Method for estimating skin condition, and method for proposing composition and / or stimulation suitable for subject

Abstract

To provide a novel method for estimating the skin condition of a subject using skin samples.SOLUTION: A method for estimating the skin condition of a subject includes performing gene expression analysis on skin samples from the subject and estimating the subject's skin condition based on the analysis results.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a method for estimating a skin condition and a method for suggesting a composition and / or stimulation suitable for a subject. [Background technology]

[0002] Conventionally, as a technique combining skin-derived samples with diagnosis, a method for diagnosing the condition of a subject using nucleic acids derived from sebum is known (Patent Document 1).

[0003] In addition, a method for testing acne skin is known in which FABP-5 in the stratum corneum of the skin is measured and compared with the measured value of FABP-5 in normal stratum corneum of the skin measured beforehand or simultaneously, and an increase in expression is used as an indicator of acne malignancy (Patent Document 2). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2018-000206 A [Patent Document 2] JP 2016-095185 A Summary of the Invention [Problem to be solved by the invention]

[0005] With the above-mentioned prior art, since nucleic acids derived from sebum are not nucleic acids found in the skin itself, it has been difficult to evaluate deep inside the skin or evaluate genes that are not secreted into sebum. In addition, since cell nuclei are enucleated in the stratum corneum, it was difficult to perform accurate genetic evaluation.

[0006] In view of the above circumstances, an object of the present invention is to provide a novel method for estimating the skin condition of a subject using the skin. Another objective of the present invention is to provide a novel method (excluding medical procedures) for suggesting a composition and / or stimulation suitable for a subject based on the subject's estimated skin condition. [Means for solving the problem]

[0007] The present invention, which solves the above-mentioned problems, provides a method for estimating a subject's skin condition, which comprises performing gene expression analysis using skin sampled from a subject, and estimating the subject's skin condition based on the results of the analysis.

[0008] In a preferred embodiment of the present invention, the gene expression analysis is carried out by PCR or transcriptome analysis using RNA extracted from the skin as a sample.

[0009] In a preferred embodiment of the invention, the gene has one or more Gene Ontologies selected from the group consisting of: [Table 1]

[0010] In a preferred embodiment of the invention, the gene has one or more Gene Ontologies selected from the group consisting of: [Table 2]

[0011] In a preferred embodiment of the invention, the gene is one or more selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL.

[0012] In a preferred embodiment of the present invention, the gene is one or more selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

[0013] In a preferred form of the invention, the skin condition is pigmentation.

[0014] In a preferred embodiment of the present invention, the method includes estimating that the skin condition is pigmentation caused by fluctuations in the expression level of one or more genes selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL.

[0015] In a preferred embodiment of the present invention, the skin condition is estimated to be pigmentation caused by fluctuations in the expression level of one or more genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

[0016] In a preferred embodiment of the present invention, the skin condition of a subject is estimated based on a comparison between the results of gene expression analysis performed on skin taken from a pigmented site of the subject and the results of gene expression analysis performed on skin taken from a non-pigmented site of the subject.

[0017] In a preferred embodiment of the present invention, the skin is skin that has been dried under normal temperature conditions.

[0018] In a preferred embodiment of the invention, the volume of skin immediately after acquisition from the subject is less than 0.05 mm 3 The following is the result.

[0019] In a preferred embodiment of the present invention, the skin includes an epidermis region including the stratum corneum, and a dermis region located at a depth of 0.2 to 0.5 mm from the outermost layer of the skin.

[0020] The present invention also provides a method for determining a skin condition of a subject by carrying out gene expression analysis using skin samples taken from the subject, and estimating the skin condition of the subject based on the results of the analysis; and a proposing step of proposing a composition and / or stimulation suitable for the subject based on the estimated skin condition (excluding medical treatment). It also includes a method (excluding medical procedures) for suggesting suitable compositions and / or stimuli to a subject.

[0021] In a preferred embodiment of the present invention, the method further comprises the steps of: applying to said subject the composition and / or stimulus proposed to said subject; the estimation step being carried out based on the results of gene expression analysis performed using the skin sampled from the subject after the application step; and a re-suggestion step of re-suggesting a composition and / or stimulation suitable for the subject (excluding medical treatment) based on the estimated skin condition.

[0022] The present invention also provides an apparatus for estimating a skin condition of a subject, comprising: a gene expression analysis result acquisition unit that acquires the results of gene expression analysis performed using skin sampled from a subject; an estimation means for estimating a skin condition of the subject based on a result of the gene expression analysis; The present invention is also an apparatus for estimating a skin condition of a subject.

[0023] The present invention also provides a suggestion device for suggesting a composition and / or a stimulus suitable for a subject, comprising: a gene expression analysis result acquisition unit that acquires the results of gene expression analysis performed using skin sampled from a subject; an estimation means for estimating a skin condition of the subject based on a result of the gene expression analysis; A suggestion means for suggesting a composition and / or stimulation suitable for the subject based on the estimated skin condition; The present invention is also directed to a device for suggesting a composition and / or stimulus suitable for a subject, comprising:

[0024] The present invention also provides a program for estimating the skin condition of a subject using gene expression levels in skin collected from the subject, the program causing a computer to execute a step of estimating the skin condition of the subject based on the gene expression levels.

