Equine induced pluripotent stem cells are responsive to inflammatory cytokines before and after differentiation into musculoskeletal cell types.
Abstract: Persistent inflammation is associated with the poor regeneration of musculoskeletal tissues. Embryonic stem cells (ESCs) have an attenuated response to inflammatory cytokines, but there are mixed reports on the response of induced pluripotent stem cells (iPSCs) to inflammation. Horses provide a relevant large animal model for studying musculoskeletal tissue diseases and the testing of novel therapies. The aim of this study was to determine if equine iPSCs are responsive to the inflammatory cytokines IL-1β, TNFα and IFN-γ in their undifferentiated state, or following differentiation into tendon and cartilage-like cells. We demonstrated that in undifferentiated iPSCs, the cytokines induce NF-κB P65 and STAT1 nuclear translocation which leads to cell death, decreased OCT4 expression and increased expression of inflammatory genes. Following differentiation towards cartilage-like cells exposure to the cytokines resulted in STAT1 nuclear translocation, changes in cartilage gene expression and increased expression of matrix metalloproteinases (MMPs) and inflammatory genes. Exposure of iPSC-derived tendon-like cells to the cytokines resulted nuclear translocation of NF-κB P65 and STAT1, altered tendon gene expression, increased MMP expression and increased expression of inflammatory genes. Equine iPSCs are therefore capable of responding to inflammatory stimulation and this may have relevance for their future clinical application.
© 2023. The Author(s).
Publication Date: 2023-08-15 PubMed ID: 37582999PubMed Central: 2195860DOI: 10.1007/s11626-023-00800-3Google 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.
- Journal Article
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 study reveals that horse-derived pluripotent stem cells are sensitive to inflammation both in their raw and derived state into musculoskeletal cell types. This becomes significant in the context of inflammatory diseases affecting musculoskeletal structures and could provide important insights for future regenerative therapies.
Research Purpose and Context
- The article presents research in the field of regenerative medicine, specifically focusing on induced pluripotent stem cells (iPSCs) derived from horses. Horses are considered a useful large animal model in studying musculoskeletal disorders and related therapies.
- Persistent inflammation is known to hinder the regeneration of musculoskeletal tissues, and the researchers aim to study the response of iPSCs to inflammatory cytokines before and after these cells have differentiated into tendon and cartilage-like cells.
Methodology
- The research focuses on how equine iPSCs respond to pro-inflammatory cytokines, specifically IL-1β, TNFα, and IFN-γ.
- These interactions were studied in their undifferentiated state (raw stem cells) and in differentiated states as tendon and cartilage-like cells.
- The induced changes including nuclear translocation of NF-κB P65 and STAT1, cell death, altered gene expression, and increased expression of matrix metalloproteinases (MMPs) and inflammatory genes were also examined.
Key Findings
- The researchers found that these inflammatory cytokines induce changes in undifferentiated iPSCs, marked by nuclear translocation of NF-κB P65 and STAT1, leading to cell death, decreased OCT4 expression, and heightened expression of inflammatory genes.
- When iPSCs were differentiated into cartilage-like cells and exposed to these cytokines, the observed changes included STAT1 nuclear translocation and altered cartilage gene and MMP expression.
- The same cytokines induced the nuclear translocation of NF-κB P65 and STAT1 and increased inflammatory gene expression in the iPSC-derived tendon-like cells.
Implications of the Research
- The results demonstrate that equine iPSCs, in both their undifferentiated state and when differentiated into musculoskeletal cell types, are capable of responding to inflammatory cytokines.
- This has potential implications for future use of these cells in a clinical setting, particularly in the treatment of inflammatory-related musculoskeletal disorders, as these cells can react and potentially exacerbate the inflammatory condition.
- Moving forward, this could influence how stem cells are used in regenerative therapy and may pave the way for developing measures to modify such responses to inflammation.
Cite This Article
APA
Palomino Lago E, Jelbert ER, Baird A, Lam PY, Guest DJ.
(2023).
Equine induced pluripotent stem cells are responsive to inflammatory cytokines before and after differentiation into musculoskeletal cell types.
In Vitro Cell Dev Biol Anim.
https://doi.org/10.1007/s11626-023-00800-3 Publication
Researcher Affiliations
- Department of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, AL9 7TA, Herts, UK.
- Department of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, AL9 7TA, Herts, UK.
- Animal Health Trust, Lanwades Park, Kentford, Newmarket, CB8 7UU, UK.
- Department of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, AL9 7TA, Herts, UK.
- Department of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, AL9 7TA, Herts, UK. djguest@rvc.ac.uk.
