Method and kit for hair DNA typing

Abstract

An artificial intelligence–based gene testing kit and software system for classifying curly, coily, and wavy hair types using a sample from the scalp containing the hair follicle and surrounding peri‑follicular extracellular matrix (ECM). A U‑shaped skin clipper collects the tissue sample, which can be analyzed using Omics and Spatial Omics techniques, including genomics, transcriptomics, proteomics, and AI‑assisted molecular profiling. Variables examined include Wnt gene family members, Frizzled receptors, co‑receptors, EDAR, SHH, macrophage phenotypes, macrophage‑secreted factors, immune proteins, and additional ECM‑related markers. These data are fed into an artificial neural network that classifies the individual's "hair genotype," which is correlated with a photographic hair‑shaft phenotype and communicated to the individual. The method provides personalized insights into genetic hair makeup and supports the development of individualized hair‑care treatments and ancestry‑related genomic databases.

Description

TITLEMETHOD AND KIT FOR HAIR DNA TYPINGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. § 119 of U.S. Provisional Application Serial No. 63 / 763,900, filed on February 26, 2025, which is incorporated herein by reference.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] Not applicable.BACKGROUND OF THE INVENTION

[0003] The present disclosure generally relates to genetic analysis and DNA kits. More specifically, the disclosure relates to an Al-based method for typing curly, coily, and wavy human scalp hair or body hair by analyzing the genetic and molecular composition of hair follicles and their surrounding three-dimensional peri-follicular extracellular matrix (ECM).

[0004] Gene sequencing technologies determine the order of the building blocks of each gene with a goal to finding “variants” and linking those variants to inherited conditions. The variant can also be linked to the biological mechanisms underlying human diseases, mainly cancer. Different technologies are used and the most common is Next-Generation Sequencing (NGS), wherein the whole human genome is analyzed by uploading a customer’s sample onto a sequencing matrix, called a sequencing chip, to generate a data library. The human samples examined can be saliva, mouth swab or blood.

[0005] Many of the currently marketed testing kits are designed for the diagnosis of cancer. In these kits, artificial intelligence (Al) software transforms complex genetic data into clear, actionable insights, empowering efficient decisions across the healthcare pathway. These kits are driving a new era of scientific progress at every stage of cancer care and drug development. Currently marketed gene-testing kits use various NGS-based gene panels — such as sixteen-gene breast cancer panels (Atila BioSystems), 117-gene breast cancer sequencing kits (BioDynamic), and the APIS subtyping kit (Biocartis). Other kits include direct-to-consumer saliva-based kits such as 23andMe, CRI Genetics, AncestryDNA, and others, which primarily test for ancestry and disease predisposition as opposed to oncology.

[0006] Unlike genetic methodologies used for diseases, existing hair typing systems classify hair using qualitative or quantitative assessment of the hair shaft’s curvature andmorphology (e.g., Andre Walker, STAM, high-throughput phenotyping, dynamic mechanical analysis). Scalp hair consists of two domains: the shaft and the hair follicle (HF). In humans, the hair shaft has enormous diversity in the degree of curvature, which is programmed from the follicle. Further, the hair fiber is a memory shape, meaning the shape imparted by the follicle is retained as the hair grows. For many years cosmetic scientists have attempted to develop hair typing systems by measuring the specific physical 3D features of human hair, including qualitative hair typing system classifying hair into Type I-IV with subtypes (a-c). Later, quantitative hair typing of STAM and modified-STAM classification was introduced. Recently, hair science was reimagined to classify curly hair phenotypes using a dynamic mechanical analyzer.

[0007] Genetic studies, including genome-wide association studies of blood or saliva, have implicated Wnt-related genes (e.g., WntlOA, Wnt7b), wherein Wnt refers to Wingless-type MMTV integration site family, and the EDAR (Ectodysplasin-A receptor) gene in hair morphology. Wnt-related genes encode pathways for protein growth factors. Macrophages, resident in the dermal tissue surrounding the hair follicle, are secreting and receiving Wnt ligands. In a published hypothesis, Al Jasim N. (2024) proposes that macrophage can curve the hair follicle and curl the hair by secreting Wnt genes and possibly acting on the APM of the hair follicle to induce fiber contraction. This hypothesis expands and innovates on existing molecular and genetic mechanisms regulating curly hair phenotypes.

