COMMENTARY

Woollard WJ, Pullabhatla V, Lorenc A, Patel VM, pathways preceding tumor formation in a mu- of yeasts within the basidiomycetes that
Butler RM, Bayega A, et al. Candidate driver rine model of T-cell lymphoma in skin. J Invest
is closely related to the smut plant
genes involved in genome maintenance and Dermatol 2011;131:1727e34.
DNA repair in Sezary syndrome. Blood pathogen Ustilago maydis and more
Yu H, Lee H, Herrmann A, Buettner R, Jove R.
2016;127:3387e97.
Revisiting STAT3 signalling in cancer: new and
distantly related to skin-infecting fungal
Wu X, Sells RE, Hwang ST. Upregulation of in- unexpected biological functions. Nat Rev species, such as Candida albicans
flammatory cytokines and oncogenic signal Cancer 2014;14:736e46. and the dermatophytes. The unique
Malassezia genus currently includes 17
species, of which M. globosa,
See related article on pg 1137 M. restricta, and M. sympodialis are
most commonly associated with
The Skin Commensal Yeast humans (Sparber and Leibund Gut-
Landmann, 2017). A common feature
Malassezia globosa Thwarts is the inability of Malassezia to
synthesize lipids due to the loss of the
Bacterial Biofilms to Benefit fatty acid synthase gene, and as a
consequence, these species mainly
colonize sebaceous skin. Comprehen-
the Host sive molecular and genetic research on
Malassezia has highlighted unique
Giuseppe Ianiri1, Joseph Heitman1 and Annika Scheynius2
features compared with other fungi,
Malassezia are abundant, lipid-dependent, commensal yeasts in the skin such as a reduction in genome size (as
microbiome that also have a pathogenic lifestyle associated with several com- small as approximately 7e9 Mbp) with
mon skin disorders. Malassezia genomes encode myriad lipases and proteases extensive loss of genes encoding hy-
thought to mediate lipid utilization and pathogenesis. Li et al. report the drolases and other enzymes involved
biochemical characterization of a unique secreted aspartyl protease produced in carbohydrate metabolism, and
by Malassezia globosa, MgSAP1, and demonstrate its active role in hindering concomitant expansion of genes
biofilm formation of the bacterium Staphylococcus aureus. Because biofilms are encoding secreted lipases and pro-
an established virulence attribute of S. aureus, this study reveals a potential teases thought to mediate lipid utiliza-
benefit to the host of the fungal aspartyl protease MgSAP1 and opens the door tion and play roles in pathogenesis
for the investigation of the roles of such molecules in microbial interactions and (Wu et al., 2015).
their possible effects on the host.
Is Malassezia a beneficial skin
Journal of Investigative Dermatology (2018) 138, 1026e1029. doi:10.1016/j.jid.2018.01.008
inhabitant?
Although Malassezia are commensal
inhabitants of the skin, they are asso-
ciated with several skin disorders,
Fungi of the genus Malassezia including pityriasis versicolor, dandruff,
A secreted aspartyl seborrheic dermatitis, atopic dermatitis
The skin of humans and animals is
colonized by numerous microorgan-
protease from (AD), and folliculitis (Saunders et al.,
isms that constitute the skin micro- Malassezia globosa 2012). It is presumed that Malassezia
strains associated with these clinical
biome. These include commensal exhibits unexpected conditions are derived from normal
bacteria and fungi that establish a hydrolytic activity
nonpathogenic interaction with the commensal strains, but this has
host, but in certain circumstances, such that acts on not been experimentally proven. As
as immune conditions within the host, Staphylococcus a result, the pathophysiology of
or intrinsic microbial imbalance, some Malassezia and the mechanisms that
aureus biofilms. govern its shift from commensal to
commensals can become pathogenic or
beneficial. The study by Li et al. (2018) pathogen are poorly understood.
