Molecular signature of a common alopecia state, associated with cell junctions

Abstract

Molecular signature of a common alopecia state, associated with cell junctions. The present invention relates to a method for the in vitro prognosis and / or diagnosis of a common alopecia state of the scalp in a subject, in particular androgenetic alopecia, comprising at least one step a) of measuring the level of expression, in a biological sample taken from an area suspected of being an alopecia area or of becoming an alopecia area in said subject, of at least one gene selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MYO6, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene selected from CTNNB1 and CTNND2.The invention also relates to an in vitro method for evaluating the effectiveness of a treatment for a common alopecia condition, a cosmetic treatment method, the use of a modulator of said gene(s) mentioned above, as well as a method for identifying a compound enabling the prevention and / or treatment of a common alopecia condition.

Description

Title of the invention: Molecular signature of a common alopecic condition, associated with cell junctions

[0001] The present invention relates to a method for prognosis and / or diagnosis of a common alopecic condition of the scalp.

[0002] Human hair represents a set of approximately 150,000 hairs. Each of them is generated by a specialized appendage of the skin, a true autonomous organ, the hair follicle. Hair growth and renewal is not a continuous process, and is determined by the activity of the hair follicles and their perifollicular matrix environment. The activity of such follicles is cyclical and essentially comprises four phases. Indeed, the follicle passes successively from a growth phase with production of the hair shaft (anagen phase), to a rapid involution phase (catagen phase), then to a resting phase with hair loss (telogen phase) which precedes a regeneration phase (neogen phase) to end again in the anagen phase. The anagen phase, the active or growth phase during which the hair grows longer, lasts several years. The very short catagen phase lasts a few weeks.The telogen phase or resting phase lasts a few months. At the end of this resting period, the hair falls out and another cycle begins. The hair is therefore constantly renewed and of the approximately 150,000 hairs in a head of hair, approximately 10% are at rest and will be replaced in the following months.

[0003] Natural hair loss or shedding can be estimated, on average, at a few hundred hairs per day for a normal physiological state. This process of permanent physical renewal undergoes a natural evolution during aging, the hair becomes finer and its cycles shorter.

[0004] However, various causes can lead to significant, temporary or permanent hair loss. Hair loss, in particular common alopecia, such as androgenetic alopecia, traction alopecia, diffuse alopecia in women (female pattern hair loss), cicatricial alopecia, alopecia induced by chemotherapy or radiotherapy, telogen effluvium, stress-related alopecia, seasonal alopecia, age-related alopecia and micro-inflammatory alopecia, is essentially due to disturbances in the hair cycle, as opposed to immune alopecias such as alopecia areata, alopecia universalis and alopecia totalis. These disturbances initially lead to a shortening of the anagen phase and a progressive thinning of the hair, then a reduction in their quantity. There is a progressive miniaturization of the bulbs, with joint isolation of these by progressive thickening of the collagen matrix of the external connective sheath. Revascularization around the hair follicle is therefore made more difficult cycle after cycle. The hair regresses, miniaturizing until it is nothing more than unpigmented down, and this phenomenon leads to a progressive impoverishment of the hair.

[0005] Areas are preferentially affected, notably the temporal or frontal gulfs in men, and in women, there is rather diffuse alopecia of the vertex.

[0006] The term alopecia also covers a whole family of hair follicle disorders that ultimately result in permanent, partial or general hair loss. This is more specifically androgenic alopecia. In a significant number of cases, premature hair loss occurs in genetically predisposed individuals; this is then androgenetic alopecia; this form of alopecia most often affects men.

[0007] It is also known that certain factors such as hormonal imbalance, physiological stress, malnutrition, can accentuate the phenomenon. In addition, hair loss or alteration can be related to seasonal phenomena.

[0008] Generally speaking, any factor influencing these processes, namely the acceleration of the frequency of cycles, the progressive miniaturization of the bulbs, the progressive thickening of the perifollicular collagen matrix, the thickening of the external connective sheath, and the reduction of vascularization, will have an effect on the growth of hair follicles.

[0009] From the above, it is understood the importance of finding new biological pathways and new biomarkers for detecting an alopecic condition of the scalp, and in particular when it is not yet visible, in order to be able to reduce and / or slow down hair loss. More particularly, there is a significant need for biomarkers specific to common alopecia, in particular androgenetic alopecia, which are differentially expressed in the hair growth zone compared to the alopecia progression zone, in order to target it appropriately in a given subject. The object of the present invention is to satisfy this need.

[0010] The Applicant has surprisingly discovered that the expression of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and possibly CTNNB1 and CTNND2, involved in the intercellular junctions of scalp and / or hair follicle cells, such as keratinocytes and fibroblasts, is decreased in the area of ​​progression of alopecia in a subject presenting with common alopecia.

[0011] Thus, the present invention has as its first subject a method for prognosis and / or in vitro diagnosis of a common alopecic condition of the scalp in a subject, in particular androgenetic alopecia, comprising at least one step a) of measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2.

[0012] The present invention also relates to an in vitro method for evaluating the efficacy of a treatment of a common alopecic condition comprising at least one step a) of measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2 before and after treatment.

[0013] Another object of the present invention is a method of cosmetic treatment for the prevention and / or treatment of a common alopecic condition of the scalp, in particular androgenetic alopecia, comprising at least the following steps: a) measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB 1 and CTNND2; b) deducing from step a) whether the scalp of said subject has a common alopecic condition; c) if the scalp is identified as having an alopecic condition common to step b) treating said scalp with a cosmetic composition making it possible to induce and / or stimulate hair growth and / or slow down hair loss.

[0014] The present invention also relates to the use of at least one modulator of the expression level of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB 1 and CTNND2, or of a cosmetic composition comprising it or them, for the prevention and / or treatment of a condition common alopecia, including androgenetic alopecia.

[0015] Another object of the present invention relates to a method for identifying a compound allowing the prevention and / or treatment of a common alopecic condition, in particular androgenetic alopecia, comprising the following steps: (a) bringing a compound to be tested into contact with a biological sample; b) measuring the level of expression, in said biological sample, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2; c) selecting a compound for which the level of expression of said at least one gene measured in step b) is increased compared to its level of expression in the absence of said compound.