[0025] The present invention also provides a suggestion program for suggesting a composition and / or stimulation suitable for a subject by utilizing gene expression levels in skin sampled from the subject, Obtaining results of gene expression analysis using skin samples taken from a subject; estimating a skin condition of the subject based on the gene expression level; A suggestion step of suggesting a composition and / or stimulation suitable for the subject based on the estimated skin condition; It is also a program for causing a computer to execute the above.

[0026] The present invention also relates to a method for treating a subject comprising: applying a composition and / or a stimulus to the subject; and an estimation step of performing gene expression analysis using the skin sampled from the subject after the application step, and estimating the skin condition of the subject based on the results of the analysis. evaluating the suitability of the composition and / or the irritation for the subject by comparing the result of the estimation step with the skin condition of the subject obtained in advance before the application step; It is also a method for evaluating a composition and / or stimulus suitable for a subject (but excluding medical procedures). Effect of the Invention

[0027] According to the present invention, the skin condition of a subject can be estimated using the skin. In addition, it is possible to suggest compositions and / or stimulations suitable for the subject based on the estimated skin condition of the subject (but excluding medical procedures). [Brief description of the drawings]

[0028] [Figure 1] FIG. 2 is a flow chart showing a specific embodiment of the estimation method of the present invention. [Diagram 2] FIG. 2 is a flow diagram showing a specific embodiment of the proposed method of the present invention. [Diagram 3] FIG. 1 is a flow diagram showing a specific embodiment of a preferred mode of the proposed method of the present invention. [Figure 4]FIG. 2 is a hardware block diagram of the estimation device of the present invention. [Diagram 5] FIG. 2 is a hardware block diagram of the proposed device of the present invention. [Figure 6] FIG. 1 shows the results of PCR analysis in <Test Example 1>. [Figure 7] FIG. 1 shows the results of RNA-seq in <Test Example 2>. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] <1> Method for estimating subject's skin condition The present invention provides a method for estimating a skin condition of a subject, which comprises performing gene expression analysis using skin sampled from the subject, and estimating the skin condition of the subject based on the results of the analysis.

[0030] The skin includes the epidermis, dermis and / or subcutaneous tissue, and includes skin fragments and skin cells. The skin preferably includes an epidermis region including the stratum corneum, and a dermis region located at a depth of 0.2 to 0.5 mm from the outermost layer of the skin.

[0031] The dermis region is preferably located at a depth of 0.20 mm or more, more preferably 0.21 mm or more, and even more preferably 0.22 mm or more from the outermost layer of the skin. Moreover, the dermis region is preferably located at a depth of 0.50 mm or less, more preferably 0.45 mm or less, and even more preferably 0.40 mm or less, from the outermost layer of the skin.

[0032] Although the method for collecting the skin is not particularly limited, it is preferable to adopt a minimally invasive collection method. Specifically, it is preferable to collect the skin using a needle with a diameter of 1 mm or less. The diameter of the needle is preferably 0.75 mm or less, more preferably 0.50 mm or less, and even more preferably 0.40 mm or less.

[0033] The amount of skin to be collected is not particularly limited, but the volume of the skin immediately after collection from the subject is 0.05 mm 3It is preferably equal to or less than 0.3 mm, and more preferably 0.3 mm 3 Less than 0.1 mm, more preferably 3 The following is the result.

[0034] The area from which the skin is taken is not particularly limited, but examples include the face, neck, back of the hand, abdomen, feet, and fingers. Preferably, the skin is taken from an area prone to skin diseases or an area that is easy to use for comparing exposed and non-exposed areas, such as the arms.

[0035] In the present invention, "gene expression analysis" refers to investigating the expression state of a gene.

[0036] The method of gene expression analysis is not particularly limited, but it is preferable that comprehensive gene expression analysis is possible. Specifically, mRNA is extracted from the skin, and after appropriate sample processing, comprehensive gene expression analysis (RNA-seq, etc.) can be performed by transcriptome analysis such as microarray or next-generation sequencer. Histological analysis (RNAscope (registered trademark) etc.) can also be used. In addition, PCR analysis or the like can be used as a method for analyzing the expression of each isolated gene.

[0037] In the present invention, "skin condition" includes skin pigmentation, skin inflammation, wrinkles, texture, sagging, rough skin, pore size, elasticity, flexibility, skin color, and the like.

[0038] Here, the size of a pore includes its area, volume, and depth.

[0039] Furthermore, the "skin condition" in the present invention can also include skin conditions based on information (parameters) regarding the moisture content of the skin, the amount of moisture evaporated from the skin, the amount of sebum on the skin, the three-dimensional appearance (volume) of the skin, the height of the cheeks, and the length of the face line.

