FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / DNA Repair (Amst) / A novel DDB2 mutation causes defective recognition of UV-ind...
DNA repair2020; 97; 103022; doi: 10.1016/j.dnarep.2020.103022

A novel DDB2 mutation causes defective recognition of UV-induced DNA damages and prevalent equine squamous cell carcinoma.

Abstract: Squamous cell carcinoma (SCC) occurs frequently in the human Xeroderma Pigmentosum (XP) syndrome and is characterized by deficient UV-damage repair. SCC is the most common equine ocular cancer and the only associated genetic risk factor is a UV-damage repair protein. Specifically, a missense mutation in horse DDB2 (T338M) was strongly associated with both limbal SCC and third eyelid SCC in three breeds of horses (Halflinger, Belgian, and Rocky Mountain Horses) and was hypothesized to impair binding to UV-damaged DNA. Here, we investigate DDB2-T338M mutant's capacity to recognize UV lesions in vitro and in vivo, together with human XP mutants DDB2-R273H and -K244E. We show that the recombinant DDB2-T338M assembles with DDB1, but fails to show any detectable binding to DNA substrates with or without UV lesions, due to a potential structural disruption of the rigid DNA recognition β-loop. Consistently, we demonstrate that the cellular DDB2-T338M is defective in its recruitment to focally radiated DNA damages, and in its access to chromatin. Thus, we provide direct functional evidence indicating the DDB2-T338M recapitulates molecular defects of human XP mutants, and is the causal loss-of-function allele that gives rise to equine ocular SCCs. Our findings shed new light on the mechanism of DNA recognition by UV-DDB and on the initiation of ocular malignancy.
Copyright © 2020 Elsevier B.V. All rights reserved.
Get Full Text
Publication Date: 2020-11-12 PubMed ID: 33276309PubMed Central: PMC8080515DOI: 10.1016/j.dnarep.2020.103022Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates a specific mutation in horses that impairs the ability to repair DNA damage caused by UV light, leading to a type of eye cancer called squamous cell carcinoma (SCC). The researchers compared the faulty DNA repair in horses with a similar situation in a human disease, Xeroderma Pigmentosum (XP), to better understand the underlying processes.

Research Context and Objectives

  • Previous studies established that a prevalent form of eye cancer in horses, SCC, is related to a mutation in the DDB2 gene. This mutation (T338M) interferes with the normal process of UV-damage repair, a mechanism the body uses to fix DNA damage from UV light exposure.
  • The researchers aimed to investigate in detail how this mutation affects the repair mechanism and how this leads to cancer, using both in vitro (lab conditions) and in vivo (within the organism) methods.
  • They also decided to compare observations from this horse mutation with similar known faults in human DNA repair linked to a disorder known as XP, where individuals are also prone to SCC.

Research Findings

  • Results showed that the mutated DDB2 in horses retains its ability to join up with another protein, DDB1, which is a normal step in the UV-damage repair process. However, the mutation impairs the next step – binding to the DNA that needs repair.
  • This incapacity to bind to damaged DNA was found to be a result of a structural disruption in a specific feature of the protein, named the DNA recognition β-loop.
  • Follow-up experiments demonstrated that the mutated DDB2 is not only unable to bind to the damaged DNA but also fails to reach chromosomes in the horse cells, which is another necessary step in the repair process.

Research Conclusion

  • The study provided concrete evidence that the DDB2 mutation in horses does indeed cause defects in the UV-damage repair process. As a result, it underscores the mutation as a decisive factor triggering the formation of SCC.
  • The researchers indicated that their findings represent a breakthrough in understanding the molecular defects behind the human condition, XP, and the development of SCC, pointing out similarities between the mutation in humans and horses.

Further Impact

  • The study also contributes significantly to the broader understanding of how DNA repair operates. It widens the knowledge about DNA recognition by UV-DDB (the complex involving DDB1 and DDB2 proteins) and how this recognition process, when it fails, can lead to ailments such as cancer.

