|
Education
·
Doctorate: 29th
September 2004 (cum laude)
Radboud University Nijmegen Medical Center, Department of Dermatology
Title of thesis: The
Role of Cystatin M/E in Epidermal Differentiation. From Tag to Function
Promotores: Prof dr J.Schalkwijk
and Prof dr P.C.M. van de Kerkhof, co-promotor: dr
W.Hendriks
·
Master's (1988 – 1992)
Laboratory School for Higher Professional Education Nijmegen (Bachelor of
Science, Biosciences)
Main subject: Molecular Biology (Molecular characterization of
protein-tyrosine phosphatases)
Professional
Positions
·
Ph.D on a NWO-Veni grant, July
2005 – July 2008 at the Laboratory of Skin Biology and Experimental
Dermatology, Radboud University Nijmegen Medical Center, supervision of
Ph.D-student (1 fte) and technician (1 fte).
·
Ph.D, July 2004 – July 2005 at
the Laboratory of Skin Biology and Experimental Dermatology, Radboud
University Nijmegen Medical Center, supervision of Ph.D-student (1 fte) and
technician (1 fte).
·
Ph.D-student within an NWO
program grant, July 2000 – July 2004 at the Department of Dermatology,
University Medical Center Nijmegen, supervision of technicians (1.5 fte)
and graduate students.
·
Research Technician, July 1992 –
July 2000, Department of Dermatology, University Medical Center Nijmegen.
Thesis
supervision and co-promotor
·
2004 – present: Legumain-cystatin M/E: a novel tissue specific
protease-antiprotease system. The role in human physiology and
pathology –
Tsing Cheng
Brief summary of research over last
five years
Following my training in molecular biology at the department of Cell
Biology at the UMCN (prof dr B.Wieringa/dr W.Hendriks), I started as a
research technician at the department of Dermatology of the UMCN. After
being involved in projects of various PhD students, I gradually got the
ambition to have my own research project, and in 2000 I was given the
opportunity to start as a PhD student within an NWO program grant (together
with the department of Lung Diseases at the LUMC) on the role of epithelial
protease inhibitors. The role of proteases and their inhibitors in barrier
function and host defense has been a major topic of my investigations over
the past years. On the basis of large-scale transcriptome analysis of human
keratinocytes, I subsequently identified the cysteine protease inhibitor
cystatin M/E as a differentiation-specific protein in human epidermis. I
found that cystatin M/E deficient mice die shortly after birth, because of
a compromised skin barrier, similar to a lethal form of a human congenital
skin disorder (ichthyosis). Recently, we have established that legumain, a
novel asparaginyl endoprotease, is the major target of cystatin M/E, and we
provided evidence for the mechanism by which deficiency of cystatin M/E
leads to disturbed cornification, impaired barrier function and
dehydration. Absence of cystatin M/E causes unrestricted activity of its
target protease legumain in hair follicles and epidermis, which is the
exact location where cystatin M/E is normally expressed. In accordance, legumain
was found to mediate the activation of TGase-3 zymogen via lysosomal
cysteine proteases in vitro. Disturbance of this
protease-antiprotease balance causes increased enzyme activity of TGase-3
that could explain the observed abnormal cornification. The evidence that I
have provided on the role of the legumain-cystatin M/E dyad in the
formation of the skin barrier could serve as a paradigm for the molecular
pathology that results from disturbance of the protease-antiprotease
balance in human skin. Finally, I have speculated how cystatin M/E
regulates legumain activity in skin, resulting in TGase-3 processing, as
these proteins are not found in the same cellular compartments. The
relevance of these findings could well extend beyond our original findings,
and will potentially lead to a new model for one of the steps in terminal
differentiation. I think that through my work on the role of the
legumain-cystatin M/E dyad I have the edge over my competitors in the
field, and that I have created a niche that may expand to other tissues
than skin. Currently we are
generating double knockout mice (cystatin M/E-/- and legumain-/-).