[0040] In the present invention, "pigmentation" refers to pigmentation symptoms caused by increased melanin production, excessive accumulation, increased expression of inflammatory factors, abnormal deposition, etc., such as age spots, dullness, melasma, freckles, sunburn, and darkening due to inflammation or irritation of the skin.

[0041] Hereinafter, a method for estimating a subject's skin condition according to the present invention will be described in detail with reference to FIG. 1, using a case where the "skin condition" according to the present invention is "age spots" as one embodiment.

[0042] First, in S11, skin is prepared from a subject. Here, skin is prepared from a site on the subject's face that has blemishes (hereinafter referred to as the blemish site) and skin is prepared from a site on the same subject's face that does not have blemishes (hereinafter referred to as the non-blemish site). As described above, the method of collecting skin and the amount of skin to be collected are not particularly limited. However, skin including the epidermis region including the stratum corneum and the dermis region located at a depth of 0.2 to 0.5 mm from the outermost layer of the skin is collected by a minimally invasive collection method, and the skin is collected by a 0.05 mm deep extraction. 3 It is preferable to collect the following:

[0043] Then, in S12, gene expression analysis is performed using the RNA extracted from the skin prepared in S11 as a sample. As described above, the method of gene expression analysis is not particularly limited, but it is preferable to perform PCR or transcriptome analysis.

[0044] Next, the expression level of a specific gene is calculated by the gene expression analysis. In the gene expression analysis, the "expression level of a gene" is calculated as an absolute value or a relative value (such as a ratio or difference from a comparative or standard expression level).

[0045] The specific gene is a gene related to a skin condition. Any gene can be the subject of analysis as long as it is related to a skin condition. In this embodiment, the specific gene is a gene associated with spots.

[0046] In a preferred embodiment of the present invention, the specific gene has one or more Gene Ontologies selected from the following group: [Table 3]

[0047] In a preferred embodiment of the present invention, the specific gene has one or more gene ontologies selected from the following group: [Table 4]

[0048] Gene Ontology is a controlled vocabulary of terms that structure and assign biological meaning to the information contained in individual genes. Each term is called a GO Term. Gene Ontology is broadly classified into three categories: Molecular Function, Cellular Component, and Biological Process, each of which has a hierarchical structure.

[0049] When inferring the meaning of altered genes obtained from comprehensive genetic analysis, gene ontology can be used to understand which genes are significantly altered, such as what functions, subcellular locations, or gene groups related to biological reaction processes.

[0050] As shown in the Examples below, gene expression analysis at S12 extracted a group of genes that showed significant changes in gene expression in the skin at the blemish site, and enrichment analysis (Biological Process) was performed. It was confirmed that epidermal differentiation (Epidermis Development) and melanin synthesis (Melanin Biosynthetic Process) were significantly increased. Therefore, gene expression analysis is performed using skin collected from a subject, and if it is found in S13 described below that the expression of genes having one or more gene ontologies selected from the above group has significantly changed, it can be inferred that the subject's skin condition is a blemish.

[0051] In a more preferred embodiment of the present invention, the number of genes having one or more gene ontologies selected from the above group is preferably three or more, more preferably five or more, more preferably nine or more, and even more preferably all of the genes.

[0052] In a preferred embodiment of the present invention, the specific gene is one or more selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL.

[0053] In a more preferred embodiment of the present invention, the specific genes are selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL, preferably three or more, more preferably five or more, more preferably nine or more, and even more preferably all of the genes.

[0054] In a preferred embodiment of the present invention, the specific genes are one or more selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

[0055] In a more preferred embodiment of the present invention, the specific genes are selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2, preferably three or more, more preferably five or more, more preferably seven or more, and even more preferably all of the genes.

[0056] In a further preferred embodiment of the present invention, the specific gene is one or more selected from TYR, TYRP1, DCT, and MITF.

[0057] Gene expression analysis is performed using skin collected from a subject, and if it is found in S13 described below that the expression of the above genes has significantly changed, the skin condition of the subject can be estimated to be blemishes.

[0058] In a more preferred embodiment of the present invention, if a significant increase in expression of one or more genes selected from TYR, TYRP1, DCT, MITF, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, and KRT10 is found, the subject's skin condition is presumed to be pigmented.

[0059] In a further preferred embodiment of the present invention, when a significant increase in expression of one or more genes selected from TYR, TYRP1, DCT, and MITF is observed, the skin condition of the subject is presumed to be pigmented.

[0060] In a more preferred embodiment of the present invention, if a significant decrease in the expression of one or more genes selected from IL1A, FLG, LORICLIN, and SCEL is found, the subject's skin condition is presumed to be pigmented.

[0061] In a more preferred embodiment of the present invention, if a significant increase in expression of one or more genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2 is found, the subject's skin condition is presumed to be a blemish.

[0062] Finally, in S13, the skin condition of the subject is estimated using the gene expression levels calculated in S12. Specifically, the gene expression levels calculated in S12 are compared with the gene expression levels in normal skin.

