○会期：平成 28 年２月 28 日（日）10:00〜18:00○会場：福岡リーセントホテル

レインボーホール：下図参照

住所：福岡市東区箱崎 2-52-1
電話：092-641-7741○主催：九州大学大学院
日本学術振興会

歯学研究院
頭脳循環を加速する戦略的国際研究ネットワーク推進プログラム

口腔から健康長寿を支えるプロジェクト推進に向けた研究拠点構築プログラム

連絡先：

西村

英紀

九州大学大学院 歯学研究院・歯学府・歯学部 歯周病学分野〒812-8582

福岡市東区馬出 3-1-1

電話：092-642-6358
ファックス：092-642-6360
E-mail：fusanori@dent.kyushu-u.ac.jp

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PROGRAM頭脳循環第二回シンポジウム
2nd Symposium:
Program for2016年2月28日■

10：00〜10：10

Opening remarks■

10：10〜11：10 Special Lecture１

Masato Hirata

Chair :

Hidefumi Maeda

Dr. Junichi Iwata：
Role of cholesterol in craniofacial development and diseases
The University of Texas Health Science Center at Houston(UTHealth) School of Dentistry Department of Diagnostic
and Biomedical Sciences■

11：20〜12：20 Special Lecture２

Chair :

Seiji Nakamura

Dr. Ophir Klein：
Sinking our teeth into development, evolution and regeneration
The Larry L. Hillblom Distinguished Professor in Craniofacial
Anomalies, Professor of Orofacial Sciences and Pediatrics,
and Chair of the Divisions of Craniofacial Anomalies andOrthodontics at the University of California, San Francisco(UCSF). Medical Director of the UCSF Craniofacial Center andDirector, UCSF Program in Craniofacial Biology.

■

13：30〜14：30 Plenary Lecture

Chair :

Masato Hirata

Dr. Yuji Mishina：
Enhanced BMP signaling in neural crest cells leads to
Craniosynostosis
University of Michigan, School of Dentistry, Department of
Biologic and Materials Sciences■

14：50〜15：30 Special Lecture３

Chair :

Naohisa Wada

Dr. Keigo Yoshizaki：
Comprehensive analysis for the functions of genes during
epithelial-mesenchymal interactions in tissue morphogenesis
Section of Orthodontics and Dentofacial Orthopedics, Facultyof Dental Science, Kyushu University, Japan■

15：30〜16：10 Reports after circulation

Chair :

Fusaniri Nishimura

Dr. Takashi Maehara：
Lesional CD4+ IFN-γ++ cytotoxic T lymphocytes in IgG4related dacryoadenitis and sialoadenitis
Section of Oral and Maxillofacial Oncology
Division of Maxillofacial Diagnostic and Surgical Sciences
Faculty of Dental Science,Kyushu University
Ragon Institute of MGH, MIT and Harvard
Harvard Medical School■

16：30〜17：30 Young researchers for next circulation

Chair :

Takayoshi Yamaza

Takao Fukuda：M olecular basis of amelogenin-induced
periodontal tissue regeneration
Sachiko Furukawa：I L-33 secreted by M2 macrophages
promotes the pathogenesis of IgG4related disease
Shusuke Iwata：S timulus repetition enhances responses of
the chorda tympani nerve to sweeteners
via endocannabinoid system
Hideki Sugii：A ctivin A reversely works between humanpre-osteoblastic cells and periodontal ligament
cells on their osteoblastic differentiation

■

17：30

Photo-taking

Closing remarks

Prof. Fusanori Nishimura【MEMO】

Plenary LectureEnhanced BMP signaling in neural crest cells leads to
Craniosynostosis
Yuji Mishina
University of Michigan, School of Dentistry, Department of Biologic and Materials
Sciences

Craniosynostosis is caused by premature cranial suture fusion resulting in abnormal facial features.
Approximately 20% of human craniosynostoses are thought to result from gene mutations altering
growth factor signaling; however, the molecular mechanisms by which these mutations cause
craniosynostosis are incompletely characterized, and the causative genes for diverse types of
syndromic craniosynostosis have yet to be identified.
Bone morphogenetic protein (BMP) signaling plays broad roles in developmental patterning, including
skull morphogenesis. We have previously reported enhanced BMP signaling through the BMP
type IA receptor (BMPR1A) in cranial neural crest cells causes craniosynostosis. We have noticed
approximately 55% of the mutants show neonatal lethality characterized by distended gastrointestinal
tract. TUNEL assay demonstrated that an increase of apoptosis in condensing mesenchymal tissues
and chondrocytes in the nasal region of the mutants. Furthermore, immunohistochemical staining
for p53 revealed an increase of p53 in mutant nasal cartilage. To rescue the phenotype, pifithrin-a, achemical inhibitor of p53, was injected to the pregnant female. The treatment resulted in preventionof apoptosis in mesenchymal cells and neonatal lethality. Furthermore, the survived mutant pups
showed suture patency strongly suggesting that enhanced BMP signaling induces p53-mediatedapoptosis leading to depletion of stem cell/progenitor population within suture mesenchyme resulting
in premature suture fusion.
To dissect the molecular mechanism of how enhanced BMP signaling leads to p53-mediatedapoptosis in our mutants, we examined the expression level of p53 and its regulatory network
for apoptosis. We found that expressions of Bax and Caspase3 were increased in the mutants;
however, p53 expression level was not changed. It is previously reported that Mdm2 ubiquitinates
p53 to prompt its degradation and SMAD1 prevents from the MDM2-mediated p53 ubiquitination.
We compared ubiquitinated p53 level between controls and the mutants by western blotting andimmunoprecipitation. We found that ubiquitination of p53 was decreased in the mutants. These
results indicate that BMP signaling induces p53-mediated apoptosis by preventing p53 ubiquitinationin early stage of chondrogenesis and thus appropriate level of BMP signaling is required for proper
craniofacial morphogenesis.

1

CURRICULUM VITAE

Bibliography
Education and Advanced Training:
1983-1986

Ph.D. University of Tokyo, Tokyo, Japan (Molecular Biology)

1981-1983

M.S. University of Tokyo, Tokyo, Japan (Molecular Biology)

1977-1981

B.S. University of Tokyo, Tokyo, Japan (Biology)

Academic Appointment:
1992-1998

Postdoctoral Fellow with Dr. Richard Behringer,
The University of Texas, M.D. Anderson Cancer Center, Texas

1998-2008	Group Head, Molecular Developmental Biology Group, Laboratory of
Reproductive and Developmental Toxicology, National Institute ofEnvironmental Health Sciences, National Institutes of Health, Research Triangle
Park, NC
2008-2014	Associate Professor, University of Michigan, School of Dentistry, Department of
Biologic and Materials Sciences, Ann Arbor, MI
2014-Present	Professor, University of Michigan, School of Dentistry, Department of Biologic
and Materials Sciences, Ann Arbor, MI

2

【MEMO】

3

Special Lecture 1

Role of cholesterol in craniofacial
development and diseases
Junichi Iwata
The University of Texas Health Science Center at Houston (UTHealth) School of
Dentistry Department of Diagnostic and Biomedical Sciences