[0016] Definitions

[0017] The genes involved in the intercellular junctions of the aforementioned scalp cells can be classified into subfamilies, as presented in Table 1 below.

[0018] [Tables 1] Genes involved in intercellular junctions CDH1 cadherin 1 Genes involved in adherens junctions (Simpson CL et al. (2011) Nat Rev Mol Cell Biol 12: 565-80). ACTB beta-actin ACTBL2 actin beta like 2 TUBB tubulin beta, class I TUBB2A tubulin beta 2A GSN gelsolin MY03B myosin-IIIb MY05B myosin-Vb MY06 myosin-VI DSG2 desmoglein 2 Genes involved in desmosomes (Simpson CL et al. (2011) Nat Rev Mol Cell Biol 12: 565-80). DSG3 desmoglein 3 DSG4 desmoglein 4 DSC2 desmocollin 2 GJB2 gap junction protein beta 2 Genes involved in gap junctions (Simpson CL et al. (2011) Nat Rev Mol Cell Biol 12: 565-80). GJA1 gap junction protein alpha 1 GJB6 gap junction protein beta 6 GJA3 gap junction protein alpha 3 TJP2 tight junction protein Genes involved in tight junctions (Simpson CL et al. (2011) Nat Rev Mol Cell Biol 12: 565-80).CLDN8 Claudine 8 CLDN10 Claudine 10 CLDN19 Claudine 19 CTNNB1 beta catenin 1 Genes involved in the interaction with CDH1 (Simpson CL et al. (2011) Nat Rev Mol Cell Biol 12: 565-80). CTNND2 catenin delta 2.

[0019] The term "common alopecic condition" means all forms of alopecia excluding immune alopecia. Preferably, the common alopecic condition is chosen from androgenetic alopecia, traction alopecia, diffuse alopecia in women (female pattern hair loss), cicatricial alopecia, alopecia induced by chemotherapy or radiotherapy, telogen effluvium, stress-related alopecia, alopecia seasonal, age-related alopecia and microinflammatory alopecia.

[0020] The term “immune alopecia” means an alopecia of autoimmune origin chosen from alopecia areata, alopecia universalis and alopecia totalis.

[0021] By "CDH1 gene" we mean here the gene encoding the cadherin 1 protein. Cadherin 1 protein is also known as epithelial cadherin (E-cadherin) or ovomorulin. This protein is found in the membrane surrounding epithelial cells, which are the cells that line the surfaces and cavities of the body. E-cadherin belongs to a family of proteins called cadherins whose function is to help neighboring cells adhere to each other (cell adhesion) to form organized tissues. It is typically described in Indra et al. (2018) Spatial and temporal organization of cadherin in punctate adhesion junctions, Proceedings of the National Academy of Sciences of the United States of America, 115(19):E4406-E4415. The human CDH1 gene sequence is typically referenced as gene ID 999 (NCBI). The human CDH1 protein sequence is typically referenced as UniProt P12830.

[0022] By "ACTB gene" is meant here the gene encoding the beta-actin protein. It represents one of the six actin isoforms identified in humans. It is expressed in non-muscle tissues and is a component of the cytoskeleton. It is involved in cell motility, structure and integrity. It is typically described in Erba HP et al. (1988), Molecular and Cellular Biology, Structure, chromosome location, and expression of the human gamma-actin gene: differential evolution, location, and expression of the cytoskeletal beta- and gamma-actin genes, 8(4):1775-1789. The sequence of the human ACTB gene is typically referenced under the gene ID number 60 (NCBI). The sequence of the human ACTB protein is typically referenced under the UniProt number P60709.

[0023] By "ACTBL2 gene" is meant here the gene encoding the protein "actin beta like 2". It is also known as kappa actin. It is expressed in non-muscle tissues and is a component of the cytoskeleton. It is involved in cell motility, structure and integrity. It is typically described in Saba Ghazanfar et al. (2017), Journal of Proteomics, volume 152, p33-40. The sequence of the human ACTBL2 gene is typically referenced under the gene ID number 345651 (NCBI). The sequence of the human ACTBL2 protein is typically referenced under the UniProt number Q562R1.

[0024] By "TUBB gene" is meant here the gene encoding the protein beta tubulin, class I. The gene is also known as TUBB5 and the protein as tubulin beta 5. This protein forms a dimer with alpha tubulin and acts as a structural component of microtubules. It is typically described in Romina Ro-maniello et al., European Journal of Medical Genetics, volume 61, p744-754. The The human TUBB gene sequence is typically referenced as gene ID 203068 (NCBI). The human TUBB protein sequence is typically referenced as UniProt P07437.

[0025] By "TUBB2A gene" is meant here the gene encoding the protein tubulin beta 2A, class IIa. The protein is also known as tubulin beta 2. This protein forms a dimer with alpha tubulin and acts as a structural component of microtubules. It is typically described in Romina Romaniello et al., European Journal of Medical Genetics, volume 61, p744-754. The sequence of the human TUBB2A gene is typically referenced under the gene ID number 7280 (NCBI). The sequence of the human TUBB2A protein is typically referenced under the UniProt number Q13885.

[0026] By "GSN gene" is meant here the gene coding for the gelsolin protein. This cytosolic protein is capable of binding to actin filaments and creating a local dislocation of these. In particular, it promotes vesicle exocytosis by local dissolution of the submembrane cytoskeleton of cells. It is typically described in Eke Gungor H et al. (2016), Allergologia & Immunopathologia, 44(3):221-5. The sequence of the human GSN gene is typically referenced under the gene ID number 2934 (NCBI). The sequence of the human GSN protein is typically referenced under the UniProt number P06396.

