Single-cell profiling reveals a potent role of quercetin in promoting hair regeneration
Qian Zhao, Yandong Zheng, Dongxin Zhao, Liyun Zhao, Lingling Geng, Shuai Ma, Yusheng Cai, Chengyu Liu, Yupeng Yan, Juan Carlos Izpisua Belmonte, Si Wang, Weiqi Zhang, Guang-Hui Liu, Jing Qu
Single-cell profiling reveals a potent role of quercetin in promoting hair regeneration
Hair loss affects millions of people at some time in their life, and safe and efficient treatments for hair loss are a significant unmet medical need. We report that topical delivery of quercetin (Que) stimulates resting hair follicles to grow with rapid follicular keratinocyte proliferation and replenishes perifollicular microvasculature in mice. We construct dynamic single-cell transcriptome landscape over the course of hair regrowth and find that Que treatment stimulates the differentiation trajectory in the hair follicles and induces an angiogenic signature in dermal endothelial cells by activating HIF-1α in endothelial cells. Skin administration of a HIF-1α agonist partially recapitulates the pro-angiogenesis and hair-growing effects of Que. Together, these findings provide a molecular understanding for the efficacy of Que in hair regrowth, which underscores the translational potential of targeting the hair follicle niche as a strategy for regenerative medicine, and suggest a route of pharmacological intervention that may promote hair regrowth.
single-cell RNA-sequencing / Que / hair follicle regeneration / endothelial cells / HIF-1α
[1] |
Aibar S, Gonzalez-Blas CB, Moerman T et al. SCENIC: single-cell regulatory network inference and clustering. Nat Methods 2017;14:1083–1086.
CrossRef
Google scholar
|
[2] |
Alonso L, Fuchs E. The hair cycle. J Cell Sci 2006;119:391–393.
CrossRef
Google scholar
|
[3] |
Anand K, Asthana P, Kumar A et al. Quercetin mediated reduction of angiogenic markers and chaperones in DLA-induced solid tumours. Asian Pac J Cancer Prev 2011;12:2829–2835.
|
[4] |
Anders S, Pyl PT, Huber W. HTSeq—a Python framework to work with high-throughput sequencing data. Bioinformatics 2015;31:166–169.
CrossRef
Google scholar
|
[5] |
Bach A, Bender-Sigel J, Schrenk D et al. The antioxidant quercetin inhibits cellular proliferation via HIF-1-dependent induction of p21WAF. Antioxid Redox Signal 2010;13:437–448.
CrossRef
Google scholar
|
[6] |
Bassino E, Gasparri F, Giannini V et al. Paracrine crosstalk between human hair follicle dermal papilla cells and microvascular endothelial cells. Exp Dermatol 2015;24:388–390.
CrossRef
Google scholar
|
[7] |
Blake JA, Baldarelli R, Kadin JA et al. Mouse Genome Database (MGD): knowledgebase for mouse-human comparative biology. Nucleic Acids Res 2021;49:D981–D987.
CrossRef
Google scholar
|
[8] |
Cai Y, Song W, Li J et al. The landscape of aging. Sci China Life Sci 2022.
CrossRef
Google scholar
|
[9] |
Chai M, Jiang M, Vergnes L et al. Stimulation of hair growth by small molecules that activate autophagy. Cell Rep 2019;27:3413–3421.e3.
CrossRef
Google scholar
|
[10] |
Cheng CC, Tsutsui K, Taguchi T et al. Hair follicle epidermal stem cells define a niche for tactile sensation. Elife 2018;7:e38883.
CrossRef
Google scholar
|
[11] |
Chovatiya G, Ghuwalewala S, Walter LD et al. High-resolution single-cell transcriptomics reveals heterogeneity of self-renewing hair follicle stem cells. Exp Dermatol 2021;30:457–471.
CrossRef
Google scholar
|
[12] |
DasGupta R, Fuchs E. Multiple roles for activated LEF/TCF transcription complexes during hair follicle development and differentiation. Development 1999;126:4557–4568.