Cite This Article

APA
Chen L, Bellone RR, Wang Y, Singer-Berk M, Sugasawa K, Ford JM, Artandi SE. (2020). A novel DDB2 mutation causes defective recognition of UV-induced DNA damages and prevalent equine squamous cell carcinoma. DNA Repair (Amst), 97, 103022. https://doi.org/10.1016/j.dnarep.2020.103022

Publication

DNA repair
ISSN: 1568-7856
NlmUniqueID: 101139138
Country: Netherlands
Language: English
Volume: 97
Pages: 103022
PII: S1568-7864(20)30282-2

Researcher Affiliations

Chen, Lu
  • Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: luchen2@stanford.edu.
Bellone, Rebecca R
  • Department of Population Health and Reproduction, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA; Veterinary Genetics Laboratory, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA.
Wang, Yan
  • Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA.
Singer-Berk, Moriel
  • Department of Population Health and Reproduction, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA; Veterinary Genetics Laboratory, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA.
Sugasawa, Kaoru
  • Biosignal Research Center, Kobe University, Kobe, Hyogo 657-8501, Japan.
Ford, James M
  • Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Genetics, Stanford University School of Medicine, Stanford, CA 94304, USA.
Artandi, Steven E
  • Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA.

MeSH Terms

  • Animals
  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / metabolism
  • Carcinoma, Squamous Cell / veterinary
  • DNA / metabolism
  • DNA / radiation effects
  • DNA Repair
  • DNA-Binding Proteins / metabolism
  • Eyelid Neoplasms / genetics
  • Eyelid Neoplasms / metabolism
  • Eyelid Neoplasms / veterinary
  • Horse Diseases / genetics
  • Horse Diseases / metabolism
  • Horses
  • Mutation, Missense
  • Nucleic Acid Conformation
  • Protein Binding
  • Ultraviolet Rays

Grant Funding

  • P30 CA124435 / NCI NIH HHS
  • R01 AG056575 / NIA NIH HHS
  • R35 CA197563 / NCI NIH HHS

Conflict of Interest Statement

Declaration of Competing Interest. RRB directs the Veterinary Genetics Laboratory (University of California-Davis), where a commercial test for the DDB2-T338M mutation in horses is offered.