[0063] In an embodiment of the present invention, gene expression analysis is performed using skin taken from a subject where there are blemishes and skin taken from the same subject where there are no blemishes, the gene expression levels calculated for each skin are compared, and the skin condition is estimated based on the comparison results. Specifically, the difference between the expression level of the gene associated with the pigmentation calculated for the skin at the pigmentation site and the expression level of the gene associated with the pigmentation calculated for the skin at the non-pination site is calculated. If the calculated value is equal to or greater than a certain value, the subject's skin condition is estimated to be a blemish caused by a change in the expression level of the gene associated with the blemish.

[0064] Here, the expression level of a gene related to a blemish in the skin of a certain estimated target site of the subject may be calculated, and the calculated expression level of the gene related to a blemish may be compared with the expression level of the gene related to a blemish in the skin of the same estimated target site of a person with normal skin. The expression level of the gene related to a blemish in the skin of a person with normal skin can be evaluated more objectively by using the average value of data from multiple people with normal skin.

[0065] In a more preferred embodiment of the present invention, the subject's skin condition is presumed to be a blemish caused by fluctuations in expression of one or more genes selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL.

[0066] In a further preferred embodiment of the present invention, the subject's skin condition is predicted to be a blemish caused by alterations in the expression of preferably three or more, more preferably five or more, more preferably nine or more, and even more preferably all, of genes selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, ITLG, EDN1, KRT5, KRT10, and SCEL.

[0067] In a more preferred embodiment of the present invention, the subject's skin condition is estimated to be a blemish caused by fluctuations in the expression level of one or more genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

[0068] In a further preferred embodiment of the present invention, the subject's skin condition is estimated to be a blemish caused by alterations in the expression of preferably three or more, more preferably five or more, more preferably seven or more, and even more preferably all, of genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

[0069] Here, in an even more preferred embodiment of the present invention, it is presumed that the blemish is caused by a change in the expression of the gene used as an indicator in S12.

[0070] In a more preferred embodiment of the present invention, the spots are presumed to be caused by increased expression of one or more genes selected from TYR, TYRP1, DCT, MITF, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, and KRT10.

[0071] In a more preferred embodiment of the present invention, the blemishes are presumed to be caused by decreased expression of one or more genes selected from IL1A, FLG, LORICLIN, and SCEL.

[0072] In a further preferred embodiment of the present invention, the blemishes are presumed to be caused by increased expression of one or more genes selected from TYR, TYRP1, DCT, and MITF.

[0073] In a more preferred embodiment of the present invention, the spots are presumed to be caused by increased expression of one or more genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

[0074] By using the above method, the skin condition of the subject can be estimated in more detail.

[0075] <2> Methods for proposing compositions and / or stimuli suitable for subjects (excluding medical procedures) The present invention includes a step of performing gene expression analysis using skin sampled from a subject, and estimating the skin condition of the subject based on the results of the analysis; A suggestion step of suggesting a composition and / or stimulation suitable for the subject based on the estimated skin condition (excluding medical treatment); The present invention also relates to a method for proposing a composition and / or a stimulus suitable for a subject (excluding medical procedures).

[0076] The composition and / or stimulation of the present invention can be applied without any particular limitation as long as it is commonly known (excluding medical procedures).

[0077] As stated above, the present invention does not include medical procedures. Here, the term "medical procedure" in the present invention refers to the procedure in which a doctor (including a person under the instructions of a doctor) performs surgery, treatment, or diagnosis on a human being. Specifically, it does not include a physician prescribing a composition and / or stimulation appropriate for a subject based on an estimated skin condition.

[0078] The compositions of the present invention may include commercially available compounds (including peptides), known compounds (including peptides), compounds obtained by combinatorial chemistry techniques, natural components derived from plants or marine organisms, and animal tissue extracts.

[0079] The extract derived from an animal or plant is a general term not only for the extract itself derived from an animal or plant, but also for a fraction of the extract, a purified fraction, and a product obtained by removing the solvent from the extract, fraction, or purified product. Examples of plant-derived extracts include extracts using wild-growing or cultivated plants, extracts using plants sold as raw materials for herbal medicines, and commercially available extracts.

[0080] The compositions of the present invention also include cosmetics and topical skin preparations that contain the above compounds, natural ingredients, and substances.

[0081] Examples of the stimulation in the present invention include face packs, massage techniques, aromas, laser light irradiation, electric pulses, ion introduction, electroporation, ultrasound, and microneedles.

[0082] Hereinafter, a method for proposing a composition and / or stimulation suitable for a subject (excluding medical procedures) when the "skin condition" according to the present invention is blemishes will be described with reference to FIG.

[0083] First, the skin condition of the subject is estimated by steps S11 to S13. <1> The description of the above is incorporated by reference.

[0084] Below, a preferred embodiment of the method of the present invention for proposing a composition and / or stimulation suitable for a subject (excluding medical procedures) will be described in detail, using as one embodiment a case in which the subject's skin condition is suspected to be a blemish.

[0085] In S14, a composition and / or stimulation suitable for the subject is suggested based on the subject's skin condition estimated in S13 (excluding medical procedures). In an embodiment of the present invention, the subject's skin condition is estimated to be blemishes, and a composition and / or stimulation that can reduce (or increase) the expression level of genes associated with blemishes is proposed (medical procedures are excluded).