The double knockout will be interesting for a number of reasons including
the possibility that cystatin M/E has functions not related to its
antiprotease activity. These double knockout mice should also give a
definitive answer to the question if the legumain-cystatin M/E balance is
truly important in the pathway that leads to a correct composition and
formation of the stratum corneum. We are also now rescuing the cystatin M/E
deficient mice (neonatal lethal) by the introduction of a transgene that
drives epidermis-specific cystatin M/E expression on a cystatin M/E null
background. By doing this we could eventually study the consequences of cystatin
M/E deficiency in other
tissues that are affected by unrestricted legumain activity.
International
activities
Conferences:
·
Invited lecture at the Institute
for Molecular Medicine and Cell Research, at the seminar “Birth, thrive and
death of normal and aberrant cells“, 7 April 2005,
Albert-Ludwigs-Universität Freiburg, Germany.
·
Invited lecture for the Deutsche
Forschungsgemeinschaft, at the Symposium “Keratinocytes – Proliferation and
Differentiation in the Epidermis”, 24 – 25 May 2004, Rheinischen
Friedrich-Wilhelms Universität Bonn, Germany.
·
Invited lecture for the
department of Dermatology at the Center for Molecular Medicine, 10 October
2003, University of Cologne, Germany.
·
Lecture in plenary session at
the 4 th meeting of the International Investigative Dermatology,
30 April – 4 May 2003, Miami Beach, Florida, USA.
·
Attendance at the 3 th
International Congress ‘Psoriasis: From Gene to Clinic', 21 – 23 November
2002, London, UK.
·
Attendance and poster
presentation at the 27 th annual meeting of the European Society
for Dermatological Research, 4 – 7 October 1997, Rome, Italy.
·
Attendance and poster
presentation at the 23 th annual meeting of the European Society
for Dermatological Research, 3 – 6 April 1993, Amsterdam, The Netherlands.
Other:
·
April – May 2004, working visit
of two months at the lab of dr I. Haase (Cologne Center for Molecular
Medicine, Germany).
Ad Hoc reviewer for:
·
Oncogene
·
Human
Molecular Genetics
·
Molecular
Biology Reports
Other
academic activities
Grants:
·
Personal Veni-grant (NWO,
919-56-117) on a project entitled “Epidermal proteases: leading the way to
skin barrier formation”.
·
Co-applicant with Prof dr J.
Schalkwijk in a project entitled “Legumain-cystatin
M/E: a novel tissue specific protease-antiprotease system. The role in
human physiology and pathology ” that was granted to us
by the Research Institute of the Radboud University Nijmegen Medical Center
(Ph.D-project 2004).
Teaching experience:
·
Guiding undergraduate students
during their 6-month master degree traineeship (1998 – 2004)
·
Participation in the course
“Medical Biotechnology” (Keuzeblok) of the Medical School of the University
of Nijmegen (1997 – 2000, 2 weeks/year, 8 students), in which I was
responsible for setting up and teaching of the practical part (DNA
diagnostics of skin diseases).
Conferences/meetings:
·
Invited lecture for the
department of Pulmonology of the Leiden University Medical Center, 18 May
2004, Leiden, The Netherlands.
·
Oral presentation at the 5
th scientific annual meeting of the “Nederlandse Vereniging voor
Experimentele Dermatologie”, 29 – 30 January 2004, Lunteren, The
Netherlands.
·
Poster presentation at the
“Science Day” of the UMCN St Radboud, 23 January 2004, Nijmegen, The
Netherlands.
·
Oral presentation at the NCMLS
Forum evening, 15 December 2003, University of Nijmegen, The Netherlands.
·
Oral presentation at the 4
th scientific annual meeting of the “Nederlandse Vereniging voor
Experimentele Dermatologie”, 30 – 31 January 2003, Lunteren, The
Netherlands.
·
Oral presentation at the “ICS
theme discussion”, 28 October 2002, University of Nijmegen, The
Netherlands.
·
Poster presentation at the
annual graduate student meeting of the Institute of Cellular Signalling,
April 2003, Papendal, The Netherlands.
·
Oral presentation at the 3
th scientific annual meeting of the “Nederlandse Vereniging voor
Experimentele Dermatologie”, 31 January – 1 February 2002, Lunteren, The
Netherlands.
·
Oral presentation at the 2
th scientific annual meeting of the “Nederlandse Vereniging voor
Experimentele Dermatologie”, 8 – 9 February 2001, Lunteren, The
Netherlands.