[0008] The extra-cellular matrix (ECM) of hair follicles contains complex networks of proteins, cellular compartments, and immune cells. Wnt proteins are secreted extracellular signaling molecules that diffuse across ECM tissue gradients, binding Frizzled (Fz) receptors and associated co-receptors to activate intracellular pathways governing embryonic patterning, morphogenesis, and hair growth cycles. Once released by the producing cell, Wnt proteins travel in the extracellular space to reach their target cell and interact with Frizzled cell-surface receptors. The complexity of signal activation increases significantly by the presence of a family of Frizzled receptors interacting with the repertoire of Wnt ligands. A combinatorial interaction of different Wnt proteins with FZ receptors lead to the activation of multiple downstream pathways.

[0009] Numerous investigative tools — including single-cell RNA-seq, scRNA-seq-based atlas mapping, immunofluorescence, interactomics, Omics, spatial transcriptomics, functional enrichment analysis, PPI network mapping, and Al-mediated high-throughput screening — have elucidated relationships among macrophages, fibroblasts, Wnt ligands, ECM structure, and hair follicle morphogenesis.

[0010] Despite substantial advances in genome-wide association studies, organ-on-chip systems, curated genetic atlases, and functional hair science, there is no existing genetic test that classifies curly, coily, or wavy hair by measuring Wnt-related molecular composition, macrophage phenotypes, ECM signaling, or peri-follicular tissue variables namely the FZD receptors located on the cell-surface of the arrector pili muscle. Having genetic data could enable tailored advice on daily haircare routine and best products for an individual to use, in hair saloon for proper haircare techniques, and in the clinic to discuss the effect of chemicals, heat, or mechanical pulling used in personal routine hair styling

[0011] Therefore, it would be advantageous to develop a gene panel targeting Wnt family members, Frizzled receptors, ED AR, SHH (Sonic Hedgehog), and immune-cell markers, and an Al-based software (algorithm) and its application for classifying hair type according to the individual’s hair-specific genotype to provide more specific guidance than qualitative and quantitative assessments.BRIEF SUMMARY

[0012] According to embodiments of the present disclosure is an Al -based software and its application in an analytical system for typing curly / coily / wavy human hair by evaluating the genetic and molecular characteristics of hair follicles and the surrounding peri-follicular ECM. A sample containing the hair follicle and surrounding dermal components is harvested using methods known in the art or a novel U-shaped skin clipper tool. The sample is preserved and delivered to a specialized laboratory for analysis via omics, spatial omics, and Al-assisted methods.

[0013] Variables tested by the kit may include: nineteen (19) human Wnt gene family members; ten (10) Frizzled receptor family members (FZD); Wnt co-receptors; Wnt-function-related genes (EDAR, SHH); macrophage phenotypes, including their Wnt secretion / uptake profiles; macrophage-secreted growth factors and immune proteins (e.g., IFN, TNF); and ECM-related molecular components and additional genetic markers.

[0014] Quantitative and qualitative data from these genetic and molecular pathways form the input to an Al algorithm that is an information processing network, such as an artificial neural network, that recognizes complex patterns, classifies data, and predicts outcomes to classify the individual’s “hair genotype.” The genotype can be linked to a photographically documented hair shaft type and is communicated to the individual for additional information. The data may also inform personalized aesthetic and clinical treatments for relaxing ormanaging curly and coily hair at the follicular level and may be incorporated into ancestry-related genomic databases.BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0015] Fig. 1 shows the anatomical structure of a tissue sample.

[0016] Fig. 2 is a chart showing different variables as inputs to a testing system.

[0017] Fig. 3 is a flowchart depicting the analysis step of the method.

[0018] Fig. 4 is a kit according to one embodiment.

