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1.Bioalternatives, Gençay, France

Samuel Guenin1, Julien Garnier1, Christine Barrault1, Nathalie Pedretti1, Sevda Cordier-Dirikoc1, François-Xavier Bernard1

androgen-sensitive sebocyte cell line.

Pharmacological effects of anti-androgens and other compounds in an

Download the full poster version

INTRODUCTION & HYPOTHESIS

We previously described the effects and behaviour of the main circulating active androgen testosterone (Testo) and its main active intracellular metabolite DHT in an androgen sensitive sebocytes (abstract X, SID, 2019). Testo and DHT readily induce 1) androgen receptor (AR) translocation, 2) expression of target genes and then 3) a lipogenic differentiation program. DHEA is not significantly transformed into Testo, had no significant androgenic (agonist), nor anti-androgenic (antagonist) effect by itself in the tested conditions.
Here we analysed the activity of reference anti-androgens (Finasteride, Dutasteride, Cyproterone acetate), at the level of their potential targets (5-alpha-reductase, AR translocation, induced transcripts) and in a functional lipid accumulation assay. We also evaluated this androgen-induced lipid accumulation assay as a potential pharmacological tool to detect potential inhibitors from both androgenic and non-androgenic origins.

The androgen sensitive sebocytes should allow to characterize both the activity and the mechanism of action of anti-androgen drugs. These anti-androgens may act at the level of androgen metabolism, AR translocation, AR nuclear activity and thus may have (or not) a functional effect on androgen-induced lipogenic differentiation of these cells. If we focus on these functional effects, other (non-androgenic) potential cell metabolism modulators may also work.

MATERIALS & MEHODS

[14C]-Testo metabolism was analysed after incubation with cell cultures at different times and extraction, chromatography/phosphorimaging2. RASD1 expression was measured by RT-qPCR1.
AR translocation was followed by immunofluorescence1. Lipid accumulation was evidenced by incubation with the fluorescent probe Bodipy®1. Automated photography and image analysis was performed using an INCell AnalyzerTM 2200 (GE Healthcare). The tested compounds are listed in fig 3.

RESULTS

1 – Effects of Dutasteride, Finasteride and CypAc on [14C]-Testo metabolism (A) and DHT production (B); 24h assay.

2 – (A) Effects of Dutasteride, Finasteride and CypAc on RASD1 expression after 4 h stimulation with DHT or Testo – (B) Effects of drugs (1µM) on AR translocation (1h assay with DHT) – Effects of drugs (1µM) on lipid accumulation (7days assay, with DHT or Testo).
Download the full version of the poster to see all the results

CONCLUSIONS

In this model,

– Dutasteride is a strong inhibitor of DHT production (1000-fold more active than Finasteride).

– CypAc strongly inhibits the transcription of AR-dependent genes induced by DHT or Testo, without interfering with AR translocation (nuclear effects).

– CypAc is not as effective as expected on the functional lipid accumulation assay. Dutasteride is active in the lipid accumulation assay even in the presence of DHT as inducer, indicating potential additional targets than 5-α-reductase. Taken together it indicates that there is no direct correlation between early inhibitory processes and the final functional effects after 7 days.

– The androgen lipid accumulation assay is suitable for high throughput testing of various compounds with various mechanisms of action, including anti-androgens, cell metabolism inhibitors, etc. Among other, mTOR inhibitors are effective. In these experimental conditions, isotretinoin, which is a reference treatment for acne does not interfere with this androgen-stimulated lipid accumulation (even shows an opposite effect).

REFERENCES

1 Barrault et al. (2015) J Steroid Biochem Mol Biol ;152:34-44
2 Bernard et al. (2000) Int J Cosmet Sci ;22:397-407

Innate immunity activation of sebocyte cells by living bacteria. Evidence of a potential immunosuppressive effect of DHT.