·
Oral
presentation at the 6 th meeting of the “NWO-werkgemeenschap
HUID en werkgroep HUID”, 18 – 19 January 1996, Groesbeek, The Netherlands.
Scholarships and
prizes:
·
Traveling fellowship from the
Journal of Cell Science, July 2004 (travel grant to cover a working visit
to Cologne).
·
Winner poster award at the
“Science Day” of the UMCN St Radboud, 23 January 2004, Nijmegen, The
Netherlands.
·
Albert M. Kligman Award (travel
grant to participate at the 4 th meeting of the International
Investigative Dermatology, 30 April – 4 May 2003, Miami Beach, Florida,
USA).
·
1 th prize for the
best lecture at the 4 th scientific annual meeting of the
“Nederlandse Vereniging voor Experimentele Dermatologie”, 30 – 31 January
2003, Lunteren, The Netherlands.
|
International
(refereed) journals
Vos,J.B., Datson,N.A.,
van Kampen,A.H., Luyf,A.C., Verhoosel,R.M., Zeeuwen,P.L.,
Olthuis,D., Rabe,K.F., Schalkwijk,J., and Hiemstra,P.S. (2005) A Molecular
signature of epithelial host defense: comparative gene expression analysis
of cultured bronchial epithelial cells and keratinocytes. BMC Genomics
(submitted).
Tjabringa,G.S., Vos,J.B.,
Olthuis,D., Ninaber,D.K., Rabe,K.F., Schalkwijk,J., Hiemstra,P.S., and Zeeuwen,P.L.
(2005) Host defense effector molecules in mucosal fluids. FEMS
Immunology and Medical Microbiology (in press).
Franssen, M.E., Zeeuwen,P.L.
, Vierwinden,G., van de Kerkhof,P.C., Schalkwijk,J., and van
Erp,P.E. (2005) Phenotypical and functional differences in germinative
subpopulations derived from normal and psoriatic epidermis. J.Invest.Dermatol. 124:373-383.
Zeeuwen,P.L. (2004) Epidermal differentiation: The role of proteases and their
inhibitors. Eur.J.Cell Biol. 83:761-773.
Zeeuwen,P.L. , Vlijmen-Willems,I.M., Olthuis,D., Johanson,H.T., Hitomi,K.,
Hara-Nishimura,I., Powers,J.C., James,K.E., Op Den Camp,H.J., Lemmens,R.,
and Schalkwijk,J. (2004) Evidence that unrestricted legumain activity is
involved in disturbed epidermal cornification in cystatin M/E deficient
mice. Hum.Mol.Genet . 13:1069-1079.
Zeeuwen,P.L. , Dale,B.A., de Jongh,G.J., Vlijmen-Willems,I.M., Fleckman,P.,
Kimball,J.R., Stephens,K., and Schalkwijk,J. (2003) The human cystatin M/E
gene ( CST6 ): exclusion as a candidate gene for harlequin
ichthyosis. J.Invest.Dermatol. 121: 65-68.
Pol,A., Pfundt,R., Zeeuwen,P.L.
, Molhuizen,H.O., and Schalkwijk,J. (2003) Transcriptional regulation
of the elafin gene in human keratinocytes. J.Invest.Dermatol. ,
120 , 301-307.
Zeeuwen,P.L. , Vlijmen-Willems,I.M., Hendriks,W., Merkx,G.F., and Schalkwijk,J.
(2002) A null mutation in the cystatin M/E gene of ichq mice
causes juvenile lethality and defects in epidermal cornification. Hum.Mol.Genet. 11: 2867-2875.
Zeeuwen,P.L. , Vlijmen-Willems,I.M., Egami,H., and Schalkwijk,J.
(2002) Cystatin M/E expression in inflammatory and neoplastic skin
disorders. Br.J.Dermatol. 147: 87-94.
van
Ruissen,F., Jansen,B.J., de Jongh,G.J., Zeeuwen,P.L. , and
Schalkwijk,J. (2002) A partial transcriptome of human epidermis. Genomics
79 :671-678.
