METHOD OF PROGNOSIS AND/OR DIAGNOSIS OF A PELLICULAR CONDITION OF THE SCALP

Abstract

The present invention relates to a method of prognosis and/or diagnosis of a pellicular condition of the scalp, based on the measurement of the quantity of the active form of transglutaminase 3. The present invention also relates to a cosmetic treatment method for the scalp, a method for identifying compounds for reducing and/or eliminating the pellicular condition. Finally, the present invention relates to a kit and the use thereof for identifying a compound for reducing and/or eliminating the pellicular condition.

Claims

1. A method of prognosis and/or diagnosis of a pellicular condition of the scalp, in a subject, said method comprising the following step: a. measuring the quantity of the active form of transglutaminase 3 (TGM3), in a scalp sample from said subject.

2. A method of cosmetic prevention and/or treatment of a scalp having a pellicular condition in a subject, said method comprising the following steps: a. measuring the quantity of the active form of TGM3, in a scalp sample from said subject; b. inferring from step a) whether said subject's scalp has a pellicular condition or whether said subject's scalp is likely to develop a pellicular condition; c. if the scalp is identified as having a pellicular condition or as being likely to develop a pellicular condition in step b), treating said scalp with a cosmetic composition for the pellicular condition.

3. A method for identifying a compound for preventing and/or reducing and/or eliminating the pellicular condition of a scalp, said method comprising the following steps: a. measuring the quantity of the active form of TGM3, in a scalp sample exposed to said candidate compound; b. comparing said quantity measured in step a) to the quantity of the active form of TGM3, in a scalp sample not exposed to said compound; c. identifying said candidate compound as a compound for preventing and/or reducing and/or eliminating the pellicular condition of a scalp when a reduction in the quantity of the active form of TGM3 in the scalp sample that was exposed to said candidate compound is detected, with respect to the quantity of the active form of TGM3 in the scalp sample that was not exposed to the candidate compound.

4. The method according to claim 3, wherein said scalp sample used in the identification method according to the invention, consists of a cell culture obtained from a skin having body hair or a scalp, cells obtained from a skin having body hair or a scalp, a reconstructed complete scalp, or a human scalp explant.

5. The method according to claim 1, wherein the scalp sample is taken by rubbing the scalp surface or using an adhesive surface.

6. The method according to claim 1, wherein the quantity of the active form of TGM3 is measured using a ligand specific for the active form of TGM3.

7. A kit comprising: a. a means for measuring in a scalp sample the quantity of the active form of TGM3, and b. an instruction leaflet.

8. The kit according to claim 7, wherein the means for measuring, in a scalp sample, the quantity of the active form of TGM3, comprises a ligand specific for the active form of TGM3.

9. The kit according to claim 7, wherein the means for measuring in a scalp sample comprises an immunochromatographic test.

10. A method for identifying a compound for preventing and/or reducing and/or eliminating the pellicular condition of a scalp which comprises using the kit according to claim 7 to measure the quantity of the active form of TGM3 in a scalp sample exposed to a candidate compound.

11. The method according to claim 2, wherein the scalp sample is taken by rubbing the scalp surface or using an adhesive surface.

12. The method according to claim 3, wherein the scalp sample is taken by rubbing the scalp surface or using an adhesive surface.

13. The method according to claim 4, wherein the scalp sample is taken by rubbing the scalp surface or using an adhesive surface.

14. The method according to claim 2, wherein the quantity of the active form of TGM3 is measured using a ligand specific for the active form of TGM3.

15. The method according to claim 3, wherein the quantity of the active form of TGM3 is measured using a ligand specific for the active form of TGM3.

16. The method according to claim 4, wherein the quantity of the active form of TGM3 is measured using a ligand specific for the active form of TGM3.

17. The method according to claim 5, wherein the quantity of the active form of TGM3 is measured using a ligand specific for the active form of TGM3.

18. The kit according to claim 8, wherein the means for measuring in a scalp sample comprises an immunochromatographic test.

19. A method for identifying a compound for preventing and/or reducing and/or eliminating the pellicular condition of a scalp which comprises using the kit according to claim 8 to measure the quantity of the active form of TGM3 in a scalp sample exposed to a candidate compound.

20. A method for identifying a compound for preventing and/or reducing and/or eliminating the pellicular condition of a scalp which comprises using the kit according to claim 9 to measure the quantity of the active form of TGM3 in a scalp sample exposed to a candidate compound.