reports a previously unknown benefi- Malassezia species are the most It is reasonable to hypothesize that
cial role for a secreted aspartyl protease abundant fungal skin inhabitant of the immune system has adapted to the
produced by the fungus Malassezia humans (Findley et al., 2013). This commensal nature of Malassezia and
globosa. genus includes a monophyletic group thus tolerates its presence on the skin,
but the immune system can also
recognize the fungus as a pathogen and
1
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, respond. Malassezia can be recognized
North Carolina, USA; and 2Science for Life Laboratory, Department of Clinical Science and Education,
Karolinska Institutet, and Sachs’ Children and Youth Hospital, Södersjukhuset, Stockholm, Sweden
by the host, either directly through
Correspondence: Annika Scheynius, Department of Clinical Science and Education, Karolinska Institutet,
membrane-bound pattern recognition
and Sachs’ Children and Youth Hospital, Södersjukhuset, SE-118 83 Stockholm, Sweden. E-mail: annika. receptors or indirectly through produc-
scheynius@ki.se tion of inflammatory metabolites
ª 2018 The Authors. Published by Elsevier, Inc. on behalf of the Society for Investigative Dermatology. released by hydrolysis of sebum by

1026 Journal of Investigative Dermatology (2018), Volume 138


Figure 1. Staphylococcus aureus biofilms are inhibited by an aspartyl protease secreted by Malassezia globosa. On the left, a schematic view illustrates
a biofilm formed on the skin by the bacterium S. aureus and the inflammation that is produced as a host response. On the right, formation of the biofilm is
inhibited by the secreted aspartyl protease MgSAP1 of M. globosa, and as a consequence, there is predicted to be reduced inflammation of the skin.
fungal lipases and proteases. These
metabolites are thought to be respon-
sible for clinical dandruff and sebor-
rheic dermatitis (White et al., 2014).
Another way in which Malassezia is
known to interact with the immune
system is through the production of
specific allergens. These include a
set of nine conserved and four
unknown proteins identified in the
M. sympodialis genome (Gioti et al.,
2013). Approximately half of adult pa-
tients with AD have allergen-specific
IgE and T-cell reactivity to Malassezia,
which are rarely observed in respiratory
allergies, suggesting a host-microbe
interaction related to the skin environ-
ment in AD (White et al., 2014). A
clinical hallmark of AD is dry, itchy skin
with increased permeability and water
loss. There is a distinct reciprocal
expression pattern of induced inflam-
matory genes and repressed lipid
metabolism genes (Sääf et al., 2008).
This results in an inhospitable environ-
ment for the lipid-dependent Malasse-
zia, and there is an increased pH level
on AD skin that stimulates the yeast to
release more allergens compared with
culture at normal skin pH (Selander
et al., 2006).
Yet another way for Malassezia to
communicate with the host is through
the release of extracellular nanosized
vesicles, designated MalaEx, that carry
allergens and can induce inflammatory
cytokine responses (Gehrmann et al.,
2011). MalaEx, which contain small
RNAs as a cargo (Rayner et al., 2017),
have the potential—like other fungal
extracellular vesicles—to deliver func-
tional mRNAs and microRNA-like
RNAs to recipient host cells, thereby
interfering with the host RNAi ma-
chinery to silence host immune genes
and cause infection (Weiberg et al.,
2013).
Although establishing the basis and
the relationship between commen-
salism and pathogenicity is a recog-
nized and active research field,
Malassezia interactions with other
microbes on the skin and their effects
on the host are largely unexplored. Li
et al. (2018) reported that a secreted
aspartyl protease produced by
M. globosa displays antibiofilm prop-
erties against Staphylococcus aureus,
which is responsible for skin and soft
tissue infections and which is
frequently associated with skin in-
fections in AD. Previous genomic
studies revealed a robust secretory
repertoire for all species belonging to
the genus Malassezia, consistent with
the importance of these hydrolytic en-
zymes for Malassezia biology and
pathophysiology (Wu et al., 2015; Zhu
et al., 2017).