[0086] In a preferred embodiment of the present invention, the subject's skin condition is presumed to be a blemish caused by altered expression of one or more genes selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL. Therefore, in this embodiment, a composition and / or a stimulation capable of decreasing (or increasing) the expression amount of the above-mentioned genes associated with spots is preferably proposed (however, medical treatment is excluded).

[0087] In a more preferred embodiment of the present invention, a composition and / or stimulation capable of reducing the expression level of one or more genes selected from TYR, TYRP1, DCT, MITF, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, and KRT10 is proposed (excluding medical procedures).

[0088] In addition, in a more preferred embodiment of the present invention, we propose a composition and / or stimulation that can increase the expression level of one or more genes selected from IL1A, FLG, LORICLIN, and SCEL (excluding medical procedures).

[0089] In a preferred embodiment of the present invention, the subject's skin condition is presumed to be a blemish caused by altered expression of one or more genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2. Therefore, in this embodiment, a composition and / or a stimulation capable of reducing the expression amount of the above-mentioned spots-related genes is preferably proposed (however, medical treatment is excluded).

[0090] As shown in the Examples below, it was confirmed that the patterns of gene expression that cause spots differ depending on the subject. Therefore, the above method makes it possible to suggest compositions and / or stimuli suitable for the subject (excluding medical procedures).

[0091] In another embodiment of the present invention, a composition and / or stimulation suitable for a subject can be selected based on the skin condition of the subject estimated by steps S11 to S13 (excluding medical procedures).

[0092] In the proposed method of the present invention, preferably, further comprising an application step of applying the composition and / or stimulation proposed to the subject to the subject (excluding medical procedures); the estimation step being carried out based on the results of gene expression analysis performed using the skin sampled from the subject after the application step; and a re-suggestion step of re-suggesting a composition and / or stimulation suitable for the subject (excluding medical treatment) based on the estimated skin condition.

[0093] A preferred embodiment of the method proposed by the present invention will now be described in detail with reference to FIG.

[0094] In the application step S21, the composition and / or stimulation proposed to the subject in S14 is applied to the site from which the subject's skin was taken in S11 (excluding medical procedures). In an embodiment of the present invention, the composition and / or stimulation suggested to the subject in S14 is applied to the blemish site of the subject from which the skin was sampled in S11 (excluding medical procedures).

[0095] The application step S21 may be performed multiple times, for example, the composition and / or stimulation may be applied once or more than once a day (excluding medical procedures) for two or more consecutive days. By periodically carrying out the application step S21, the effectiveness of the composition and / or stimulation on the target site can be accurately evaluated.

[0096] Next, using a method similar to S11 to S13, gene expression analysis is performed (S22) using skin collected from the site where the composition and / or stimulation was applied in the application step S21, and the skin condition of the subject is estimated (S23). In an embodiment of the present invention, gene expression analysis is performed (S22) using skin collected from the spot site to which the composition and / or stimulus was applied in the application step S21, and the skin condition of the subject is estimated (S23).

[0097] Then, in S24, a composition and / or stimulation suitable for the subject is again suggested in a manner similar to that of S14 (with the exception of medical procedures). According to the present invention, a composition and / or stimulation suitable for a subject can be suggested with greater accuracy (excluding medical procedures).

[0098] In another embodiment of the present invention, the subject's skin condition estimated in S23 may be compared to the subject's skin condition estimated in S13. By the above comparison, the suitability of the composition and / or stimulation applied to the subject in S21 for the subject can be evaluated.

[0099] <3> Apparatus for estimating subject's skin condition The present invention is also an apparatus for estimating a subject's skin condition, comprising a gene expression analysis result acquisition unit that acquires the results of gene expression analysis using skin collected from the subject, and an estimation means that estimates the skin condition of the subject based on the results of the gene expression analysis.

[0100] Hereinafter, the estimation device of the present invention will be described with reference to a hardware block diagram (FIG. 4). <1> The present invention is an apparatus for carrying out the method for estimating the skin condition of a subject described in the above item. <1> The explanation in the previous section also applies to the following estimation device.

[0101] The estimation device 1 of the present invention comprises a gene expression analysis result acquisition unit 11 that acquires the results of gene expression analysis performed using skin collected from a subject, and an estimation means 131 that estimates the skin condition of the subject based on the results of the gene expression analysis.

[0102] As shown in FIG. 4, the estimation device 1 includes a gene expression analysis result acquisition unit 11 that acquires the results of gene expression analysis using skin collected from a subject, a ROM (Read Only Memory) 12 that has a storage means 121, a CPU (Central Processing Unit) 13 that has an estimation means 131 that estimates the skin condition of the subject, and an estimation result display unit 14.

[0103] The gene expression analysis result acquisition unit 11 acquires the results of gene expression analysis using skin sampled from a subject. Specifically, it acquires data on the expression level of a specific gene calculated as a result of the gene expression analysis.

