Abe, R.

Akiyama, M.

Long, H.

McMillan, J. R.

Miyamura, Y.

Qiao, H.

Shibaki, A.

Shimizu, H.

English

Elsevier - Publisher Connector 

Abstracts 

Plikus, M.

Baker, Ruth E.

Mayer, J.

Maini, P. K.

Chuong, C.

ORA - Oxford University Research Archive

BMB activity defines refractory and competent hair follicle populations during the propagation of regenerative waves

Brans, Richard

Yao, Feng

Publikationsserver der RWTH Aachen University

Vaccination with CJ9-gD, a replication-defective and dominant-negative herpes simplex virus type 1 (HSV-1), induces protective immunity against genital herpes

Plikus, M. V.

Mathematical Institute Eprints Archive

ABSTRACTS
637
Adult hair follicle stem cell compartment changes in androgenetic alopecia demonstrate
maintenance of progenitor stem cells with loss of descendant CD200high A6 integrinhigh
expressing cells
LA Garza,1 M Lee,1 Y Liu,1,4 D Stanton,2 L Carrasco,2 J Tobias3 and G Cotsarelis1 1
Department of Dermatology, University of Pennsylvania School of Medicine, Philadelphia,
PA, 2 Department of Oral and Maxillofacial Surgery, University of Pennsylvania, Philadelphia,
PA, 3 Bioinformatics Core, University of Pennsylvania, Philadelphia, PA and 4 Merck &
Company, Wales, PA
The status of adult stem cell compartments in tissue specific disease has not been thoroughly
addressed. We tested the hypothesis that hair follicle stem cells might be depleted in andro-
genetic alopecia (AGA), which is characterized by drastic miniaturization of the hair follicle.
To compare hair follicle stem cell numbers between paired haired and bald scalp samples
from the same individuals, we used flow cytometry to quantitate cell cycle, cell size, and
expression of CYTOKERATIN 15 (KRT15), FOLLISTATIN (FST), CD200 and alpha-6 integrin.
We found a gradient of stem cell characteristics, as defined by a high degree of KRT15 and
FST expression, cellular quiescence and small cell size. This gradient is not grossly altered
between haired and bald scalp, and stem cells are maintained in bald scalp. However, a spe-
cific CD200 high alpha-6-integrin high population, which has characteristics of early stem
cell progeny, is lost in bald scalp. Consistent with the loss of the immunosuppressive CD200
protein, array based expression profiling demonstrates significant increases in inflammation
associated genes in androgenetic alopecia. Previous reports of CD200 loss leading to alope-
cia in mouse models suggest that AGA may be exacerbated or caused by CD200 downregu-
lation in the human hair follicle stem cell compartment.
639
BMP activity defines refractory and competent hair follicle populations during the propa-
gation of regenerative waves
M Plikus,1 R Baker,2 J Mayer,1 R Maxson,1 P Maini2 and C Chuong1 1 Pathology, University
of Southern California, Los Angeles, CA and 2 Oxford, Oxford, United Kingdom
Hair cycle can be regulated by molecular oscillation within the follicle (autonomously) or by
dynamic changes in the inter-follicular environment (non-autonomously). While human hair
follicles largely cycle autonomously, cycling of mouse hair follicles is affected by the status
of adjacent follicles. Thus hair cycle progression can advance like a transverse wave on the
surface of the skin. Observations of the dynamic hair cycle domains on the skin of wild type
and Msx2-/- mice show that each hair cycle domain is made of an initiation site, a propagat-
ing wave, and boundaries. Analyses of patterns of wave propagation and boundary formation
led us to identify a novel stage in telogen. In the first stage of telogen, hair follicles are refrac-
tory to the anagen spreading wave, thus forming a sharp domain boundary. Refractory telo-
gen is characterized by multiple site expression of Bmp4/Bmp2 intra-follicularly and inter-
follicularly. These Bmps sustain BMP reporter activity within follicles throughout refractory
stage of telogen. Telogen follicles become competent to respond to anagen initiation signals
after Bmp 2/4 loss. Hair plucking can induce anagen in competent follicles, but not refrac-
tory ones. In KRT14-NOG mice there is drastic shortening of the refractory telogen, and fast
anagen re-entry. Individual follicle cycles faster and the wave dynamics of follicle populations
are altered. Transplantation of a piece of KRT14-NOG skin to normal skin microenvironment
results in restoration of Bmp signaling activity and refractory telogen. Our data identifies two
new stages of telogen, refractory and competent. They are defined by the ability of telogen
follicles to re-enter anagen, and based on the on-and-off switches of Bmp2/4 expression within
and surround the follicle. This study presents a novel population approach to the study of hair
cycling, and gives a more systematic and integrative explanation to the hair follicle behaviors
observed in classical literatures.
641
Reactive oxygen species (ROS)-mediated androgen-inducible TGF-β1 regulation in dermal
papilla cells
H Yoo, Y Kang, M Sung, S Kim, O Kwon, K Kim, H Eun and K Cho Department of
Dermatology, Seoul National University, College of Medicine, Laboratory of Cutaneous Aging
and Hair Research, Clinical Research Institute, Seoul National University Hospital and
Institute of Dermatological Science, Seoul National University, Seoul, South Korea
Intracellular signaling roles of reactive oxygen species (ROS) generated in response to andro-
gen hormone in hair follicle dermal papilla cells are not well defined. To assess the correla-
tion of ROS (hydrogen peroxide) known to increase in aging process to hair loss, the rat vib-
rissae dermal papilla cell line (DP-6) overexpressed with AR was investigated to evaluate the
role of ROS on androgen-induced increase of TGF-β1 secretion. The AR stably-transfected DP-
6 cells were incubated with synthetic androgen, R1881. Intracellular production of ROS
markedly increased with R1881 treatment in DP-6 cells, which was measured by flow cytom-
etry and laser scanning confocal microscopy. Moreover, androgen-inducible TGF-β1 was sig-
nificantly suppressed by several species of ROS scavenger or inhibitors. Luciferase reporter
assays showed suppression of TGF-β1 promoter signaling by ROS scavengers. In conclusion,