RNAseq analysis revealed that 2 of
the 15 predicted aspartyl proteases of
M. globosa, encoded by MGL_3328
and MGL_1932, were highly expressed
in vitro during growth in both rich and
minimal media. Through biochemical
analyses, Li et al. (2018) characterized
a specific aspartyl protease activity,
revealing strong cleavage of specific
substrates (RPKPYAvWM and
RPKPVEvWR), increasing activity in the
first 16 hours of growth, and a pH
optimum between 4 and 5. Mass
spectrometry and trypsin in-gel digest
analyses identified MGL_1932 as the
major aspartyl protease in culture, and it
was named MgSAP1 (from M. globosa
COMMENTARY
secreted aspartyl protease 1). Further
analyses demonstrated that MgSAP1
has a cleavage preference adjacent
to aromatic residues, in particular
tryptophan, which distinguishes it from
other known proteases.
Li et al. then aimed to establish if the
secreted protease MgSAP1 was (1)
produced in situ on human skin, (2) had
any role in Malassezia biology and/or
pathophysiology, and (3) could poten-
tially mediate interactions with existing
skin microflora. MgSAP1 was found to
be expressed on the faces of 17 of 18
healthy male and female volunteers,
indicating its ubiquitous production.
Among the skin microflora, S. aureus
coexists in many sites with M. globosa
(Oh et al., 2014) and produces protein
A (SpA), which is essential for biofilm
formation, a major S. aureus virulence
factor (Archer et al., 2011). Because
SpA is rich in lysine and aromatic resi-
dues, the authors hypothesized that it
could be hydrolyzed by MgSAP1, thus
impacting S. aureus pathogenicity.
Strikingly, MgSPA1 rapidly degraded
recombinant SpA in vitro and strongly
reduced the in situ volume of S. aureus
biofilm formation without affecting
bacterial viability. A representation of
MgSAP1-mediated biofilm destruction
is shown in Figure 1.
This study raises questions about the
specific roles of Malassezia proteases, as
their common in situ expression has
been correlated with their involvement
in pathogenesis (White et al., 2014). This
predicted function as virulence factors is
likely based on the role of analogous
proteases in more well-studied fungi that
can live on the skin such as the derma-
tophytes and C. albicans. In fact, their
www.jidonline.org 1027
 COMMENTARY
genome comparisons reveal a signifi-
cant expansion of genes predicted to
encode proteases in Malassezia, der-
matophytes, and C. albicans, suggesting
an important role of proteases in evolu-
tion and adaptation on the skin. In
C. albicans, secreted aspartyl proteases
have long been recognized as virulence
factors that act by promoting adhesion
to, invasion of, and damage to epithelial
cells and tissues and by inducing the
secretion of proinflammatory cytokines
independently from their proteolytic
activity (Pietrella et al., 2010). With
respect to dermatophytes, these fungi
secrete an abundance of proteases for
multiple functions, such as adherence
on the skin, keratin digestion for pene-
tration, and modulation of cell meta-
bolism and the host immune system (de
Hoog et al., 2017; White et al., 2014).
The study by Li et al. (2018) repre-
sents a paradigmatic example of a
fungal-bacterial interaction that is
mediated by a secreted molecule and
that potentially results in a beneficial
effect for the host. Fungal and bacterial
interactions are common, although
they are understudied and technically
challenging to elucidate. However,
there is an increasing awareness of their
biological and pathophysiological
importance (Peleg et al., 2010). As an
example, Graham et al. (2017) recently
described an interesting interaction
between the bacterium Enterococcus
faecalis and C. albicans. These two
microbes occupy overlapping niches in
the mammalian microbiome and
display antagonistic activity resulting in
reduced virulence for both microbes. In
C. albicans, this reduced virulence was
attributed to the reduction of hyphal
development and biofilm formation
due to the activity of the secreted
E. faecalis bacteriocin, EntV (Graham
et al., 2017).