[0104] The storage means 121 stores correlations between skin conditions and genes. For example, it stores that the expression changes of genes such as TYR, TYRP1, DCT, and MITF are correlated with blemishes.

[0105] The estimation means 131 estimates the skin condition of the subject based on the gene expression analysis results acquired by the gene expression analysis result acquisition unit 11 and the correlation between the skin condition and genes stored in the storage means 121.

[0106] The estimation result display unit 14 is a display that displays the result estimated by the estimation means 131.

[0107] The estimation device 1 of the present invention configured as described above can easily estimate the skin condition of a subject by performing gene expression analysis using skin sampled from the subject.

[0108] <4> Apparatus for suggesting compositions and / or stimuli suitable for subjects The present invention provides a suggestion device that suggests a composition and / or stimulus suitable for a subject, comprising: A gene expression analysis result acquisition unit that acquires a result of a gene expression analysis performed using skin sampled from a subject, and an estimation means that estimates a skin condition of the subject based on the result of the gene expression analysis; A suggestion means for suggesting a composition and / or stimulation suitable for the subject based on the estimated skin condition; Equipped with It is also a device for suggesting compositions and / or stimuli appropriate for a subject.

[0109] The following description will be given with reference to the hardware block diagram (FIG. 5) of the proposed device of the present invention. <2> The present invention is an apparatus for carrying out the method for proposing a composition and / or a stimulus suitable for a subject described in the item (excluding medical procedures). <2> The explanation in the previous section also applies to the following estimation device.

[0110] The proposed device 2 of the present invention comprises a gene expression analysis result acquisition unit 21 that acquires the results of gene expression analysis performed using skin collected from a subject, an estimation means 231 that estimates the skin condition of the subject based on the results of the gene expression analysis, and a proposal means 232 that proposes a composition and / or stimulation suitable for the subject based on the estimated skin condition.

[0111] As shown in Figure 5, the proposal device 2 has a gene expression analysis result acquisition unit 21 that acquires the results of gene expression analysis using skin collected from a subject, a ROM (Read Only Memory) 22 that has a first memory means 221 and a second memory means 222, a CPU (Central Processing Unit) 23 that has an estimation means 231 that estimates the skin condition of the subject and a proposal means 232 that proposes a composition and / or stimulation (excluding medical procedures) suitable for the subject, and a proposal result display unit 24.

[0112] The gene expression analysis result acquisition unit 21 acquires the results of gene expression analysis using skin sampled from a subject. Specifically, it acquires data on the expression level of a specific gene calculated as a result of the gene expression analysis.

[0113] The first storage means 221 stores correlations between skin conditions and genes. For example, it stores that the expression changes of genes such as TYR, TYRP1, DCT, and MITF are correlated with blemishes.

[0114] The estimation means 231 estimates the skin condition of the subject based on the gene expression analysis result acquired by the gene expression analysis result acquisition unit 21 and the correlation between the skin condition and the gene stored in the first storage means 221 .

[0115] The second storage means 222 stores a composition and / or a stimulus that can decrease (or increase) the expression level of a gene, for example, that laser toning decreases the expression levels of all of the TYR, TYRP1, and DCT genes, that 4-n-butylresorcinol (Rucinol (registered trademark)) can suppress the activity of TYR and TYRP1, and the like.

[0116] The suggestion means 232 suggests a composition and / or stimulation suitable for the subject based on the skin condition of the subject estimated by the estimation means 231 and the composition and / or stimulation stored in the second memory means 222 (excluding medical procedures).

[0117] The proposal result display unit 24 is a display that displays the results proposed by the proposal means 232 .

[0118] The proposed device 2 of the present invention configured in this manner can easily propose a composition and / or stimulation suitable for the subject by performing gene expression analysis using skin sampled from the subject.

[0119] <5> A program to estimate the subject's skin condition The present invention also relates to a subject's skin condition estimation program that causes a computer to execute the subject's skin condition estimation method. <4> In order to correspond to each means in the CPU included in the estimation device described in the section 1, the description will be given while using the reference symbols in FIG.

[0120] The estimation program of the present invention is a program for estimating the skin condition of a subject using gene expression levels in skin collected from the subject, and causes a computer to execute a step of estimating the skin condition of the subject based on the gene expression levels.

[0121] <6> A program for suggesting compositions and / or stimuli suitable for subjects The present invention also relates to a program for suggesting a composition and / or a stimulus suitable for a subject, which program causes a computer to execute the method for suggesting a composition and / or a stimulus suitable for a subject. <4> In order to correspond to each means in the CPU included in the proposed device described in the section, the explanation will be given while adding the reference numerals in FIG.

[0122] The proposal program of the present invention is a proposal program that uses gene expression levels in skin collected from a subject to propose a composition and / or stimulation suitable for the subject, and causes a computer to execute the following steps: obtaining the results of gene expression analysis using skin collected from the subject; estimating the skin condition of the subject based on the gene expression levels; and proposing a composition and / or stimulation suitable for the subject based on the estimated skin condition. EXAMPLES

[0123] <Test Example 1> Examination of gene expression analysis method (RNA-seq) First, skin samples were prepared from the blemished area (corresponding to the pigmented area in the present invention) and non-blemished area on the face of the subject. 3 After being collected from the subjects, the samples were stored at room temperature in Eppendorf tubes and then refrigerated.