CrossRef
Google scholar
|
[13] |
Driskell RR, Jahoda CA, Chuong CM et al. Defining dermal adipose tissue. Exp Dermatol 2014;23:629–631.
CrossRef
Google scholar
|
[14] |
Fang J, Yang J, Wu X et al. Metformin alleviates human cellular aging by upregulating the endoplasmic reticulum glutathione peroxidase 7. Aging Cell 2018;17:e12765.
CrossRef
Google scholar
|
[15] |
Fang X, Jiang M, Zhou M et al. Elucidating the developmental dynamics of mouse stromal cells at single-cell level. Life Med 2022.
CrossRef
Google scholar
|
[16] |
Foitzik K, Lindner G, Mueller-Roever S et al. Control of murine hair follicle regression (catagen) by TGF-beta1 in vivo. FASEB J 2000;14:752–760.
CrossRef
Google scholar
|
[17] |
Fuchs E. Scratching the surface of skin development. Nature 2007;445:834–842.
CrossRef
Google scholar
|
[18] |
Geng L, Liu Z, Wang S et al. Low-dose quercetin positively regulates mouse healthspan. Protein Cell 2019a;10:770–775.
CrossRef
Google scholar
|
[19] |
Geng L, Liu Z, Zhang W et al. Chemical screen identifies a geroprotective role of quercetin in premature aging. Protein Cell 2019b;10:417–435.
CrossRef
Google scholar
|
[20] |
Gilhar A, Etzioni A, Paus R. Alopecia areata. N Engl J Med 2012;366:1515–1525.
CrossRef
Google scholar
|
[21] |
Glossmann HH, Lutz OMD. Metformin and aging: a review. Gerontology 2019;65:581–590.
CrossRef
Google scholar
|
[22] |
Greco V, Chen T, Rendl M et al. A two-step mechanism for stem cell activation during hair regeneration. Cell Stem Cell 2009;4:155–169.
CrossRef
Google scholar
|
[23] |
Gunnarsson AP, Christensen R, Li J et al. Global gene expression and comparison between multiple populations in the mouse epidermis. Stem Cell Res 2016;17:191–202.
CrossRef
Google scholar
|
[24] |
Hafemeister C, Satija R. Normalization and variance stabilization of single-cell RNA-seq data using regularized negative binomial regression. Genome Biol 2019;20:296.
CrossRef
Google scholar
|
[25] |
Hammond NL, Headon DJ, Dixon MJ. The cell cycle regulator protein 14-3-3sigma is essential for hair follicle integrity and epidermal homeostasis. J Invest Dermatol 2012;132:1543–1553.
CrossRef
Google scholar
|
[26] |
He X, Memczak S, Qu J et al. Single-cell omics in ageing: a young and growing field. Nat Metab 2020;2:293–302.
CrossRef
Google scholar
|
[27] |
Hirota K. HIF-alpha Prolyl Hydroxylase inhibitors and their implications for biomedicine: a comprehensive review. Biomedicines 2021;9:468.
CrossRef
Google scholar
|
[28] |
Horsley V, Aliprantis AO, Polak L et al. NFATc1 balances quiescence and proliferation of skin stem cells. Cell 2008;132:299–310.
CrossRef
Google scholar
|
[29] |
Hsu YC, Fuchs E. A family business: stem cell progeny join the niche to regulate homeostasis. Nat Rev Mol Cell Biol 2012;13:103–114.
CrossRef
Google scholar
|
[30] |
Hsu YC, Li L, Fuchs E. Emerging interactions between skin stem cells and their niches. Nat Med 2014a;20:847–856.
CrossRef
Google scholar
|
[31] |
Hsu YC, Li L, Fuchs E. Transit-amplifying cells orchestrate stem cell activity and tissue regeneration. Cell 2014b;157:935–949.
CrossRef
Google scholar
|
[32] |
Jahangir S, Hosseini S, Mostafaei F et al. 3D-porous beta-tricalcium phosphate-alginate-gelatin scaffold with DMOG delivery promotes angiogenesis and bone formation in rat calvarial defects. J Mater Sci Mater Med 2018;30:1.