[0027] By “MY03B gene” is meant here the gene coding for the protein myosin-IIIb. It belongs to the myosin family. Myosins are actin-activated ATPases that move along actin filaments in the cell. It is typically described in Andréa C. Dosé and Beth Burnside (2002), Genomics, 79(5):621-4. The human MY03B gene sequence is typically referenced as gene ID 140469 (NCBI). The human MY03B protein sequence is typically referenced as UniProt Q8WXR4.

[0028] By “MY05B gene” is meant here the gene coding for the protein myosin-Vb. It belongs to the myosin family. Myosins are ATPases, activated by actin, moving along actin filaments in the cell. Myosin Vb helps determine the position of various components within cells (cell polarity). Myosin Vb also plays a role in moving cell membrane components into the cell for recycling. It is typically described in Sonal et al. (2014), PLoS Genet. Sep 18;10(9):el004614. The human MY05B gene sequence is typically referenced as gene ID 4645 (NCBI). The human MY05B protein sequence is typically referenced as UniProt Q9ULV0.

[0029] By "MY06 gene" is meant here the gene coding for the protein myosin-VI. It belongs to the myosin family. Myosins are ATPases, activated by actin, moving along actin filaments in the cell. Myosin VI plays a role in the transport of intracellular vesicles and organelles. It is typically described in Lister Ida et al. (2004), EMBO J. Apr 21;23(8):1729-38. The human MY06 gene sequence is typically referenced as gene ID 4646 (NCBI). The human MY06 protein sequence is typically referenced as UniProt Q9UM54.

[0030] By "DSG2 gene", "DSG3 gene" and "DSG4 gene" are meant here the genes coding respectively for the proteins desmoglein 2, desmoglein 3, desmoglein 4. They belong to the cadherin family of cell adhesion molecules, in particular the desmoglein subfamily. Desmogleins are calcium-binding transmembrane glycoprotein components of desmosomes, cell-cell junctions between epithelial, myocardial and other cell types. They are typically described in Wu Hong et al. (2003), J Invest Dermatol. Jun; 120(6): 1052-7 and Masayuki Amagai et al. (2012), J Invest Dermatol. Mar; 132(3 Pt 2):776-84. doi: 10.1038 / jid.2011.390. The human DSG2 gene sequence is typically referenced as gene ID 1829 (NCBI). The human DSG2 protein sequence is typically referenced as UniProt Q14126. The human DSG3 gene sequence is typically referenced as gene ID 1830 (NCBI). The human DSG3 protein sequence is typically referenced as UniProt P32926. The human DSG4 gene sequence is typically referenced as gene ID 147409 (NCBI). The sequence of the human DSG4 protein is typically referenced under the UniProt number Q86SJ6.

[0031] By "DSC2" gene is meant here the gene encoding the protein desmocollin 2. Desmocollin 2. This protein is found in many tissues, although it appears to be particularly important in cardiac muscle and skin.Desmocollin-2 is a major component of specialized structures called desmosomes. These structures help hold neighboring cells together, providing strength and stability to tissues. It is typically described in King Ian et al. (1995), J Invest Dermatol. Sep;105(3):314-21. The human DSC2 gene sequence is typically referenced as gene ID 1824 (NCBI). The human DSC2 protein sequence is typically referenced as UniProt Q02487.

[0032] By "GJB2 gene" is meant here the gene coding for the protein "gap junction protein beta 2". This protein is more commonly known as connexin 26. Connexin 26 is a member of the connexin protein family. Connexin proteins form channels called "gap junctions" that allow the transport of nutrients, charged atoms (ions) and signaling molecules between adjacent cells. The size of the "gap junctions" and the types of particles that pass through them are determined by the particular connexin proteins that make up the channel. Gap junctions made with connexin 26 transport potassium ions and certain small molecules. It is typically described in Iguchi et al. (2003), Experimental dermatology;12(3):283-8. The human GJB2 gene sequence is typically referenced as gene ID 2706 (NCBI). The human GJB2 protein sequence is typically referenced as UniProt P29033.

[0033] By "GJA1 gene" is meant here the gene coding for the protein "gap junction protein alpha 1". This protein is more commonly known as connexin 43. Connexin 43 is one of 21 connexin proteins. Connexins play a role in cell-to-cell communication by forming channels, or "gap junctions", between cells. "Gap junctions" allow the transport of nutrients, charged particles (ions) and other small molecules that carry the necessary communication signals between cells. In addition, connexin 43 attaches (binds) several signaling molecules that can relay communication signals within the cell. Connexin 43 is found in many tissues such as the eyes, skin, bones, ears, heart and brain, and it plays a role in their development and function. It is typically described in Salomon D et al.(1994), The Journal of Investigative Dermatology; 103(2):240-7. The human GJA1 gene sequence is typically referenced as gene ID 2697 (NCBI). The human GJA1 protein sequence is typically referenced as UniProt P17302.

[0034] By "GJB6 gene" is meant here the gene encoding the protein "gap junction protein beta 6". This protein is more commonly known as connexin 30. Connexin 30 is a member of the connexin protein family. Connexin proteins form channels called "gap junctions" that allow the transport of nutrients, charged atoms (ions), and signaling molecules between adjacent cells. The size of the "gap junction" and the types of particles that pass through it are determined by the particular connexin proteins that make up the channel. "Gap junctions" made with connexin 30 transport potassium ions and certain small molecules. Connexin 30 is found in several different tissues in the body, including the brain, inner ear, skin (especially the palms of the hands and soles of the feet), hair follicles, and nails. It is typically described in Salomon D et al.(1994), The Journal of Investigative Dermatology; 103(2):240-7. The human GJB6 gene sequence is typically referenced as gene ID 10804 (NCBI). The human GJB6 protein sequence is typically referenced as UniProt 095452.

[0035] By "GJA3 gene" is meant here the gene coding for the protein "gap junction protein alpha 3". This protein is more commonly called connexin 46. Connexin 46 is a member of the connexin protein family. The proteins of Connexin proteins form channels called gap junctions that allow the transport of nutrients, charged atoms (ions), and signaling molecules between adjacent cells. The size of the gap junction and the types of particles that pass through it are determined by the particular connexin proteins that make up the channel. It is typically described in Salomon D et al. (1994), The Journal of Investigative Dermatology;103(2):240-7. The human GJA3 gene sequence is typically referenced as gene ID 2700 (NCBI). The human GJA3 protein sequence is typically referenced as UniProt Q9Y6H8.