Zeeuwen,P.L. , Vlijmen-Willems,I.M., Jansen,B.J., Sotiropoulou,G.,
Curfs,J.H., Meis,J.F., Janssen,J.J., van Ruissen,F., and Schalkwijk,J.
(2001) Cystatin M/E expression is restricted to differentiated epidermal
keratinocytes and sweat glands: a new skin-specific proteinase inhibitor
that is a target for cross-linking by transglutaminase. J.Invest.Dermatol.
116: 693-701.
Jansen,B.J.,
van Ruissen,F., de Jongh,G., Zeeuwen,P.L. , and
Schalkwijk,J. (2001) Serial analysis of gene expression in differentiated
cultures of human epidermal keratinocytes. J.Invest.Dermatol. 116:
12-22.
Castelijns,F.A.,
Ezendam,J., Latijnhouwers,M.A., Vlijmen-Willems,I.M., Zeeuwen,P.L. ,
Gerritsen,M.J., van de Kerkhof,P.C., and van Erp,P.E. (1998) Epidermal cell
kinetics by combining in situ hybridization and immunohistochemistry. Histochem.J.
30 :869-877.
Wingens,M.,
van Bergen,B.H., Hiemstra,P.S., Meis,J.F., Vlijmen-Willems,I.M., Zeeuwen,P.L.
, Mulder,J., Kramps,H.A., van Ruissen,F., and Schalkwijk,J. (1998)
Induction of SLPI (ALP/HUSI-I) in epidermal keratinocytes. J.Invest.Dermatol.
111 :996-1002.
Kuijpers,A.L.,
Pfundt,R., Zeeuwen,P.L. , Molhuizen,H.O., Mariman,E.C.,
van de Kerkhof,P.C., and Schalkwijk,J. (1998) SKALP/elafin gene
polymorphisms are not associated with pustular forms of psoriasis. Clin.Genet.
, 54 , 96-101.
Zeeuwen,P.L. , Hendriks,W., de Jong,W.W., and Schalkwijk,J. (1997)
Identification and sequence analysis of two new members of the SKALP/elafin
and SPAI-2 gene family. Biochemical properties of the transglutaminase
substrate motif and suggestions for a new nomenclature. J.Biol.Chem. 272:
20471-20478.
Kuijpers,A.L.,
Bergers,M., Siegenthaler,G., Zeeuwen,P.L. , van de
Kerkhof,P.C., and Schalkwijk,J. (1997) Skin-derived antileukoproteinase
(SKALP) and epidermal fatty acid-binding protein (E-FABP): two novel
markers of the psoriatic phenotype that respond differentially to topical
steroid. Acta Derm.Venereol. 77 :14-19.
Kuijpers,A.L.,
Zeeuwen,P.L. , de Jongh,G.J., van de Kerkhof,P.C.,
Alkemade,H.A., and Schalkwijk,J. (1996) Skin-derived antileukoproteinase
(SKALP) is decreased in pustular forms of psoriasis. A clue to the
pathogenesis of pustule formation? Arch.Dermatol.Res. 288:
641-647.
Pfundt,R.,
van Ruissen,F., Vlijmen-Willems,I.M., Alkemade,H.A., Zeeuwen,P.L. ,
Jap,P.H., Dijkman,H., Fransen,J., Croes,H., van Erp,P.E., and Schalkwijk,J.
(1996) Constitutive and inducible expression of SKALP/elafin provides
anti-elastase defense in human epithelia. J.Clin.Invest . 98:
1389-1399.
van
Ruissen,F., de Jongh,G.J., Zeeuwen,P.L. , van Erp,P.E.,
Madsen,P., and Schalkwijk,J. (1996) Induction of normal and psoriatic
phenotypes in submerged keratinocyte cultures. J.Cell Physiol. 168:
442-452.
Hendriks,W.,
Schepens,J., Brugman,C., Zeeuwen,P.L. , and Wieringa,B.
(1995) A novel receptor-type protein tyrosine phosphatase with a single
catalytic domain is specifically expressed in mouse brain. Biochem.J. 305:
499-504.
