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TOWARDS THE GENERATION OF A LASER CAPTURE MICRODISSECTION-BASED HUMAN ANAGEN HAIR FOLLICLE TRANSCRIPTOME ATLAS

ABSTRACT: While this is available for murine HFs, systematic characterization of the transcriptome of specific human hair follicle (HF) compartments remains a major unmet challenge in human hair research. Here, we have aimed to initiate the generation of such an atlas to reveal HF compartment-specific transcriptome. We performed laser capture microdissection (LCM) on 8 selected compartments, i.e. bulge outer root sheath (ORS), suprabulbar ORS, inner root sheath (IRS), precortical hair matrix (HM), germinative HM, dermal papilla (DP), the lower dermal cup (DC), and the connective tissue sheath (CTS), of human anagen VI scalp HFs, and analysed these by RNA-seq. Principal component analysis (PCA) was used to validate the distinctness of the microdissected compartments. Biological replicates were clearly separated by HF compartments and a high degree of similarity of the distinct transcriptional profiles of each examined compartment was seen. Afterwards, we matched well-known signature genes to the correct HF compartments. For example, VCAN (versican) and ALPL (alkaline phosphatase) were correctly expressed in the DP, the lower DC, and suprabulbar CTS, while TCHH (trichohyalin) was solely found in the proximal IRS, and KRT85 (Keratin 85) and KRT31 (Keratin 31) were almost exclusively expressed in the precortical HM. These transcriptomic data were used to generate an easily accessible database for identifying and defining compartment-specific gene expression profiles, which can be ultimately made publicly accessible (“hair follicle MONasterium InteraCtive atlas – hairMONic”). This human HF transcriptomic map and the associated interactive database will support the identification of HF compartment-specific signature genes, and facilitates the development of compartment-specific, therapeutic interventions for the management of human HF disorders.

ORGANISM(S): Homo sapiens

PROVIDER: GSE272333 | GEO | 2025/11/12

REPOSITORIES: GEO

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Project description:Transcriptome profiling reveals different expression pattern in baseline group(after isolated hair follicles---HFs,collect samples immediately), 0G group(sham irradiation, collect organ-cultured HFs for 3 days),and 5G group(5G dose irradiation, collect organ-cultured HFs for 3 days) in ex vivo organ-cultured HFs. Hair greying or canities is one of the earliest phenomena in aging. A worldwide survey based on a large sample of human subjects showed that men grey faster than women, but in distinct scalp regions, and the populations of Asian and African descent had less grey hair than those of the same age of Caucasian descent. The average grey hair age of whites is 34 ± 9.6 years in Caucasians, while 43.9 ± 10.3 years in African. And then clinical observation demonstrates that by the age of 50 in 50% of all men in senile canities, 50% of all hair follicles (HFs) have lost their pigment. In addition, an increasing number of studies found that hair greying acts as an important predictor of aging-associated pathology, including Alzheimer’s disease, Parkinson’s disease and cardiovascular disease, etc. Grey hair is both visible and non-lethal, and it makes this phenotype a unique model system for investigating mechanisms that contribute to tissue aging and therapeutic strategies to combat this process. In humans and mice, age-related grey hair is attributed to the loss of melanocyte stem cells (MeSCs), precursor cells, and melanocytes, or their reduced or restricted functions, which is induced by a multitude of exogenous and endogenous challenges, such as radiation, inflammation, psycho-emotional stress, or gene mutation (BCL-2 deficiency), etc. Of which, ionizing radiation (IR) produces multiple clusters of double-strand breaks that mediate cell cycle arrest, apoptosis, and DNA repair, and successfully induces grey hair in mice. The accelerating process of aging by IR may provide an avenue to create a grey hair model. Although there is a mouse model of grey hair, the existence of species differences also limits the translational potential of the mice HF models for human applications. Recently, Rachmin et al demonstrated that IR could result to ectopically pigmented melanocytes (EPM) in outer root sheath (ORS) followed hair greying and speculated that this may provide a tool to prevent or reverse hair greying in humans. They proved that different genotoxicity can induce premature differentiation of MeSCs and/or amelanotic cells. However, they didn’t determine the appropriate dose of irradiation for creating the hair grey model and not discuss the mechanisms of forming the model. Hence, this study focuses on establishing a suitable ex vivo human grey HF model by IR, demonstrating the series change of grey hair and exploring the possible mechanisms of the model. In our study, parameters about senescence and melanotic features in our model are almost in line with essential traits of human grey hair in previous studies. In addition,to explore the profound mechanism of the irradiated model, we performed transcriptome analysis for baseline, 0G and 5G groups. Then, three pairwise comparisons were established---0G vs 5G, baseline vs 5G and baseline vs 0G. This profiling results be used to understand the different molecular mechanism of creating ex vivo grey HFs model,and those provided a possibility for anti-aging treatment.In summary, our results indicate that the ex vivo grey HFs by irradiation can perfectly replace elderly grey hair and establish a potential preclinical aging model for pharmacological and mechanism studies.

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TOWARDS THE GENERATION OF A LASER CAPTURE MICRODISSECTION-BASED HUMAN ANAGEN HAIR FOLLICLE TRANSCRIPTOME ATLAS

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