[0036] By "TJP2 gene" is meant here the gene encoding the protein "tight junction protein". This protein is more commonly called zonula occluden 2 which is a member of the membrane-associated guanylate kinase homolog family. The encoded protein functions as a component of the tight junction barrier in epithelial and endothelial cells and is required for proper assembly of tight junctions. It is typically described in Brandner Johanna et al. (2003), Archives of dermatological research;295(5):211-21. The human TJP2 gene sequence is typically referenced under the gene ID number 9414 (NCBI). The human TJP2 protein sequence is typically referenced under the UniProt number Q9UDY2.

[0037] By "CLDN8 gene", "CLDN10 gene", and "CLDN19 gene" are meant here the genes encoding the proteins Claudin 8, Claudin 10 and Claudin 19, respectively. Claudins are integral membrane proteins and components of tight junction strands. Tight junction strands serve as a physical barrier to prevent solutes and water from freely passing through the paracellular space between epithelial or endothelial cell sheets, and also play essential roles in maintaining cell polarity and signal transductions. They are typically described in Ashikari Daisaku et al. (2017), Cancer Science Jul; 108(7): 1386-1393, Günzel Dorothee et al. (2009), J Cell Science. May 15;122(Pt 10):1507-17, Konrad Martin et al. (2006), Am J Hum Genet. Nov;79(5):949-57. The human CLDN8 gene sequence is typically referenced as gene ID 9073 (NCBI).The human CLDN8 protein sequence is typically referenced as UniProt P56748. The human CLDN10 gene sequence is typically referenced as Gene ID 9071 (NCBI). The human CLDN10 protein sequence is typically referenced as UniProt P78369. The human CLDN19 gene sequence is typically referenced as Gene ID 149461 (NCBI). The human CLDN19 protein sequence is typically referenced as UniProt Q8N6F1.

[0038] By “CTNNB1 gene” is meant here the gene coding for the protein catenin beta 1.

[0039] This protein is present in many types of cells and tissues, where it is found mainly at junctions connecting neighboring cells (adherent junctions). Beta-catenin plays an important role in cell adhesion and intercellular communication. It is typically described in Hamburg Emily et al. (2012), J Invest Dermatol. Oct;132(10):2469-2472. The human CTNNB1 gene sequence is typically referenced as gene ID 1499 (NCBI). The human CTNNB1 protein sequence is typically referenced as UniProt P35222.

[0040] By "CTNND2 gene" is meant here the gene coding for the protein catenin delta 2. This protein is active in the nervous system, where it participates in cell adhesion and plays a role in cell movement. It is typically described in Lu Qun et al. (1999), J Cell Biol. Feb 8;144(3):519-32. The sequence of the human DSG4 gene is typically referenced under the gene ID number 1501 (NCBI). The sequence of the human CTNND2 protein is typically referenced under the UniProt number Q9UQB3.

[0041] In the context of the invention, the Gene ID (NCBI) and UniProt references cited above are those which were available on February 27, 2020.

[0042] By "expression level of gene X" is meant here the mRNA encoded by said gene X or the protein encoded by said gene X. The expression level of gene X can therefore be measured by quantifying the corresponding mRNA or protein. In a particular embodiment, said expression level of gene X is the mRNA encoded by said gene X.

[0043] Preferably, the level of expression corresponds to the concentration or quantity of the expression product (mRNA and / or protein).

[0044] The level of the expression product of a gene X can be measured by any technique well known to those skilled in the art. In particular, when the expression product is a protein, the level of the expression product can be measured by means of immunological assays such as ELISA assays, immunofluorescent assays (IFA), radioimmunological assays (RIA), competitive binding tests or Western Blots. When the expression product is an mRNA, the level of the expression product can be measured by RT-PCR, qRT-PCR, ddPCR (Droplet Digital PCR), by sequencing, for example by NGS (Next generation sequencing) type sequencing or by ddSEQTM single cell isolator sequencing.

[0045] By "in vitro diagnostic method of a common alopecic condition" is meant here a method for determining whether a subject is affected by a common alopecic condition.

[0046] By "in vitro prognostic method of a common alopecic condition" is meant here a method for determining whether a subject is at risk of being affected by a common alopecic condition.

[0047] The expression "prevention of a common alopecic condition" here means a prophylactic or preventive treatment of a common alopecic condition, which consists of preventing or delaying the onset of a common alopecic condition, in particular hair loss in the context of androgenetic alopecia.

[0048] The expression "treatment of a common alopecic condition" here means a treatment which consists of reducing, inhibiting, eliminating a common alopecic condition, in particular reducing, inhibiting, eliminating hair loss and / or promoting hair growth in the context of androgenetic alopecia.

[0049] For the purposes of the present invention, by "area suspected of being an alopecic area", we mean for example an area of ​​the scalp of a subject showing a loss of hair follicle density, in particular perceived or measured, for example: thinning hair, abnormal hair loss, thinning of the hair.

[0050] For the purposes of the present invention, the term “area suspected of becoming an alopecic area” means, for example, an area of ​​the scalp of a subject that has been an alopecic area.

[0051] By "subject" is meant a human being, preferably aged 17 to 80 years, preferably 20 to 70 years, 23 to 66 years. Preferably, the subject is a man. Detailed description of the invention

[0052] Diagnostic method

[0053] The subject of the present invention is a method for prognosis and / or in vitro diagnosis of a common alopecic condition of the scalp in a subject, in particular androgenetic alopecia, comprising at least one step a) of measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2.

[0054] Advantageously, the biological sample in step a) may be a scalp biopsy and / or a biopsy of one or more hair follicle(s), even better a biopsy of one or more hair follicle(s) such as at least one follicular transplant unit, in particular with a diameter of approximately 1 mm.

[0055] Preferably, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB1 and CTNND2.