Abstract
Cholesterol is 1) important as a structural lipid in cellular membranes to maintain membrane
structural integrity and fluidity; 2) abundant in specialized microdomains called lipid rafts, which
are organizing centers for the assembly of signaling molecules; 3) an obligatory biogenic precursor
for steroid hormones, oxysterols, vitamin D, and bile acids. In addition, recent studies indicate thatcholesterol can control a cell signaling activity. Therefore, the proper amount of cellular cholesterol
is crucial for a wide variety of biological processes, including bone formation and salivary glands
homeostasis. This talk will cover two topics related to cholesterol metabolic disorders; craniofacialbone formation and saliva production.
It has been largely unknown how disturbances in cholesterol production lead to bone abnormalities.
In this study, we investigated the effects of cholesterol metabolism on craniofacial bone formation
using mouse genetic approaches. So far, we discovered that mice with cholesterol metabolic
aberrations altered: 1) calvaria bone formation; 2) osteoblast differentiation; 3) the formation of primarycilia (a microtubule-based organelle that functions in sensory and signaling pathways); 5) cell signalingcascades in osteoblasts.
It has long been appreciated that individuals with metabolic syndromes and abnormal diets (highcholesterol diets, poor nutrition, etc.) are at risk of chronic dry mouth (a.k.a. xerostomia). Hypofunctionof the salivary glands results in severe periodontitis, dental caries, and oral ulcers, as well as difficultyin swallowing, tasting, chewing and speaking and substantially decreased quality of life. Despite suchimportant physiological functions, we know very little about the regulatory mechanism of saliva
exocytosis because of the lack of animal models for xerostomia and of tools for tracking exocytosis. In
this study, we investigated the association between high cholesterol and xerostomia. The mice witha high cholesterol level displayed xerostomia with the accumulation of secretory vesicles in acinar
cells of the salivary glands. We found that autophagic machinery (non-autophagic roles of autophagyrelated molecules) was compromised in the salivary glands.
Thus, the role of cholesterol is diverse in craniofacial development and diseases. Taking advantage of
our animal models, we are trying to determine new roles of cholesterol and its related molecules. Ourfindings will provide new insights into the role of cholesterol metabolism and lead to innovations in
the prevention, diagnosis, and treatment of cholesterol-related disorders.
4

CURRICULUM VITAE

Biography
Dr. Iwata graduated from Kyushu University School of Dentistry in 2000, and received his Ph.D. at
Kyushu University Graduate School of Dentistry in 2004. He carried out his Ph.D. training under
Dr. Kenji Yamamoto, a well-established investigator in the fields of protease and molecular biology.
He analyzed the properties of cathepsin E using knockout and transgenic mice as well as opticaltechniques, and found that cathepsin E could induce anti-angiogenic factors. He then worked as an
Assistant Professor at Juntendo University Medical School through 2004 to 2007. He worked witha number of collaborators in the field of autophagy which is a major protein degradation system.
They investigated a variety of genetic mouse models from 2004 to 2007. Their characterization ofdevelopmental defects in autophagy knockout mice inspired his interest in developmental regulation.
To expand this interest, he joined the laboratory of Dr. Yang Chai, a well-established investigatorin the fields of transforming growth factor beta (TGF β ) signaling and craniofacial development,
at University of Southern California School of Dentistry from 2007 to 2013. They discovered ectopic
noncanonical TGF β signaling in the absence of TGF β receptor type II and identified downstream
target molecules of this altered TGF β signaling. In his laboratory at The University of Texas
Health Center at Houston (UTHealth) School of Dentistry from August 2013, his group focuses onunderstanding molecular and cellular mechanisms that cause developmental defects, and work closelywith researchers from the Medical as well as the Dental School. As a DDS-PhD scientist, Dr. Iwata isvery pleased to contribute to the craniofacial and dental research community. He is named on morethan 30 peer-reviewed publications.
Selected Publications
1．Iwata J, Ezaki J, Komatsu M, Yokota S, Ueno T, Tanida I, Chiba T, Tanaka K, and Kominami E. (2006)
Excess peroxisomes are degraded by autophagic machinery in mammals. J. Biol. Chem., 281, 403541.
2. Sou Y*, Waguri S*, Iwata J*, Ueno T, Fujimura T, Hara T, Sawada N, Yamada A, Mizushima
N, Uchiyama Y, Kominami E, Tanaka K, Komatsu M. (2008). The Atg8 conjugation system is
indispensable for proper development of autophagic isolation membranes in mice. Mol. Biol. Cell,
19(11):4762-75. (*These authors contributed equally to this work.)
3. Iwata J, Hosokawa R, Sanchez-Lara PA, Urata M, Slavkin H, and Chai Y. (2010) Transforming
growth factor-beta regulates basal transcriptional regulatory machinery to control cell proliferationand differentiation in cranial neural crest–derived osteoprogenitor cells. J. Biol. Chem., 285(7):4975-82.
4. Iwata J, Tung L, Urata M, Hacia JG, Suzuki A, Ramenzoni L, Chaudhry O, Parada C, SanchezLara PA, and Chai Y. (2012) FGF9–PITX2 pathway mediates TGF β signaling to regulate cellproliferation in palatal mesenchyme during mouse palatogenesis. J. Biol. Chem., 287(4):2353-63.
5. Iwata J, Hacia JG, Suzuki A, Sanchez-Lara PA, Urata M, and Chai Y. (2012) Modulation of noncanonical TGF- β signaling prevents cleft palate in Tgfbr2 mutant mice. J. Clin. Invest., 122(3):87385.
6. Pelikan RC*, Iwata J*, Suzuki A, Chai Y, and Hacia JG. (2013) Identification of candidate downstream
5

targets of TGF β signaling during palate development by genome-wide transcript profiling. J. Cell
Biochem., 114:796-807. (*These authors contributed equally to this work.)
7. Iwata J, Suzuki A, Pelikan RC, Ho TV, Sanchez-Lara PA, Urata M, Dixon MJ, and Chai Y. (2013)
Smad4–Irf6 genetic interaction and TGF β –mediated IRF6 signaling cascade are crucial for palatalfusion in mice. Development, 140(6):1220-30.
8. Song ZC*, Liu C*, Iwata J*, Gu SP, Suzuki A, Sun C, He W, Shu R, Li L, Chai Y, and Chen YP. (2013)
Mice with Tak1-deficiency in neural crest lineage exhibit cleft palate associated with abnormal
tongue development. J. Biol. Chem., 288(15):10440-50. (*These authors contributed equally to thiswork.)
9. Iwata J, Suzuki A, Pelikan RC, Ho TV, and Chai Y. (2013) Cranial neural crest cells regulate tongue
muscle formation via TGF β –mediated BMP and FGF signaling. J. Biol. Chem. 288:29760-70.
10. Iwata J, Suzuki A, Pelikan RC, Ho TV, Sanchez-Lara PA, and Chai Y. (2014) Modulation of lipid
metabolic defects rescues cleft palate in Tgfbr2 mutant mice. Hum. Mol. Genetics, 23(1):182-93.
11. Iwata J, Suzuki A, Yokota T, Ho TV, Pelikan RC, Urata M, Sanchez-Lara P, and Chai Y. (2014) TGF
β regulates epithelial–mesenchymal interactions through WNT signaling activity to control muscle
development in the soft palate. Development, 14 (4):909-17.
12. Ho TV*, Iwata J*, Ho HA, Grimes WC, Park S, Sanchez-Lara PA, and Chai Y. (2015) Integration ofcomprehensive 3D microCT and signaling analysis reveals differential regulatory mechanisms of
craniofacial bone development. Dev. Biol. 400(2):180-90. (*These authors contributed equally to thiswork.)
13. Suzuki A, Pelikan RC, and Iwata J. (2015) WNT/ β -catenin signaling regulates multiple steps ofmyogenesis by regulating step-specific targets. Mol. Cell. Biol. 35(10):1763-76.

6

Special Lecture 2

Sinking our teeth into development, evolution andregeneration
Ophir David Klein, MD, PhD
The Larry L. Hillblom Distinguished Professor in Craniofacial Anomalies, Professorof Orofacial Sciences and Pediatrics, and Chair of the Divisions of Craniofacial
Anomalies and Orthodontics at the University of California, San Francisco (UCSF).
Medical Director of the UCSF Craniofacial Center and Director, UCSF Program in Craniofacial Biology.

