intracellular cholesterol homeostasis. In EGFR-positive carcinoma cell, inhibition of SC4MOL or NSDHL induces upregulation of two canonical LXR targets: the ATP-binding cassette transporter A1 and G1 (ABCA1 and ABCG1). In keeping with the LXR activation, expression of low density lipoprotein receptor (LDLR) is markedly reduced in SC4MOL or NSDHL-deficient cells. These effects were completely reversible with inactivation of LXR alpha, or via inactivation of an upstream enzyme, CYP51A1, which eliminated MAS. Inactivation of SC4MOL and NSDHL sterol pathway enzymes and upregulation of cholesterol efflux via ABCA1 depleted cholesterol from cellular membranes as evidenced by marked increase of nuclear form of SREBP2. Direct manipulation of SREBP, LXR or LDLR produced concordant synergistic effects with anti-EGFR drugs in carcinoma cell lines suggesting novel combinatorial strategies to treat EGFR-positive carcinomas. In in vivo experiments, SC4MOL or NSDHL-depleted A431 carcinoma xenografts showed exquisite sensitivity to cetuximab. This phenotype was associated with upregulation of LXR target ABCA1 and loss of LDLR. Conditional inactivation of a “floxed” Nsdhl allele via K14-Cre transgene triggered ABCA1 expression in the skin, arrested keratinocytes proliferation and caused lethality in newborn male pups. The anti-proliferative effect of NSDHL deficiency was tested in vivo against KRAS-driven tumors. Skin tumors induced by KRAS in transgenic LSL-KRasG12D mice are EGFR-dependent. Tamoxifen-inducible Tg(K5-CreERTam) mice were crossed with LSL-KRasG12D;NsdhlloxP/loxP and tumors induced by oral tamoxifen administration. Both, NsdhlloxP/Y (NSDHL-null) males carrying only the “floxed” allele and NsdhlloxP/+ heterozygous mosaic females formed skin tumors at similar rate. However, the growth of tumors in NSDHL-null males was completely abrogated. In sum, specific sterol metabolites play a fundamental role in regulating activity of oncogenic EGFR. Inhibition of SC4MOL or NSDHL is a highly effective strategy to counteract oncogenic signaling in carcinomas with activated EGFR-KRAS axis. Citation Format: Linara Gabitova, Andrey Gorin, Diana Restifo, Dong-Hua Yang, David Cunningham, Gail E. Herman, Igor A. Astsaturov. Meiosis activating sterols counteract KRas-driven epithelial carcinogenesis via an LXR-dependent mechanism. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2448. doi:10.1158/1538-7445.AM2014-2448