[0019] Fig. 5 is a flowchart of the method, according to one embodiment.DETAILED DESCRIPTION

[0020] According to embodiments of the disclosure is a method of typing a person’s hair genotype. Parts of the method can be implemented as software. Specifically, disclosed is a laboratory-based DNA / RNA genetic test kit 100 (as shown in Fig. 4) and associated testing method 200 for classifying human scalp hair, including curly, coily, wavy, frizzy, and kinky hair types, based on genetic makeup and molecular composition derived from a three-dimensional perifollicular dermis and perifollicular extracellular matrix sample. A biological sample is collected from scalp skin containing hair and at least one intact hair follicle, using a purpose-designed skin clipper 110 or punch, and is placed into a preservative solution and container 111 supplied with the kit 100 according to provided instructions before being shipped to a specialized laboratory for analysis. The preservative solution may include nucleic acid stabilization buffers or similar chemicals. Once the sample is collected, it is tested according to the following method 200, which may include: sample collection 201, analysis 202, and genetic study 203. Fig. 5 is a flowchart depicting the method 200. Alternatively, a sample can be collected using an extracted (i.e. pulled-out) hair follicle or a blood sample to measure Wnt gene family members.

[0021] OVEVIEW

[0022] Prior to molecular analysis, the hair shaft associated with the sampled follicle is photographed and morphologically typed using state-of-the-art imaging techniques to enable subsequent correlation with genetic data. After morphological typing, the extracellular matrix and enclosed hair follicle are analyzed to identify genetic and molecular pathways regulating hair follicle curvature utilizing artificial intelligence tools and algorithms, including artificial neural networks. The analysis includes, but is not limited to, assessment of one or more members of the Wnt gene family, Frizzled receptor family members located in the extracellularmatrix, hair follicle, or arrector pili muscle, macrophage cell phenotypes and associated Wnt signaling activity, and additional sub-variables such as Wnt co-receptors, Wnt-related genes including ED AR and SHH, macrophage-secreted growth factors, immune proteins, and other genes present within the extracellular matrix.

[0023] The artificial intelligence methodologies employed may include omics-based approaches such as genomics, transcrip tomics, proteomics, and / or spatial sequencing, including qualitative and quantitative evaluation of Wnt signaling components, Frizzled receptors, macrophage phenotypes, immune markers, and tissue-specific proteins known in the art. Input data generated from these genetic and molecular analyses are processed by a trained algorithm to classify hair follicle genotype and to link the genotype with the corresponding hair shaft image, thereby producing a hair type classification that is communicated to the individual.

[0024] The testing method 200 may be performed on scalp skin tissue containing hair follicles or, alternatively, on skin obtained from other hair-bearing areas of the body. The kit 100 further includes a tag or label having a unique identification number, shipping and return materials, an instruction manual, a U-shaped skin clipper 110 having a sharpened edge configured to push, pinch, and pick the skin sample, an additional skin swab, and a plaster.

[0025] The method 200 applies artificial intelligence software, algorithms, and tools known in the art in an aesthetic and cosmetic genomics context to provide individuals with new information regarding their personal hair genetic makeup and the biological basis of hair uniqueness. The resulting classification supports the development of personalized hair treatments and daily care recommendations for use in salons, clinics, or in-person consultations, wherein personalization may include adjustment of active ingredient concentration, treatment duration, or use of combination therapies.

[0026] In one embodiment, the method 200 further includes a structured study design stage 203 in which a well-defined study subject is identified and phenotypic data are collected using current knowledge and state-of-the-art techniques for hair shaft classification. Photographs of the hair shaft are obtained and classified according to one or more of the previously described methods, and the resulting phenotypic classification is correlated, via software, with genetic testing results to generate a novel “genetic curly hair type.”

[0027] SAMPLE COLLECTION (201)

[0028] Sample collection 201 includes obtaining tissue from the participant’s hair follicle and surrounding dermis and its extracellular matrix, from which both DNA and genotypic information are derived. The sample is obtained using the skin clipper 110, which resembles a nail clipper and may include a U-shaped sharpened edge configured to punch, pick, and retainthe tissue sample for placement into a preservative fluid, although alternative sampling techniques, such as follicular unit punching methods used in hair transplant procedures, may also be employed or simple pulled-out hair follicles. The diameter and / or volume of the perifollicular dermal tissue including the extracellular matrix surrounding the hair follicle is measured, as Wnt genes and related signaling molecules diffuse into surrounding tissue and establish concentration gradients that influence biological function.