An important challenge facing medicine today is the development of strategies for organ regeneration
and repair. The tooth provides a model that allows us to integrate investigations into organogenesis,
stem cells and evolution with studies into therapeutic approaches. I will present data from our recent
work focusing on development and renewal of the rodent incisor, which unlike human teeth grows
throughout the life of the animal. The growth of this organ, like that of the skin, gastrointestinal tract,
hematopoietic system, and others, is dependent on the continuous generation of progeny from stem
cells that have the capacity to self-renew as well as to give rise to the required differentiated cell
types. In addition, I will discuss new mechanisms of tooth formation as well as how evolutionarily
informative transitions can be experimentally reproduced to provide development-based expectations
for evolutionary studies. I will conclude with some connections between the evolutionary studies andtherapies for human disease.

7

CURRICULUM VITAE

Name:

Ophir David Klein, MD, PhD

Position:

Professor
Orofacial Sciences & Pediatrics Schools of Dentistry & Medicine
Larry L. Hillblom Distinguished Professor in Craniofacial Anomalies Chair, Divisionof Craniofacial Anomalies
Chair, Division of Orthodontics
Director, Program in Craniofacial Biology

EDUCATION
1989 - 1993

University of California,
Berkeley

B.A.

Cum Laude,
Spanish(Research Advisor: Daniel
Koshland, Ph.D.)

1993 - 2000

Yale University Schoolof Medicine

M.D.

1995 - 1999

Yale University Schoolof Medicine

Ph.D.

Genetics(Thesis Advisor: Daniel
DiMaio, M.D., Ph.D.)

2000 - 2003

Yale University Schoolof Medicine

Resident

Pediatrics

2003 - 2007

University of California,
San Francisco

Resident

MedicalGenetics(Research Advisor: Gail
Martin, Ph.D.)

HONORS AND AWARDS
1989

Chancellor's Scholarship

University of California, Berkeley

1993

Medical Scientist Training Program

National Institutes of Health

1999

Dean's Distinguished Thesis
Commendation

Yale University Graduate School

2004

Fellow

American Academy of Pediatrics

2004

Fellow

Pediatric Scientist Development Program

2005

David W. Smith Research Award

Western Society for Pediatric Research

2005

Fellow Research Award

Society for Pediatric Research

2005

Young Investigator Research Grant

American Academy of Pediatrics Section
on Genetics and Birth Defects

2008

Young Investigator

United States Bone and Joint Decade

2008

Charles E. Culpeper Scholar

Goldman Philanthropic Partnerships

2008

New Faculty Award

California Institute of Regenerative
Medicine8

2008Elected Member

Society for Pediatric Research

2009

Basil O'Connor Award

March of Dimes

2009Elected Member

Western Society for Pediatric Research

2009

Harold M. Frost Young Investigator
Award

American Society for Bone and Mineral
Research2010

New Innovator Award

National Institutes of Health2013

Physician Scientist Award

California Institute of Regenerative
Medicine

2013

Elected Member

American Society for Clinical Investigation

2013

Outstanding Faculty MentorshipAward

UCSF Graduate Students' Association2014

Larry L. Hillblom Distinguished Professor
in Craniofacial Anomalies

2014

Fellow

American Association for the
Advancement of Science (AAAS)2015

E. Mead Johnson Award

Society for Pediatric Research

KEYWORDS/AREAS OF INTEREST
Developmental biology, stem cell biology, clinical genetics, receptor tyrosine kinase signaling,
craniofacial, tooth, intestinal stem cells

SERVICE TO PROFESSIONAL PUBLICATIONS
2007 - present A
 d hoc referree for American Journal of Medical Genetics, Archives of Oral Biology,
Birth Defects Research Part A, BMC Developmental Biology, BMC Musculoskeletal
Disorders, Cancer Research, Cell Proliferation, Cell Reports, Cell Stem Cell, Cells
Tissues Organs, Developmental Biology, Developmental Cell, Developmental
Dynamics, Development, e Cells and Materials, European Journal of Oral Sciences,
Frontiers in Physiology, Human Molecular Genetics, International Journal of
Developmental Biology, Journal of Cellular and Molecular Medicine, Journal of Dental
Research, Journal of Experimental Medicine, Journal of Experimental Zoology B,
Mammalian Genome, Mechanisms of Development, Odontology, Oral Diseases, Nature
Biotechnology, Nature, Nature Cell Biology, Nature Communications, Nucleic
Acids Research, Odontology, Oral Diseases, PLoSONE, PNAS, Stem Cells Development,
Stem Cell Research.
 ditorial Board, Frontiers in Craniofacial Biology 2011 - present Section Editor,
2010 - present E
Dentistry 3000
2012 - present E ditorial Board, Molecular Genetics and Genomic Medicine 2013 - 2013
Editor, PNAS
2013 - present E ditorial Board, International Journal of Oral Science9

G u e s t

2014 - 2015

Guest Editor, Journal of Dental Research, Special Issue on Craniofacial Stem Cells

PEER REVIEWED PUBLICATIONS
1.

Lee ME, Dyer DH, Klein OD, Bolduc JM, Stoddard BL, Koshland DE. Mutational analysis of thecatalytic residues lysine 230 and tyrosine 160 in the NADP(+)-dependent isocitrate dehydrogenasefrom Escherichia coli. Biochemistry. 1995 Jan 10; 34(1):378-84. PMID: 7819221
2.

Klein O, Polack GW, Surti T, Kegler-Ebo D, Smith SO, DiMaio D. Role of glutamine 17 of thebovine papillomavirus E5 protein in platelet-derived growth factor beta receptor activation and celltransformation. J Virol. 1998 Nov; 72(11):8921-32. PMID: 9765437. PMCID: PMC110309
3.

Surti T, Klein O, Aschheim K, DiMaio D, Smith SO. Structural models of the bovine papillomavirus

E5 protein. Proteins. 1998 Dec 1; 33(4):601-12. PMID: 9849943
4.

Klein O, Kegler-Ebo D, Su J, Smith S, DiMaio D. The bovine papillomavirus E5 protein requiresa juxtamembrane negative charge for activation of the platelet-derived growth factor beta
receptor and transformation of C127 cells. J Virol. 1999 Apr; 73(4):3264-72. PMID: 10074180. PMCID:
PMC104090
5.

Raga F, Bonilla-Musoles F, Casañ EM, Klein O, Bonilla F. Assessment of endometrial volume

by three-dimensional ultrasound prior to embryo transfer: clues to endometrial receptivity. Hum
Reprod. 1999 Nov; 14(11):2851-4. PMID: 10548635
6.

Klein OD, Cotter PD, Albertson DG, Pinkel D, Tidyman WE, Moore MW, Rauen KA. Prader-

Willi syndrome resulting from an unbalanced translocation: characterization by array comparative
genomic hybridization. Clin Genet. 2004 Jun; 65(6):477-82. PMID: 15151506
7.

Klein OD, Backstrand K, Cotter PD, Marco E, Sherr E, Slavotinek A. Case report: Y;6translocation with deletion of 6p. Clin Dysmorphol. 2005 Apr; 14(2):93-6. PMID: 15770132
8.

Ahn CP, Lachman RS, Cox VA, Blumberg B, Klein OD. Brachydactylic multiple delta phalanges

plus syndrome. Am J Med Genet A. 2005 Sep 15; 138(1):41-4. PMID: 16092122
9.

Klein OD, Cotter PD, Schmidt AM, Bick DP, Tidyman WE, Albertson DG, Pinkel D, Rauen KA.