[0124] Next, RNA was extracted from the prepared skin using a Picopure kit (Thermo Fisher).

[0125] The quality of the extracted RNA was then confirmed using Bioanalyzer (Agilent). After that, samples with a DV200 value of 60% or more were subjected to RNA-seq according to the protocol of the SMART-seq Stranded kit (Takara Bio Inc.). The results are shown in Tables 5 and 6.

[0126] [Table 5]

[0127] [Table 6]

[0128] As shown in Tables 5 and 6, RNA-seq confirmed changes in the expression of genes known to be related to pigmentation. By comprehensively evaluating these gene expression changes, the skin condition of the subject can be estimated with high accuracy.

[0129] (Enrichment analysis) As a result of the above RNA-seq, a group of genes that showed significant changes in gene expression in the skin taken from the spots were extracted, and biological process (BP) was searched for by gene ontology (GO) enrichment analysis using the public web tool Enrichr (https: / / maayanlab.cloud / Enrichr / ). The results are shown in Tables 7 and 8.

[0130] [Table 7]

[0131] [Table 8]

[0132] Enrichment analysis confirmed that epidermal differentiation (Epidermis Development) and melanin synthesis (Melanin Biosynthetic Process) were significantly increased.

[0133] By comprehensively evaluating the expression changes of genes having one or more gene ontologies selected from Tables 7 and 8 above, the skin condition of a subject can be estimated with high accuracy.

[0134] (PCR analysis) First, RNA was extracted from the skin in the same manner as in Test Example 1.

[0135] The extracted RNA was analyzed using real-time PCR to evaluate the expression of TYR, TYRP1, DCT, and MITF genes, which are known to be involved in melanin synthesis. The results are shown in Figure 6.

[0136] As shown in Figure 6, PCR analysis confirmed that the expression of genes involved in melanin synthesis, namely TYR, TYRP1, DCT, and MITF, was increased in the skin sampled from the spots. By comprehensively evaluating the changes in gene expression, the skin condition of the subject can be estimated with high accuracy.

[0137] <Test Example 2> Estimation of subject's skin condition First, RNA was extracted from the facial skin of subject A and subject B using the same method as in Test Example 1.

[0138] The extracted RNA was subjected to RNA-seq to evaluate the expression of TYR, TYRP1, and DCT genes in the same manner as in Test Example 1. The results are shown in FIG.

[0139] As shown in Figure 7, it was confirmed that the expression of the TYR, TYRP1, and DCT genes was all increased in the skin sampled from the pigmented area of ​​subject A. On the other hand, it was confirmed that the expression of the TYR and TYRP1 genes was increased in the skin sampled from the pigmented area of ​​subject B, but the expression of the DCT gene was only slightly increased.

[0140] Based on the above results, it is presumed that subject A's spots are caused by increased expression of the TYR, TYRP1, and DCT genes, while subject B's spots are presumed to be caused by increased expression of the TYR and TYRP1 genes.

[0141] <Test Example 3> Selection and / or suggestion of compositions and / or stimuli (excluding medical procedures) suitable for the subject Based on the results of Test Example 2, it is considered preferable to select and / or propose to subject A a composition and / or stimulation (excluding medical procedures) that reduces the expression levels of all of the TYR, TYRP1, and DCT genes, and to select and / or propose to subject B a composition and / or stimulation that reduces the expression levels of the TYR and TYRP1 genes to a minimum extent (excluding medical procedures).

[0142] Here, laser toning is known to reduce the expression levels of all of the TYR, TYRP1, and DCT genes (Reference 1). Therefore, laser toning can be selected and / or proposed as a composition and / or stimulation suitable for subject A (excluding medical procedures). On the other hand, it is known that 4-n-butylresorcinol (Rucinol®) can suppress the activity of TYR and TYRP1 (Reference 2). Therefore, 4-n-butylresorcinol (Rucinol®) can be selected and / or proposed as a composition and / or stimulation suitable for subject B (excluding medical procedures).

[0143] As described above, based on the estimation results of the subject's skin condition, a composition and / or stimulation suitable for the subject can be selected and / or suggested (excluding medical procedures). [Industrial Applicability]

[0144] According to the present invention, it is possible to estimate the skin condition of a subject, select and / or suggest a composition and / or stimulation suitable for the subject (excluding medical procedures), and further select and / or suggest again a composition and / or stimulation suitable for the subject (excluding medical procedures).

