BACKGROUND: Evidence for aspirin's chemopreventative properties on colorectal cancer (CRC) is substantial, but its mechanism of action is not well-understood. We combined a proteomic approach with Mendelian randomization (MR) to identify possible new aspirin targets that decrease CRC risk. METHODS: Human colorectal adenoma cells (RG/C2) were treated with aspirin (24 hours) and a stable isotope labeling with amino acids in cell culture (SILAC) based proteomics approach identified altered protein expression. Protein quantitative trait loci (pQTLs) from INTERVAL (N = 3,301) and expression QTLs (eQTLs) from the eQTLGen Consortium (N = 31,684) were used as genetic proxies for protein and mRNA expression levels. Two-sample MR of mRNA/protein expression on CRC risk was performed using eQTL/pQTL data combined with CRC genetic summary data from the Colon Cancer Family Registry (CCFR), Colorectal Transdisciplinary (CORECT), Genetics and Epidemiology of Colorectal Cancer (GECCO) consortia and UK Biobank (55,168 cases and 65,160 controls). RESULTS: Altered expression was detected for 125/5886 proteins. Of these, aspirin decreased MCM6, RRM2, and ARFIP2 expression, and MR analysis showed that a standard deviation increase in mRNA/protein expression was associated with increased CRC risk (OR: 1.08, 95% CI, 1.03-1.13; OR: 3.33, 95% CI, 2.46-4.50; and OR: 1.15, 95% CI, 1.02-1.29, respectively). CONCLUSIONS: MCM6 and RRM2 are involved in DNA repair whereby reduced expression may lead to increased DNA aberrations and ultimately cancer cell death, whereas ARFIP2 is involved in actin cytoskeletal regulation, indicating a possible role in aspirin's reduction of metastasis. IMPACT: Our approach has shown how laboratory experiments and population-based approaches can combine to identify aspirin-targeted proteins possibly affecting CRC risk.

Behavioral and Epidemiology Research Group American Cancer Society Atlanta Georgia

Broad Institute of Harvard and MIT Cambridge Massachusetts

Cancer Epidemiology Division Cancer Council Victoria Melbourne Victoria Australia

Center for Gastrointestinal Biology and Disease University of North Carolina Chapel Hill North Carolina

Center for Public Health Genomics University of Virginia Charlottesville Virginia

Centre for Epidemiology and Biostatistics Melbourne School of Population and Global Health The University of Melbourne Melbourne Victoria Australia

Centre for Research in Biosciences The Faculty of Health and Applied Sciences The University of the West of England Bristol United Kingdom

Channing Division of Network Medicine Brigham and Women's Hospital and Harvard Medical School Boston Massachusetts

CIBER Epidemiología y Salud Pública Madrid Spain

Clalit National Cancer Control Center Haifa Israel

Clinical and Translational Epidemiology Unit Massachusetts General Hospital and Harvard Medical School Boston Massachusetts

Colorectal Oncogenomics Group Department of Clinical Pathology The University of Melbourne Parkville Victoria Australia

Department of Clinical Genetics Karolinska University Hospital Stockholm Sweden

Department of Clinical Sciences Faculty of Medicine University of Barcelona Barcelona Spain

Department of Community Medicine and Epidemiology Lady Davis Carmel Medical Center Haifa Israel

Department of Epidemiology Harvard T H Chan School of Public Health Harvard University Boston Massachusetts

Department of Epidemiology Johns Hopkins Bloomberg School of Public Health Baltimore Maryland

Department of Epidemiology School of Public Health and Institute of Health and Environment Seoul National University Seoul South Korea

Department of Epidemiology University of Washington School of Public Health Seattle Washington

Department of Family Medicine University of Virginia Charlottesville Virginia

Department of General Surgery University Hospital Rostock Rostock Germany

Department of Health Science Research Mayo Clinic Scottsdale Arizona

Department of Immunology and Infectious Diseases Harvard T H Chan School of Public Health Harvard University Boston Massachusetts

Department of Internal Medicine University of Utah Salt Lake City Utah

Department of Medicine and Epidemiology University of Pittsburgh Medical Center Pittsburgh Pennsylvania

Department of Medicine Memorial Sloan Kettering Cancer Center New York New York

Department of Medicine Samuel Oschin Comprehensive Cancer Institute Cedars Sinai Medical Center Los Angeles California

Department of Medicine University of North Carolina School of Medicine Chapel Hill North Carolina