CrossRef
Google scholar
|
[33] |
Jave-Suarez LF, Winter H, Langbein L et al. HOXC13 is involved in the regulation of human hair keratin gene expression. J Biol Chem 2002;277:3718–3726.
CrossRef
Google scholar
|
[34] |
Ji J, Ho BS, Qian G et al. Aging in hair follicle stem cells and niche microenvironment. J Dermatol 2017;44:1097–1104.
CrossRef
Google scholar
|
[35] |
Jin S, Guerrero-Juarez CF, Zhang L et al. Inference and analysis of cell-cell communication using CellChat. Nat Commun 2021;12:1088.
CrossRef
Google scholar
|
[36] |
Joost S, Annusver K, Jacob T et al. The molecular anatomy of mouse skin during hair growth and rest. Cell Stem Cell 2020;26, 441–457.e7 e447.
CrossRef
Google scholar
|
[37] |
Kadaja M, Keyes BE, Lin M et al. SOX9: a stem cell transcriptional regulator of secreted niche signaling factors. Genes Dev 2014;28:328–341.
CrossRef
Google scholar
|
[38] |
Kageyama T, Chun YS, Fukuda J. Hair follicle germs containing vascular endothelial cells for hair regenerative medicine. Sci Rep 2021;11:624.
CrossRef
Google scholar
|
[39] |
Kanehisa M, Goto S. KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res 2000;28:27–30.
CrossRef
Google scholar
|
[40] |
Kim BK, Yoon SK. Hairless down-regulates expression of Msx2 and its related target genes in hair follicles. J Dermatol Sci 2013;71:203–209.
CrossRef
Google scholar
|
[41] |
Kim D, Langmead B, Salzberg SL. HISAT: a fast spliced aligner with low memory requirements. Nat Methods 2015;12:357–360.
CrossRef
Google scholar
|
[42] |
Kim J, Kim SR, Choi YH et al. Quercitrin stimulates hair growth with enhanced expression of growth factors via activation of MAPK/CREB signaling pathway. Molecules 2020;25:4004.
CrossRef
Google scholar
|
[43] |
Kobayashi Y, Yonehara S. Novel cell death by downregulation of eEF1A1 expression in tetraploids. Cell Death Differ 2009;16:139–150.
CrossRef
Google scholar
|
[44] |
Kondo S, Schutte BC, Richardson RJ et al. Mutations in IRF6 cause Van der Woude and popliteal pterygium syndromes. Nat Genet 2002;32:285–289.
CrossRef
Google scholar
|
[45] |
Kulkarni AS, Gubbi S, Barzilai, N. Benefits of Metformin in attenuating the hallmarks of aging. Cell Metab 2020;32:15–30.
CrossRef
Google scholar
|
[46] |
Lee DH, Lee YJ. Quercetin suppresses hypoxia-induced accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) through inhibiting protein synthesis. J Cell Biochem 2008;105:546–553.
CrossRef
Google scholar
|
[47] |
Li G, Tang X, Zhang S et al. SIRT7 activates quiescent hair follicle stem cells to ensure hair growth in mice. EMBO J 2020;39:e104365.
CrossRef
Google scholar
|
[48] |
Li J, Zheng Y, Yan P et al. A single-cell transcriptomic atlas of primate pancreatic islet aging. Natl Sci Rev 2021;8:nwaa127.
CrossRef
Google scholar
|
[49] |
Li KN, Jain P, He CH et al. Skin vasculature and hair follicle cross-talking associated with stem cell activation and tissue homeostasis. Elife 2019;8:e45977.
CrossRef
Google scholar
|
[50] |
Li KN, Tumbar T. Hair follicle stem cells as a skin-organizing signaling center during adult homeostasis. EMBO J 2021;40:e107135.
CrossRef
Google scholar
|
[51] |
Liu F, Zhang X, Peng Y et al. miR-24 controls the regenerative competence of hair follicle progenitors by targeting Plk3. Cell Rep 2021;35:109225.
CrossRef
Google scholar
|
[52] |
Liu Z, Li W, Geng L et al. Cross-species metabolomic analysis identifies uridine as a potent regeneration promoting factor. Cell Discov 2022;8:6.
CrossRef
Google scholar
|
[53] |
Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 2014;15:550.
CrossRef
Google scholar
|
[54] |
Ma S, Sun S, Geng L et al. Caloric restriction reprograms the single- cell transcriptional landscape of rattus norvegicus aging. Cell 2020;180:984–1001.e22.
CrossRef
Google scholar
|
[55] |
Ma S, Sun S, Li J et al. Single-cell transcriptomic atlas of primate cardiopulmonary aging. Cell Res 2021;31:415–432.
CrossRef
Google scholar
|
[56] |
Ma S, Wang S, Ye Y et al. Heterochronic parabiosis induces stem cell revitalization and systemic rejuvenation across aged tissues. Cell Stem Cell 2022;29:990–1005.e10.
CrossRef
Google scholar
|
[57] |
Martins Cardoso R, Creemers E, Absalah S et al. Hypercholesterolemia in young adult APOE(-/-) mice alters epidermal lipid composition and impairs barrier function. Biochim Biophys Acta Mol Cell Biol Lipids 2019;1864:976–984.
CrossRef
Google scholar
|
[58] |
Matsumura H, Mohri Y, Binh NT et al. Hair follicle aging is driven by transepidermal elimination of stem cells via COL17A1 proteolysis. Science 2016;351:aad4395.
CrossRef
Google scholar
|
[59] |
Mecklenburg L, Tobin DJ, Muller-Rover S et al. Active hair growth (anagen) is associated with angiogenesis. J Invest Dermatol 2000;114:909–916.
CrossRef
Google scholar
|
[60] |
Medkour Y, Dakik P, McAuley M et al. Mechanisms underlying the essential role of mitochondrial membrane lipids in yeast chronological aging. Oxid Med Cell Longev 2017;2017:2916985.
CrossRef
Google scholar
|
[61] |
Modlich U, Kaup FJ, Augustin HG. Cyclic angiogenesis and blood vessel regression in the ovary: blood vessel regression during luteolysis involves endothelial cell detachment and vessel occlusion. Lab Invest 1996;74:771–780.
|
[62] |
Mootha VK, Lindgren CM, Eriksson KF et al. PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet 2003;34:267–273.
CrossRef
Google scholar
|
[63] |
Morfin R, Guiraud JM, Ducouret B et al. [Evidence of 5 alpha-androstane-3 beta, 6 alpha, 17 beta-triol and of 5 alpha-androstane-3 beta, 7 alpha, 17 beta-triol in the anterior pituitary of the prepuberal male rate]. C R Seances Acad Sci D 1979;288:437–440.
|
[64] |
Muller-Rover S, Handjiski B, van der Veen C et al. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages. J Invest Dermatol 2001;117:3–15.
CrossRef
Google scholar
|
[65] |
Niemann C, Watt FM. Designer skin: lineage commitment in postnatal epidermis. Trends Cell Biol 2002;12:185–192.
CrossRef
Google scholar
|
[66] |
Orasan MS, Roman II, Coneac A et al. Hair loss and regeneration performed on animal models. Clujul Med 2016;89:327–334.
CrossRef
Google scholar
|
[67] |
Oshimori N, Fuchs E. Paracrine TGF-beta signaling counterbalances BMP-mediated repression in hair follicle stem cell activation. Cell Stem Cell 2012;10:63–75.
CrossRef
Google scholar
|
[68] |
Phillips TG, Slomiany WP, Allison R. Hair loss: common causes and treatment. Am Fam Physician 2017;96:371–378.
|
[69] |
Plikus MV, Mayer JA, de la Cruz D et al. Cyclic dermal BMP signalling regulates stem cell activation during hair regeneration. Nature 2008;451:340–344.
CrossRef
Google scholar
|
[70] |
Porter RM. Mouse models for human hair loss disorders. J Anat 2003;202:125–131.