Interstitial deletion of chromosome 12q: genotype-phenotype correlation of two patients utilizing
array comparative genomic hybridization. Am J Med Genet A. 2005 Nov 1; 138(4):349- 54. PMID:
16200635
10. Klein OD, Minowada G, Peterkova R, Kangas A, Yu BD, Lesot H, Peterka M, Jernvall J, Martin

GR. Sprouty genes control diastema tooth development via bidirectional antagonism of epithelialmesenchymal FGF signaling. Dev Cell. 2006 Aug; 11(2):181-90. PMID: 16890158. PMCID: PMC2847684
11. Klein OD, Cotter PD, Moore MW, Zanko A, Gilats M, Epstein CJ, Conte F, Rauen KA. Interstitial10

deletions of chromosome 6q: genotype-phenotype correlation utilizing array CGH. Clin Genet. 2007
Mar; 71(3):260-6. PMID: 17309649
12.
13. Klein OD, Lyons DB, Balooch G, Marshall GW, Basson MA, Peterka M, Boran T, Peterkova R,

Martin GR. An FGF signaling loop sustains the generation of differentiated progeny from stem cells
in mouse incisors. Development. 2008 Jan; 135(2):377-85. PMID: 18077585. PMCID: PMC2555979
14. Metzger RJ, Klein OD, Martin GR, Krasnow MA. The branching programme of mouse lungdevelopment. Nature. 2008 Jun 5; 453(7196):745-50. PMID: 18463632. PMCID: PMC2892995
15. Klein OD*, Kostiner DR, Weisiger K, Moffatt E, Lindeman N, Goodman S, Tuchman M, Packman

S. Acute fatal presentation of ornithine transcarbamylase deficiency in a previously healthy male.
Hepatol Int. 2008 Sep; 2(3):390-4. PMID: 19669271. PMCID: PMC2716892
16. Kalfa N, Liu B, Klein O, Audran F, Wang MH, Mei C, Sultan C, Baskin LS. Mutations of CXorf6

are associated with a range of severities of hypospadias. Eur J Endocrinol. 2008 Oct; 159(4):453-8.
PMID: 18635673
17. Kalfa N, Liu B, Klein O, Wang MH, Cao M, Baskin LS. Genomic variants of ATF3 in patients withhypospadias. J Urol. 2008 Nov; 180(5):2183-8; discussion 2188. PMID: 18804813
18. Catón J, Luder HU, Zoupa M, Bradman M, Bluteau G, Tucker AS, Klein O, Mitsiadis TA. Enamel-

free teeth: Tbx1 deletion affects amelogenesis in rodent incisors. Dev Biol. 2009 Apr 15; 328(2):493-505.
PMID: 19233155. PMCID: PMC2880856
19. Peterkova R*, Churava S, Lesot H, Rothova M, Prochazka J, Peterka M, Klein OD*. Revitalizationof a diastemal tooth primordium in Spry2 null mice results from increased proliferation anddecreased apoptosis. J Exp Zool B Mol Dev Evol. 2009 Jun 15; 312B(4):292- 308. PMID: 19127536.
PMCID: PMC2880865
20.

Boran T, Peterkova R, Lesot H, Lyons DB, Peterka M, Klein OD. Temporal analysis of ectopic

enamel production in incisors from sprouty mutant mice. J Exp Zool B Mol Dev Evol. 2009 Jul 15;
312B(5):473-85. PMID: 19101957. PMCID: PMC2837846
21. Charles C, Lazzari V, Tafforeau P, Schimmang T, Tekin M, Klein O*, Viriot L.* Modulation of Fgf3

dosage in mouse and men mirrors evolution of mammalian dentition. Proc Natl Acad Sci U S A. 2009
Dec 29; 106(52):22364-8. PMID: 20018768. PMCID: PMC2799759
22. Kettunen P, Kivimäe S, Keshari P, Klein OD, Cheyette BN, Luukko K. Dact1-3 mRNAs exhibit

distinct expression domains during tooth development. Gene Expr Patterns. 2010 Feb- Mar; 10(23):140-3. PMID: 20170752. PMCID: PMC2849867
23.

B§Prochazka J§, Pantalacci S, Churava S, Rothova M, Lambert A, Lesot H, Klein O, Peterka M,

Laudet V, Peterkova R. Patterning by heritage in mouse molar row development. Proc Natl Acad Sci
11

U S A. 2010 Aug 31; 107(35):15497-502. PMID: 20709958. PMCID: PMC2932592
24.

Ahn Y, Sanderson BW, Klein OD, Krumlauf R. Inhibition of Wnt signaling by Wise (Sostdc1)

and negative feedback from Shh controls tooth number and patterning. Development. 2010 Oct;137(19):3221-31. PMID: 20724449
25.

Jelin A, Perry H, Hogue J, Oberoi S, Cotter PD, Klein OD. Clefting in trisomy 9p patients:

genotype-phenotype correlation using microarray comparative genomic hybridization. J Craniofac
Surg. 2010 Sep; 21(5):1376-9. PMID: 20856024
26. Parsa S, Kuremoto K, Seidel K, Tabatabai R, Mackenzie B, Yamaza T, Akiyama K, Branch J, Koh

CJ, Al Alam D, Klein OD*, Bellusci S*. Signaling by FGFR2b controls the regenerative capacity ofadult mouse incisors. Development. 2010 Nov; 137(22):3743-52. PMID: 20978072. PMCID: PMC3049274
27.

Seidel K, Ahn CP, Lyons D, Nee A, Ting K, Brownell I, Cao T, Carano RA, Curran T, Schober

M, Fuchs E, Joyner A, Martin GR, de Sauvage FJ*, Klein OD*. Hedgehog signaling regulates thegeneration of ameloblast progenitors in the continuously growing mouse incisor. Development. 2010
Nov; 137(22):3753-61. PMID: 20978073. PMCID: PMC3049275
28. Milunsky JM, Maher TM, Zhao G, Wang Z, Mulliken JB, Chitayat D, Clemens M, Stalker HJ,

Bauer M, Burch M, Chénier S, Cunningham ML, Drack AV, Janssens S, Karlea A, Klatt R, Kini U,
Klein O, Lachmeijer AM, Megarbane A, Mendelsohn NJ, Meschino WS, Mortier GR, Parkash S, Ray
CR, Roberts A, Roberts A, Reardon W, Schnur RE, Smith R, Splitt M, Tezcan K, Whiteford ML,
Wong DA, Zori R, Lin AE. Genotype-phenotype analysis of the branchio-oculo-facial syndrome. Am J
Med Genet A. 2011 Jan; 155A(1):22-32. PMID: 21204207
29.

Jheon AH, Mostowfi P, Snead ML, Ihrie RA, Sone E, Pramparo T, Attardi LD, Klein OD. PERP

regulates enamel formation via effects on cell-cell adhesion and gene expression. J Cell Sci. 2011 Mar
1; 124(Pt 5):745-54. PMID: 21285247. PMCID: PMC3039019
30. Hovorakova M, Prochazka J, Lesot H, Smrckova L, Churava S, Boran T, Kozmik Z, Klein O,

Peterkova R, Peterka M. Shh expression in a rudimentary tooth offers new insights into developmentof the mouse incisor. J Exp Zool B Mol Dev Evol. 2011 Jul 15; 316(5):347-58. PMID: 21455944
31. Hochheiser H, Aronow BJ, Artinger K, Beaty TH, Brinkley JF, Chai Y, Clouthier D, Cunningham

ML, Dixon M, Donahue LR, Fraser SE, Hallgrimsson B, Iwata J, Klein O, Marazita ML, Murray JC,
Murray S, de Villena FP, Postlethwait J, Potter S, Shapiro L, Spritz R, Visel A, Weinberg SM, Trainor
PA. The FaceBase Consortium: a comprehensive program to facilitate craniofacial research. Dev Biol.
2011 Jul 15; 355(2):175-82. PMID: 21458441. PMCID: PMC3440302
32.

Petersen CI, Jheon AH, Mostowfi P, Charles C, Ching S, Thirumangalathu S, Barlow LA, Klein

OD. FGF signaling regulates the number of posterior taste papillae by controlling progenitor fieldsize. PLoS Genet. 2011 Jun; 7(6):e1002098. PMID: 21655085. PMCID: PMC3107195

12

33.