our study shows for the first time that androgen-induced TGF-β1 regulation would be medi-
ated by ROS and prevented by antioxidants or ROS inhibitors in hair follicle dermal papilla
cells. We suggest that antioxidants could be one of the candidates to control androgen-medi-
ated pattern hair loss.
640
A role of CYLD in hair cycling mouse
K Moriwaki,1 D Tsuruta,1 K Sugawara,1 H Kobayashi,1 R Massoumi2 and M Ishii1 1
Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan and 2
Molecular Medicine, MaxPlanck-Institute for Biochemistry, Martinsried, Germany
CYLD is a tumor suppressor gene mutated in familial cylindromatosis, an autosomal-dominant
condition that predisposes to multiple skin tumors. The gene product, CYLD, is a deubiquiti-
nating enzyme recently implicated in modulation of the NF-κB pathway. Recent report by Mas-
soumi R. et al (2006) described that CYLD protein is prominently expressed in the inner root
sheath of normal human scalp hair follicles. Thereupon we considered CYLD might have some
effect in normal hair cycle thru NF-κB signaling. To investigate a role of CYLD during hair
cycling, we adopted C57BL/6 hair cycling model mouse, and characterized changes in their
expression by semiquantitative RT-PCR and western immunoblot and immunohistochemistry
for protein. We found that CYLD at mRNA level increased from anagen stage to catagen
stage, as well as protein level in immunobolt. In immunohistochemistry, CYLD protein is
expressed in surrounding bulge areas. Moreover, we assessed whether CYLD regulate hair
growth using in vitro hair culture. Recombinant CYLD inhibited hair growth. In conclusion,
our results indicate that CYLD function on conversion from anagen stage to catagen stage.
638
Keratinocyte-specific expression of c-myb is dependent on growth and differentiation state
JM Luna,1 ML Clarke,2 J Frampton2 and A Engelhard1 1 Department of Dermatology, College
of Physicians and Surgeons, Columbia University, New York, NY and 2 Institute of Biomedical
Research, University of Birmingham Medical School, Edgbaston, United Kingdom
Many genes with a role first defined in hematopoiesis have recently been shown to define
stem cell niches and regulate essential cellular processes in the skin. In hematopoietic cells,
c-myb expression is required for proliferation but prohibitive of differentiation. Viral trans-
duction of truncated c-myb causes leukemia in chickens, and murine c-myb null mutation
is lethal at e15, due to the failure of hematopoiesis. Here, we examined the expression pat-
tern of the c-myb transcription factor in keratinocytes to determine if it might also regulate
cell growth and differentiation in the skin. Upon release from serum starvation, absolute quan-
tification of c-myb RNA levels demonstrated cell cycle-regulated expression in cultured human
keratinocytes. Temporal studies of c-myb RNA levels during calcium-induced differentiation
defined downregulation of c-myb expression as a late marker of keratinocyte differentiation.
Both results are consistent with the expression pattern of c-myb in hematopoietic cell lines.
Furthermore, immunohistochemistry on normal murine skin revealed that c-myb protein is
present not only in the proliferating basal layer of the interfollicular epidermis, but also in
the suprabasal layer. In the hair follicle, c-myb is expressed in a hair cycle-dependent man-
ner. During late anagen, c-myb is expressed in the hair matrix and the outer root sheath and
is absent from the inner root sheath, cortex and dermal papillae. This expression pattern
extends into early catagen, but c-myb nuclear staining is not detected in telogen. The tem-
poro-spatial expression pattern of c-myb suggests that it is a novel regulator of keratinocyte
proliferation and differentiation in the epidermis and hair follicle. Conditional ablation in the
epidermis is ongoing, and is expected to reveal in vivo functions as well as insights into down-
stream transcriptional target genes.
642
Global microarray analysis of the follicular and glandular types of epidermal differentiation:
a role for BMP signaling in suppressing trans-differrentiation of the foot pad epidermis towards
folliculogenesis
AA Sharov,1 TY Sharova,1 AN Mardaryev,1,2 AV Sargsyan1 and VA Botchkarev1 1
Dermatology, Boston University School of Medicine, Boston, MA and 2 Medical Biosciences,
School of Life Sciences, University of Bradford, Bradford, United Kingdom
During skin development, epithelium and mesenchyme interact with each other and give raise
the formation of epidermis and a variety of appendages (hairs, nails, glands). To characterize
global gene expression profiles associated with distinct types of epidermal differentiation (fol-
licular versus glandular), epithelial buds of the hair follicles (HFs) and sweat glands (SWGs),
as well as the matrix and epithelium of fully-developed HFs and SWGs were obtained from
embryonic and adult mouse foot pads and ventral skin using laser capture microdissection.
Epithelial buds of the HFs and SWGs showed expression of the bud-specific adhesion mole-
cules, signaling/transcription components, as well as appendage-specific markers such as
HF-specific keratins (Krt1-c29, Krt2-6g) or SWG-specific ion channels (Clcn3, Bcng-3a). Fully
developed SWGs were characterized by strong downregulation of over 4000 genes com-
pared to the epidermis and by upregulation of SWG-specific genes involved in regulation of
the ion exchange/water metabolism, while HFs showed less prominent differences in gene
expression versus the epidermis. In addition, SWG buds and fully developed SWGs showed
strongly reduced expression of Bmp7 and Smad1/4 compared to the epidermis and HFs.
Interestingly, K14-Noggin and Bmpr-Ib knockout mice showed ectopic formation of the HFs
in foot pads accompanied by decreased expression of Engrailed 1, a potent repressor of the
dorsal phenotype in ventral epidermis. These data suggest that HFs and SWGs are character-
ized by markedly different gene expression profiles, which may underlie fundamental differ-
ences in their functions, and that BMP signaling suppresses trans-differentiation of the foot pad
epidermis towards folliculogenesis at least in part via stimulating the Engrailed 1 expression.
www.jidonline.org   S107