Grant Funding
- S21-1022-1061 / Petplan Charitable Trust
- vet/prj/792 / Horserace Betting Levy Board
References
This article includes 60 references
- Allen M, Svensson L, Roach M, Hambor J, McNeish J, Gabel CA. Deficiency of the stress kinase p38alpha results in embryonic lethality: characterization of the kinase dependence of stress responses of enzyme-deficient embryonic stem cells.. J Exp Med 191:859–870.
- Avella CS, Ely ER, Verheyen KL, Price JS, Wood JL, Smith RK. Ultrasonographic assessment of the superficial digital flexor tendons of National Hunt racehorses in training over two racing seasons.. Equine Vet J 41:449–454.
- Badley EM, Rasooly I, Webster GK. Relative importance of musculoskeletal disorders as a cause of chronic health problems, disability, and health care utilization: findings from the 1990 Ontario Health Survey.. J Rheumatol 21:505–514.
- Baird A, Dominguez Falcon N, Saeed A, Guest DJ. Biocompatible Three-Dimensional Printed Thermoplastic Scaffold for Osteoblast Differentiation of Equine Induced Pluripotent Stem Cells.. Tissue Eng Part C Methods 25:253–261.
- Baird A, Lindsay T, Everett A, Iyemere V, Paterson YZ, McClellan A, Henson FMD, Guest DJ. Osteoblast differentiation of equine induced pluripotent stem cells.. Biol Open 7:bio033514.
- Bavin EP, Smith O, Baird AE, Smith LC, Guest DJ. Equine Induced Pluripotent Stem Cells have a Reduced Tendon Differentiation Capacity Compared to Embryonic Stem Cells.. Front Vet Sci 2:55.
- Bertuglia A, Pagliara E, Grego E, Ricci A, Brkljaca-Bottegaro N. Pro-inflammatory cytokines and structural biomarkers are effective to categorize osteoarthritis phenotype and progression in Standardbred racehorses over five years of racing career.. BMC Vet Res 12:246.
- Chen B, Gurung C, Guo J, Kwon C, Guo YL. Pluripotent stem cells are insensitive to the cytotoxicity of TNFα and IFNγ.. Reproduction 160:547–560.
- Chen GY, Hwang SM, Su HJ, Kuo CY, Luo WY, Lo KW, Huang CC, Chen CL, Yu SH, Hu YC. Defective antiviral responses of induced pluripotent stem cells to baculoviral vector transduction.. J Virol 86:8041–8049.
- Chen LL, Yang L, Carmichael GG. Molecular basis for an attenuated cytoplasmic dsRNA response in human embryonic stem cells.. Cell Cycle 9:3552–3564.
- Correia-Álvarez E, Gómez E, Martín D, Carrocera S, Pérez S, Otero J, Peynot N, Giraud-Delville C, Caamaño JN, Sandra O, Duranthon V, Muñoz M. Expression and localization of interleukin 1 beta and interleukin 1 receptor (type I) in the bovine endometrium and embryo.. J Reprod Immunol 110:1–13.
- D’Angelo W, Chen B, Gurung C, Guo YL. Characterization of embryonic stem cell-differentiated fibroblasts as mesenchymal stem cells with robust expansion capacity and attenuated innate immunity.. Stem Cell Res Ther 9:278.
- D’Angelo W, Gurung C, Acharya D, Chen B, Ortolano N, Gama V, Bai F, Guo Y. The Molecular Basis for the Lack of Inflammatory Responses in Mouse Embryonic Stem Cells and Their Differentiated Cells.. J Immunol (Baltimore Md. 1950) 198:2147–2155.
- Dakin SG, Buckley CD, Al-Mossawi MH, Hedley R, Martinez FO, Wheway K, Watkins B, Carr AJ. Persistent stromal fibroblast activation is present in chronic tendinopathy.. Arthritis Res Ther 19:16.
- Dakin SG, Coles M, Sherlock JP, Powrie F, Carr AJ, Buckley CD. Pathogenic stromal cells as therapeutic targets in joint inflammation.. Nat Rev Rheumatol 14:714–726.
- Dakin SG, Dudhia J, Smith RK. Science in brief: resolving tendon inflammation. A New Perspect. Equine Vet J 45:398–400.
- Dakin SG, Dudhia J, Smith RK. Resolving an inflammatory concept: the importance of inflammation and resolution in tendinopathy.. Vet Immunol Immunopathol 158:121–127.
- Dakin SG, Newton J, Martinez FO, Hedley R, Gwilym S, Jones N, Reid HAB, Wood S, Wells G, Appleton L, Wheway K, Watkins B, Carr AJ. Chronic inflammation is a feature of Achilles tendinopathy and rupture.. Br J Sports Med 52:359–367.
- Demine S, Schiavo AA, Marín-Cañas S, Marchetti P, Cnop M, Eizirik DL. Pro-inflammatory cytokines induce cell death, inflammatory responses, and endoplasmic reticulum stress in human iPSC-derived beta cells.. Stem Cell Res Ther 11:7.
