Rapidly cycling Lgr5+stem cells are exquisitely sensitive to extrinsic dietary factors that modulate colon cancer risk.
The majority of colon tumors are driven by aberrant Wnt signaling in intestinal stem cells, which mediates an efficient route toward initiating intestinal cancer. Natural lipophilic polyphenols and long-chain polyunsaturated fatty acids (PUFAs) generally suppress Wnt- and NF-κB- (nuclear factor-κ light-chain enhancer of activated B-cell) related pathways. However, the effects of these extrinsic agents on colonic leucine-rich repeat-containing G-protein-coupled receptor 5-positive (Lgr5+) stem cells, the cells of origin of colon cancer, have not been documented to date. Therefore, we examined the effect of n-3 PUFA and polyphenol (curcumin) combination on Lgr5+stem cells during tumor initiation and progression in the colon compared with an n-6 PUFA-enriched control diet. Lgr5-EGFP-IRES-creERT2knock-in mice were fed diets containing n-6 PUFA (control), n-3 PUFA, n-6 PUFA+curcumin or n-3 PUFA+curcumin for 3 weeks, followed by 6 azoxymethane (AOM) injections, and terminated 17 weeks after the last injection. To further elucidate the effects of the dietary bioactives at the tumor initiation stage, Lgr5+stem cells were also assessed at 12 and 24 h post AOM injection. Only n-3 PUFA+curcumin feeding reduced nuclear β-catenin in aberrant crypt foci (by threefold) compared with control at the progression time point. n-3 PUFA+curcumin synergistically increased targeted apoptosis in DNA-damaged Lgr5+stem cells by 4.5-fold compared with control at 12 h and maximally reduced damaged Lgr5+stem cells at 24 h, down to the level observed in saline-treated mice. Finally, RNAseq analysis indicated that p53 signaling in Lgr5+stem cells from mice exposed to AOM was uniquely upregulated only following n-3 PUFA+curcumin cotreatment. These novel findings demonstrate that Lgr5+stem cells are uniquely responsive to external dietary cues following the induction of DNA damage, providing a therapeutic strategy for eliminating damaged Lgr5+stem cells to reduce colon cancer initiation.
Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.|Department of Cellular and Molecular Medicine, Texas A&M Health Science Center, College Station, TX, USA.
Eunjoo
E
Kim
Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.|Center for Translational Environmental Health Research, Texas A&M University, College Station, TX, USA.
Laurie A
LA
Davidson
Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.|Department of Epidemiology and Biostatistics, Texas A&M Health Science Center, College Station, TX, USA.
Roger S
RS
Zoh
Department of Veterinary Pathobiology, Texas A&M University, College Station, TX, USA.
Martha E
ME
Hensel
Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.|Center for Translational Environmental Health Research, Texas A&M University, College Station, TX, USA.
Michael L
ML
Salinas
Vegetable Crop Improvement Center, Texas A&M University, College Station, TX, USA.
Bhimanagouda S
BS
Patil
Vegetable Crop Improvement Center, Texas A&M University, College Station, TX, USA.
Guddadarangavvanahally K
GK
Jayaprakasha
Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.|Center for Translational Environmental Health Research, Texas A&M University, College Station, TX, USA.
Evelyn S
ES
Callaway
Center for Translational Environmental Health Research, Texas A&M University, College Station, TX, USA.
Clinton D
CD
Allred
Center for Translational Environmental Health Research, Texas A&M University, College Station, TX, USA.|Vegetable Crop Improvement Center, Texas A&M University, College Station, TX, USA.
Nancy D
ND
Turner
Department of Veterinary Pathobiology, Texas A&M University, College Station, TX, USA.
Brad R
BR
Weeks
Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.|Center for Translational Environmental Health Research, Texas A&M University, College Station, TX, USA.|Vegetable Crop Improvement Center, Texas A&M University, College Station, TX, USA.|Department of Microbial Pathogenesis and Immunology, Texas A&M Health Science Center, College Station, TX, USA.
Robert S
RS
Chapkin
D058739:Aberrant Crypt Foci / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001397:Azoxymethane; D063646:Carcinogenesis / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002273:Carcinogens; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D018890:Chemoprevention; D003106:Colon / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology*; D003474:Curcumin / Q000494:pharmacology; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects; D015254:DNA Modification Methylases / Q000378:metabolism; D045643:DNA Repair Enzymes / Q000378:metabolism; D004032:Diet; D015525:Fatty Acids, Omega-3; D005395:Fish Oils / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D012038:Regeneration / Q000187:drug effects; D012307:Risk Factors; D015398:Signal Transduction / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
D002273:Carcinogens; D015525:Fatty Acids, Omega-3; D005395:Fish Oils; C493597:Lgr5 protein, mouse; D043562:Receptors, G-Protein-Coupled; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D051176:beta Catenin; D049452:Green Fluorescent Proteins; D015254:DNA Modification Methylases; C528331:MGMT protein, mouse; D045643:DNA Repair Enzymes; D003474:Curcumin; D001397:Azoxymethane