Inventors: Ritsuko Ehama, Jiro Kishimoto, Ritsuro Ideta, Tsutomu Soma; Shiseido Company, Ltd.
Publication date: 2008-04-17
Invention summary:
The present invention relates to a method for regenerating and/or forming hair follicles by attempting to suppress expression of a gene having the ability to inhibit hair follicle formation, and a method for culturing hair follicle dermal papilla cells by suppressing expression of that gene. Moreover, the present invention relates to a method for regenerating and/or forming hair follicles by attempting to accelerate expression of a gene having the ability to induce hair follicle formation, and a method for culturing hair follicle dermal papilla cells by up-regulating expression of that gene.
Abstract:
It is intended to provide a method of regenerating follicles which comprises inhibiting the expression of one or more genes capable of inhibiting follicle formation that are selected from the group consisting of S100a6, Ctgf, Gsto1, Gas6, Klf2, Thbs1 and Thbd, or promoting the expression of one or more genes capable of inducing follicle formation that are selected from the group consisting of Tgfbi, Gas1, Thbs2, Ifi202A, Bmp7, Efna1, Efna3, Cidea, Serping1, MS1, Irf6, Fmod and Fxyd4.
First Claim:
A method for regenerating hair follicles by suppressing expression of one or a plurality of genes having the ability to inhibit hair follicle formation selected from the group consisting of S100 calcium binding protein A6 gene (S100a6), connective tissue growth factor gene (Ctgf), glutathione-S-transferase omega 1 gene (Gsto1), growth arrest-specific 6 gene (Gas6), Kruppel-like factor 2 (Klf2), thrombospongin 1 (Thbs1) and thrombomodulin (Thbd).
Background:
There is a high demand for attempting to compensate for hair loss accompanying aging and so forth, and typically not only the use of hair tonics, but also hair transplants and other procedures are performed at health care institutions and so forth. On the other hand, there are considerable expectations, albeit somewhat excessive, being placed on regenerative medical technologies in the form of advanced medicine as an alternative to conventional organ transplants accompanying technical breakthroughs attributable to recent progress in stem cell research, a serious shortage of donors due to problems with tissue compatibility and so forth, and requirements based on ethical perspectives such as the issue of assessing brain death. Thus, attention unlike that in the past is being focused on hair follicle regenerative research as a model organ of regenerative medicine.
Research on the follicle formation mechanism in the developmental stage has been conducted comparatively extensively, and hair follicles have been determined to be formed as a result of a complex interaction attributable to signal transmission between epithelial cells (epidermal cells) and mesenchymal cells (hair follicle dermal papilla cells or DPC) lying directly beneath (R. Pause, et al., N. Engl. J. Med., 341, 491-497, 1999; K. S. Stenn, et al., Physiol. Rev. 81, 449-494, 2001; S. E. Miller, et al., J. Invest. Dermatol., 118, 216-225, 2002). In addition, once hair follicles have been formed, they undergo a cyclic regeneration in which anagen, catogen and telogen are repeated. Although numerous growth factors, cytokines, hormones, neuropeptides and other physiologically active substances are known to be involved in the regulation thereof, these physiologically active substances do not necessarily coincide with those involved in the mechanism of the developmental stage of hair follicle formation.
Based on the results of mouse hair follicle reconstitution experiments using nude mice, it has been determined that both epithelial cells and mesenchymal cells are essential for hair follicle regeneration, and that hair follicle regeneration is not induced unless a fixed number of cells are present (J. Kishimoto, et al., Proc. Natl. Acad. Sci., 96, 7336-7341, 1999). Moreover, although the regeneration of chimeric hair follicles comprised of mouse DPC and human epithelial cells has been suggested to be possible (Japanese Patent Application No. 2004-048332; Ehama, et al., 26th Annual Meeting of the Molecular Biology Society of Japan, Abstracts, 2PC-024, 2003), it is still not possible to regenerate completely human hair follicles. One of the reasons for this is that it is difficult to obtain an adequate amount of human DPC having the ability to induce hair follicle-formation to be used for transplant.
Although cells such as DP expressing versican have been shown to specifically have the ability to induce hair follicle-formation under specific conditions (J. Kishimoto, et al., Proc. Natl. Acad. Sci., 96, 7336-7341, 1999), there are many aspects of the phenomenon of hair follicle induction at the molecular level which remain unknown.
Summary:
An object of the present invention is to identify factors which regulate hair follicle-formation induction in mouse DPC, which is known to have the ability to induce hair follicle-formation, in order to find factors to enable activation of human DPC by attempting to establish a method for promoting human hair follicle regeneration by controlling the action of molecules regulating hair follicle formation or hair follicle regeneration.
The inventors of the present invention found that, although the ability to induce hair follicle-formation by hair follicle dermal papilla cells is lost as a result of culturing thereof, in the case of culturing those cells at a predetermined high density, the induction ability thereof tends to be retained. When hair follicle dermal papilla cells were cultured under high-density (specifically, 3 to 7.times.10.sup.5 cells/cm.sup.2) and low-density (specifically, 5 to 9.times.10.sup.4 cells/cm.sup.2) conditions to investigate expressed genes, the inventors of the present invention found that expression of the following specific genes is specifically up-regulated in hair follicle dermal papilla cells which were unable to form hair follicles as a result of culturing under low-density conditions, while also found that expression of the following specific genes is specifically up-regulated in hair follicle dermal papilla cells which formed hair follicles as a result of culturing under high-density conditions. |
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