Dr.

Deepika Bali
(Associate Professor)
Deptt. Of Periodontoloy &
Oral Implantology
D.A.V (C) Dental College,
Yamuna Nagar
PERIODONTITIS

A chronic
inflammatory
disease
modified by Host factors
diabetes, Host
multiple risk
smoking, susceptibility
factors
genetics in form of
genetic
variation
Genetic factors in Periodontitis and their
potential influences
HISTORY
MODERN SCIENCE OF GENETICS

➢ Gregor Mendel – (Austrian Monk)

▪ Mendel’s factor Gregor Johann
Mendel

▪ Discrete units called genes are responsible for

inheritance

▪ Alternate forms are Alleles.

DEFINITION
CHROMOSOME: A Nuclear structure containing
Gene
Human genetic studies by:

• Pedigree analysis: 30 – 40,000 genes.

DNA-
Bases:
Purines (9 membered)– A,G

Pyrimidines (6membered)- C,T

U(RNA)
 CHARGAFF’S RULE

G=C
A=T

Complementary Paired Bases

A+G=T+C
A+T/G+C

Constant for given species

• Introns: Non coding region of DNA.

• Exon: Protein coding region of

DNA.
STR: Short Tandem Repeat 2-6 BP
• VNTR: A variable non tandem repeat is a location in a genome
where a short nucleotide sequence is organized as a tandem
repeat (10-60bp). Variation between different allele is caused by
a difference in no. of repeat units that results in alleles that are
of different lengths and are known as length polymorphism.

• STR: Short sequence of DNA normally of length 2-5 bp

• Gene: Basic unit of heredity that occupies a
specific locus on a chromosome.

• Major Disease Gene: Causative Gene.

• Modifying Disease Gene: Contributes to

susceptibility and severity of disease
Genes

Constitutive- Expressed all time

Non-constitutive- Expressed when products needed
Structural – Code for polypeptides and RNAs
Regulator-Regulate functional units in DNA
Operator- Act as a switch to turn on or off the
transcription.

Promotor – Sites where RNA polymerase binds.

Terminator – Suspend transcription.

Non functional – Which replicate but do not code for

any substance.

Jumping – Repetitive DNA sequences change their

position in the DNA.

Modifier – Influence the traits which are controlled

by other pairs of genes.
• Allele: one of several possible alternative form of
a gene caused by small or large differences in,
DNA Sequence within or near the gene.

• N-allele: less than 99% frequency

• R-allele: more than 1% frequency

• Minor Allele Frequency (MAF) : Proportion of least
frequent allele in a population( 0 – 50%).

• > 5% = common variant

• 1 – 5% = rare
• < 1% = mutation
• Autosomal Dominant: DNA variation in a gene located on
an autosome that has dominant effect.
• Autosomal Recessive: Recessive effect.

• Homozygous: Presence of identical allele at specific

positions.
• Heterozygous: Presence of 2 different allele at specific
position.
•Genetic Code: consecutive nucleotide
triplet ( codons) in RNA, DNA.

• Genotype: The genetic make up of an

organism.

• Phenotype: Observable characteristics

displayed by an organism.
• Polymorphism: It exist when 2 or more different phenotypes exist
with in different individuals of same populations.

• SNP: A polymorphism in a gene caused by change in single

nucleotide in DNA sequence. SNP is considered when is in ≥ 1% of
population.

• Hereditary: Transmission of characters, resemblances as

well as variation from one generation to next.
• Variation: Differences shown by the individual of a
species and also by offsprings of same parents.

• Mutation: Change in DNA sequence.

• Genotype Relative Risk: Ratio of risk of disease

between individuals with and without genotype.
• Linkage: Tendency for certain gene to be transmitted from
parent to child together because they are located close on
same chromosome.

• Linkage Disequilibrium: Occurrence of specific allele at

different locations in DNA that are relatively close to each
other more often than would be expected by chance alone.
• Gene Expression: The process by which information in a
gene is used via transcription and translation. Difference in
gene expression affect phenotype.

• GWAS: Investigates genetic variation across the entire

genome simultaneously with the aim of identifying
genetics association related to a trait or disease.
 DIAGNOSIS

Indirect:
Diagnosis based on
Direct : probands
Mutant gene is • Segregation
diagnosed Analysis
• Twin Studies
• LinkageAnalysis
• Association Studies
DISEASES

• Monogenic ( HUNTINGTON disease)

• Oligogenic

• Polygenic (multifactorial)
 Genetics and
Periodontal diseases

Major disease gene Modifying disease

Gene
Syndromes Associated with
Aggressive Periodontitis Associated with
Chronic Periodontitis
 SEGREGATION ANALYSIS

• Observed pattern of disease compared with those expected under

various models of inheritance to select the best fitting model.

• Penetrance- partial / full.

• Heterogenicity- different cause of disease.

Less power to distinguish b/w genetic effects and unmeasured

environmental causes of disease. (transmission of pathogenic org.
within families.)
TWIN STUDIES

• Compare concordance rate of MZ and DZ twins.