References

This article includes 40 references
  1. Gillet LC, Schärer OD. Molecular mechanisms of mammalian global genome nucleotide excision repair.. Chem Rev 2006 Feb;106(2):253-76.
    pubmed: 16464005doi: 10.1021/cr040483fgoogle scholar: lookup
  2. Hauer MH, Gasser SM. Chromatin and nucleosome dynamics in DNA damage and repair.. Genes Dev 2017 Nov 15;31(22):2204-2221.
    pmc: PMC5769766pubmed: 29284710doi: 10.1101/gad.307702.117google scholar: lookup
  3. Hwang BJ, Chu G. Purification and characterization of a human protein that binds to damaged DNA.. Biochemistry 1993 Feb 16;32(6):1657-66.
    pubmed: 8431446doi: 10.1021/bi00057a033google scholar: lookup
  4. Tang J, Chu G. Xeroderma pigmentosum complementation group E and UV-damaged DNA-binding protein.. DNA Repair (Amst) 2002 Aug 6;1(8):601-16.
    pmc: PMC2894533pubmed: 12509284doi: 10.1016/s1568-7864(02)00052-6google scholar: lookup
  5. Scrima A, Konícková R, Czyzewski BK, Kawasaki Y, Jeffrey PD, Groisman R, Nakatani Y, Iwai S, Pavletich NP, Thomä NH. Structural basis of UV DNA-damage recognition by the DDB1-DDB2 complex.. Cell 2008 Dec 26;135(7):1213-23.
    pmc: PMC2676164pubmed: 19109893doi: 10.1016/j.cell.2008.10.045google scholar: lookup
  6. Matsumoto S, Cavadini S, Bunker RD, Grand RS, Potenza A, Rabl J, Yamamoto J, Schenk AD, Schübeler D, Iwai S, Sugasawa K, Kurumizaka H, Thomä NH. DNA damage detection in nucleosomes involves DNA register shifting.. Nature 2019 Jul;571(7763):79-84.
    pmc: PMC6611726pubmed: 31142837doi: 10.1038/s41586-019-1259-3google scholar: lookup
  7. Sugasawa K. The xeroderma pigmentosum group C protein complex and ultraviolet-damaged DNA-binding protein: functional assays for damage recognition factors involved in global genome repair.. Methods Enzymol 2006;408:171-88.
    pubmed: 16793369doi: 10.1016/S0076-6879(06)08011-6google scholar: lookup
  8. Fischer ES, Scrima A, Böhm K, Matsumoto S, Lingaraju GM, Faty M, Yasuda T, Cavadini S, Wakasugi M, Hanaoka F, Iwai S, Gut H, Sugasawa K, Thomä NH. The molecular basis of CRL4DDB2/CSA ubiquitin ligase architecture, targeting, and activation.. Cell 2011 Nov 23;147(5):1024-39.
    pubmed: 22118460doi: 10.1016/j.cell.2011.10.035google scholar: lookup
  9. Sugasawa K, Okuda Y, Saijo M, Nishi R, Matsuda N, Chu G, Mori T, Iwai S, Tanaka K, Tanaka K, Hanaoka F. UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex.. Cell 2005 May 6;121(3):387-400.
    pubmed: 15882621doi: 10.1016/j.cell.2005.02.035google scholar: lookup
  10. Fassihi H, Sethi M, Fawcett H, Wing J, Chandler N, Mohammed S, Craythorne E, Morley AM, Lim R, Turner S, Henshaw T, Garrood I, Giunti P, Hedderly T, Abiona A, Naik H, Harrop G, McGibbon D, Jaspers NG, Botta E, Nardo T, Stefanini M, Young AR, Sarkany RP, Lehmann AR. Deep phenotyping of 89 xeroderma pigmentosum patients reveals unexpected heterogeneity dependent on the precise molecular defect.. Proc Natl Acad Sci U S A 2016 Mar 1;113(9):E1236-45.
    pmc: PMC4780618pubmed: 26884178doi: 10.1073/pnas.1519444113google scholar: lookup
  11. Strafuss AC. Squamous cell carcinoma in horses.. J Am Vet Med Assoc 1976 Jan 1;168(1):61-2.
    pubmed: 1245449
  12. Lassaline M, Cranford TL, Latimer CA, Bellone RR. Limbal squamous cell carcinoma in Haflinger horses.. Vet Ophthalmol 2015 Sep;18(5):404-8.
    pubmed: 25312447doi: 10.1111/vop.12229google scholar: lookup
  13. Bellone RR, Liu J, Petersen JL, Mack M, Singer-Berk M, Drögemüller C, Malvick J, Wallner B, Brem G, Penedo MC, Lassaline M. A missense mutation in damage-specific DNA binding protein 2 is a genetic risk factor for limbal squamous cell carcinoma in horses.. Int J Cancer 2017 Jul 15;141(2):342-353.
    pubmed: 28425625doi: 10.1002/ijc.30744google scholar: lookup
  14. Singer-Berk M, Knickelbein KE, Vig S, Liu J, Bentley E, Nunnery C, Reilly C, Dwyer A, Drögemüller C, Unger L, Gerber V, Lassaline M, Bellone RR. Genetic risk for squamous cell carcinoma of the nictitating membrane parallels that of the limbus in Haflinger horses.. Anim Genet 2018 Oct;49(5):457-460.