Charles C, Hovorakova M, Ahn Y, Lyons DB, Marangoni P, Churava S, Biehs B, Jheon A, Lesot

H, Balooch G, Krumlauf R, Viriot L, Peterkova R, Klein OD. Regulation of tooth number by fine-tuning
levels of receptor-tyrosine kinase signaling. Development. 2011 Sep; 138(18):4063- 73. PMID: 21862563.
PMCID: PMC3160100
34.

Jheon AH, Li CY, Wen T, Michon F, Klein OD. Expression of microRNAs in the stem cell nicheof the adult mouse incisor. PLoS One. 2011; 6(9):e24536. PMID: 21931743. PMCID: PMC3169592
35.

Tian H, Biehs B, Warming S, Leong KG, Rangell L, Klein OD*, de Sauvage FJ*. A reserve

stem cell population in small intestine renders Lgr5-positive cells dispensable. Nature. 2011 Oct 13;478(7368):255-9. PMID: 21927002. PMCID: PMC4251967
36. Iraqi FA, … Broman KW, Hallgrimsson B, Klein O, Weinstock G, Warren WC, Yang YV, Schwartz

D. The genome architecture of the Collaborative Cross mouse genetic reference population. Genetics.
2012 Feb; 190(2):389-401.
37.

Purcell P*, Jheon A, Vivero MP, Rahimi H, Joo A, Klein OD*. Spry1 and spry2 are essential fordevelopment of the temporomandibular joint. J Dent Res. 2012 Apr; 91(4):387-93. PMID: 22328578.
PMCID: PMC3310757
38. Kyrylkova K, Kyryachenko S, Biehs B, Klein O, Kioussi C, Leid M. BCL11B regulates epithelial

proliferation and asymmetric development of the mouse mandibular incisor. PLoS One. 2012;
7(5):e37670. PMID: 22629441. PMCID: PMC3358280
39.

Li CY, Cha W, Luder HU, Charles RP, McMahon M, Mitsiadis TA, Klein OD. E-cadherin

regulates the behavior and fate of epithelial stem cells and their progeny in the mouse incisor. Dev
Biol. 2012 Jun 15; 366(2):357-66. PMID: 22537490
40. Juuri E, Saito K, Ahtiainen L, Seidel K, Tummers M, Hochedlinger K, Klein OD, Thesleff I, Michon

F. Sox2+ stem cells contribute to all epithelial lineages of the tooth via Sfrp5+ progenitors. Dev Cell.
2012 Aug 14; 23(2):317-28. PMID: 22819339. PMCID: PMC3690347
41. Justice CM, Yagnik G, Kim Y, Peter I, Jabs EW, Erazo M, Ye X, Ainehsazan E, Shi L,

Cunningham ML, Kimonis V, Roscioli T, Wall SA, Wilkie AO, Stoler J, Richtsmeier JT, Heuzé Y,
Sanchez-Lara PA, Buckley MF, Druschel CM, Mills JL, Caggana M, Romitti PA, Kay DM, Senders C,
Taub PJ, Klein OD, Boggan J, Zwienenberg-Lee M, Naydenov C, Kim J, Wilson AF, Boyadjiev SA. A
genome-wide association study identifies susceptibility loci for nonsyndromic sagittal craniosynostosis
near BMP2 and within BBS9. Nat Genet. 2012 Nov 18; 44(12):1360-4.
42.

Chavez MG, Yu W, Biehs B, Harada H, Snead ML, Lee JS, Desai TA, Klein OD. Characterizationof dental epithelial stem cells from the mouse incisor with two-dimensional and three-dimensional
platforms. Tissue Eng Part C Methods. 2013 Jan; 19(1):15-24. PMID: 22742471. PMCID: PMC3522131
43. Wan M, Gao B, Sun F, Tang Y, Ye L, Fan Y, Klein OD, Zhou X, Zheng L. microRNA miR- 34a
13

regulates cytodifferentiation and targets multi-signaling pathways in human dental papilla cells. PLoS
One. 2012; 7(11):e50090. PMID: 23226240. PMCID: PMC3511455
44. Goodwin AF, Oberoi S, Landan M, Charles C, Groth J, Martinez A, Fairley C, Weiss LA, Tidyman

WE, Klein OD, Rauen KA. Craniofacial and dental development in cardio-facio- cutaneous syndrome:
the importance of Ras signaling homeostasis. Clin Genet. 2013 Jun; 83(6):539-44. PMID: 22946697.
PMCID: PMC4115672
45. Griffin JN, Compagnucci C, Hu D, Fish J, Klein O, Marcucio R, Depew MJ. Fgf8 dosage

determines midfacial integration and polarity within the nasal and optic capsules. Dev Biol. 2013 Feb
1; 374(1):185-97. PMID: 23201021
46.

Felfly H, Klein OD. Sprouty genes regulate proliferation and survival of human embryonic stem

cells. Sci Rep. 2013; 3:2277. PMID: 23880645. PMCID: PMC3721083
47. Plaks V, Brenot A, Lawson DA, Linnemann JR, Van Kappel EC, Wong KC, de Sauvage F, Klein

OD, Werb Z. Lgr5-expressing cells are sufficient and necessary for postnatal mammary glandorganogenesis. Cell Rep. 2013 Jan 31; 3(1):70-8. PMID: 23352663. PMCID: PMC3563842
48. Yamada Y, Nakamura S, Ito K, Umemura E, Hara K, Nagasaka T, Abe A, Baba S, Furuichi Y,

Izumi Y, Klein OD, Wakabayashi T. Injectable bone tissue engineering using expanded mesenchymal
stem cells. Stem Cells. 2013 Mar; 31(3):572-80. PMID: 23225744. PMCID: PMC4043220
49. Juuri E, Jussila M, Seidel K, Holmes S, Wu P, Richman J, Heikinheimo K, Chuong CM, Arnold K,

Hochedlinger K, Klein O, Michon F, Thesleff I. Sox2 marks epithelial competence to generate teeth inmammals and reptiles. Development. 2013 Apr; 140(7):1424-32. PMID: 23462476. PMCID: PMC3596986
50. Lagronova-Churava S, Spoutil F, Vojtechova S, Lesot H, Peterka M, Klein OD, Peterkova R. The

dynamics of supernumerary tooth development are differentially regulated by Sprouty genes. J Exp
Zool B Mol Dev Evol. 2013 Jul; 320(5):307-20. PMID: 23606267
51.

Jones KB, Goodwin AF, Landan M, Seidel K, Tran DK, Hogue J, Chavez M, Fete M, Yu W,

Hussein T, Johnson R, Huttner K, Jheon AH, Klein OD. Characterization of X-linked hypohidrotic
ectodermal dysplasia (XL-HED) hair and sweat gland phenotypes using phototrichogram analysis
and live confocal imaging. Am J Med Genet A. 2013 Jul; 161A(7):1585-93. PMID: 23687000. PMCID:
PMC4414120
52.

Biehs B, Hu JK, Strauli NB, Sangiorgi E, Jung H, Heber RP, Ho S, Goodwin AF, Dasen JS,

Capecchi MR, Klein OD. BMI1 represses Ink4a/Arf and Hox genes to regulate stem cells in therodent incisor. Nat Cell Biol. 2013 Jul; 15(7):846-52. PMID: 23728424. PMCID: PMC3735916
53. Cao H, Jheon A, Li X, Sun Z, Wang J, Florez S, Zhang Z, McManus MT, Klein OD, Amendt BA.

The Pitx2:miR-200c/141:noggin pathway regulates Bmp signaling and ameloblast differentiation.
Development. 2013 Aug; 140(16):3348-59. PMID: 23863486. PMCID: PMC3737717
14

54.