• Noack (1940)

• MZ twins same characters

• Concordance rate – percentage of twin pairs in which

both twins are affected

• Heritability – 50% half variance due to genetic variance.

 Minnesota and Virginia group

• Twins reared together and apart .

• Gingivitis, PD, CAL – Genetic variation 38-82%

.
• CR- ↑ MZ than DZ
Twin Intra class Correlation for Mean Attachment Loss in
Sub groups of different gender and race.
MICHALOWICZ (2000)
LINKAGE STUDIES

• To map allele to specific regions on chromosomes

• Recombination or cross over events- proportional to

distance b/w two allele.

• Low statistical power for complete diseases.

• Set of families
Unaffected Affected
Models of inheritance

(mode of inheritance, frequency of marker allele, disease penetrance ,only

major gene diagnosed)

LOD score: marker and disease allele linked versus not linked
Recombination frequency & qualitative traits. Linkage present
within 20-30 CM
1cM=app. 106nucleotide bases
ASSOCIATION STUDIES/ LINKAGE
DISEQUILIBRIUM

• Non random association of alleles at two or more loci.

• Loci are in LD when frequency of alleles is higher or lower than

expected.

• To test for association

• Frequency of alleles at given locus is compared between

subjects with disease/case and healthy controls.

• Alleles- different for different population.

Method Objective
Twin studies • To assess relative contributions of genes and
environment to a disease trait.
• If a disease has high heritability, identical
twins will be more likely to be either both
affected or unaffected.
Segregation analysis • To study the pattern of disease transmission
(autosomal, X- linked, dominant, recessive,
complex, multi locus or random environmental)
Linkage analysis • To localize the gene for a trait to a specific
chromosomal location
Limitations
The base assumption is complicated :
• When a genetic mutation does not have
complete penetrance, environmental factors
such as smoking may lead to disease
development.
• No good results in a polygenic disease that is
caused by alterations in multiple genes.
• Cannot find a specific gene responsible for a
disease trait.
• Segregation analysis are comparisons
between two models of transmission
• Only the first step in determining the
approximate location of a gene of interest.
• Subsequent tests are required to identify the
gene mutations/variants responsible for a
disease trait.
Method Objective Limitations

Candidate gene approach; • Genes are based on their known or presumed • Requires some knowledge of the candidate

hypothesis-testing approach function. gene.

• Requires some knowledge of the candidate

gene.

Genome-wide association • To investigate genetic variation across the • The results need to be further validated by a

study (GWAS); hypothesis- entire genome simultaneously cohort with a candidate gene
• To identify allelic variants associated with a
generating approach
trait or disease of interest.

• Performed as an open-ended study with no

candidate gene in mind.
GENETIC DISORDERS AND AGGRESSIVE
PERIODONTITIS

• Inherited and genetic disorders having gene mutation affect risk

forAP.

• Resultant proteins/ biochemical defects.

Affect the function of phagocytic immune cells.

• Affects structure of epithelia/ CT of teeth

 Disorder
Leucocyte adhesion deficiency type I
Leucocyte adhesion deficiency type II
Acatalasia
Chronic and Cyclic Neutropenia
Chediak higashi syndrome
Ehler donlos syndrome(EDS)
Papillon lefevre syndrome
hypophosphatasia
Trisomy 21
Prepubertal periodontitis (non- syndromic)
Kindler Syndrome
Protein or Tissue defects
CD 18(β-2 integrin of LFA- leukocyte function associated
antigen molecule.
CD 15(neutrophil ligand for E and P selectins); inborn
error in fucose metabolism (chromosome21)
Catalase Enzyme
Unknown
Abnormal transport of vesicles to and from neutrophil
lysosomes caused by mutation in lysosomal trafficking regulator
gene( LYST)WW
Type III collagen for EDS type IV unknown for EDS type
VIII
Cathepsin C( di peptidylaminopeptidase) CTSC gene
Tissue non specific alkaline phosphatase.
Multiple, vertical trisomic regions atleast 5Mb
(megabase) long.
Cathepsin-C
Defect in actin- extracellular matrix linkage caused by loss of
function in KIND-1
Papillon-Lefevre Syndrome
Leukocyte Adhesion Deficiency Type I
• Hypophosphatasia ( 1p36-1p34 )Tissue non specific Alk
Phosphatase

• PLS- 11(11q14 – 11q21) mutation in cathepsin C gene.

• Haim Munk Syndrome- Cathepsin C

• Down’s Syndrome – Chromosome 21

• Ehler Danlos – 11 type AD,AR, X linked.

SEGREGATION ANALYSIS

• Mode of Inheritance: Dominant & Recessive.

• Melnick 197 – X linked female probands

• Saxen 1980- AR (Finnish population)

• Boughman 1988 - AD

• Marazita (1994) largest JP family study

• 227 probands with AgP

• Hart (1991)- affected females more attributed to bias

U.S Study- AA & Caucasian families
• Schenkein 1994 – distinguished etiologies of LAP &
GAP.