[0056] In another preferred embodiment, said at least one gene is selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally selected from CTNNB1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0057] In this embodiment, said at least one gene is preferably chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, and optionally the group consisting of the genes CTNNB1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0058] In another embodiment, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06 and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0059] According to another embodiment, said at least one gene is chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0060] In another embodiment, said at least one gene is selected from the group consisting of the genes GJB2, GJA1, GJB6, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0061] According to another embodiment, said at least one gene is chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB1 and CTNND2.

[0062] Whatever the embodiment, said prognostic and / or diagnostic method may comprise at least one step a) of measuring the level of expression of at least two, three, four or five of the aforementioned genes.

[0063] The method according to the invention may further comprise the following steps: b) comparing the level of said at least one gene measured in step a) with a control; (c) based on the comparison of step (b), determining whether the scalp of said subject has a common alopecic condition.

[0064] The term "compare" refers to determining whether the expression level of said at least one gene is essentially identical to or different from a control. Preferably, the expression level of said at least one gene is considered different from a control if the observed difference is statistically significant. If the difference is not statistically significant, the expression level of said at least one gene and the control are essentially identical.

[0065] Based on this comparison, it can be determined whether a subject's scalp exhibits a common alopecic condition.

[0066] Thus, said scalp is confirmed as having a common alopecic condition when the level of expression of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2, is decreased compared to a control level.

[0067] Preferably, said scalp is confirmed as having a common alopecic condition when the expression level of at least one gene selected from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB1 and CTNND2, is decreased compared to a control level.

[0068] In another preferred embodiment, said scalp is confirmed as having a common alopecic condition when the expression level of at least one gene selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally at least one gene selected from CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19, is decreased compared to a control level.

[0069] In this mode, said scalp is confirmed as having a common alopecic condition when preferably the level of expression of at least one gene selected from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, and optionally the group consisting of the genes CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19, is decreased compared to a control level.

[0070] In another embodiment, said scalp is confirmed as having a common alopecic condition when the expression level of at least one gene selected from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB 1 and CTNND2, is decreased compared to a control level.

[0071] According to another embodiment, said scalp is confirmed as having a common alopecic condition when the level of expression of at least one gene chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2, is decreased compared to a control level.

[0072] In another embodiment, said scalp is confirmed as having a common alopecic condition when the expression level of at least one gene selected from the group consisting of the genes GJB2, GJA1, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2, is decreased compared to a control level.

[0073] According to another embodiment, said scalp is confirmed as having a common alopecic condition when the level of expression of at least one gene chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB 1 and CTNND2, is decreased compared to a control level.

[0074] By "decreased level" is meant here a level statistically significantly decreased compared to the control.

[0075] In a first embodiment, the “control” or “control level” is an average value determined by measuring the level of expression of at least one of said genes in a biological sample from a non-alopecic area in a population of subjects, for example a population of subjects presenting common alopecia, in particular androgenetic alopecia.

[0076] In a second embodiment, the “control” or “control level” is determined by measuring the level of expression of at least one of said genes in a biological sample from a non-alopecic area of ​​the same subject as that of step a).

[0077] In vitro method for evaluating the effectiveness of a treatment

[0078] The present invention also relates to an in vitro method for evaluating the efficacy of a treatment of a common alopecic condition comprising at least one step a) of measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB 1 and CTNND2 before and after treatment.

[0079] Preferably, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB 1 and CTNND2.

[0080] In another preferred embodiment, said at least one gene is selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally selected from CTNNB1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0081] In this embodiment, said at least one gene is preferably chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, and optionally the group consisting of the genes CTNNB1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0082] In another embodiment, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06 and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0083] According to another embodiment, said at least one gene is chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0084] In another embodiment, said at least one gene is selected from the group consisting of the genes GJB2, GJA1, GJB6, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0085] According to another embodiment, said at least one gene is chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB1 and CTNND2.

[0086] Whatever the embodiment, said in vitro method for evaluating the effectiveness of a treatment may comprise at least one step a) of measuring the level of expression of at least two, three, four or five of the aforementioned genes.

[0087] By the evaluation method as described, a given treatment will be considered effective for the treatment of a common alopecic condition when the expression level of at least one gene selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene selected from CTNNB1 and CTNND2, is increased after treatment compared to at the level of expression of said at least one gene before treatment.

[0088] By "increased level" is meant here a level statistically significantly increased compared to the level of expression of said at least one gene before treatment.

[0089] Preferably, a given treatment will be considered effective for the treatment of a common alopecic condition when the expression level of at least one gene selected from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB1 and CTNND2, is increased compared to a control level.

[0090] In another preferred embodiment, a given treatment will be considered effective for the treatment of a common alopecic condition when the expression level of at least one gene selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally at least one gene selected from CTNNB1, CTNND2, MY06, GJB6, CLDN10, and CLDN19, is increased relative to a control level.

[0091] In this mode, a given treatment will be considered effective for the treatment of a common alopecic condition when preferably the expression level of at least one gene selected from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8,, and optionally the group consisting of the genes CTNNB1, CTNND2, MY06, GJB6, CLDN10, and CLDN19, is increased compared to a control level.

[0092] In another embodiment, a given treatment will be considered effective for the treatment of a common alopecic condition when the expression level of at least one gene selected from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2, is increased compared to a control level.

[0093] According to another embodiment, a given treatment will be considered effective for the treatment of a common alopecic condition when the level of expression of at least one gene chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2, is increased compared to a control level.

[0094] In another embodiment, a given treatment will be considered as effective for the treatment of a common alopecic condition when the expression level of at least one gene selected from the group consisting of the genes GJB2, GJA1, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2, is increased compared to a control level.

[0095] According to another embodiment, a given treatment will be considered effective for the treatment of a common alopecic condition when the level of expression of at least one gene chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB1 and CTNND2, is increased compared to a control level.

[0096] A treatment will be considered to have no effect if the expression levels of the chosen gene, before and after treatment, are substantially identical, or if the observed differences are not significant.