Oishi PE, Klein OD, Keller RL. Developing physician-scientists in the fields of neonatology andpediatric critical care medicine: an effort to formulate a departmental policy. J Pediatr. 2013 Sep;
163(3):616-7.e1. PMID: 23973233
55. Zheng L, Warotayanont R, Stahl J, Kunimatsu R, Klein O, Denbesten PK, Zhang Y. Inductive

ability of human developing and differentiated dental mesenchyme. Cells Tissues Organs. 2013;
198(2):99-110. PMID: 24192998
56.

Goodwin AF, Tidyman WE, Jheon AH, Sharir A, Zheng X, Charles C, Fagin JA, McMahon M,

Diekwisch TG, Ganss B, Rauen KA, Klein OD. Abnormal Ras signaling in Costello syndrome (CS)negatively regulates enamel formation. Hum Mol Genet. 2014 Feb 1; 23(3):682-92. PMID: 24057668
57. Sun F, Wan M, Xu X, Gao B, Zhou Y, Sun J, Cheng L, Klein OD, Zhou X, Zheng L. Crosstalk

between miR-34a and Notch Signaling Promotes Differentiation in Apical Papilla Stem Cells (SCAPs).
J Dent Res. 2014 Apr 7; 93(6):589-595. PMID: 24710391
58. Mues G, Bonds J, Xiang L, Vieira AR, Seymen F, Klein O, D’Souza RN. The WNT10A gene in

ectodermal dysplasias and selective tooth agenesis. Am J Med Genet A. 2014 Oct; 164(10):2455-60.
PMID: 24700731. PMCID: PMC4167166
59.

Chavez MG, Hu J, Seidel K, Li C, Jheon A, Naveau A, Horst O, Klein OD. Isolation and culture ofdental epithelial stem cells from the adult mouse incisor. J Vis Exp. 2014; (87). PMID: 24834972
60.

Ching ST, Cunha GR, Baskin LS, Basson MA, Klein OD. Coordinated activity of Spry1 and Spry2is required for normal development of the external genitalia. Dev Biol. 2014 Feb 1; 386(1):1-11. PMID:
24361260
61. Zhao H, Feng J, Seidel K, Shi S, Klein O, Sharpe P, Chai Y. Secretion of shh by a neurovascularbundle niche supports mesenchymal stem cell homeostasis in the adult mouse incisor. Cell Stem Cell.
2014 Feb 6; 14(2):160-73. PMID: 24506883
62.

Mendelsohn BA, Jeng LL, Oberoi S, Klein OD. Dental findings in 14q terminal deletion syndrome.

Clin Dysmorphol. 2014 Apr; 23(2):60-2. PMID: 24535103
63.

Goodwin AF, Oberoi S, Landan M, Charles C, Massie JC, Fairley C, Rauen KA, Klein OD.

Craniofacial and dental development in Costello syndrome. Am J Med Genet A. 2014 Jun; 164(6):142530. PMID: 24668879
64. Michailovici I, Harrington HA, Azogui HH, Yahalom-Ronen Y, Plotnikov A, Ching S, Stumpf

MP, Klein OD, Seger R, Tzahor E. Nuclear to cytoplasmic shuttling of ERK promotes differentiationof muscle stem/progenitor cells. Development. 2014 Jul; 141(13):2611-20. PMID: 24924195. PMCID:
PMC4067960

15

65. Castillo D, Seidel K, Salcedo E, Ahn C, de Sauvage FJ, Klein OD*, Barlow LA*. Induction of ectopic

taste buds by SHH reveals the competency and plasticity of adult lingual epithelium. Development.
2014 Aug; 141(15):2993-3002. PMID: 24993944. PMCID: PMC4197660
66. Harjunmaa E, Seidel K, Häkkinen T, Renvoisé E, Corfe IJ, Kallonen A, Zhang ZQ, Evans AR,

Mikkola ML, Salazar-Ciudad I, Klein OD*, Jernvall J*. Replaying evolutionary transitions from thedental fossil record. Nature. 2014 Aug 7; 512(7512):44-8. PMID: 25079326. PMCID: PMC4252015
67.

Goodwin AF, Larson JR, Jones KB, Liberton DK, Landan M, Wang Z, Boekelheide A,

Langham M, Mushegyan V, Oberoi S, Brao R, Wen T, Johnson R, Huttner K, Grange DK, Spritz
RA, Hallgrímsson B*, Jheon AH, Klein OD*. Craniofacial morphometric analysis of individuals with
X-linked hypohidrotic ectodermal dysplasia. Mol Genet Genomic Med. 2014 Sep; 2(5):422- 9. PMID:
25333067. PMCID: PMC4190877
68. Jheon AH, Prochazkova M, Sherman M, Manoli DS, Shah NM, Carbone L, Klein O. Spontaneous

emergence of overgrown molar teeth in a colony of Prairie voles (Microtus ochrogaster). Int J Oral
Sci. 2015 Mar; 7(1):23-6. PMID: 25634121.
69. Dye BR, Hill DR, Ferguson MA, Tsai YH, Nagy MS, Dyal R, Wells JM, Mayhew CN, Nattiv R,

Klein OD, White ES, Deutsch GH, Spence JR. In vitro generation of human pluripotent stem cell
derived lung organoids. Elife. 2015 March 24; 4. PMID: 25803487. PMCID: PMC4370217
70.

Tian H, Biehs B, Chiu C, Siebel CW, Wu Y, Costa M, de Sauvage FJ*, Klein OD*. Opposing

activities of notch and wnt signaling regulate intestinal stem cells and gut homeostasis. Cell Rep. 2015
Apr 7; 11(1):33-42. PMID: 25818302. PMCID: PMC4394041
71.

Thys A, Vandenberghe P, Hague P, Klein OD, Erneux C, Vanderwinden JM. Hyperplasia ofinterstitial cells of cajal in sprouty homolog 4 deficient mice. PLoS One. 2015 Apr 29; 10(4):e0124861.
PMID: 25923139. PMCID: PMC4414615
72. Zarate YA, Perry H, Ben-Omran T, Sellars EA, Stein Q, Almureikhi M, Simmons K, Klein O, Fish

J, Feingold M, Douglas J, Kruer MC, Si Y, Mao R, McKnight D, Gibellini F, Retterer K, Slavotinek
A. Further supporting evidence for the SATB2-associated syndrome found through whole exome
sequencing. Am J Med Genet A. 2015 May; 167(5):1026-32. PMID: 25885067.
73.

Tapaltsyan V, Eronen JT, Lawing AM, Sharir A, Janis C, Jernvall J*, Klein OD*. Continuously

Growing Rodent Molars Result from a Predictable Quantitative Evolutionary Change over 50 Million
Years. Cell Rep. 2015 May 5; 11(5):673-80. PMID: 25921530. PMCID: PMC4426059
74.

Wu D, Mandal S, Choi A, Anderson A, Prochazkova M, Perry H, Gil-Da-Silva-Lopes VL, Lao R,

Wan E, Tang PL, Kwok PY, Klein O, Zhuan B, Slavotinek AM. DLX4 is associated with orofacial
clefting, and abnormal jaw development. Hum Mol Genet. 2015 May 7. pii: ddv167. [Epub ahead ofprint] PMID: 25954033

16

75.

Finkbeiner SR, Hill DR, Altheim CH, Dedhia PH, Taylor MJ, Tsai YH, Chin AM, Mahe MM,

Watson CL, Freeman JJ, Nattiv R, Thomson M, Klein OD, Shroyer NF, Helmrath MA, Teitelbaum
DH, Dempsey PJ, Spence JR. Transcriptome-wide Analysis Reveals Hallmarks of Human Intestine
Development and Maturation In Vitro and In Vivo Stem Cell Reports. 2015 Jun 3; pii: S22136711(15)00122-8. doi: 10.1016/j.stemcr.2015.04.010. [Epub ahead of print] PMID: 26050928. PMCID:
PMC4471827
76.