Oxford University Research Archive

RWTH Publications

Oxford University Research Archive (ORA)

Kindler syndrome (KS) is a rare disorder leading to keratinocyte fragility, poikiloderma and palmar hyperkeratosis (OMIM173650). Defects in the actin/focal contact associated protein kindlin-1 encoded by the gene KIND1 have been shown to underlie this disease. We assessed the function of kindlin-1 in human keratinocytes and fibroblasts using RNAi knockdown. RNAi treatment reduced KIND1 message by 72% and 63% in cultured keratinocytes and fibroblasts, respectively. RNAi treatment reduced cultured keratinocyte but not fibroblast cell adhesion to plastic. Furthermore, kindlin-1 depletion reduced keratinocyte (but not fibroblast) survival by increasing rates of adherent keratinocyte apoptosis, as detected by TUNEL staining. Kindlin-1 depleted keratinocytes demonstrated an initial delay in cell spreading and filopodia formation (as observed by scanning EM) but were associated with some focal adhesion / contact assembly (as visualized by vinculin staining). Keratinocyte (but not fibroblast) migration was increased after kindlin-1 depletion. Taken together our data suggest that kindlin-1 plays important roles in keratinocyte cell-substrate adhesion, spreading, migration and in keratinocyte survival in vitro. These observations may explain the loss of keratinocyte-basement membrane adhesion in the pathogenesis of KS induced skin fragility. Furthermore, reductions in keratinocyte survival and/or increased rates of apoptotic cell death may underlie the early-onset poikiloderma observed in KS cases

University of Queensland eSpace

Kindlin-1 is important in human keratinocyte cell adhesion, spreading, migration and protection from apoptosis in vitro

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BMB activity defines refractory and competent hair follicle populations during the propagation of regenerative waves

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Elsevier - Publisher Connector

Publikationsserver der RWTH Aachen University

Mathematical Institute Eprints Archive

Oxford University Research Archive

RWTH Publications

Oxford University Research Archive (ORA)

University of Queensland eSpace