Department of Molecular Biology of Cancer Institute of Experimental Medicine of the Czech Academy of Sciences Prague Czech Republic

Department of Molecular Medicine and Surgery Karolinska Institutet Stockholm Sweden

Department of Preventive Medicine and USC Norris Comprehensive Cancer Center Keck School of Medicine University of Southern California Los Angeles California

Department of Preventive Medicine Chonnam National University Medical School Gwangju Korea

Department of Preventive Medicine Keck School of Medicine University of Southern California Los Angeles California

Department of Public Health and Primary Care University of Cambridge Cambridge United Kingdom

Department of Radiation Sciences Oncology Unit Umeå University Umeå Sweden

Department of Surgery Chonnam National University Hwasun Hospital and Medical School Hwasun Korea

Department of Surgical Sciences Uppsala University Uppsala Sweden

Discipline of Genetics Memorial University of Newfoundland St John's Canada

Division of Cancer Epidemiology and Genetics National Cancer Institute National Institutes of Health Bethesda Maryland

Division of Cancer Epidemiology German Cancer Research Center Heidelberg Germany

Division of Clinical Epidemiology and Aging Research German Cancer Research Center Heidelberg Germany

Division of Epidemiology Department of Medicine Vanderbilt Ingram Cancer Center Vanderbilt Epidemiology Center Vanderbilt University School of Medicine Nashville Tennessee

Division of Gastroenterology Massachusetts General Hospital and Harvard Medical School Boston Massachusetts

Division of Human Genetics Department of Internal Medicine The Ohio State University Comprehensive Cancer Center Columbus Ohio

Division of Human Nutrition and Health Wageningen University and Research Wageningen the Netherlands

Division of Preventive Oncology German Cancer Research Center Heidelberg Germany

Division of Research Kaiser Permanente Northern California Oakland California

Faculty of Medicine and Biomedical Center in Pilsen Charles University Pilsen Czech Republic

Gastroenterology Department Hospital Clínic Institut d'Investigacions Biomèdiques August Pi i Sunyer University of Barcelona Barcelona Spain

Genetic Medicine and Family Cancer Clinic The Royal Melbourne Hospital Parkville Victoria Australia

German Cancer Consortium Heidelberg Germany

Huntsman Cancer Institute and Department of Population Health Sciences University of Utah Salt Lake City Utah

Institute of Biology and Medical Genetics 1st Faculty of Medicine Charles University Prague Czech Republic

Institute of Cancer Research Department of Medicine 1 Medical University Vienna Vienna Austria

Institute of Environmental Medicine Karolinska Institutet Stockholm Sweden

Institute of Medical Biometry and Informatics University of Heidelberg Heidelberg Germany

Jeonnam Regional Cancer Center Chonnam National University Hwasun Hospital Hwasun Korea

Leeds Institute of Cancer and Pathology University of Leeds Leeds United Kingdom

Lunenfeld Tanenbaum Research Institute Mount Sinai Hospital University of Toronto Toronto Ontario Canada

Massachusetts General Hospital and Harvard Medical School Clinical and Translational Epidemiology Unit Boston Massachusetts

Medical Research Council Integrative Epidemiology Unit Bristol Medical School University of Bristol Bristol United Kingdom

Nutrition and Metabolism Section International Agency for Research on Cancer World Health Organization Lyon France

ONCOBEL Program Bellvitge Biomedical Research Institute L'Hospitalet de Llobregat Barcelona Spain

Oncology Data Analytics Program Catalan Institute of Oncology IDIBELL L'Hospitalet de Llobregat Barcelona Spain

Precision Medicine School of Clinical Sciences at Monash Health Monash University Clayton Victoria Australia

Proteomics Facility Faculty of Life Sciences University of Bristol Bristol United Kingdom

Public Health Sciences Division Fred Hutchinson Cancer Research Center Seattle Washington

Ruth and Bruce Rappaport Faculty of Medicine Technion Israel Institute of Technology Haifa Israel

School of Cellular and Molecular Medicine University of Bristol Bristol United Kingdom

School of Public Health University of Washington Seattle Washington

University Medical Centre Hamburg Eppendorf University Cancer Centre Hamburg Hamburg Germany

University of Hawaii Cancer Center Honolulu Hawaii

University of Melbourne Centre for Cancer Research Victorian Comprehensive Cancer Centre Parkville Victoria Australia

University of Southern California Preventative Medicine Los Angeles California

Wallenberg Centre for Molecular Medicine Umeå University Umeå Sweden

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