Tapaltsyan V, Charles C, Hu J, Mindell D, Ahituv N, Wilson GM, Black BL, Viriot L, Klein OD.

Identification of novel Fgf enhancers and their role in dental evolution. Evol Dev. 2015 Jun
18. doi: 10.1111/ede.12132. [Epub ahead of print]
76.

Marangoni P, Charles C, Tafforeau P, Laugel-Haushalter V, Joo A, Bloch-Zupan A, Klein OD,

Viriot L. Phenotypic and evolutionary implications of modulating the ERK-MAPK cascade using thedentition as a model. Sci Rep. 2015 June 30; 5:11658. PMID: 26123406. PMCID: PMC4485067
77.

Jheon AH*, Prochazkova M, Meng B, Wen T, Lim YJ, Naveau A, Espinoza R, Cox TC, Sone ED,

Ganss B, Siebel CW, Klein OD*. Inhibition of Notch Signaling During Mouse Incisor Renewal Leads toEnamel Defects. J Bone Miner Res. 2015 Jul 14. [Epub ahead of print] PMID: 26179131(* = co-senior authors)

17

Special Lecture 3

Comprehensive analysis for the functions of genes during epithelialmesenchymal interactions in tissue morphogenesis
Keigo Yoshizaki
Section of Orthodontics and Dentofacial Orthopedics, Faculty of Dental Science,
Kyushu University, Japan

Epithelial-mesenchymal (E-M) interactions play pivotal roles in the morphogenesis of various organs,such as tooth, lung, kidney, submandibular grand (SMG) and hair. In previous study, we found thatthe transcriptional coactivator, Mediator 1 (Med1), conditional deficient mice showed an ectopic hairgeneration in incisor (Yoshizaki et al., PLoS One 2014). This result indicated that the cell fate of toothand hair could be switched by alteration of specific transcriptional programs. In E-M interactions,
thickening of epithelial layer forming lamina, as the first event, is followed by bud formation. We
hypothesized that the critical genes for E-M interactions are expressed in every organs during theseearly developmental stages. To identify the specific genes for E-M interactions, we performed Cap
Analysis of Gene Expression (CAGE) for comprehensive analysis, using the early developmental
stages of tooth, lung, kidney, SMG and hair, and performed bioinformatics analysis. CAGE is genomewide transcription starting detection technology, which is capable of determining the transcriptional
start sites (TSS) by sequencing 5’ end of single-strand cDNA. We finally identified 42 tooth specific
TSS and 19 TSS commonly peaked during various developing organs as the candidate genes criticalfor E-M interactions. In this presentation, I would like to discus about the candidate genes regulating
E-M interaction and the possibility of tooth regeneration.

18

CURRICULUM VITAEEDUCATION
1998-2004

DDS, Kyushu University School of Dentistry, Fukuoka, Japan

2004-2008

PhD, Dental Science, Kyushu University School of Dentistry, Fukuoka, JapanEMPLOYMENT HISTORY
2008-2009

Resident, Kyushu University Hospital, Fukuoka, Japan

2009-2011	Research Fellow, the Japan Society for the Promotion of Science (JSPS), Fukuoka,Japan
2010-2013	Postdoctoral Fellow, Laboratory of Cell and Developmental Biology, NIDCR, NIH,
Bethesda, MD, USA
2013-present	Assistant professor, Section of Orthodontics and Dentofacial Orthopedics, Division of
Oral Health, Growth and Development, Faculty of Dental Science, Kyushu University,
Japan

19

【MEMO】

20

Reports after circulation

Lesional CD4 + IFN- γ+ + cytotoxic T lymphocytes in IgG4-related
dacryoadenitis and sialoadenitis
Takashi Maehara
Section of Oral and Maxillofacial Oncology
Division of Maxillofacial Diagnostic and Surgical Sciences
Faculty of Dental Science,Kyushu University
Ragon Institute of MGH, MIT and Harvard
Harvard Medical School

Background: IgG4-related disease (IgG4-RD) is a chronic, systemic, inflammatory condition of
unknown etiology characterized by storiform fibrosis and tumefactive lesions. We have recently
described clonally expanded circulating CD4 + cytotoxic T lymphocytes (CTLs) in IgG4-RD that
secrete IL-1b and TGFb1 in affected tissues. However, the contributions of CD4+CTLs to IgG4-RD
pathogenesis remain to be fully defined.
Objective: To examine the pathogeneis of IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS) andto determine whether CD4+ CTLs in IgG4-RD lesions secrete IFN-γ.
Methods: Gene expression was analyzed by DNA microarray in submandibular glands (SMGs) fromfive patients with IgG4-DS, three with chronic sialoadenitis (CS) and three healthy controls. Infiltrating
CD4+ CTLs were examined by quantitative multi-color imaging in tissue samples from eleven patientswith IgG4-DS, ten with Sjögren’s syndrome (SS), five with CS, and five healthy controls.
Results: In IgG4-DS tissues, nine genes associated with CD4 + CTLs were overexpressed. The
expression of granzyme A mRNA was significantly higher in samples from patients with IgG4RD compared to corresponding tissues from SS and healthy controls. Quantitative imaging showedthat infiltrating CD4+GranzymeA+ CTLs were more abundant in IgG4-DS patients than in the othergroups. The ratio of CD4+GranzymeA+ CTLs in submandibular salivary glands (SMGs) from IgG4DS patients correlated with serum IgG4 concentrations and the number of affected organs. The
percentage of CD4+GranzymeA+IFN-γ+ CTLs in SMGs was significantly higher in IgG4-DS than in
SS and CS patients.
Conclusions: The pathogenesis of IgG4-DS is associated with tissue infiltration by CD4+GranzymeA+
CTLs that secrete IFN-γ.

21

Young researchers
for next circulation

Molecular basis of amelogenin-induced periodontal tissue regeneration
Takao Fukuda, Terukazu Sanui, Kyosuke Toyoda, Urara Tanaka,
Kensuke Yamamichi, Ryo Atomura, Hajime Akiyama,
and Fusanori Nishimura

Department of Periodontology, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu
University, Japan

Amelogenin, the major component of enamel matrix derivative (Straumann® Emdogain), is wellrecognized in periodontology. It is used in periodontal surgery to regenerate cementum, periodontal
ligament, and alveolar bone. However, the precise molecular mechanisms underlying periodontal
regeneration are still unclear. Although the involvement of other enamel matrix proteins in theregeneration process cannot be excluded, recent studies have demonstrated that recombinantamelogenin induced the in vivo regeneration of all tooth-supporting tissues. In order to gain further
insight into how amelogenin induces periodontal tissue regeneration, we conducted a protein
interaction screen using recombinant full-length amelogenin (rM180) as bait. Using this proteomic
screen we identified glucose-regulated protein 78 (Grp78) as an amelogenin-binding protein, present inboth the cytosolic and membrane-enriched fractions of osteoblastic cells.
Mesenchymal stem cell-derived periodontal ligament stem cells (PDLSCs) play pivotal roles insuccessful periodontal regeneration. Thus, we evaluated the biological interaction between amelogenin
and Grp78, and its effect on cellular responses in human PDLSCs by using the cell line 1-17. Confocal
co-localization experiments revealed the internalization of recombinant amelogenin (rM180) via binding
to cell surface Grp78. Microarray analysis indicated that rM180 and Grp78 regulate the expression
profiles of cell migration-associated genes in 1-17 cells. Our study revealed that Grp78 is essential forenhancing amelogenin-induced cell migration in 1-17 cells. Furthermore, we demonstrated that Rac1
activation and lamellipodia formation are critical steps in amelogenin-induced cell migration.
In conclusion, our study demonstrates that the biological interaction of Grp78 with amelogeninenhances amelogenin-induced cell migration in PDLSCs. Although the effect of the biologicalinteraction of amelogenin with Grp78 on osteogenic differentiation must be further explored, ourfindings provide a better understanding of the molecular basis of amelogenin-induced periodontal
regeneration.