Conclusions
The study of Li et al. (2018) reveals that
the commensal and sometimes patho-
genic fungus M. globosa produces an
aspartyl protease, unique in its evolu-
tionary trajectory and substrate speci-
ficity, which impacts a recognized
virulence attribute of a commensal and
pathogenic bacteria, thus potentially
resulting in unexpected benefits for the
host. This finding underscores the
importance of in vivo studies to
1028 Journal of Investigative Dermatology (2018), Volume 138
determine the function of unknown
proteins during microbial interactions,
and the beneficial or detrimental
impact that they might have on the
host. For example, whether the char-
acterized MgSAP1 protease also plays a
role in Malassezia pathogenesis as
described for other aspartyl proteases is
still unknown. There is a crucial need to
develop and establish reliable
host-pathogen models to study how
Malassezia interacts with both cells in
the skin and different immune cells,
and with other microbes that live on the
skin, as well as the induced immune
responses in robust mouse and
humanized-mouse models of infection.
This coupled with the recent develop-
ment of tools for targeted and inser-
tional mutagenesis (Celis et al., 2017;
Ianiri et al., 2016) will help to charac-
terize the mechanisms that establish the
commensal, pathogenic, and beneficial
lifestyles of Malassezia species on the
skin.
CONFLICT OF INTEREST
AS has co-authored large collaborative publica-
tions with the senior author (Tom Dawson) of the
present article commented on herein, published
in 2013 (Gioti et al., 2013), 2014 (White et al.,
2014), 2015 (Wu et al., 2015), and 2017 (Zhu
et al., 2017). This did not influence the views
presented here with respect to the Commentary
published in the Journal of Investigative Derma-
tology. JH has co-authored large collaborative
publications with the senior author (Tom Dawson)
of the article discussed, published in 2013 (Gioti
et al., 2013), 2015 (Wu et al., 2015), and 2017
(Zhu et al., 2017). This did not influence the views
presented here with respect to the article pub-
lished in the Journal of Investigative Dermatology.
AS has research support from the Swedish Research
Council Medicine, the Cancer and Allergy Foun-
dation, the ChAMP (Centre for Allergy Research
Highlights Asthma Markers of Phenotype) con-
sortium that is funded by the Swedish Foundation
for Strategic Research, the Karolinska Institutet,
AstraZeneca and Science for Life Laboratory Joint
Research Collaboration. AS is a member in the Joint
Steering Committee for the Human Translational
Microbiome Program at SciLifeLab/Karolinska
Institutet together with Ferring Pharmaceuticals,
Switzerland, during 2016e2018, outside the scope
of this work and without any financial support. GI
states no conflict of interest.
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 COMMENTARY
White TC, Findley K, Dawson TL Jr, Scheynius A, on human skin. PLoS Genet 2015;11: within the lamina lucida); dystrophic EB
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Kennell JC, Sun S, et al. Proteogenomics pro-
Wu G, Zhao H, Li C, Rajapakse MP, Wong WC, duces comprehensive and highly accurate and Kindler syndrome with multiple
Xu J, et al. Genus-wide comparative protein-coding gene annotation in a complete levels of blistering. The treatment of
genomics of Malassezia delineates its phy- genome assembly of Malassezia sympodialis. these different forms of EB relies pri-
logeny, physiology, and niche adaptation Nucleic Acids Res 2017;45:2629e43.
marily on prevention of trauma and in-
fections, and there is no specific cure for
See related articles on pgs 1157 and 1228 this, currently intractable group of dis-
orders. However, over the past decade,
and especially over the past few years,
The Conundrum of Allogeneic there has been considerable progress in
understanding the mechanistic details
Bone Marrow Transplantation leading from the genetic defects into
blistering phenotypes, and preclinical
for Epidermolysis Bullosa work has suggested a number of poten-
tial treatment approaches, some of
Jouni Uitto1 which have been extended to the level
of early clinical trials.