- Diederichs S, Klampfleuthner FAM, Moradi B, Richter W. Chondral Differentiation of Induced Pluripotent Stem Cells Without Progression Into the Endochondral Pathway.. Front Cell Dev Biol 7:270.
- Dyson SJ. Medical management of superficial digital flexor tendonitis: a comparative study in 219 horses (1992–2000).. Equine Vet J 36:415–419.
- Gardner AK, Kelly CS, van Eps AW, Burns TA, Watts MR, Belknap JK. Mitogen-activated kinase pathway activation in epidermal lamellae in the acute stages of carbohydrate overload laminitis models and the effect of regional deep hypothermia on signalling pathways.. Equine Vet J 48:633–640.
- Glaser DE, Gower RM, Lauer NE, Tam K, Blancas AA, Shih AJ, Simon SI, McCloskey KE. Functional characterization of embryonic stem cell-derived endothelial cells.. J Vasc Res 48:415–428.
- Goodrich LR, Nixon AJ. Medical treatment of osteoarthritis in the horse - A review.. Vet J 171:51–69.
- Guest DJ, Allen WR. Expression of cell surface antigens and embryonic stem cell pluripotency genes in equine blastocysts.. Stem Cells Dev 16:789–795.
- Guo YL, Carmichael GG, Wang R, Hong X, Acharya D, Huang F, Bai F. Attenuated Innate Immunity in Embryonic Stem Cells and Its Implications in Developmental Biology and Regenerative Medicine.. Stem Cells 33:3165–3173.
- Hagman S, Mäkinen A, Ylä-Outinen L, Huhtala H, Elovaara I, Narkilahti S. Effects of inflammatory cytokines IFN-γ, TNF-α and IL-6 on the viability and functionality of human pluripotent stem cell-derived neural cells.. J Neuroimmunol 331:36–45.
- Hosaka Y, Kirisawa R, Yamamoto E, Ueda H, Iwai H, Takehana K. Localisation of cytokines in tendinocytes of the superficial digital flexor tendon in the horse.. J Vet Med Sci 64:945–947.
- Hyvärinen T, Hagman S, Ristola M, Sukki L, Veijula K, Kreutzer J, Kallio P, Narkilahti S. Co-stimulation with IL-1β and TNF-α induces an inflammatory reactive astrocyte phenotype with neurosupportive characteristics in a human pluripotent stem cell model system.. Sci Rep 9:16944.
- Kamm JL, Nixon AJ, Witte TH. Cytokine and catabolic enzyme expression in synovium, synovial fluid and articular cartilage of naturally osteoarthritic equine carpi.. Equine Vet J 42:693–699.
- Kasembeli MM, Bharadwaj U, Robinson P, Tweardy DJ. Contribution of STAT3 to inflammatory and fibrotic diseases and prospects for its targeting for treatment.. Int J Mol Sci 19(8):2299.
- Lam KH, Parkin TD, Riggs CM, Morgan KL. Descriptive analysis of retirement of Thoroughbred racehorses due to tendon injuries at the Hong Kong Jockey Club (1992–2004).. Equine Vet J 39:143–148.
- Lee J, Smeriglio P, Chu CR, Bhutani N. Human iPSC-derived chondrocytes mimic juvenile chondrocyte function for the dual advantage of increased proliferation and resistance to IL-1β.. Stem Cell Res Ther 8:244.
- Li Y, Wu T, Liu S. Identification and distinction of tenocytes and tendon-derived stem cells.. Front Cell Dev Biol 9:629515.
- Liu T, Zhang L, Joo D, Sun S-C. NF-κB signaling in inflammation.. Signal Transduct Target Ther 2:17023.
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real time quantitative PCR and the 2 Method.. Methods 25:402–408.
- McClellan A, Evans R, Sze C, Kan S, Paterson Y, Guest D. A novel mechanism for the protection of embryonic stem cell derived tenocytes from inflammatory cytokine interleukin 1 beta.. Sci Rep 9:2755.
- McClellan A, Paterson YZ, Paillot R, Guest DJ. Equine Fetal, Adult, and Embryonic Stem Cell-Derived Tenocytes Are All Immune Privileged but Exhibit Different Immune Suppressive Properties In Vitro.. Stem Cells Dev 28:1413–1423.
- Morita W, Dakin SG, Snelling SJB, Carr AJ. Cytokines in tendon disease: A Systematic Review.. Bone Joint Res 6:656–664.
- Morita W, Snelling SJ, Dakin SG, Carr AJ. Profibrotic mediators in tendon disease: a systematic review.. Arthritis Res Ther 18:269.
- Morita W, Snelling SJB, Wheway K, Watkins B, Appleton L, Carr AJ, Dakin SG. ERK1/2 drives IL-1β-induced expression of TGF-β1 and BMP-2 in torn tendons.. Sci Rep 9:19005.