Description

FIGURES

[0115] FIG. 1 shows the linear regression based on the logarithm of the mean TGM3 measurement with respect to the overall score of the pellicular condition. The logarithm of the mean TGM3 measurements is on the y-axis and the dandruff score is on the x-axis.

[0116] FIG. 2 shows the ROC curve of the use of the logarithm of the quantity of TGM3 to detect pellicular conditions of the scalp.

EXAMPLES

Example 1: Development of Recombinant Human Antibodies Targeted Against TGM3 and Selection of a Pair of Antibodies

[0117] Highly specific recombinant human antibodies against TGM3 were generated using Bio-Rad AbD Serotec HuCAL technology. In brief, a HuCAL bank containing 4.510.sup.10 antibodies, which are presented on phage particles, was screened with 200 g of the active form of human recombinant TGM3 protein. Three amplification cycles were performed in E. coli using an auxiliary phase to enrich the antibody phage selected. The DNA of the selected antibody phages was collected and subcloned in a FabFab-A_FSx2 formatbivalent Fab fusion antibodybacterial alkaline phosphatase expression vector, tracked by FLAG and a His6 marker. After transformation and expression in E. coli, the DNA of the positive clones was selected and sequenced to identify unique antibody clones. Each unique clone was expressed in E. coli and purified by affinity chromatography. The affinity and the specificity of each antibody was evaluated by direct ELISA, using a recombinant TGM3 and using non-related antigens and markers.

Example 2: Sandwich Test Based on TGM3 Antibody

[0118] Microfluidic Cartridges

[0119] Personalized TGM3 microfluidic cartridges were used to measure the quantity of TGM3 in soluble protein extracts from stratum corneum samples collected using sterile swabs from the scalp of individuals with and without dandruff. The recombinant capture antibody was fixed inside microfluidic grooves (NanoEnTek as described in the published patent FR3023486). A standard curve, from 1 to 170 ng/ml, was plotted using a recombinant TGM3 (active form). The proteins from the sample on the sterile swab were solubilized in a buffer (50 mM Tris, 150 mM NaCl, pH 8.0), and 35 l was injected inside the microfluidic cartridges. Detection was performed using fluorescent beads coated with an anti-TGM3 detection antibody.

Example 3: Clinical Study

[0120] Study Design

[0121] This is a single-center, randomized, double-blind comparative study, with 3 parallel groups and under medical supervision. A period of 2 treatment-free weeks is followed by 4 weeks of treatments. The treatments consist of the use of a shampoo. Three shampoos are tested and each condition is monitored on a sample of 20 subjects.

[0122] The subjects selected for the study are healthy male or female volunteers, aged from 18 to 60 years inclusive, whose hair length is greater than 2 cm and preferably shorter than shoulder length; having a moderate to severe pellicular condition of the scalp; usually using shampoo 3 times/week and seeking to shampoo three times per week during the study;

[0123] Evaluation Criteria

[0124] The primary criterion is the overall score of the pellicular condition of the scalp: evaluation of adherent and non-adherent dandruff according to an ordinal scale ranging from 0 to 5 (total dandruff score ranging from 0 to 10), at baseline and after 4 weeks of treatment.

[0125] Analysis of Results

[0126] The TGM3 protein concentration was converted into a logarithmic scale.

[0127] The volunteers were classified as dandruff + or according to their overall score of the pellicular condition (adherent+non-adherent dandruff) of the entire scalp area by following these rules:

Dandruff +=(overall score of pellicular condition)>2.5

Dandruff =(overall score of pellicular condition)<1

[0128] Linear Regression

[0129] FIG. 1 represents the linear regression based on the mean TGM3 measurement with respect to the overall score of the pellicular condition.

[0130] The analysis suggests a good correlation between the TGM3 concentration and the overall score of the pellicular condition with a determination coefficient R.sup.2 of 0.968.

[0131] ROC Curve

[0132] The receiver operating characteristic, or ROC curve, is a graphical plot that illustrates the diagnostic ability of a binary classifier system as its discrimination threshold is varied. The rate of true positives is also known as the sensitivity. The rate of false positives is also known as the probability of false alarm and can be calculated as (1 specificity).

[0133] FIG. 2 represents the ROC curve of the use of the logarithm of the quantity of TGM3 to detect pellicular conditions.

[0134] This analysis makes it possible to determine the threshold that differentiates the two groups (with and without dandruff) the most and the sensitivity and specificity of the biomarker.

[0135] Based on the best sensitivity and specificity of the ROC curve, the threshold for separating the two groups is here 10.44 ng/ml of TGM3 (i.e. 2.346 on a logarithmic scale, corresponding to the third point from the left in FIG. 2).