[0097] When comparing the expression levels of more than one gene, the treatment is considered effective for the treatment of a common alopecic condition, including androgenetic alopecia, if, for the majority of the genes tested, and preferably for all the genes tested individually, the treatment is considered effective. For the other genes selected, the treatment should preferably have no effect, but not the opposite effect.

[0098] According to another implementation, the method for evaluating the effectiveness of a treatment of a common alopecic condition comprises the diagnosis of a common alopecic condition according to the method of the invention, before and after treatment, and the comparison of the differences in expression levels observed between a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area and a biological sample from a non-alopecic area.

[0099] According to this implementation of the evaluation of the effectiveness of a treatment, the treatment is considered effective if the difference between the expression levels of the chosen gene(s) in said biological sample from an area suspected of being an alopecic area or of becoming an alopecic area and said biological sample from a non-alopecic area is less after treatment compared to what it was before the treatment.

[0100] When the expression levels of more than one gene are compared, the effectiveness of the treatment is preferably concluded when for the majority of the genes chosen, and preferably for all the genes chosen, taken individually, it can be concluded that treatment is effective.

[0101] Preferably, the comparison of the expression levels of the chosen gene(s) is carried out on a biological sample chosen from a scalp biopsy and / or a biopsy of one or more hair follicle(s), even better a biopsy of one or more hair follicle(s) such as at least one transplant follicular unit, in particular with a diameter of approximately 1 mm.

[0102] The treatment considered, evaluated by the method of the invention, is not limited to a particular type of treatment. It may be a treatment by one or more compounds of natural and / or synthetic origin, a natural extract, in particular an essential oil, a nucleic acid, a protein complex or any other molecule or combination of molecules. It may also be a treatment by physical means such as waves, in particular electromagnetic waves. It is preferably a topical treatment, but it is also envisaged to evaluate the effectiveness of treatments administered orally, by injection, or by any other means of administration.

[0103] The treatment tested may aim to reduce or inhibit hair loss linked to a common alopecic condition, in particular linked to androgenetic alopecia and / or to promote hair growth.

[0104] Particularly preferred treatments within the scope of this invention are cosmetic treatments, more particularly cosmetic topical treatments.

[0105] By the methods of the invention, it is also possible to evaluate the effectiveness of the combination of several treatments. It is indeed possible to evaluate combinations which best restore the expression levels of one, several or all of the genes of the invention, as they are expressed within a biological sample from a non-alopecic area.

[0106] By the evaluation methods described above, it is possible to evaluate the effectiveness of a new treatment envisaged, or also to quantify or qualify the effectiveness of already existing treatments against common alopecia, in particular androgenetic alopecia.

[0107] In this way, combinations of treatments that may be particularly effective, synergistic or complementary can also be considered.

[0108] Cosmetic treatment method

[0109] The present invention also relates to a method for the cosmetic treatment of a common alopecic condition of the scalp, in particular androgenetic alopecia, comprising at least the following steps: a) measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2; b) deducing from step a) whether the scalp of said subject has a common alopecic condition; c) if the scalp is identified as having an alopecic condition common to step b) treating said scalp with a cosmetic composition making it possible to induce and / or stimulate hair growth and / or slow down hair loss.

[0110] Preferably, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB1 and CTNND2.

[0111] In another preferred embodiment, said at least one gene is selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally selected from CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0112] In this embodiment, said at least one gene is preferably chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, and optionally the group consisting of the genes CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0113] In another embodiment, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06 and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB 1 and CTNND2.

[0114] According to another embodiment, said at least one gene is chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0115] In another embodiment, said at least one gene is selected from the group consisting of the genes GJB2, GJA1, GJB6, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB 1 and CTNND2.

[0116] According to another embodiment, said at least one gene is chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB1 and CTNND2.

[0117] Whatever the embodiment, said cosmetic treatment method may comprise at least one step a) of measuring the level of expression of at least two, three, four or five of the aforementioned genes.

[0118] Use of modulator

[0119] The present invention also relates to the use of at least one modulator of the expression level of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2, or of a cosmetic composition comprising it or them, for the prevention and / or treatment of a common alopecic condition, in particular androgenetic alopecia.

[0120] Preferably, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB1 and CTNND2.

[0121] In another preferred embodiment, said at least one gene is selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally selected from CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0122] In this embodiment, said at least one gene is preferably chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, and optionally the group consisting of the genes CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0123] In another embodiment, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06 and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB 1 and CTNND2.

[0124] According to another embodiment, said at least one gene is chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0125] In another embodiment, said at least one gene is chosen from the group consisting of the genes GJB2, GJA1, GJB6, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0126] According to another embodiment, said at least one gene is chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB1 and CTNND2.

[0127] Whatever the embodiment, said modulator can modulate the level of expression of at least two, three, four or five of the aforementioned genes.

[0128] The term "cosmetic composition" means a composition comprising a physiologically acceptable medium, that is to say a medium compatible with the skin, in particular the skin of the scalp. According to a particular embodiment, the pH of the cosmetic composition is between 4 and 7.5, in particular between 4.5 and 7, and in particular between 4.7 and 6.5.

[0129] More particularly, said physiologically acceptable medium may comprise water and / or one or more water-miscible organic solvents which may be chosen from linear or branched C1-C6 monoalcohols such as ethanol, isopropanol, tert-butanol or n-butanol; polyols such as glycerol, propylene glycol, hexylene glycol (or 2-methyl-2,4-pentanediol), and polyethylene glycols; polyol ethers such as dipropylene glycol monomethyl ether; and mixtures thereof.

[0130] Preferably, the composition has a water content ranging from 20% to 95% by weight, even better from 40% to 90% by weight relative to the total weight of the composition.

[0131] Advantageously, the composition comprises one or more water-miscible organic solvents in a content ranging from 0.5% to 25% by weight, preferably from 5% to 20% by weight, even better from 10% to 15% by weight relative to the total weight of the composition.

[0132] A composition containing said modulator may preferably be administered topically.

[0133] The composition may further comprise other compounds different from the modulators of the aforementioned genes but known for their anti-hair loss / regrowth activity.