[0029] Fig. 1 depicts the novel 3-dimensional peri-follicular spatial tissue of human skin of the scalp, or dermal tissue. This dermal tissue and perifollicular extracellular matrix (ECM) is the anatomical location of the human scalp to be tested. Fig. 1 demonstrates the molecular and cellular composition of the dermis and ECM, wherein: the black-colored parts are the classic hair follicle structural parts including the young soft hair shaft that is topped by the stem cells (SC) thar are Wnt secreting cells. The hair is encircled by 3 layers including the inner root sheath (IRS); the outer root sheath (ORS); and the dermal sheath (Dermal S). The anatomical and functional parts relevant to the testing method 200 are included within the dermis, wherein the cells are the macrophages (Macro), the gene family and subfamily are Wnt (Wnt), and the receptors are the Frizzled receptor family (Frz). The ECM extends upwards to the dermal-epidermal junction (DEJ). The arrector pili muscle of the HF (APM) is embedded in the dermis. An alternative sample for Wnt gene family member detection can be obtained from a pulled-out hair with its surrounding ECM and dermal tissue. Or alternatively yje Wnt gene family member testing can be conducted on blood samples.

[0030] ANALYSIS (202)

[0031] Once the sample is collected, the sample is examined for its gene makeup including which type and optionally the quantity of gene family members and what gene interactions are occurring with local receptors and cells. The outputs of this gene analysis form a gene panel or library that can be used as an input to an Al program, such as an artificial neural network that identifies patterns, classifies information, and makes predictions of hair types. The data generated can be used for the development of personalized / precision treatment to relax curly and coily hair from the roots for a healthier scalp and hair shaft. By using Al-based software for analysis, patterns can be identified without a strict threshold for any individual marker analyzed.

[0032] As shown in Fig. 2, during the analysis step 202, 3 layers of variables are used in classifying curly / wavy hair types located specifically in the dermis surrounding the hair follicle. As noted in Fig. 1, the dermis is a complex network of proteins and other molecules and cells, wherein biological markers form the input data for the algorithm of the method 200.The main variables in total or part of them are placed in row #1 and they are the major input for the neural network software and include 19 members of Wnt gene family, 10 members of the Frizzled receptor family, and the 6 phenotypes of tissue specific macrophages secreting and receiving different family members of Wnt (genes). Row #2 contains sub-variables of Wnt function-related genes (EDAR and SHH) and 5 co-receptors for Wnt gene ligands. Row #3 data are related to macrophage-secreted growth factors and immune proteins, and possibly other related genes. These variables form the neural network of the software used in classifying curly hair genotypes and find an association between them and single type of curly (i.e. curly, coily, and wavy) hair shaft and possibly correlate the value to the physical type characteristics with the genetic testing (visual correlation). The Al-based software may be trained on existing libraries of genetic data associated with the biomarkers and other input variables discussed above. Validation occurs through visual correlation or human interpretation.

[0033] By way of further detail, the Al-based gene analysis includes one or more variables, whether single or multiple, that are analyzed in each sample, including assessment of Wnt family genes comprising the nineteen known human Wnt members, quantitative measurement of Wnt gene expression levels, evaluation of functionally related genes including EDAR and SHH, detection of Frizzled family receptors expressed on the outer root sheath, dermal sheath, and particularly on the arrector pili muscle embedded within the dermis, assessment of Wnt co-receptors, and characterization of macrophage cell populations having tissue-specific genotypes. Macrophages are evaluated due to their ability to secrete and receive Wnt ligands, their role in fetal hair morphogenesis, their secretion of growth factors, and their production of functional immune proteins, including interferons and tumor necrosis factor, which have receptors on the arrector pili muscle.