    pubmed: 29999543doi: 10.1111/age.12695google scholar: lookup
  15. Knickelbein KE, Lassaline ME, Bellone RR. Limbal squamous cell carcinoma in a Rocky Mountain Horse: Case report and investigation of genetic contribution.. Vet Ophthalmol 2019 Mar;22(2):201-205.
    pubmed: 30238589doi: 10.1111/vop.12612google scholar: lookup
  16. Singer-Berk MH, Knickelbein KE, Lounsberry ZT, Crausaz M, Vig S, Joshi N, Britton M, Settles ML, Reilly CM, Bentley E, Nunnery C, Dwyer A, Lassaline ME, Bellone RR. Additional Evidence for DDB2 T338M as a Genetic Risk Factor for Ocular Squamous Cell Carcinoma in Horses.. Int J Genomics 2019;2019:3610965.
    pmc: PMC6766160pubmed: 31637255doi: 10.1155/2019/3610965google scholar: lookup
  17. Yeh JI, Levine AS, Du S, Chinte U, Ghodke H, Wang H, Shi H, Hsieh CL, Conway JF, Van Houten B, Rapić-Otrin V. Damaged DNA induced UV-damaged DNA-binding protein (UV-DDB) dimerization and its roles in chromatinized DNA repair.. Proc Natl Acad Sci U S A 2012 Oct 9;109(41):E2737-46.
    pmc: PMC3478663pubmed: 22822215doi: 10.1073/pnas.1110067109google scholar: lookup
  18. Fitzgerald DJ, Berger P, Schaffitzel C, Yamada K, Richmond TJ, Berger I. Protein complex expression by using multigene baculoviral vectors.. Nat Methods 2006 Dec;3(12):1021-32.
    pubmed: 17117155doi: 10.1038/nmeth983google scholar: lookup
  19. Nichols AF, Ong P, Linn S. Mutations specific to the xeroderma pigmentosum group E Ddb- phenotype.. J Biol Chem 1996 Oct 4;271(40):24317-20.
    pubmed: 8798680doi: 10.1074/jbc.271.40.24317google scholar: lookup
  20. Rapić-Otrin V, Navazza V, Nardo T, Botta E, McLenigan M, Bisi DC, Levine AS, Stefanini M. True XP group E patients have a defective UV-damaged DNA binding protein complex and mutations in DDB2 which reveal the functional domains of its p48 product.. Hum Mol Genet 2003 Jul 1;12(13):1507-22.
    pubmed: 12812979doi: 10.1093/hmg/ddg174google scholar: lookup
  21. Beecher M, Kumar N, Jang S, Rapić-Otrin V, Van Houten B. Expanding molecular roles of UV-DDB: Shining light on genome stability and cancer.. DNA Repair (Amst) 2020 Oct;94:102860.
    pmc: PMC7873659pubmed: 32739133doi: 10.1016/j.dnarep.2020.102860google scholar: lookup
  22. He YJ, McCall CM, Hu J, Zeng Y, Xiong Y. DDB1 functions as a linker to recruit receptor WD40 proteins to CUL4-ROC1 ubiquitin ligases.. Genes Dev 2006 Nov 1;20(21):2949-54.
    pmc: PMC1620025pubmed: 17079684doi: 10.1101/gad.1483206google scholar: lookup
  23. Hwang BJ, Toering S, Francke U, Chu G. p48 Activates a UV-damaged-DNA binding factor and is defective in xeroderma pigmentosum group E cells that lack binding activity.. Mol Cell Biol 1998 Jul;18(7):4391-9.
    pmc: PMC109023pubmed: 9632823doi: 10.1128/MCB.18.7.4391google scholar: lookup
  24. Wittschieben BØ, Iwai S, Wood RD. DDB1-DDB2 (xeroderma pigmentosum group E) protein complex recognizes a cyclobutane pyrimidine dimer, mismatches, apurinic/apyrimidinic sites, and compound lesions in DNA.. J Biol Chem 2005 Dec 2;280(48):39982-9.
    pubmed: 16223728doi: 10.1074/jbc.M507854200google scholar: lookup
  25. Ghodke H, Wang H, Hsieh CL, Woldemeskel S, Watkins SC, Rapić-Otrin V, Van Houten B. Single-molecule analysis reveals human UV-damaged DNA-binding protein (UV-DDB) dimerizes on DNA via multiple kinetic intermediates.. Proc Natl Acad Sci U S A 2014 May 6;111(18):E1862-71.
    pmc: PMC4020048pubmed: 24760829doi: 10.1073/pnas.1323856111google scholar: lookup
  26. Kornberg RD. The molecular basis of eukaryotic transcription.. Proc Natl Acad Sci U S A 2007 Aug 7;104(32):12955-61.
    pmc: PMC1941834pubmed: 17670940doi: 10.1073/pnas.0704138104google scholar: lookup
  27. Fitch ME, Nakajima S, Yasui A, Ford JM. In vivo recruitment of XPC to UV-induced cyclobutane pyrimidine dimers by the DDB2 gene product.. J Biol Chem 2003 Nov 21;278(47):46906-10.