[0145] Reference 1: JEKim et al., Histopathological study of the treatment of melasma lesions using a low-fluence Q-switched 1064-nm neodymium:yttrium-aluminium-garnet laser, Clinical and Experimental Dermatology, 2013, 38(2), 167-171 Reference 2: Takayuki Katagiri, Inhibitory effect of 4-n-butylresorcinol (Rucinol (registered trademark)) on melanin production and its effectiveness on human pigmentation, 2001, 35(1), 42-49 [Explanation of symbols]

[0146] 1 Estimation device 11 Gene Expression Analysis Results Acquisition Department 12 ROM 121 Memory means 13 CPU 131 Estimation means 14 Estimation result display section 2 Proposed device 21 Gene Expression Analysis Results Acquisition Department 22 ROM 221 Primary memory means 222 Second storage means 23 CPU 231 Estimation means 232 Proposal means 24 Proposal result display area

Claims

1. A method for estimating a skin condition of a subject, comprising performing gene expression analysis using skin sampled from the subject, and estimating the skin condition of the subject based on the results of the analysis.

2. The method according to claim 1 , wherein the gene expression analysis is carried out by PCR or transcriptome analysis using RNA extracted from the skin as a sample.

3. The method according to claim 1 or 2, wherein the gene has one or more gene ontologies selected from the following group: 【Table 1】

4. The method according to claim 1 or 2, wherein the gene has one or more gene ontologies selected from the following group: 【Table 2】

5. The method of claim 1 or 2, wherein the gene is one or more selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL.

6. The method of claim 1 or 2, wherein the gene is one or more selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

7. The method according to claim 1 , wherein the skin condition is pigmentation.

8. The method of claim 7, further comprising estimating that the skin condition is pigmentation caused by a change in the expression level of one or more genes selected from TYR, TYRP1, DCT, MITF, IL1A, FLG, LORICLIN, KIT, EDNRB, CCR2, FGFR1, MIA, TRPM1, MLANA, PMEL, FGF7, FGFR2, KITLG, EDN1, KRT5, KRT10, and SCEL.

9. The method of claim 7, further comprising estimating that the skin condition is pigmentation caused by a change in the expression level of one or more genes selected from G0S2, HOXD, SLC11A1, PITX1, CRNN, HOXD8, TSBP1, and SOD2.

10. The estimation step includes: The results of gene expression analysis using skin samples taken from subjects in pigmented areas, The method according to claim 1 or 2, further comprising estimating the skin condition of the subject based on a comparison with the results of gene expression analysis performed using a non-pigmented site of skin taken from the subject.

11. The method according to claim 1 or 2, wherein the skin is skin dried under room temperature conditions.

12. The volume of skin immediately after acquisition from the subject was 0.05 mm ３ The estimation method according to claim 1 or 2, wherein:

13. The method according to claim 12, wherein the skin includes an epidermis region including a stratum corneum and a dermis region located at a depth of 0.2 to 0.5 mm from the outermost layer of the skin.

14. An estimation step of performing gene expression analysis using skin sampled from a subject and estimating the skin condition of the subject based on the results of the analysis; A suggestion step of suggesting a composition and / or stimulation suitable for the subject based on the estimated skin condition; A method for suggesting a composition and / or stimulation suitable for a subject (excluding medical procedures).

15. and applying to the subject the composition and / or stimulus proposed to the subject. the estimation step being carried out based on the results of gene expression analysis performed using skin tissue collected from the subject after the application step; A suggestion method (excluding medical procedures), comprising a re-suggestion step of re-suggesting a composition and / or stimulation suitable for the subject based on the estimated skin condition.

16. An apparatus for estimating a skin condition of a subject, comprising: a gene expression analysis result acquisition unit that acquires the results of gene expression analysis performed using skin sampled from a subject; an estimation means for estimating a skin condition of the subject based on a result of the gene expression analysis; An apparatus for estimating a subject's skin condition comprising:

17. A suggestion device that suggests a composition and / or stimulus suitable for a subject, a gene expression analysis result acquisition unit that acquires the results of gene expression analysis performed using skin sampled from a subject; an estimation means for estimating a skin condition of the subject based on a result of the gene expression analysis; A suggestion means for suggesting a composition and / or a stimulus suitable for the subject based on the estimated skin condition; An apparatus for suggesting a composition and / or stimulus suitable for a subject, comprising:

18. A program for estimating a skin condition of a subject by using gene expression levels in skin sampled from the subject, estimating a skin condition of the subject based on the gene expression level; A program for causing a computer to execute the following.

19. A suggestion program for suggesting a composition and / or stimulation suitable for a subject by using gene expression levels in skin collected from the subject, Obtaining results of gene expression analysis using skin samples taken from a subject; estimating a skin condition of the subject based on the gene expression level; A suggestion step of suggesting a composition and / or a stimulus suitable for the subject based on the estimated skin condition; A program for causing a computer to execute the following.

20. applying a composition and / or stimulus to the subject; and an estimation step of performing gene expression analysis using the skin sampled from the subject after the application step, and estimating the skin condition of the subject based on the results of the analysis. evaluating the suitability of the composition and / or the irritation for the subject by comparing the result of the estimation step with the skin condition of the subject obtained in advance before the application step; A method for evaluating a composition and / or stimulus suitable for a subject (but excluding medical procedures).

Images