Hendriks,W.,
Schepens,J., Bachner,D., Rijss,J., Zeeuwen,P.L. ,
Zechner,U., Hameister,H., and Wieringa,B. (1995) Molecular cloning of a
mouse epithelial protein-tyrosine phosphatase with similarities to
submembranous proteins. J.Cell Biochem. 59: 418-430.
Kremer,H.,
Zeeuwen,P.L. , McLean,W.H., Mariman,E.C., Lane,E.B., van
de Kerkhof,P.C., Ropers,H.H., and Steijlen,P.M. (1994) Ichthyosis bullosa of
Siemens is caused by mutations in the keratin 2e gene. J.Invest.Dermatol.
103: 286-289.
Alkemade,J.A.,
Molhuizen,H.O., Ponec,M., Kempenaar,J.A., Zeeuwen,P.L. ,
de Jongh,G.J., Vlijmen-Willems,I.M., van Erp,P.E., van de Kerkhof,P.C., and
Schalkwijk,J. (1994) SKALP/elafin is an inducible proteinase inhibitor in
human epidermal keratinocytes. J.Cell Sci. 107: 2335-2342.
Molhuizen,H.O.,
Zeeuwen,P.L. , Olde,W.D., Geurts,v.K., and Schalkwijk,J.
(1994) Assignment of the human gene encoding the epidermal serine
proteinase inhibitor SKALP (PI3) to chromosome region 20q12-q13. Cytogenet.Cell
Genet. 66: 129-131.
Molhuizen,H.O.,
Alkemade,H.A., Zeeuwen,P.L. , de Jongh,G.J., Wieringa,B.,
and Schalkwijk,J. (1993) SKALP/elafin: an elastase inhibitor from cultured
human keratinocytes. Purification, cDNA sequence, and evidence for
transglutaminase cross-linking. J.Biol.Chem. 268: 12028-12032.
Schepens,J.,
Zeeuwen,P.L. , Wieringa,B., and Hendriks,W. (1992)
Identification and typing of members of the protein-tyrosine phosphatase
gene family expressed in mouse brain. Mol.Biol.Rep. 16: 241-248.
National
(refereed) journals
Schalkwijk,J.,
Kuijpers,A.L., Alkemade,J.A., Molhuizen,H.O., Zeeuwen,P.L. ,
van Vlijmen,I.M., de Jongh, G.J., en van de Kerkhof,P.C. (1994) De rol van
skin-derived antileukoprotease (SKALP) in de pathogenese van pustuleuze
psoriasis. Nederlands Tijdschrift voor Dermatologie &
Venereologie 4: 57 – 58.
Other
Published abstracts
(first and last author only):
Schalkwijk,J. van Vlijmen-Willems,I.M., Olthuis,D., and Zeeuwen,P.L.
(2004) The cystatin M/E – legumain balance: a regulator of
epidermal cornification. J.Invest.Dermatol. 123:A1 (abstract for
the 34 th annual meeting of the European Society for
Dermatological Research, 9 – 11 September 2004, Vienna, Austria).
Zeeuwen,P.L. , van Vlijmen-Willems,I.M., Hendriks,W.,
Merkx,G.F., and Schalkwijk,J. (2003) Cystatin M/E deficiency in mice causes
faulty cornification and impaired barrier function. J.Invest.Dermatol. 121:218
(abstract for the 4 th meeting of the International
Investigative Dermatology, 30 April – 4 may 2003, Miami Beach, Florida,
USA).
Zeeuwen,P.L. , Hendriks,W., de Jong,W.W., and Schalkwijk,J.
(1997) Identification and sequence analysis of two new members of the
SKALP/elafin protein family; biochemical properties of the transglutaminase
substrate motif. J.Invest.Dermatol. 109:428 (abstract for the 27
th annual meeting of the European Society for Dermatological
Research, 4 – 7 October 1997, Rome, Italy).
Zeeuwen,P.L. , Molhuizen,H.O.,
Geurts van Kessel,A.H., and Schalkwijk,J. (1993) Chromosomal localization
of the human SKALP/Elafin gene. J.Invest.Dermatol. 100:454. (abstract for
the 23 th annual meeting of the European Society for
Dermatological Research, 3 – 6 April 1993, Amsterdam, The Netherlands).
|
