The Thyroid Hormone Analogue KB2115 (Eprotirome) Prolongs Human Hair Growth (Anagen) Ex Vivo
2016; Elsevier BV; Volume: 136; Issue: 8 Linguagem: Inglês
10.1016/j.jid.2016.03.033
ISSN1523-1747
AutoresAttila Oláh, Jennifer Gherardini, Marta Bertolini, Jérémy Chéret, L. Ponce, Jennifer E. Kloepper, Tamás Bı́ró, Michael Soeberdt, Christoph Abels, Ralf Paus,
Tópico(s)Growth Hormone and Insulin-like Growth Factors
ResumoPremature termination of the active growth phase of the human hair follicle (HF) cycle (anagen) lies at the base of most clinically important hair loss disorders. Therefore, the development of effective, topically applied, and safe anagen-prolonging agents constitutes one of the principle challenges in translational human hair research (Paus, 2006Paus R. Therapeutic strategies for treating hair loss.Drug Discovery Today: Ther Strateg. 2006; 3: 101-110Crossref Scopus (58) Google Scholar, Paus et al., 2014Paus R. Langan E.A. Vidali S. Ramot Y. Andersen B. Neuroendocrinology of the hair follicle: principles and clinical perspectives.Trends Mol Med. 2014; 20: 559-570Abstract Full Text Full Text PDF PubMed Scopus (87) Google Scholar, Qi and Garza, 2014Qi J. Garza L.A. An overview of alopecias.Cold Spring Harb Perspect Med. 2014; 4: a013615Crossref Scopus (44) Google Scholar). Previously, we had shown that thyroid hormones (THs) significantly prolong the anagen phase and retard the onset of apoptosis-driven HF involution (catagen) (Gáspár et al., 2010Gáspár E. Hardenbicker C. Bodó E. Wenzel B. Ramot Y. Funk W. et al.Thyrotropin releasing hormone (TRH): a new player in human hair-growth control.FASEB J. 2010; 24: 393-403Crossref PubMed Scopus (61) Google Scholar, van Beek et al., 2008van Beek N. Bodó E. Kromminga A. Gáspár E. Meyer K. Zmijewski M.A. et al.Thyroid hormones directly alter human hair follicle functions: anagen prolongation and stimulation of both hair matrix keratinocyte proliferation and hair pigmentation.J Clin Endocrinol Metab. 2008; 93: 4381-4388Crossref PubMed Scopus (98) Google Scholar), strongly supporting the concept that TH receptors (TRs) are promising targets for treating HF cycle-based hair loss disorders (Billoni et al., 2000Billoni N. Buan B. Gautier B. Gaillard O. Mahé Y.F. Bernard B.A. Thyroid hormone receptor beta1 is expressed in the human hair follicle.Br J Dermatol. 2000; 142: 645-652Crossref PubMed Scopus (94) Google Scholar, Slominski et al., 2002Slominski A. Wortsman J. Kohn L. Ain K.B. Venkataraman G.M. Pisarchik A. et al.Expression of hypothalamic-pituitary-thyroid axis related genes in the human skin.J Invest Dermatol. 2002; 119: 1449-1455Abstract Full Text Full Text PDF PubMed Scopus (131) Google Scholar). However, because of their potential systemic adverse effects, such as cardiovascular effects (Devereaux and Tewelde, 2014Devereaux D. Tewelde S.Z. Hyperthyroidism and thyrotoxicosis.Emerg Med Clin North Am. 2014; 32: 277-292Abstract Full Text Full Text PDF PubMed Scopus (62) Google Scholar, Jameson, 2013Jameson J.L. Harrison's Endocrinology. 3rd ed. McGrawHill, New York2013Google Scholar) and overstimulation of mitochondrial activity (Cioffi et al., 2013Cioffi F. Senese R. Lanni A. Goglia F. Thyroid hormones and mitochondria: with a brief look at derivatives and analogues.Mol Cell Endocrinol. 2013; 379: 51-61Crossref PubMed Scopus (67) Google Scholar), administration of classic TR agonists, especially those that fully target the myocardially expressed TR-α (Mullur et al., 2014Mullur R. Liu Y.-Y. Brent G.A. Thyroid hormone regulation of metabolism.Physiol Rev. 2014; 94: 355-382Crossref PubMed Scopus (1046) Google Scholar), are difficult to justify for non–life-threatening hair loss disorders. Therefore, in the current study we asked whether a synthetic TR-modulator, KB2115 (eprotirome), which shows higher affinity for TR-β than for TR-α (Shiohara et al., 2012Shiohara H. Nakamura T. Kikuchi N. Ozawa T. Nagano R. Matsuzawa A. et al.Discovery of novel indane derivatives as liver-selective thyroid hormone receptor β (TRβ) agonists for the treatment of dyslipidemia.Bioorg Med Chem. 2012; 20: 3622-3634Crossref PubMed Scopus (23) Google Scholar) and thus is less likely to lead to cardiovascular adverse effects than thyroxine (T4) (Ladenson et al., 2010Ladenson P.W. Kristensen J.D. Ridgway E.C. Olsson A.G. Carlsson B. Klein I. et al.Use of the thyroid hormone analogue eprotirome in statin-treated dyslipidemia.N Engl J Med. 2010; 362: 906-916Crossref PubMed Scopus (270) Google Scholar), may also prolong anagen. This was tested in microdissected, organ-cultured human scalp HFs in serum-free medium and without addition of natural THs (Kloepper et al., 2010Kloepper J.E. Sugawara K. Al-Nuaimi Y. Gáspár E. van Beek N. Paus R. Methods in hair research: how to objectively distinguish between anagen and catagen in human hair follicle organ culture.Exp Dermatol. 2010; 19: 305-312Crossref PubMed Scopus (100) Google Scholar, Langan et al., 2015Langan E.A. Philpott M.P. Kloepper J.E. Paus R. Human hair follicle organ culture: Theory, application and perspectives.Exp Dermatol. 2015; 24: 903-911Crossref PubMed Scopus (107) Google Scholar). The study was approved by the Ethics Committee of the University of Lübeck and the University of Münster (reference nos. 06-109, 2014-041-b-N, 2015-602-f-S), and written informed consent was obtained from the patients (see Supplementary Materials and Methods and Supplementary Figure S1a–f online). First, we probed whether KB2115 stimulated mitochondrial function in the HF epithelium by assessing how low concentrations (1 and 100 pmol/L) of KB2115 influence the expression of mitochondrially encoded cytochrome C oxidase I (MTCO1; a key enzyme of respiratory complex IV). We had previously documented that the immunoreactivity intensity of MTCO1 correlates well with respiratory complex IV activity and even mitochondrial biogenesis (Knuever et al., 2012Knuever J. Poeggeler B. Gáspár E. Klinger M. Hellwig-Burgel T. Hardenbicker C. et al.Thyrotropin-releasing hormone controls mitochondrial biology in human epidermis.J Clin Endocrinol Metab. 2012; 97: 978-986Crossref PubMed Scopus (27) Google Scholar, Poeggeler et al., 2010Poeggeler B. Knuever J. Gáspár E. Bíró T. Klinger M. Bodó E. et al.Thyrotropin powers human mitochondria.FASEB J. 2010; 24: 1525-1531Crossref PubMed Scopus (28) Google Scholar, Vidali et al., 2014Vidali S. Knuever J. Lerchner J. Giesen M. Bíró T. Klinger M. et al.Hypothalamic-pituitary-thyroid axis hormones stimulate mitochondrial function and biogenesis in human hair follicles.J Invest Dermatol. 2014; 134: 33-42Abstract Full Text Full Text PDF PubMed Scopus (59) Google Scholar); moreover, classical THs up-regulate complex IV activity and MTCO1 immunoreactivity in human HF epithelium (van Beek et al., 2008van Beek N. Bodó E. Kromminga A. Gáspár E. Meyer K. Zmijewski M.A. et al.Thyroid hormones directly alter human hair follicle functions: anagen prolongation and stimulation of both hair matrix keratinocyte proliferation and hair pigmentation.J Clin Endocrinol Metab. 2008; 93: 4381-4388Crossref PubMed Scopus (98) Google Scholar). This showed that KB2115 did not significantly alter MTCO1 immunoreactivity (see Supplementary Figure S2a–b online), suggesting that the tested picomolar concentrations may not exert major effects on energy metabolism. Preliminary results from microarray analysis (data not shown), followed up by quantitative, real-time PCR for three selected TH-responsive, mitochondrial biology-related genes (peroxisome proliferator-activated receptor gamma coactivator 1 [PGC1]-α and -β) (Lesmana et al., 2016Lesmana R. Sinha R.A. Singh B.K. Zhou J. Ohba K. Wu Y. et al.Thyroid hormone stimulation of autophagy is essential for mitochondrial biogenesis and activity in skeletal muscle.Endocrinology. 2016; 157: 23-38Crossref PubMed Scopus (62) Google Scholar, Weitzel et al., 2003Weitzel J.M. Hamann S. Jauk M. Lacey M. Filbry A. Radtke C. et al.Hepatic gene expression patterns in thyroid hormone-treated hypothyroid rats.J Mol Endocrinol. 2003; 31: 291-303Crossref PubMed Scopus (55) Google Scholar) and mitochondrial creatine kinase 1A (CMKT1A), showed that KB2115 induced significantly smaller (if any) up-regulation of these genes in human HFs compared with T4 (see Supplementary Figure S3 online). This supports the concept that KB2115 is less likely to exert unwanted systemic, mitochondrial activity-related adverse effects than T4 (see Supplementary Discussion 1 online for details). Therefore, we studied the effects of these (presumably "mitochondrially safe") concentrations of KB2115 on the spontaneous anagen-catagen transition of human scalp HFs ex vivo. Quantitative hair cycle histomorphometry (Kloepper et al., 2010Kloepper J.E. Sugawara K. Al-Nuaimi Y. Gáspár E. van Beek N. Paus R. Methods in hair research: how to objectively distinguish between anagen and catagen in human hair follicle organ culture.Exp Dermatol. 2010; 19: 305-312Crossref PubMed Scopus (100) Google Scholar) showed that KB2115 treatment significantly increased the percentage of anagen HFs compared with vehicle controls (Figure 1a). Given that KB2115 thus effectively countered the tendency of organ-cultured HFs to spontaneously enter into catagen (Kloepper et al., 2010Kloepper J.E. Sugawara K. Al-Nuaimi Y. Gáspár E. van Beek N. Paus R. Methods in hair research: how to objectively distinguish between anagen and catagen in human hair follicle organ culture.Exp Dermatol. 2010; 19: 305-312Crossref PubMed Scopus (100) Google Scholar, Langan et al., 2015Langan E.A. Philpott M.P. Kloepper J.E. Paus R. Human hair follicle organ culture: Theory, application and perspectives.Exp Dermatol. 2015; 24: 903-911Crossref PubMed Scopus (107) Google Scholar), our findings support the concept that KB2115 may prevent or reduce premature catagen entry leading to telogen effluvium in vivo. The anagen-prolonging effect of KB2115 was supported by the finding that the percentage of Ki67+ (i.e., proliferating) cells in the hair matrix was also significantly increased, whereas the percentage of apoptotic (TUNEL+) hair matrix cells remained unchanged (Figure 1b–c). KB2115 also slightly but nonsignificantly increased hair bulb melanin content compared with vehicle control HFs, as assessed by quantitative Masson-Fontana histochemistry (Figure 2a–b). Moreover, KB2115 significantly increased the intrafollicular activity of tyrosinase in situ (Han et al., 2002Han R. Baden H.P. Brissette J.L. Weiner L. Redefining the skin's pigmentary system with a novel tyrosinase assay.Pigment Cell Res. 2002; 15: 290-297Crossref PubMed Scopus (44) Google Scholar) (Figure 2c–d), the rate-limiting enzyme of melanogenesis (Slominski et al., 2005Slominski A. Wortsman J. Plonka P.M. Schallreuter K.U. Paus R. Tobin D.J. Hair follicle pigmentation.J Invest Dermatol. 2005; 124: 13-21Abstract Full Text Full Text PDF PubMed Scopus (377) Google Scholar). None of the tested KB2115 concentrations significantly altered the histochemically detected hair bulb melanin content or tyrosinase activity in situ compared with controls when exclusively anagen VI HFs were compared with each other (data not shown); therefore, this pigmentary effect of KB2115 most likely reflects prolonged anagen-associated intrafollicular melanogenesis (Han et al., 2002Han R. Baden H.P. Brissette J.L. Weiner L. Redefining the skin's pigmentary system with a novel tyrosinase assay.Pigment Cell Res. 2002; 15: 290-297Crossref PubMed Scopus (44) Google Scholar, Kloepper et al., 2010Kloepper J.E. Sugawara K. Al-Nuaimi Y. Gáspár E. van Beek N. Paus R. Methods in hair research: how to objectively distinguish between anagen and catagen in human hair follicle organ culture.Exp Dermatol. 2010; 19: 305-312Crossref PubMed Scopus (100) Google Scholar). However, further scrutiny is needed to determine whether long-term administration of KB2115 may also stimulate HF pigmentation in a hair cycle-independent manner and thus may also exert "anti-graying" actions. Finally, we asked whether and how KB2115 affects the two best-studied human hair cycle-regulatory growth factors: transforming growth factor-β2, which promotes catagen, and IGF-1, which maintains anagen (Fischer et al., 2014Fischer T.W. Herczeg-Lisztes E. Funk W. Zillikens D. Bíró T. Paus R. Differential effects of caffeine on hair shaft elongation, matrix and outer root sheath keratinocyte proliferation, and transforming growth factor-β2/insulin-like growth factor-1-mediated regulation of the hair cycle in male and female human hair follicles in vitro.Br J Dermatol. 2014; 171: 1031-1043Crossref PubMed Scopus (42) Google Scholar, Gáspár et al., 2010Gáspár E. Hardenbicker C. Bodó E. Wenzel B. Ramot Y. Funk W. et al.Thyrotropin releasing hormone (TRH): a new player in human hair-growth control.FASEB J. 2010; 24: 393-403Crossref PubMed Scopus (61) Google Scholar, Hibino and Nishiyama, 2004Hibino T. Nishiyama T. Role of TGF-beta2 in the human hair cycle.J Dermatol Sci. 2004; 35: 9-18Abstract Full Text Full Text PDF PubMed Scopus (164) Google Scholar, Inui and Itami, 2013Inui S. Itami S. Androgen actions on the human hair follicle: perspectives.Exp Dermatol. 2013; 22: 168-171Crossref PubMed Scopus (98) Google Scholar). This showed that KB2115 significantly decreased transforming growth factor-β2 protein expression in the outer root sheath (Figure 2e–f). Surprisingly, intrafollicular IGF-1 protein expression was slightly decreased (see Supplementary Figure 4a–b online), suggesting that KB2115 may prolong anagen in an IGF-1–independent manner. That the synthetic TR-modulator KB2115, which is expected to show reduced adverse effects compared with T4, promotes human HF growth ex vivo corroborates the recognized anagen-prolonging effect of endogenous THs (Billoni et al., 2000Billoni N. Buan B. Gautier B. Gaillard O. Mahé Y.F. Bernard B.A. Thyroid hormone receptor beta1 is expressed in the human hair follicle.Br J Dermatol. 2000; 142: 645-652Crossref PubMed Scopus (94) Google Scholar, van Beek et al., 2008van Beek N. Bodó E. Kromminga A. Gáspár E. Meyer K. Zmijewski M.A. et al.Thyroid hormones directly alter human hair follicle functions: anagen prolongation and stimulation of both hair matrix keratinocyte proliferation and hair pigmentation.J Clin Endocrinol Metab. 2008; 93: 4381-4388Crossref PubMed Scopus (98) Google Scholar). The apparently much lower impact of KB2115 than of T4 on mitochondrial activity parameters in human HF keratinocytes in situ suggests a favorable profile of systemic adverse effects, which may be further reduced by using a HF-targeting topical application vehicle (Vogt and Blume-Peytavi, 2014Vogt A. Blume-Peytavi U. Selective hair therapy: bringing science to the fiction.Exp Dermatol. 2014; 23: 83-86Crossref PubMed Scopus (17) Google Scholar, Vogt et al., 2015Vogt A. Hadam S. Deckert I. Schmidt J. Stroux A. Afraz Z. et al.Hair follicle targeting, penetration enhancement and Langerhans cell activation make cyanoacrylate skin surface stripping a promising delivery technique for transcutaneous immunization with large molecules and particle-based vaccines.Exp Dermatol. 2015; 24: 73-75Crossref Scopus (31) Google Scholar). Also unlike T4 (van Beek et al., 2008van Beek N. Bodó E. Kromminga A. Gáspár E. Meyer K. Zmijewski M.A. et al.Thyroid hormones directly alter human hair follicle functions: anagen prolongation and stimulation of both hair matrix keratinocyte proliferation and hair pigmentation.J Clin Endocrinol Metab. 2008; 93: 4381-4388Crossref PubMed Scopus (98) Google Scholar), KB2115 did not increase HF pigmentation under the current assay conditions. Our ex vivo pilot data suggest that KB2115 is a promising candidate for the future treatment of hair loss disorders characterized by premature catagen entry, such as various forms of telogen effluvium and androgenetic alopecia (Paus, 2006Paus R. Therapeutic strategies for treating hair loss.Drug Discovery Today: Ther Strateg. 2006; 3: 101-110Crossref Scopus (58) Google Scholar, Qi and Garza, 2014Qi J. Garza L.A. An overview of alopecias.Cold Spring Harb Perspect Med. 2014; 4: a013615Crossref Scopus (44) Google Scholar), ideally after topical application. Thus, KB2115 may serve as a lead compound for the development of other hair growth-promoting agents with reduced toxicity compared with T4. However, given that systemic KB2115 treatment may lead to cartilage damage in dogs (Sjouke et al., 2014Sjouke B. Langslet G. Ceska R. Nicholls S.J. Nissen S.E. Öhlander M. et al.Eprotirome in patients with familial hypercholesterolaemia (the AKKA trial): a randomised, double-blind, placebo-controlled phase 3 study.Lancet Diabetes Endocrinol. 2014; 2: 455-463Abstract Full Text Full Text PDF PubMed Scopus (74) Google Scholar), further rigorous toxicological and pharmacokinetic screening is required before KB2115 is applied topically to the human scalp (see Supplementary Discussion 2 online for details). Attila Oláh: http://orcid.org/0000-0003-4122-5639 This study was supported by an industry research grant (see Acknowledgements), and two of the authors (MS and CA) are employees of the sponsor. AO was employed by the sponsor between February 1, 2014, and January 31, 2015, and RP has previously served in a consultancy function for the sponsor. MS and CA are named as inventors on a patent claiming the use of eprotirome in the prevention and/or treatment of hair disorders. This work was supported by Dr. August Wolff GmbH & Co. KG Arzneimittel (Bielefeld, Germany). Download .pdf (.84 MB) Help with pdf files Supplementary Data
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