Title: A universal chemical proteomic platform to uncover the impact of dynamic protein S-acylation in cancer
Funder: Cancer Research UK

Abstract: *Background Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with a 5-year survival rate under 5%, a figure which has remained largely unchanged for 40 years, making it a high research priority for CRUK. The zDHHC class of 23 integral membrane proteins undertake regulated and reversible long-chain fatty S-acylation of thousands of cysteine residues across 10% of the mammalian proteome, with S-palmitoylation (C16) predominating. zDHHCs have been linked to high-risk cancers, including melanoma, colorectal, non-small cell lung cancer (NSCLC), and leukaemia, and our preliminary data from an in vivo functional genomic screen demonstrate an absolute requirement for zDHHC20 in PDAC metastasis to the lung. Furthermore, zDHHC20 expression is highly correlated with poor PDAC survival in the clinic, supporting the hypothesis that zDHHC20-mediated S-acylation may be a therapeutic vulnerability in PDAC. *Aims & Methods Fully characterising the role of any specific zDHHC and its protein substrates in cancer presents a formidable unsolved challenge, and although many important oncogenes are known to be S-acylated the cognate zDHHCs remain poorly defined in many cases. We will develop a new physical science-based technology platform to control the activity and identify the substrates of any specific zDHHC in a relevant cancer context, exploiting novel chemical tools coupled to advanced mass spectrometry. We will determine the role of zDHHC20 and specific zDHHC20 substrates in driving PDAC metastasis in vivo, exemplifying a universal approach for identification of zDHHC therapeutic vulnerabilities and pathway redundancies, and providing an experimental framework for future zDHHC cancer drug target validation and biomarker discovery. *How the results of this research will be used We will deliver a suite of validated chemical proteomic technologies which can be readily implemented in any cancer biology laboratory, structural understanding and chemical technology to underpin future target validation studies for zDHHCs in cancer, and important insights into the role of zDHHC20 and its bona fide substrates in PDAC metastasis. The first universal platform to systematically investigate zDHHC biology in any disease context will enable discovery of future therapeutic intervention strategies and biomarkers, and knowledge-driven selection of (combinations of) zDHHCs and zDHHC-mediated pathways for therapeutic validation.

Lay abstract: *Background Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with a 5-year survival rate under 5%, a figure which has remained largely unchanged for 40 years, making it a high research priority for CRUK. The zDHHC class of 23 integral membrane proteins undertake regulated and reversible long-chain fatty S-acylation of thousands of cysteine residues across 10% of the mammalian proteome, with S-palmitoylation (C16) predominating. zDHHCs have been linked to high-risk cancers, including melanoma, colorectal, non-small cell lung cancer (NSCLC), and leukaemia, and our preliminary data from an in vivo functional genomic screen demonstrate an absolute requirement for zDHHC20 in PDAC metastasis to the lung. Furthermore, zDHHC20 expression is highly correlated with poor PDAC survival in the clinic, supporting the hypothesis that zDHHC20-mediated S-acylation may be a therapeutic vulnerability in PDAC. *Aims & Methods Fully characterising the role of any specific zDHHC and its protein substrates in cancer presents a formidable unsolved challenge, and although many important oncogenes are known to be S-acylated the cognate zDHHCs remain poorly defined in many cases. We will develop a new physical science-based technology platform to control the activity and identify the substrates of any specific zDHHC in a relevant cancer context, exploiting novel chemical tools coupled to advanced mass spectrometry. We will determine the role of zDHHC20 and specific zDHHC20 substrates in driving PDAC metastasis in vivo, exemplifying a universal approach for identification of zDHHC therapeutic vulnerabilities and pathway redundancies, and providing an experimental framework for future zDHHC cancer drug target validation and biomarker discovery. *How the results of this research will be used We will deliver a suite of validated chemical proteomic technologies which can be readily implemented in any cancer biology laboratory, structural understanding and chemical technology to underpin future target validation studies for zDHHCs in cancer, and important insights into the role of zDHHC20 and its bona fide substrates in PDAC metastasis. The first universal platform to systematically investigate zDHHC biology in any disease context will enable discovery of future therapeutic intervention strategies and biomarkers, and knowledge-driven selection of (combinations of) zDHHCs and zDHHC-mediated pathways for therapeutic validation.

Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS