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134 Pages·2013·3.04 MB·English

by  Lea Alexander

Differential Gene Expression of Inflammatory Signaling in Localized
Aggressive Periodontitis

BY
WHITNEY DUNN WEINER
Specialty Certificate in Periodontics, University of Illinois at Chicago
College of Dentistry, 2013
D.D.S., University of Iowa College of Dentistry, 2010
B.A., The Colorado College, 2005

THESIS
Submitted as partial fulfillment of the requirements
for the degree of Master of Science in Oral Sciences
in the Graduate College of the
University of Illinois at Chicago, 2013

Chicago, Illinois

Thesis Committee

Dr. Thomas C. Hart, Chair
Dr. Joseph V. Califano, East Carolina University
Dr. Christopher G. Engeland
This thesis is dedicated to my parents, Greg and Wendy Dunn, and my
husband, S. Eliot Weiner, for their unconditional support and encouragement.
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ACKNOWLEDGEMENTS
I would like to thank my mentor, Dr. Thomas Hart, for his patience,
unwavering support, confidence, unparalleled knowledge, and constant
encouragement. He provided guidance and helped me achieve my research
goals.
I would also like to thank my thesis committee members, Dr. Califano and
Dr. Engeland, for their support and assistance. Additionally, I would like to thank
Dr. Shujuan Guo for all of her hard work culturing cells and isolating RNA.
This thesis would not have been possible without the encouragement and
flexibility from my full-time faculty in the Department of Post-Graduate
Periodontics and University of Illinois at Chicago College of Dentistry.
Particularly, a special thanks to my program director, Dr. Saba Khan, and my
clinical director, Dr. Praveen Gajendrareddy. .

WDW
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TABLE OF CONTENTS

CHAPTER PAGE

1. INTRODUCTION
1.1 Background …………………………………………………… 1
1.2 Significance …………………………………………………… 8
1.3 Specific Aims …………………………………………………… 9
1.4 Hypotheses …………………………………………………… 10

2. REVIEW OF LITERATURE
2.1 Classification of Periodontal Diseases…………………………. 11
2.2 Prevalence of Periodontitis………………………………………. 13
2.3 The Genetic Basis of AP……………………………………… 14
2.4 Role of the Early Innate Immune Response in Periodontitis... 32

3. METHODOLOGY
3.1 Study Design …………………………………………………… 49
3.2 Materials and Methods ……………………………………….. 49
3.3 Statistical Analysis …….……………………………………….. 52
3.4 IRB/ACC Approval …….……………………………………….. 52

4. RESULTS
4.1 Results ………………………………………………………….. 53

5. DISCUSSION
5.1 Discussion ………………………………………………………. 62
5.2 Limitations of the Study ……………………………………….. 72
5.3 Future Research ……………………………………………….. 74

6. CONCLUSION ………………………………………………………... 77

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TABLE OF CONTENTS (continued)

PAGE

CITED LITERATURE ……………………………………………….. 78

APPENDICES
APPENDIX A …………………………………………………. 96
APPENDIX B …………………………………………………. 97
APPENDIX C …………………………………………………. 98
APPENDIX D ………………………………………………… 99
APPENDIX E …………………………………………………. 101
APPENDIX F …………………………………………………. 102
APPENDIX G …………………………………………………. 105
APPENDIX H …………………………………………………. 106
APPENDIX I …………………………………………………. 107
APPENDIX J …………………………………………………. 108
APPENDIX K …………………………………………………. 111

VITA ………………………………………………………………….. 113

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LIST OF TABLES
TABLE PAGE

I. SINGLE NUCLEOTIDE POLYMORPHISMS REPORTED TO BE
ASSOCIATED WITH AP……………………………………………….. 37
II. 20 GENES ASSOCIATED WITH THE EARLY INNATE IMMUNE
RESPONSE AND THE PRIMARY FUNCTION OF EACH GENE
ACCORDING TO STRING DATABASE (VERSION 9.0) …………… 43

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LIST OF FIGURES

FIGURE PAGE

1. STRING analysis (version 9.0) of 20 candidate early innate immune
response genes indicating directionality and mode of action……….. 48

2. Gene Expression Following Stimulation with IL-1β…………………... 54

3. Gene Expression Following Stimulation with IL-1β…………………... 56

4. Gene Expression Following Stimulation with PgLPS………………… 57

5. Gene Expression Following Stimulation with PgLPS………………… 58

6. Gene Expression Following Stimulation with AaLPS………………… 59

7. Gene Expression Following Stimulation with AaLPS……………….... 60

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LIST OF ABBREVIATIONS

AaLPS Actinobacillus actinomycetemcomitans LPS

AAP American Academy of Periodontology

ACTB actin, beta

AD Autosomal Dominant

AP Aggressive Periodontitis

APCS Amyloid P component, serum

AR Autosomal Recessive

C3 Complement component 3

CASP1 Caspase 1, apoptosis-related cysteine
peptidase

CD4 CD4 molecule

CD8a CD8a molecule

CD14 CD14 molecule

CD40 CD40 molecule, TNF receptor superfamily
member 5

CCL2 Chemokine (C-C motif) ligand 2
Also called Monocyte chemoattractant protein-
1(MCP-1)

CCL3 Chemokine (C-C motif) ligand 3

CDC Centers for Disease Control

CTSC Cathepsin C

CP Chronic Periodontitis

CRP C-reactive protein, pentraxin-related

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LIST OF ABBREVIATIONS (continued)

CSF2 Colony stimulating factor 2 (granulocyte-
macrophage)

CXCL8 Interleukin-8 (IL-8)

DDX58 DEAD (Asp-Glu-Ala-Asp) box polypeptide 58

DEFB1 Defensin, beta 1

DNAS DNA Services

EOP Early Onset Periodontitis

FAM5C Family with sequence similarity 5, member C

FCGR Fc fragment of IgG, gamma receptor

FPR Formyl peptide receptor

GAP Generalized Aggressive Periodontitis

GJP Generalized Juvenile Periodontitis

GWAS Genome Wide Association Studies

HGP Human Genome Project

HLA Major histocompatibility complex, class II

HLA-DRA Major histocompatibility complex, class II, DR
alpha

IFNA1 interferon, alpha 1

IFNA2 interferon, alpha 2

IFNG Interferon gamma

IL1 Interleukin 1

IL1A Interleukin 1 alpha (IL-1α)
IL1B Interleukin 1 beta (IL-1β)

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LIST OF ABBREVIATIONS (continued)

IL1RN Interleukin 1 receptor antagonist

IL2 Interleukin 2

IL4 Interleukin 4

IL6 Interleukin 6

IL8 Interleukin 8

IL10 Interleukin 10

IL12 Interleukin 12

IL13 Interleukin 13

IL17 Interleukin 17

IL18 Interleukin 18

IRAK1 Interleukin-1 receptor-associated kinase 1

IRB Institutional Review Board

IRF3 Interferon regulatory factor 3

IRF7 Interferon regulatory factor 7

ITGAM Integrin, alpha M (complement component 3
receptor 3 subunit)

JE Junctional Epithelium

JP Juvenile Periodontitis

LAD Leukocyte Adhesion Deficiency

LAP Localized Aggressive Periodontitis

LD Linkage Disequilibrium

LJP Localized Juvenile Periodontitis

LPS Lipopolysaccharide
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