Therapeutic Targeting Of Keratin 17 Functional Domains And Nuclear Export Uncover Therapeutic Vulnerabilities Of Pancreatic Cancer.

The purpose of this study is to launch a novel biomarker-based targeted therapy that may result in improved precision and efficacy for the treatment of pancreatic ductal adenocarcinoma (PDAC). We discovered that Keratin 17 (K17), an oncofetal intermediate filament and nuclear oncoprotein expressed in 50% of PDAC cases, is a biomarker of the most aggressive and treatment-resistant form of PDAC. We set out to validate K17 as a druggable target and to identify K17-targeting mechanisms. In pre-clinical mouse models, animals bearing K17+ PDACs display the shortest survival, suggesting that K17 drives tumor aggression and should be explored as a potential therapeutic target. We used the following two approaches aimed to inhibit K17 nuclear shuttle functions, in order to identify therapeutic strategies: 1) Targeting K17 functional domains: By protein-sequence modeling analyses and site-directed mutagenesis, we identified functional domains in K17 (K17-FDs) that bind to, shift the subcellular localization of, and promote the degradation of nuclear tumor suppressors (e.g. p27). Using SILAC mass spectrometry of nuclear proteomes from isogenic human cells, we found that 80% of the nuclear proteome is altered by K17, that half of these proteins encode the domain targeted by the K17-FDs, and that a large proportion of these proteins are involved in gene expression. Animals harboring tumors with mutated K17-FDs lived two-times longer than control animals bearing K17+ PDACs, suggesting that the K17-FDs may be therapeutic targets. We are currently testing small molecule and peptide inhibitors to target the K17-FDs in vitro and in vivo, as a discovery approach to design small-molecule inhibitors of K17. 2) Targeting K17 nuclear export: K17 impacts the nuclear export of ~50% of the proteome and depends on exportin-1 (XPO1). We found that K17+ PDAC cells were more sensitive to Selective Inhibitor of Nuclear Export (SINE) therapy, leading to a 2-fold increase in cell death compared to control cells, suggesting that nuclear export by K17 in PDAC cells could be therapeutically targeted. SINE therapy is currently FDA-approved for treatment of other cancer types and repurposing for the treatment of K17-positive PDACs may result in enhanced therapeutic efficacy. In conclusion, these studies provide the basis for the development of novel biomarker-based therapeutic approaches using small-molecule inhibitors, to target K17 active sites or the pathways altered by this protein in PDAC. Citation Format: Cindy V. Leiton, Chun-Hao Pan, Ji Dong Bai, Lucia Roa-Pena, Sruthi Babu, Alex Penson, Nashaat Turkman, Richard Moffitt, Markus Seeliger, David Talmage, Kenneth R. Shroyer, Luisa F. Escobar-Hoyos. Therapeutic targeting of keratin 17 functional domains and nuclear export uncover therapeutic vulnerabilities of pancreatic cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2020 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2020;80(22 Suppl):Abstract nr PO-040.