22 May 2019/in Acne, Microbiome cutané, Sebaceous gland regulation, Skin microbiome, Skin microbiome /by Vanessa

Acne is a skin pathology targeting the pilosebaceous unit…sebum hypersecretion and bacterial infection. We studied the effects of living bacteria on the induction of an innate immune response in a sebocyte cell line.

Read more
https://www.bioalternatives.com/wp-content/uploads/2019/05/190527_Poster1_petit.png 368 655 Vanessa https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Vanessa2019-05-22 14:37:542019-11-12 17:58:17Innate immunity activation of sebocyte cells by living bacteria. Evidence of a potential immunosuppressive effect of DHT.

Pharmacological effects of anti-androgens and other compounds in an androgen-sensitive sebocyte cell line.

22 May 2019/in Acne, Sebaceous gland regulation /by Vanessa

We analysed the activity of reference anti-androgens (Finasteride, Dutasteride, Cyproterone acetate), at the level of their potential targets (5-alpha-reductase, AR translocation, induced transcripts) and in a functional lipid accumulation assay. We also evaluated this androgen-induced lipid accumulation assay as a potential pharmacological tool to detect potential inhibitors from both androgenic and non-androgenic origins.

Read more
https://www.bioalternatives.com/wp-content/uploads/2019/05/190527_Poster3_petit.png 368 655 Vanessa https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Vanessa2019-05-22 14:37:122019-11-12 17:36:48Pharmacological effects of anti-androgens and other compounds in an androgen-sensitive sebocyte cell line.

Androgens activate lipogenesis through an AKT-independent mTOR pathway stimulation and a limitation of autophagy in an androgen-sensitive sebocyte cell line.

22 May 2019/in Acne, Peau grasse, hyperséborrhée et séborégulation, Sebaceous gland regulation /by Vanessa

Androgens are key regulators of sebaceous function… A link between mTOR and androgen signaling has previously been reported…as well as a link between these processes and lipid synthesis…Here we aimed at confirming that DHT-induced lipid synthesis/accumulation is at least in part dependent on mTOR activation in the SEBO662AR cell line and that the autophagic process is consequently modified by the androgenic treatment.

Read more
https://www.bioalternatives.com/wp-content/uploads/2019/05/190527_Poster4_petit.png 368 655 Vanessa https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Vanessa2019-05-22 14:37:042019-11-12 17:39:16Androgens activate lipogenesis through an AKT-independent mTOR pathway stimulation and a limitation of autophagy in an androgen-sensitive sebocyte cell line.
Various forms of acne vulgaris

Acne vulgaris: Origins, symptoms and current treatments

29 November 2017/in Acne /by Julie

Acne vulgaris is a chronic inflammatory disease of the skin which affects most adolescents and is the cause of over half of all dermatology consultations. This pathology, which is characterized by a hypersecretion of sebum, can be the cause of physical complications and may also lead to psychological distress.

Read more
https://www.bioalternatives.com/wp-content/uploads/2017/11/Acné-généralité-image-à-la-une-_-EN.jpg 368 655 Julie https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Julie2017-11-29 09:31:592019-11-12 14:28:03Acne vulgaris: Origins, symptoms and current treatments

Acne vulgaris: physiopathology and cell mechanisms

29 November 2017/in Acne, Sebaceous gland regulation /by Julie

Acne Vulgaris is a chronic inflammatory pathology located in the region of the pilosebaceous follicle. It occurs following a high production of sebum linked to hormonal (androgynous) or environmental (pollution) malfunctioning and to the colonization of the pilosebaceous follicle by certain bacteria (P. acnes).
Bioalternatives has developed a panel of innovative assays to discover and evaluate compounds for the treatment of acne.

Read more
https://www.bioalternatives.com/wp-content/uploads/2017/11/Image-à-la-une.jpg 368 655 Julie https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Julie2017-11-29 09:30:252019-09-13 07:40:20Acne vulgaris: physiopathology and cell mechanisms

Physiology and functions of the sebaceous gland

28 November 2017/in Acne, Sebaceous gland regulation, Skin barrier and hydration /by Julie

The sebaceous gland is an organ located in the dermis. Its role is to synthesise and secrete sebum which is a component of the hydrolipidic film. The purpose of sebum is to protect the skin from external aggression and dehydration. Sebum also maintains the suppleness of the skin and hair.

Read more
https://www.bioalternatives.com/wp-content/uploads/2017/11/Sébocytes-image-à-la-une.jpg 368 701 Julie https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Julie2017-11-28 09:24:342019-11-07 15:45:07Physiology and functions of the sebaceous gland

Androgens induce sebaceous differentiation in sebocyte cells expressing a stable functional androgen receptor

9 April 2015/in Acne, Cell and tissue engineering, Sebaceous gland regulation /by Guillaume

In order to study the impact of active androgens in sebocytes, we constructed a stable human sebocyte cell line derived from SEBO662 [17] constitutively expressing a fully functional AR. In these SEBO662 AR+ cells, dihydrotestosterone (DHT) induced AR nuclear translocation and the strong modulation of a set of transcripts (RASD1, GREB1…) known to be androgen-sensitive in other androgenic cells and tissues.

Read more
https://www.bioalternatives.com/wp-content/uploads/2016/03/mw_androgens-induce-sebaceous-differentiation-in-sebocyte.jpg 368 655 Guillaume https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Guillaume2015-04-09 15:47:492019-11-13 11:16:58Androgens induce sebaceous differentiation in sebocyte cells expressing a stable functional androgen receptor

Immortalized sebocytes SEBO662 can spontaneously differentiate into a sebaceous-like phenotype when cultured as a 3D epithelium

24 January 2012/in Acne, Cell and tissue engineering, Sebaceous gland regulation /by Guillaume

SEBO662 multilayers spontaneously differentiate into a sebaceous-like structure as shown by the strong polarized expression of the late sebaceous marker EMA, the overexpression of some lipogenic markers and lipid production on the upper side of the epithelium.

Read more
https://www.bioalternatives.com/wp-content/uploads/2016/05/mw_Immortalized-sebocytes.jpg 368 655 Guillaume https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Guillaume2012-01-24 10:17:152019-11-13 11:28:55Immortalized sebocytes SEBO662 can spontaneously differentiate into a sebaceous-like phenotype when cultured as a 3D epithelium
skin inflammation

Keratinocytes as targets for cytokines in skin inflammation

20 November 2008/in Acne, Atopic dermatitis, Dermatology, Psoriasis /by Guillaume

Current knowledge about the effects of different cytokine families on keratinocytes, and more particularly theirinvolvement in skin inflammation and in the development of inflammatory skin diseases such as psoriasis

Read more
https://www.bioalternatives.com/wp-content/uploads/2016/05/mw_Keratinocytes-as-targets-for-cytokines.jpg 368 655 Guillaume https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Guillaume2008-11-20 14:50:532019-05-31 09:36:35Keratinocytes as targets for cytokines in skin inflammation
5-alpha-reductase

Expression of type 1 5-alpha-reductase and metabolism of testosterone in reconstructed human epidermis

10 December 2000/in Acne, Dermatology /by Guillaume

We analysed the potential of a human epidermal tissue reconstituted by cell culture (RHE, provided by SkinEthic Laboratories, Nice, France) as a model for assessing 5-alphaR activity.

Read more
https://www.bioalternatives.com/wp-content/uploads/2016/05/mw_Expression-of-type-5alpha-reductase.jpg 368 655 Guillaume https://www.bioalternatives.com/wp-content/uploads/2019/09/Logo-Bioalternatives-150dpisite-web2-300x74.png Guillaume2000-12-10 13:36:372018-10-24 22:15:34Expression of type 1 5-alpha-reductase and metabolism of testosterone in reconstructed human epidermis

 

 

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Mail: info.us[@]bioalternatives.com

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Tel. +33 (0) 5 49 36 11 37
Mail: info[@]bioalternatives.com

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Cosmetic

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