    pubmed: 12944386doi: 10.1074/jbc.M307254200google scholar: lookup
  28. Katsumi S, Kobayashi N, Imoto K, Nakagawa A, Yamashina Y, Muramatsu T, Shirai T, Miyagawa S, Sugiura S, Hanaoka F, Matsunaga T, Nikaido O, Mori T. In situ visualization of ultraviolet-light-induced DNA damage repair in locally irradiated human fibroblasts.. J Invest Dermatol 2001 Nov;117(5):1156-61.
    pubmed: 11710927doi: 10.1046/j.0022-202x.2001.01540.xgoogle scholar: lookup
  29. Mori T, Nakane M, Hattori T, Matsunaga T, Ihara M, Nikaido O. Simultaneous establishment of monoclonal antibodies specific for either cyclobutane pyrimidine dimer or (6-4)photoproduct from the same mouse immunized with ultraviolet-irradiated DNA.. Photochem Photobiol 1991 Aug;54(2):225-32.
    pubmed: 1780359doi: 10.1111/j.1751-1097.1991.tb02010.xgoogle scholar: lookup
  30. Dutto I, Cazzalini O, Stivala LA, Prosperi E. An improved method for the detection of nucleotide excision repair factors at local UV DNA damage sites.. DNA Repair (Amst) 2017 Mar;51:79-84.
    pubmed: 28185850doi: 10.1016/j.dnarep.2017.01.005google scholar: lookup
  31. Guerrero-Santoro J, Levine AS, Rapić-Otrin V. Co-localization of DNA repair proteins with UV-induced DNA damage in locally irradiated cells.. Methods Mol Biol 2011;682:149-61.
    pubmed: 21057927doi: 10.1007/978-1-60327-409-8_12google scholar: lookup
  32. Scovassi AI, Prosperi E. Analysis of proliferating cell nuclear antigen (PCNA) associated with DNA.. Methods Mol Biol 2006;314:457-75.
    pubmed: 16673899doi: 10.1385/1-59259-973-7:457google scholar: lookup
  33. Rapić-Otrin V, McLenigan MP, Bisi DC, Gonzalez M, Levine AS. Sequential binding of UV DNA damage binding factor and degradation of the p48 subunit as early events after UV irradiation.. Nucleic Acids Res 2002 Jun 1;30(11):2588-98.
    pmc: PMC117178pubmed: 12034848doi: 10.1093/nar/30.11.2588google scholar: lookup
  34. Luijsterburg MS, Goedhart J, Moser J, Kool H, Geverts B, Houtsmuller AB, Mullenders LH, Vermeulen W, van Driel R. Dynamic in vivo interaction of DDB2 E3 ubiquitin ligase with UV-damaged DNA is independent of damage-recognition protein XPC.. J Cell Sci 2007 Aug 1;120(Pt 15):2706-16.
    pubmed: 17635991doi: 10.1242/jcs.008367google scholar: lookup
  35. Knickelbein KE, Lassaline ME, Singer-Berk M, Reilly CM, Clode AB, Famula TR, Michau TM, Bellone RR. A missense mutation in damage-specific DNA binding protein 2 is a genetic risk factor for ocular squamous cell carcinoma in Belgian horses.. Equine Vet J 2020 Jan;52(1):34-40.
    pubmed: 30903710doi: 10.1111/evj.13116google scholar: lookup
  36. Fitch ME, Cross IV, Ford JM. p53 responsive nucleotide excision repair gene products p48 and XPC, but not p53, localize to sites of UV-irradiation-induced DNA damage, in vivo.. Carcinogenesis 2003 May;24(5):843-50.
    pubmed: 12771027doi: 10.1093/carcin/bgg031google scholar: lookup
  37. Aboussekhra A, Biggerstaff M, Shivji MK, Vilpo JA, Moncollin V, Podust VN, Protić M, Hübscher U, Egly JM, Wood RD. Mammalian DNA nucleotide excision repair reconstituted with purified protein components.. Cell 1995 Mar 24;80(6):859-68.
    pubmed: 7697716doi: 10.1016/0092-8674(95)90289-9google scholar: lookup
  38. Fujiwara Y, Inase A, Kawasaki Y, Yoshikawa S, Iwai S. Characterization of the binding of distamycin A to damaged DNA: a comparison with the DNA recognition of the human DDB protein.. Nucleic Acids Symp Ser (Oxf) 2006;(50):235-6.
    pubmed: 17150904doi: 10.1093/nass/nrl117google scholar: lookup
  39. Walrath JC, Hawes JJ, Van Dyke T, Reilly KM. Genetically engineered mouse models in cancer research.. Adv Cancer Res 2010;106:113-64.
    pmc: PMC3533445pubmed: 20399958doi: 10.1016/S0065-230X(10)06004-5google scholar: lookup
  40. Artandi SE, DePinho RA. Telomeres and telomerase in cancer.. Carcinogenesis 2010 Jan;31(1):9-18.
    pmc: PMC3003493pubmed: 19887512doi: 10.1093/carcin/bgp268google scholar: lookup