22

IL-33 secreted by M2 macrophages promotes
the pathogenesis of IgG4-related disease
Sachiko Furukawa 1 , Masafumi Moriyama 1, 2 , Akihiko Tanaka 1 ,Takashi Maehara 1 , Jun-Nosuke Hayashida 1 , Miho Ohta 1 , Masaki Yamauchi 1 ,Noriko Ishiguro 1 , Mizuki Sakamoto 1 , and Seiji Nakamura 1 .
1Section of Oral and Maxillofacial Oncology, Division of Maxillofacial Diagnostic
and Surgical Sciences, Faculty of Dental Science, Kyushu University, Japan

2

OBT Research Center, Faculty of Dental Science, Kyushu University, Japan

Objectives.
IgG4-related disease (IgG4-RD) is characterized by elevated serum IgG4 and marked infiltrationof IgG4-positive cells in multiple organs. Regarding immunological aspects, it has been suggested
that helper T type 2 (Th2) immune responses play a major role in IgG4 production of IgG4-RD.
Interleukin-33 (IL-33) is a recently described cytokine that is secreted by damaged epithelial cells,
macrophages (M Φ s) and dendritic cells (DCs) and potently activates Th2 immune responses. Here,
we assessed the expression of IL-33 and the producing cells in the submandibular glands (SMGs) from
IgG4-RD patients compared with those from Sjögren’s syndrome and controls.
Patients and methods.
SGs from patients with IgG4-RD (n=7), Sjögren’s syndrome (n=10), and healthy subjects (n=10) werescreened for 1) expression of IL-33, ST2, Th1 cytokine (IFN- γ ), Th2 cytokines (IL-4 and IL-13), IL33 producing cells including M Φ detected by CD68 (M1+M2 M Φ ) and CD163 (M2 M Φ ) and DCs
detected by CD11c (myeloid DC) and CD123 (plasmacytoid DC) by immunohistochemical staining; 2)
the relationship between mRNA expression of IL-33 and cytokines; 3) the co-localization of IL-33 andIL-33-producing cells by double immunofluorescence staining; 4) the population of IL-33-producing cellby flow cytometry.
Results.
mRNA expression of IL-33, ST2, IL-4, and IL-13 in IgG4-RD was significantly higher than that in
the other groups. Moreover, mRNA expression of IL-33 was positively correlated with that of Th2
cytokines only in IgG4-RD and didn’t show positive or negative correlation with IFN- γ . IL-33
was detected in/around epithelial cells in all the groups, while was strongly detected in infiltrating
lymphocytes around ectopic germinal centers only in IgG4-RD. Both of CD68 and CD163-positive cells
almost merged with IL-33. In addition, flow cytometric analysis revealed that IL-33+ cells were mainly
contituted by CD68+CD163+ M2 M Φ s in IgG4-RD patients.
Conclusion. These results suggest that IL-33 produced by M2 M Φ s might contribute to thepathogenesis of IgG4-RD via aberrant activation of Th2 immune responses.

23

Stimulus repetition enhances responses of the chorda tympani nerve tosweeteners via endocannabinoid system
Shusuke Iwata 1 , Keiko Yasumatsu 1,2 , Ryusuke Yoshida 1 ,Noriatsu Shigemura 1 , Yuzo Ninomiya 1,2

12

Oral Neurosci, Grad Sch of Dent Sci, Kyushu Univ, Japan

R&D Center for Taste and Odor Sensing, Kyushu Univ, Japan

2-Arachidonoylglycerol (2-AG), one of endocannabinoids (eCBs), enhances responses of sweet-sensitive
taste cells via activation of the cannabinoid receptor 1 (CB1). We recently found that diacylglycerol
lipase (DAGL), which synthesizes 2-AG through hydrolyses of diacylglycerol by in the GPCRphospholipase cascade, is expressed in T1R3-positive taste cells. This suggests the possibility thatthe activation of sweet taste receptor T1R2/T1R3 by sweeteners may lead to generation of 2-AG
and subsequent enhancement of sweet taste responses. In order to assess this possibility, here, weinvestigated potential effects of stimulus repetition with a single taste compound on responses of themouse chorda tympani (CT) nerve. The results indicated that CT nerve responses to sweeteners but
not to other tastants were gradually increased by stimulus repetition. Such effect was not observedin T1R3 knockout mice and was inhibited by CB1 blocker AM251. Intriguingly, intraperitoneal
injection with Angiotensin II (AngII), which is a major mediater of body fluid and sodium homeostasis,enhanced such increase of sweet taste response by stimulus repetition. CV11974, AngII type1 receptor
antagonist, inhibited the AngII’s effects. Taken together, our results suggest that the repeated T1R2/
T1R3 activation leads to generation of 2-AG, which enhances sweet taste responses by acting on CB1
expressed in sweet sensitive taste cells. AngII may cooperatively act on this eCB sweet-enhancing
effect.

24

Activin A reversely works between human pre-osteoblastic cells and periodontal
ligament cells on their osteoblastic differentiation
Hideki Sugii 1 , Atsushi Tomokiyo 2 , Sayuri Hamano 1 , Daigaku Hasegawa 2 ,Shinichiro Yoshida 1 , Suguru Serita 1 , Hiroyuki Mizumachi 1 ,Hiromi Mitarai 1 , Aoi Nozu 1 , Marina Ogawa 1 , Mai Sonoda 1 ,Naohisa Wada 3 and Hidefumi Maeda 1,2
1

Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu
University, 2 Department of Endodontology, Kyushu University Hospital 2 , and Division ofgeneral oral clinic, Kyushu University Hospital 3

Objectives : Activin A (ActA) that is a member of the TGF- β superfamily and is composed ofa dimer of inhibin β a. Our recent report suggested that ActA exerted an inhibitory effect on
osteoblastic differentiation of human periodontal ligament cells (HPDLCs) and could be applied asa therapeutic factor that promotes healing of periodontal ligament (PDL) tissue. In this study, we
have examined the effect on osteoblastic differentiation of human pre-osteoblastic cells (Saos2) andcompared the signal pathway between HPDLCs and Saos2 treated with ActA.
Materials and Methods : We purchased Saos2 from RIKEN (Saitama, Japan) and used HPDLCs
that were isolated from a healthy third molar from a 23-year-old male. These cells were cultured
with or without ActA in the presence or absence of CaCl2 that was used for inducing osteoblastic
differentiation. The cells were then subjected to Alizarin red S staining and von Kossa staining. Gene
expressions were evaluated by semi-quantitative or quantitative RT-PCR, and protein expressions
were investigated by western blotting and immunocytochemical analyses.
Results : Immunocytochemical analysis with an anti-ActA antibody revealed positive staining in
Saos2. Expression of Activin receptors (ALK4, IIa and IIb) was detected in Saos2. ActA promoted
mineralization and expression of bone-related genes in Saos2. The effect of ActA on osteoblastic
differentiation of Saos2 was inhibited by an anti-TGF- β 1 neutralizing antibody, but the effect of TGFβ 1 was not affected by an anti-ActA neutralizing antibody. Western blotting data showed that ActA
up-regulated the phosphorylation of Smad1/5/8 in Saos2, whereas promoted the phosphorylation of
Smad2/3 in HPDLCs.
Discussion and Conclusion : Our results showed that ActA and its receptors were expressed in
Saos2 and that promoted osteoblastic differentiation of Saos2. It was suggested that ActA promoted
osteoblastic differentiation of Saos2 through phosphorylation of Smad1/5/8 pathway, while inhibited
osteoblastic differentiation of HPDLCs through phosphorylation of Smad2/3 pathway. Thus, ActA
may reversely work between these cells through activation of different Smad pathway.

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