Epidermolysis bullosa is a heterogeneous group of heritable blistering disorders One of the first early clinical trials
with considerable morbidity and mortality. Currently, there is no effective for the systemic treatment of EB
treatment or cure for epidermolysis bullosa, but bone marrow transplantation consisted of allogeneic whole bone
has been suggested to improve the clinical presentation and quality of life of marrow transplantation (BMT), reported
some patients with the recessive dystrophic subtype of epidermolysis bullosa. In in 2010, in patients with severe recessive
this issue, two studies (Hünefeld et al., and Egawa and Kabashima) address the dystrophic EB (RDEB) (Wagner et al.,
issue whether bone marrow transplantation could be applied to patients with 2010). More than 40 children with
epidermolysis bullosa simplex with intraepidermal blistering. Utilizing a either RDEB or junctional EB have now
desmoglein-3 mouse model (Dsg3
/
) or keratin 5-specific reporter mice, the undergone BMT worldwide. Although
investigators show that transplanted bone marrow-derived cells migrate to the the results of the clinical experience
skin of bone marrow transplantation recipient mice, but these cells fail to involving the full cohort of these patients
transdifferentiate into epidermal keratinocytes, and there was no improvement have yet to be published, it appears that
in the clinical manifestations of the Dsg3
/
mice. Thus, further preclinical beneficial clinical response, but not
experimentation, possibly using mouse models that more faithfully recapitulate cure, can be noted in some, but not all,
the epidermolysis bullosa simplex phenotype, is advisable before commencing children with RDEB after BMT. Although
clinical trials of bone marrow transplantation for epidermolysis bullosa simplex. experience with BMT in junctional EB is
Journal of Investigative Dermatology (2018) 138, 1029e1031. doi:10.1016/j.jid.2017.12.009 limited, this approach does not appear
to have any therapeutic value for this
type of EB (Hammersen et al., 2016;
Epidermolysis bullosa (EB), a group influenced by the topographic expres- Hook et al., 2017). In this issue of the
of heritable blistering disorders, pre- sion of the affected genes within the Journal, two complementary studies
sents with a spectrum of phenotypic cutaneous basement membrane zone, now ask if BMT might be applicable to
severity, with the primary manifesta- the types and combinations of the EBS characterized by intraepidermal
tions relating to skin and mucous mutations, and their consequences at blistering (Egawa and Kabashima, 2018;
membrane fragility (Fine et al., 2014). the mRNA and protein levels (Uitto Hünefeld et al., 2018). As a disease
In the most severe forms, patients et al., 2016; Vahidnezhad et al., 2018). model, Hünefeld and coworkers
affected with this disease die within a Based on ultrastructural demonstration employ a desmoglein-3 knockout mouse
few weeks or months of life, whereas in of the level of blistering within the (Dsg3
/
) that demonstrates blistering
the milder forms, affected individuals skin and clinical presentations, EB has exclusively within the epidermis and
exhibit life-long blistering that does been divided into four broad categories: mucosal membranes, clinically mani-
not affect longevity. This spectrum of EB simplex (EBS) with intraepidermal festing with perioral erosions and a run-
phenotypic severity reflects the fact blistering; junctional EB with tissue ted phenotype, and no significant
that mutations in as many as 20 distinct separation within the dermal-epidermal increase in mortality in the first 4e6
genes can underlie the EB phenotype, basement membrane itself (primarily months of life. BMT was performed at
8e10 weeks after the birth using bone
1
marrow cells from enhanced green
Department of Dermatology and Cutaneous Biology, Sidney Kimmel Medical College, Thomas
Jefferson University, Philadelphia, Pennsylvania, USA
fluorescent protein (EGFP) expressing
Correspondence: Jouni Uitto, Department of Dermatology and Cutaneous Biology, Sidney Kimmel
wild-type Dsg3þ/þ mice that can be used
Medical College at Thomas Jefferson University, 233 S. 10th Street, Suite 450 BLSB, Philadelphia, to track the engraftment and migration of
Pennsylvania 19107, USA. E-mail: Jouni.Uitto@Jefferson.edu the transplanted cells. Ten weeks after
ª 2017 The Author. Published by Elsevier, Inc. on behalf of the Society for Investigative Dermatology. BMT, the distribution of donor cells was

www.jidonline.org 1029