- Nakajima T, Ikeya M. Development of pluripotent stem cell-based human tenocytes.. Dev Growth Differ 63:38–46.
- Nakao S, Ogata Y, Shimizu-Sasaki E, Yamazaki M, Furuyama S, Sugiya H. Activation of NFκB is necessary for IL-1β-induced cyclooxygenase-2 (COX-2) expression in human gingival fibroblasts.. Mol Cell Biochem 209:113–118.
- Oreff GL, Fenu M, Vogl C, Ribitsch I, Jenner F. Species variations in tenocytes’ response to inflammation require careful selection of animal models for tendon research.. Sci Rep 11:12451.
- Quattrocelli M, Giacomazzi G, Broeckx SY, Ceelen L, Bolca S, Spaas JH, Sampaolesi M. Equine-Induced Pluripotent Stem Cells Retain Lineage Commitment Toward Myogenic and Chondrogenic Fates.. Stem Cell Rep 6:55–63.
- Rajan S, Ye J, Bai S, Huang F, Guo YL. NF-kappaB, but not p38 MAP kinase, is required for TNF-alpha-induced expression of cell adhesion molecules in endothelial cells.. J Cell Biochem 105:477–486.
- Ribitsch I, Baptista PM, Lange-Consiglio A, Melotti L, Patruno M, Jenner F, Schnabl-Feichter E, Dutton LC, Connolly DJ, van Steenbeek FG, Dudhia J, Penning LC. Large animal models in regenerative medicine and tissue engineering: to Do or Not to Do.. Front Bioeng Biotechnol 8:972.
- Ribitsch I, Oreff GL, Jenner F. Regenerative medicine for equine musculoskeletal diseases.. Animals (Basel) 11(1):234.
- Russell J, Matika O, Russell T, Reardon RJM. Heritability and prevalence of selected osteochondrosis lesions in yearling Thoroughbred horses.. Equine Vet J 49:282–287.
- Saitta B, Passarini J, Sareen D, Ornelas L, Sahabian A, Argade S, Krakow D, Cohn DH, Svendsen CN, Rimoin DL. Patient-derived skeletal dysplasia induced pluripotent stem cells display abnormal chondrogenic marker expression and regulation by BMP2 and TGFβ1.. Stem Cells Dev 23:1464–1478.
- Saraf A, Rampoldi A, Chao M, Li D, Armand L, Hwang H, Liu R, Jha R, Fu H, Maxwell JT, Xu C. Functional and molecular effects of TNF-α on human iPSC-derived cardiomyocytes.. Stem Cell Res 52:102218.
- Sheth U, Wasserstein D, Jenkinson R, Moineddin R, Kreder H, Jaglal SB. The epidemiology and trends in management of acute Achilles tendon ruptures in Ontario Canada.. Bone Joint J 99-B:78–86.
- Shrestha K, Gilkerson JR, Stevenson MA, Flash ML. Drivers of exit and outcomes for Thoroughbred racehorses participating in the 2017–2018 Australian racing season.. PLoS One 16:e0257581.
- Sidney LE, Kirkham GR, Buttery LD. Comparison of osteogenic differentiation of embryonic stem cells and primary osteoblasts revealed by responses to IL-1beta, TNF-alpha, and IFN-gamma.. Stem Cells Dev 23:605–617.
- Smith EJ, Beaumont RE, McClellan A, Sze C, Palomino Lago E, Hazelgrove L, Dudhia J, Smith RKW, Guest DJ. Tumour necrosis factor alpha, interleukin 1 beta and interferon gamma have detrimental effects on equine tenocytes that cannot be rescued by IL-1RA or mesenchymal stromal cell-derived factors.. Cell Tissue Res 391:523–544.
- van der Kraan PM. The Interaction between Joint Inflammation and Cartilage Repair.. Tissue Eng Regen Med 16:327–334.
- Williams RB, Harkins LS, Wood JLN. Racehorse injuries, clinical problems and fatalities recorded on British racecourses from flat racing and National Hunt racing during 1996, 1997 and 1998.. Equine Vet J 33:478–486.
- Zampetaki A, Xiao Q, Zeng L, Hu Y, Xu Q. TLR4 expression in mouse embryonic stem cells and in stem cell-derived vascular cells is regulated by epigenetic modifications.. Biochem Biophys Res Commun 347:89–99.
- Zampetaki A, Zeng L, Xiao Q, Margariti A, Hu Y, Xu Q. Lacking cytokine production in ES cells and ES-cell-derived vascular cells stimulated by TNF-alpha is rescued by HDAC inhibitor trichostatin A.. Am J Physiol Cell Physiol 293:C1226–C1238.
- Zhang Y, Jordan JM. Epidemiology of osteoarthritis.. Clin Geriatr Med 26:355–369.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
