Fibroblasts from the Human Skin Dermo-Hypodermal Junction are Distinct from Dermal Papillary and Reticular Fibroblasts and from Mesenchymal Stem Cells and Exhibit a Specific Molecular Profile Related to Extracellular Matrix Organization and Modeling.

Human skin dermis contains fibroblast subpopulations in which characterization is crucial due to their roles in extracellular matrix (ECM) biology. This study investigates the properties of fibroblasts localized at the frontier of deep dermis and hypodermis, i.e., dermo-hypodermal junction fibroblasts (F-DHJ), which were compared to intermediate reticular dermis (Fr) and superficial papillary dermis (Fp) fibroblasts. F-DHJ differed from Fr and Fp cells in their wider potential for differentiation into mesodermal lineages and in their absence of contractility when integrated in a three-dimensional dermal equivalent. The transcriptomic profile of F-DHJ exhibited specificities in the expression of genes involved in ECM synthesis-processing and "tissue skeleton" organization. In accordance with transcriptome data, ECM proteins, notably Tenascin C, distributions differed between the reticular dermis and the dermo-hypodermal junction areas, which was documented in normal adult skin. Finally, genome-wide transcriptome profiling was used to evaluate the molecular proximity of F-DHJ with the two dermal fibroblast populations (Fp and Fr) and with the mesenchymal stem cells (MSCs) corresponding to five tissue origins (bone marrow, fat, amnion, chorion, and cord). This comparative analysis classified the three skin fibroblast types, including F-DHJ, as a clearly distinct group from the five MSC sample origins.

Transcriptome profiling of human papillary and reticular fibroblasts from adult interfollicular dermis pinpoints the 'tissue skeleton' gene network as a component of skin chrono-ageing.

Interactions between extracellular matrix (ECM) and fibroblasts are essential for maintaining dermis integrity, and are subject to ageing. The ötissue skeleton' network connects ECM to the nucleus and DNA, impacting nuclear shape and gene expression. In a previous Mech Ageing Dev publication, we have presented a transcriptomic study of papillary (Fp) and reticular (Fr) fibroblasts, with a main focus on Fp ageing. As shown here, ageing affects ötissue skeleton' transcripts, even more clearly in Fr than in Fp. Accordingly, using circular index measurement, we show that nuclear shape is affected by ageing in both cell fractions.

Genome-wide profiling of adult human papillary and reticular fibroblasts identifies ACAN, Col XI α1, and PSG1 as general biomarkers of dermis ageing, and KANK4 as an exemplary effector of papillary fibroblast ageing, related to contractility.

Deciphering the characteristics of dermal fibroblasts is critical to further understand skin ageing. We have conducted a genome-wide transcriptomic characterization of papillary (Fp) and reticular (Fr) fibroblasts extracted from human skin samples corresponding to younger and older adult ages. From this screen, biomarkers suitable for the assessment of chronological ageing were identified, and extrapolated to the context of photo-damaged skin. In particular, KANK4, ACAN, Col XI α1, and PSG1, were expressed at an increased level in both chronologically-aged and photo-damaged skin. Notably, analysis focused on Fp identified significant transcriptional signatures associated with ageing, which included transcripts related to extracellular matrix, focal adhesion points, and cytoskeleton, thus suggesting functional consequences on tissue structure. At a cellular level, an increased contractility was identified as a property of aged Fp. Accordingly, further investigations were conducted on the KN motif and ankyrin repeat-containing protein 4 (KANK4) to explore its possible function as an original effector involved in the acquisition of aged properties in Fp, notably their increased contractility. We show that KANK4 down-modulation using siRNA led to increased Rho pathway activity, thereby reducing their contractility. As a proof-of-principle, the present study shows that targeting KANK4 was efficient to attenuate aged Fp characteristics.