[0034] To conduct the analysis 202, each perifollicular dermis and / or hair follicle sample is processed, prepared, or purified using standard laboratory methods, and the tissue sampmle gene contents are subjected to customized chip-based beads by techniques known in the art, and scanned using automated laboratory instrumentation to detect the main variables for the neural networking that are described in Fig. 2 and selected markers of genetic variation, including single nucleotide polymorphisms (SNP). In one embodiment, a Curly Hair DNA Chip, as known in the art, comprises beads configured to bind genetic material, including DNA, RNA, and / or proteins, and to generate detectable signals, such as fluorescence, readable by the instrumentation, the specific testing method not being a limiting factor. In an alternative embodiment, the invention may be implemented using a Hair-on-Chip or organ-on-chipplatform, in which relevant cells and protein molecules are cultured within a fluidic medium to study molecular function and signaling crosstalk within the tissue.

[0035] The sample undergoes Omics and Spatial Omics analysis, including genomics, transcriptomics, proteomics, single-cell RNA-seq, and any other available gene-testing technologies. The Curly Hair DNA Chip is loaded with beads for SNP detection using fluorescence or mass-spectrometry-based readouts.

[0036] STUDY (203)

[0037] As step 203, an optional study subject adds the phenotypic hair type classification data of their hair shaft using state-of-the-art photographic and classification techniques. These data will later be linked to the genetic typing results produced during the analysis 202 to generate an integrated “genetic curly hair type.”

[0038] RESULTS

[0039] Results communicated to the individual are described as an “individual hair genotype,” which may include identification of which Wnt gene family members are present within the dermis of the sampled hair follicle, their relative quantities, the receptors engaged in ligand binding, and whether functionally related molecular interactions are observed. This individual hair genotype constitutes a new form of personal genetic information and may be incorporated into ancestry-related databases, including African ancestry databases, and into existing kits, chips, and genetic libraries. Quality control procedures are applied to identify and remove potential errors or genotyping artifacts, followed by statistical analysis of the enumerated variables and correlation of those variables with hair shaft photographs and phenotypic typing to generate a result suitable for communication to the individual. Data generated by the method 200 are further used to support the development of personalized or precision treatments directed to relaxing curly or coily hair from the roots, promoting a healthier scalp and hair shaft.

[0040] As described herein, a "genotype chip," also called a DNA microarray, is a specialized laboratory tool used to identify an individual's specific DNA sequence, particularly related to Wnt gene family members, by attaching numerous microscopic DNA probes to a solid surface, allowing for simultaneous analysis of many genetic markers at once; essentially, it is a high-throughput method for determining an individual's genotype at multiple locations across their genome.

[0041] Further, as used herein, “hair genotype” may include gene expression in addition to SNPs, expression signatures, receptor profiles, and other biological markers.

[0042] Certain embodiments include a Curly Hair-on-Chip system utilizing organ-on-chip microfluidic platforms containing cultured cells and ECM components for studying Wnt signaling, macrophage interactions, and APM-mediated follicular curvature regulation. Here, the word CHIP means Curly Hair Inheritance Patterns, and chip also refers to an Al chip as known in the art possibly made of silicon or other material and are impregnated with specifically selected gene panel.

[0043] Fig. 3 depicts a flowchart of the steps of handling the sample and producing a result to be communicated to the participant / customer classifying their “genetic hair type” and subtypes. The test is providing new information about an individual’s hair gene makeup that can be used to advise haircare treatment and daily care routines. As shown in Fig. 3, the method 200 answers several questions during the processing and analysis of the dermal tissue surrounding the hair follicle sample. The questions to be answered are qualitative and quantitative including: Which Wnt member? How much is present? Which receptor is involved in the crosstalk between the gene ligand and the receptor family? and possibly, What time during the hair growth (telogen) cycle? During the analysis step 202, laboratory artificial intelligence testing investigations are performed, including functional genomics of the cells involved (macrophages, APM and ORD and DS), immunofluorescence of tissue sections, and Volcano Plot and / or interactome and other techniques known in the art.

[0044] When used in this specification and claims, the terms "comprises" and "comprising" and variations thereof mean that the specified features, steps, or integers are included. The terms are not to be interpreted to exclude the presence of other features, steps or components.

[0045] The invention may also broadly consist in the parts, elements, steps, examples and / or features referred to or indicated in the specification individually or collectively in any and all combinations of two or more said parts, elements, steps, examples and / or features. In particular, one or more features in any of the embodiments described herein may be combined with one or more features from any other embodiment(s) described herein.