[0134] The support can be of various natures depending on the type of composition considered.

[0135] With regard more particularly to the compositions intended for external topical administration, these may be aqueous, hydroalcoholic or oily solutions, solutions or dispersions of the lotion or serum type, emulsions of liquid or semi-liquid consistency of the milk type, obtained by dispersion of a fatty phase in an aqueous phase (O / W) or vice versa (W / O), or of suspensions or emulsions, of soft, semi-solid or solid consistency, of the cream type, aqueous or anhydrous gel, microemulsions, microcapsules, microparticles, or vesicular dispersions of ionic and / or non-ionic type.

[0136] These compositions are prepared according to the usual methods.

[0137] These compositions can in particular constitute cleaning creams, protection, treatment or care, lotions, gels or mousses for the care of the scalp.

[0138] They can be used for the scalp in the form of solutions, creams, gels, emulsions, mousses or even in the form of aerosol compositions also containing a pressurized propellant.

[0139] A topical composition according to the invention can advantageously be formulated in any galenic form suitable for hair care, in particular in the form of a hair lotion, hair gel, shampoo, conditioner, conditioner, hair cream or gel, hairspray, styling lotion, treatment lotion, dyeing composition (in particular oxidation) possibly in the form of a coloring shampoo, a restructuring lotion for hair, a perm composition, an antiparasitic shampoo, or a treatment shampoo, in particular anti-seborrheic, a scalp care product, in particular anti-irritant, anti-aging, restructuring.

[0140] When a composition of the invention is an emulsion, the proportion of the fatty phase may range from 5 to 80% by weight, and preferably from 10 to 50% by weight relative to the total weight of the composition. The oils, emulsifiers and co-emulsifiers used in the composition in the form of an emulsion are chosen from those conventionally used in the cosmetic and / or dermatological field. The emulsifier and the co-emulsifier may be present, in the composition, in a proportion ranging from 0.3 to 30% by weight, and preferably from 0.5 to 20% by weight relative to the total weight of the composition.

[0141] When the composition of the invention is an oily solution or gel, the fatty phase can represent more than 90% of the total weight of the composition.

[0142] In a known manner, the galenic forms dedicated to topical administration may also contain adjuvants customary in the cosmetic, pharmaceutical and / or dermatological field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, perfumes, fillers, filters, odor absorbers, additional anti-hair loss or regrowth active agents other than the modulators of the expression of the aforementioned genes and coloring materials. The quantities of these different adjuvants are those conventionally used in the field considered, and for example from 0.01 to 20% of the total weight of the composition. These adjuvants, depending on their nature, may be introduced into the phase fatty and / or in the aqueous phase.

[0143] Method for identifying an active compound

[0144] The present invention further relates to a method for identifying a compound for the prevention and / or treatment of a common alopecic condition, in particular androgenetic alopecia, comprising the following steps: (a) bringing a compound to be tested into contact with a biological sample; b) measuring the level of expression, in said biological sample, of at least one gene chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2; c) selecting a compound for which the level of expression of said at least one gene measured in step b) is increased compared to its level of expression in the absence of said compound.

[0145] By "increased level" is meant here a statistically significantly increased level compared to the level of expression of said at least one gene in the absence of said compound.

[0146] Preferably, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB1 and CTNND2.

[0147] In another preferred embodiment, said at least one gene is selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJA3, TJP2, CLDN8 and optionally selected from CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0148] In this embodiment, said at least one gene is preferably chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CTNNB 1, CTNND2, MY06, GJB6, CLDN10, and CLDN19.

[0149] In another embodiment, said at least one gene is chosen from the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06 and optionally the group consisting of the genes DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB 1 and CTNND2.

[0150] According to another embodiment, said at least one gene is chosen from the group consisting of the genes DSG2, DSG3, DSG4, DSC2, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0151] In another embodiment, said at least one gene is selected from the group consisting of the genes GJB2, GJA1, GJB6, GJA3, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, TJP2, CLDN8, CLDN10, CLDN19, CTNNB1 and CTNND2.

[0152] According to another embodiment, said at least one gene is chosen from the group consisting of the genes TJP2, CLDN8, CLDN10, CLDN19, and optionally the group consisting of the genes CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MYO3B, MYO5B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, CTNNB1 and CTNND2.

[0153] Whatever the embodiment, said method for identifying a compound may comprise at least one step b) of measuring the level of expression of at least two, three, four or five of the aforementioned genes.

[0154] Advantageously, the biological sample is chosen from a scalp biopsy, an in vitro model of reconstructed scalp, a biopsy of one or more hair follicle(s), an in vitro model of reconstructed hair follicle, even better a biopsy of one or more hair follicle(s) such as at least one follicular transplant unit, in particular with a diameter of approximately 1 mm.

[0155] The expression “at least one” is equivalent to “one or more”.

[0156] The expressions "between ... and..." and "ranging from ... to ...", "at least..." or "at most" must be understood inclusively, unless otherwise specified.

[0157] The following examples and figures are provided for illustrative purposes and do not limit the scope of the invention. Figures

[0158] [Fig-1]: Identification of alopecic (vertex) and non-alopecic (occiput) areas.

[0159] [Fig.2]: Sampling of a follicular unit from an alopecic area (vertex).

[0160] [Fig.3]: Follicular unit taken from an alopecic area (vertex). Examples

[0161] A - Characterization of a common alopecic condition

[0162] A transcriptomic study by high-throughput sequencing (RNA-sequencing) was carried out on a category of 10 volunteers suffering from grade 4 androgenetic alopecia according to the Hamilton scale (age group 23-66 years) by dissecting follicular units in the anagen phase from biopsies taken from the two areas of their scalp. : the alopecic area at the vertex and the non-alopecic area at the occipital level (Figures 1, 2 and 3). Ten follicular units per area and per individual were isolated. A total of 20 samples of 10 follicular units each were thus obtained.