CrossRef
Google scholar
|
[71] |
Qiu X, Mao Q, Tang Y et al. Reversed graph embedding resolves complex single-cell trajectories. Nat Methods 2017;14:979–982.
CrossRef
Google scholar
|
[72] |
Ramos PM, Miot HA. Female pattern hair loss: a clinical and pathophysiological review. An Bras Dermatol 2015;90:529–543.
CrossRef
Google scholar
|
[73] |
Schneider MR, Schmidt-Ullrich R, Paus R. The hair follicle as a dynamic miniorgan. Curr Biol 2009;19:R132–R142.
CrossRef
Google scholar
|
[74] |
Shan H, Geng L, Jiang X et al. Large-scale chemical screen identifies Gallic acid as a geroprotector for human stem cells. Protein Cell 2021.
CrossRef
Google scholar
|
[75] |
Shi J, Yu T, Song K et al. Dexmedetomidine ameliorates endotox-in-induced acute lung injury in vivo and in vitro by preserving mitochondrial dynamic equilibrium through the HIF-1a/HO-1 signaling pathway. Redox Biol 2021;41:101954.
CrossRef
Google scholar
|
[76] |
Simon M, Emmrich S, Seluanov A et al. A hairy tale: SIRT7 safeguards skin stem cells during aging. EMBO J 2020;39:e106294.
CrossRef
Google scholar
|
[77] |
Son MJ, Jeong JK, Kwon Y et al. A novel and safe small molecule enhances hair follicle regeneration by facilitating metabolic reprogramming. Exp Mol Med 2018;50:1–15.
CrossRef
Google scholar
|
[78] |
Soukas AA, Hao H, Wu L. Metformin as anti-aging therapy: is it for everyone? Trends Endocrinol Metab 2019;30:745–755.
CrossRef
Google scholar
|
[79] |
Stuart T, Butler A, Hoffman P et al. Comprehensive integration of single- cell data. Cell 2019;177:1888–1902.e21.
CrossRef
Google scholar
|
[80] |
Su R, Gong G, Zhang L et al. Screening the key genes of hair follicle growth cycle in Inner Mongolian Cashmere goat based on RNA sequencing. Arch Anim Breed 2020;63:155–164.
CrossRef
Google scholar
|
[81] |
Subramanian A, Tamayo P, Mootha VK et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005;102:15545–15550.
CrossRef
Google scholar
|
[82] |
Sugaya K, Hirobe T. Exposure to gamma-rays at the telogen phase of the hair cycle inhibits hair follicle regeneration at the anagen phase in mice. Int J Radiat Biol 2014;90:127–132.
CrossRef
Google scholar
|
[83] |
Tian D, Qin Q, Li M et al. Homocysteine impairs endothelial cell barrier function and angiogenic potential via the progranulin/ EphA2 pathway. Front Pharmacol 2020;11:614760.
CrossRef
Google scholar
|
[84] |
Wang L, Liu J, Liu H et al. The secret of youth - how is systemic rejuvenation achieved at the single cell level? Life Med 2022.
CrossRef
Google scholar
|
[85] |
Wang L, Siegenthaler JA, Dowell RD et al. Foxc1 reinforces quiescence in self-renewing hair follicle stem cells. Science 2016;351:613–617.
CrossRef
Google scholar
|
[86] |
Wang S, Yao X, Ma S et al. A single-cell transcriptomic landscape of the lungs of patients with COVID-19. Nat Cell Biol 2021;23:1314–1328.
CrossRef
Google scholar
|
[87] |
Wang S, Zheng Y, Li J et al. Single-cell transcriptomic atlas of primate ovarian aging. Cell 2020;180:585–600.e19.
CrossRef
Google scholar
|
[88] |
Weger N, Schlake T. Igf-I signalling controls the hair growth cycle and the differentiation of hair shafts. J Invest Dermatol 2005;125:873–882.
CrossRef
Google scholar
|
[89] |
Wikramanayake TC, Villasante AC, Mauro LM et al. Prevention and treatment of alopecia areata with quercetin in the C3H/HeJ mouse model. Cell Stress Chaperones 2012;17:267–274.