Citations

This article has been cited 6 times.
  1. Fukuoka H, Gali HE, Bu JJ, Sella R, Afshari NA. Ultraviolet light exposure and its penetrance through the eye in a porcine model. Int J Ophthalmol 2023;16(2):172-177.
    doi: 10.18240/ijo.2023.02.02pubmed: 36816219google scholar: lookup
  2. Crausaz M, Launois T, Smith-Fleming K, McCoy AM, Knickelbein KE, Bellone RR. DDB2 Genetic Risk Factor for Ocular Squamous Cell Carcinoma Identified in Three Additional Horse Breeds. Genes (Basel) 2020 Dec 5;11(12).
    doi: 10.3390/genes11121460pubmed: 33291392google scholar: lookup
  3. Dagenais A, Juette T, Benoit-Biancamano MO, Vanore M. Reducing Recurrence in Equine Corneolimbal SCC: Outcomes of Adjunctive Cisplatin Biodegradable Bead Therapy. Vet Sci 2026 Jan 12;13(1).
    doi: 10.3390/vetsci13010076pubmed: 41600732google scholar: lookup
  4. Quatember H, Nell B, Richter B, Rigler D, Dolezal M, Sykora S, Wallner B. Studying the Impact of the DDB2 T338M Missense Mutation on the Development of Equine Squamous Cell Carcinoma and Sarcoid. Animals (Basel) 2025 Mar 22;15(7).
    doi: 10.3390/ani15070911pubmed: 40218305google scholar: lookup
  5. Truchon AR, Chase EE, Stark AR, Wilhelm SW. The diel disconnect between cell growth and division in Aureococcus is interrupted by giant virus infection. Front Microbiol 2024;15:1426193.
    doi: 10.3389/fmicb.2024.1426193pubmed: 39234538google scholar: lookup
  6. Waters KL, Spratt DE. New Discoveries on Protein Recruitment and Regulation during the Early Stages of the DNA Damage Response Pathways. Int J Mol Sci 2024 Jan 30;25(3).
    doi: 10.3390/ijms25031676pubmed: 38338953google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.