[0163] Total RNA was isolated from these samples with the RNeasy kit (Qiagen, Germantown, MD, USA). RNA purity and integrity were assessed on the Agilent 2100 Bioanalyzer with the RNA 6000 NanoLabChip kit (Agilent, Palo Alto, CA, USA) and the Nanodrop spectrophotometer. All samples had RIN values ​​> 7.

[0164] For library preparation and RNA sequencing, 75 ng of total RNA was reverse transcribed into complementary DNA (cDNA) of the first strand of RNA using a mixture of random and poly-dT primers, and omitting the DNase treatment step. Synthesis of the second strand, using a nucleotide analog for retention, generated the double-stranded cDNA. The latter was fragmented into pieces of 200 to 500 bases. Then, end repair was performed to generate a blunt-ended double-stranded cDNA, followed by ligation of indexing adapters, strand selection via targeted degradation by a nucleotide analog, and reduction of ribosomal RNA content. Finally, cDNA libraries were created by 18-cycle PCR enrichment. Equimolar amounts of each library were sequenced on a NextSeq 500, for 75-base paired-end sequencing.

[0165] The results are presented in the following Table 2:

[0166] [Tables2] Genes Gene names Adjusted p-value Fold change (FC) Log2-FC % of subjects with a FC < -1.25 Adherens junctions CDH1 cadherin 1 3.90E-03 -1.30 -0.36 70 ACTB beta-actin l.18E-03 -1.28 -0.36 70 ACTBL2 actin beta like 2 l.26E-03 -4.91 -2.30 100 TUBB tubulin beta, class I l.36E-02 -1.24 -0.31 70 TUBB2A tubulin beta 2A, class IIa 8.23E-03 -1.47 -0.55 80 GSN gelsolin 7.75E-03 -1.34 -0.42 80 MY03B myosin-IIIb 2.86E-03 -1.65 -1.65 90 MY05B myosin-Vb 5.36E-03 -1.39 -0.47 70 MY06 myosin-VI 6.24E-03 -1.26 -0.33 70 Desmosomes DSG2 desmoglein 2 4.89E-02 -1.40 -0.48 60 DSG3 desmoglein 3 l,56E-03 -1.37 -0.45 70 DSG4 desmoglein 4 4.66E-04 -4.68 -2.23 100 DSC2 desmocollin 2 2.98E-05 -1.72 -0.78 100 Gap junctions GJB2 gap junction protein beta 2 7.09E-04 -1.86 -0.89 90 GJA1 gap junction protein alpha 1 l,97E-05 -1.78 -0.83 100 GJB6 gap junction protein beta 6 2.59E-05 -2.54 -1.34 100 GJA3 gap junction protein alpha 3 6.24E-05 -2.86 -1.51 100 Tight junctions TJP2 tight junction protein l,41E-03 -1.33 -0.41 100 CLDN8 Claudine 8 l,66E-02 -1.54 -0.62 90 CLDN10 Claudine 10 4.09E-02 -1.53 ​​-0.62 70 CLDN19 Claudine 19 2.80E-03 -3.66 -1.87 90 , CDH1 interaction genes CTNNB1 beta catenin 1 l,63E-05 -1.65 -0.73 100 CTNND2 delta catenin 2 2.10E-03 -2.05 -1.03 90

[0167] The above results show that at least 60% of the selected alopecic volunteers present the differentiating expression criteria making it possible to highlight a significant imbalance of transcripts coding for intercellular junctions which make up the hair follicle (Table 2). All of these transcripts or genes are specifically overexpressed in growing hair follicles, and consequently specifically underexpressed in alopecic hair follicles.

Claims

Claims

1. In vitro diagnostic method for androgenetic alopecia, comprising at least one step a) of measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene involved in the intercellular junctions of the cells of the scalp and / or the hair follicle chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2.

2. A method according to claim 1, further comprising the following steps: b) comparing the level of said at least one gene measured in step a) with a control; c) based on the comparison of step b), determining whether the scalp of said subject exhibits androgenetic alopecia.

3. Method according to claim 1 or 2, wherein said scalp is confirmed as exhibiting androgenetic alopecia when the expression level of at least one gene involved in the intercellular junctions of scalp and / or hair follicle cells selected from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene selected from CTNNB1 and CTNND2, measured in step a) as is decreased compared to a control level.

4. In vitro method for evaluating the efficacy of a treatment for androgenetic alopecia comprising at least one step a) of measuring the level of expression, in a biological sample from an area suspected of being an alopecic area or of becoming an alopecic area in said subject, of at least one gene involved in the intercellular junctions of the cells of the scalp and / or the hair follicle chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2 before and after treatment.

5. Method according to claim 4, wherein the treatment is considered effective for the treatment of androgenetic alopecia when the level of expression of at least one gene involved in the intercellular junctions of the cells of the scalp and / or the hair follicle chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2, measured in step a) is increased after treatment compared to the level of expression of said at least one gene before treatment.

6. Cosmetic composition comprising one or more modulator(s) of the expression level of at least one gene involved in the intercellular junctions of the cells of the scalp and / or the hair follicle chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2, for use in the prevention and / or treatment of androgenetic alopecia.

7. Method for identifying a compound for the prevention and / or treatment of androgenetic alopecia, comprising the following steps: a) bringing a compound to be tested into contact with a biological sample b) measuring the level of expression, in said biological sample, of at least one gene involved in the intercellular junctions of the cells of the scalp and / or the hair follicle chosen from CDH1, ACTB, ACTBL2, TUBB, TUBB2A, GSN, MY03B, MY05B, MY06, DSG2, DSG3, DSG4, DSC2, GJB2, GJA1, GJB6, GJA3, TJP2, CLDN8, CLDN10, CLDN19, and optionally of at least one gene chosen from CTNNB1 and CTNND2; c) selecting a compound for which the level of expression of said at least one gene measured in step b) is increased compared to its level of expression in the absence of said compound.

8. Method according to claim 7, wherein the biological sample is selected from a scalp biopsy, an in vitro model of reconstructed scalp, a biopsy of one or more hair follicle(s), an in vitro model of reconstructed hair follicle.

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