CrossRef
Google scholar
|
[90] |
Wilson WJ, Poellinger L. The dietary flavonoid quercetin modulates HIF-1 alpha activity in endothelial cells. Biochem Biophys Res Commun 2002;293:446–450.
CrossRef
Google scholar
|
[91] |
Wu JH, Yan ZW, Husile
CrossRef
Google scholar
|
[92] |
Xie Y, Chen D, Jiang K et al. Hair shaft miniaturization causes stem cell depletion through mechanosensory signals mediated by a Piezo1-calcium-TNF-alpha axis. Cell Stem Cell 2022;29:70–85.e6.
CrossRef
Google scholar
|
[93] |
Xu Z, Chen D, Hu Y et al. Anatomically distinct fibroblast subsets determine skin autoimmune patterns. Nature 2022;601:118–124.
CrossRef
Google scholar
|
[94] |
Xu Z, Wang W, Jiang K et al. Embryonic attenuated Wnt/beta-catenin signaling defines niche location and long-term stem cell fate in hair follicle. Elife 2015;4:e10567.
CrossRef
Google scholar
|
[95] |
Yan P, Li Q, Wang L et al. FOXO3-engineered human ESC-derived vascular cells promote vascular protection and regeneration. Cell Stem Cell 2019;24:447–461.e8.
CrossRef
Google scholar
|
[96] |
Yano K, Brown LF, Detmar M. Control of hair growth and follicle size by VEGF-mediated angiogenesis. J Clin Invest 2001;107:409–417.
CrossRef
Google scholar
|
[97] |
Yu Z, Jiang K, Xu Z et al. Hoxc-dependent mesenchymal niche heterogeneity drives regional hair follicle regeneration. Cell Stem Cell 2018;23:487–500.e6.
CrossRef
Google scholar
|
[98] |
Yuan Q, Bleiziffer O, Boos AM et al. PHDs inhibitor DMOG promotes the vascularization process in the AV loop by HIF-1a up-regulation and the preliminary discussion on its kinetics in rat. BMC Biotechnol 2014;14:112.
CrossRef
Google scholar
|
[99] |
Zhang C, Smalley I, Emmons MF et al. Noncanonical EphA2 signaling is a driver of tumor-endothelial cell interactions and metastatic dissemination in BRAF inhibitor resistant melanoma. J Invest Dermatol 2021a;141:840–851.e4.
CrossRef
Google scholar
|
[100] |
Zhang H, Li J, Ren J et al. Single-nucleus transcriptomic landscape of primate hippocampal aging. Protein Cell 2021b;12:695–716.
CrossRef
Google scholar
|
[101] |
Zhang W, Zhang S, Yan P et al. A single-cell transcriptomic landscape of primate arterial aging. Nat Commun 2020a;11:2202.
CrossRef
Google scholar
|
[102] |
Zhang Z, Dai Q, Zhang Y et al. Design of a multifunctional biomaterial inspired by ancient Chinese medicine for hair regeneration in burned skin. ACS Appl Mater Interfaces 2020b;12:12489–12499.
CrossRef
Google scholar
|
[103] |
Zhou T, Kiran M, Lui KO et al. Decoding liver fibrogenesis with single- cell technologies. Life Med 2022.
CrossRef
Google scholar
|
[104] |
Zhou Y, Zhou B, Pache L et al. Metascape provides a biologist-oriented resource for the analysis of systems-level datasets. Nat Commun 2019;10:1523.
CrossRef
Google scholar
|
[105] |
Zou X, Dai X, Mentis A-FA et al. From monkey single-cell atlases into a broader biomedical perspective. Life Med 2022. doi:10.1093/ lifemedi/lnac028
|
[106] |
Zou Z, Long X, Zhao Q et al. A single-cell transcriptomic atlas of human skin aging. Dev Cell 2021;56:383–397.e8.
CrossRef
Google scholar
|
/
〈 | 〉 |