[0046] Protection may be sought for any features disclosed in any one or more published documents referenced herein in combination with the present disclosure. Although certain example embodiments of the invention have been described, the scope of the appended claims is not intended to be limited solely to these embodiments. The claims are to be construed literally, purposively, and / or to encompass equivalents.

Claims

CLAIMSWhat is claimed is:

1. A method of conducting curly hair genetic typing comprising:collecting a biological sample from a subject, the biological sample comprising a hair follicle and surrounding perifollicular dermis;analyzing the biological sample to detect genetic and molecular markers associated with hair follicle curvature , andgenerating, based on the genetic and molecular makers, a hair genotype classification.

2. The method of claim 1, wherein the genetic and molecular markers comprise at least one of:Wnt gene family members, Frizzled receptor family members, macrophage phenotypes, Wnt co-receptors, Wnt-function-related genes including ED AR and SHH, macrophage-secreted growth factors and immune proteins; and ECM-related genetic markers.

3. The method of claim 1 or 2, further comprising:associating the hair genotype classification with a morphological assessment.

4. The method of claim 1, wherein results from analyzing biological markers are used as input layers for Al -based software that analyzes the input layer and produces a genotype output.

5. The method of claim 4, wherein the genotype output is communicated to a customer.

6. The method of claim 1, wherein collecting a sample comprises pulling-out a hair with its surrounding dermal tissue and extracellular matrix.

7. The method of claim 1, wherein collecting a sample comprises obtaining a blood sample.

8. The method of claim 1, wherein the hair genotype classification is used to develop a personalized hair-relaxing treatment from the root of a hair follicle.

9. The method of claim 1, wherein analyzing the biological tissue sample comprises performing at least one omics-based technique selected from genomics, transcriptomics, proteomics, single-cell RNA sequencing, or spatial transcriptomics.

10. A laboratory gene test kit for typing human hair based on genetic and molecular analysis of a tissue sample containing a hair follicle, comprising:a sampling tool configured to obtain the tissue sample, wherein the sample includes a hair follicle and at least one of a dermis and extracellular matrix surrounding the hair follicle;a preservative container; andan Al-based analytical software configured to classify hair type according to genetic and molecular pathways regulating hair-follicle curvature.

11. The kit of claim 10, wherein the sampling tool comprises a U-shaped skin clipper with a sharp edge configured to push-pinch-pick the sample.

12. The kit of claim 10, wherein the preservative container comprises a solution suitable for preserving at least one of DNA, RNA, proteins, or immune markers present in the tissue sample.

13. The kit of claim 10, wherein the Al -based analytical software is configured to analyze at least one of Wnt gene family members, Frizzled receptor family members, macrophage phenotypes, Wnt co-receptors, Wnt-function-related genes including ED AR and SHH, macrophage phenotype, macrophage-secreted growth factors and immune proteins; and ECM-related genetic markers.

14. The kit of claim 10, wherein the Al-based analytical software comprises:an artificial neural network that receives gene-specific and variable data about the tissue sample as an input layer including quantitative and qualitative data from gene and molecular analyses,wherein an output layer is a person-specific hair genotype, and wherein the Al-based analytical software optionally correlates the data to a photographed hair-shaft phenotype.

15. The kit of claim 10, wherein the tissue sample comprises:three-dimensional perifollicular dermis and extracellular matrix (ECM).

16. A computer-implemented method for genotyping human hair using artificial intelligence, comprising:receiving, by an artificial intelligence system, molecular and genetic data derived from a biological tissue sample comprising a hair follicle and surrounding perifollicular dermis and / or extracellular matrix;processing the molecular and genetic data using a trained machine-learning model to identify patterns in hair-follicle-related signaling pathways; and outputting a hair genotype classification corresponding to a predicted hair-type phenotype.

17. The computer-implemented method of claim 16, wherein the trained machine-learning model comprises an artificial neural network.

18. The computer- implemented method of claim 16, wherein the molecular and genetic data include quantitative or qualitative measurements of Wnt ligand expression, Frizzled receptor expression, macrophage-mediated signaling activity, or extracellular-matrix composition and their functional interactions.

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