• 1AgP + 2IgG = GAP. Responsible allele

• 1AgP + 1IgG = LAP. Less robust IgG2 response to

LPS.
LINKAGE STUDIES

• Boughman 1986- Ist study to report linkage b/w AP and

gene localised on 4q chromosome near DGI gene

• Saxen 1984= finnish population study found that it was

unlikely AP linked to HLA antigen

Li Y et al 2004- found LAP marker linked to 1q 25

chromosome with LOD score 3.48
 GENES ASSOCIATED WITH AGGRESSIVE
PERIODONTITIS RISK

Polymorphism Gene

• IL-1A(+4845)IL-1B(+3954) • IL-1 gene

IL-4 promoter and introns • IL-4 gene

polymorphism
• Fc receptor
FcãRIIIb-NA2 allele FcãRIIIa- gene
158F polymorphism
 • Gc locus • Unknown
chromosome 4q
• N-FMLP
• FMLP receptor polymorphism

• VDR polymorphism • Vit-D receptor

polymorphism
• GLT6D1 Chromosome
9q343 • Polymorphism
ASSOCIATION STUDIES

• HLA, MHC I, II .

• MHC Class II- DM, DQ, DR, DP asociated with CD4

receptors.

• MHC I- antigen via CD8 receptors.

• MHC III- Compliment.

• HLA- A, B,C – MHC I.

• MHC II- HLA- DPA1, DPB1, HLA-DQC, HLA-DQB,
DRA, DRB.

• HLA- A2 –protective less

in AP

• HLA- D Ag associated b/w PD and Type I DM DR4

Increases risk of AP.
CHRONIC PERIODONTITIS

Twin Study

• Noack 1940- PD condition of identical twin same.

• Minnesota, Virginia.- studied to know host gene affected

composition of oral microbiota.

(Moore)

• Adults- no effect ( Michalowicz).

Association study
 EARLY IMPLANT LOSS
Author Gene Polymorphism studied

Campos et al. 2004 TNF- α (G-308A)

Campos et al. 2005a IL -2 (-330)
IL-6 (-174)
Campos et al. 2005b IL-1A (-889)
IL-1B (+3953)
IL-1B (-511)
IL-1RN (intron 2)
Leite et al. 2008 MMP-1 (-1607)
MMP-1 (-519)
Santos et al. 2004 MMP-1 (-1607)
MMP-9 (C-1562T)
 Peri-implantitis
Author Gene polymorphism

Cury et al. 2007 Cury et al. 2009 TNF- α (-308)

De Boever 2006 TNF- α (-308)
IL- 1A (-889)
Dirschnabei et al 2010 Dos Santos et al IL- 1B (+3953) IL- 1B (C-511T)
2004 TGF-β1 (C-509T) TGF-β1 (G-800A)
IL- 1A (-899)
Feloutzis et al 2003 Gruica et al 2004 IL-1B (+3954)
IL-1A (+4845)
Hamdy & Ebrahem 2011 IL-1B (+3954)
Jansson et al 2005 IL- 1A(-889)
IL-1A (-889)
Jaworska- Zaremba et al 2008 IL-1B (+3953)
IL- 1A (-889)
IL-1B (+3953)
IL-1B (-511)
Lachmann et al IL- 1A (-899) IL- 1B(+3954) IL-1A (-889)
IL-1B (+3953)
Laine et al 2006 IL- 1B (-511)
IL- 1RN (intron 2) IL- 1B(+3954)
Montes et al 2009 IL- 1R (intron 2)
IL- 1A (-889)
Rogers et al 2002 Wilson &Nunn 1999 IL- 1B (+3953)
Alvim- Pereira et al IL- 1 genotype
Vitamin D receptor Tag1 (rs731236)
 Gene Coded protein

CTR Calcitonin receptor

BMP4 Bone morphogenetic protein
IL1A Interleukin- 1α
IL1B Interleukin- 1β
IL1RN Interleukin-1 receptor antagonist
IL2 Interleukin-2 Interleukin-6
IL6 Matrix metalloproteinase-1 Matrix metalloproteinase-9
MMP1 Transforming growth factor-β Tumor necrosis factor-α
MMP9
TGFB1
TNFA
EPIGENETICS
• Epi (on top off / in addition to)

• Stable heritable phenotype resulting from changes in

chromosome without alteration in DNA.

• Occurs by
A) DNA methylation
B) histone modification
C) gene regulation by non coding RNA.
 In Periodontitis

• During inflammation at biofilm gingival interface

• Epigenetics- role of gene environment interaction on

disease phenotype:
A) occurs more than genetics
B) reversible by pharmacological agents.
Clinical application of epigenetics

• Explain why same phenotype responds differently to

reaction.

• As effect of epigenetics is reversible, epidrugs are new

treatment modality.
Epidrugs

• DNA methyltransferase (DNMT) inhibitors

• Histone deacetylases (HDACs) inhibitors

• Histone methyltransferase enhancers

• HATi (Histone acetyltransferase inhibitor)
• Anacardic (cashew)
• Garcinal (kokum)
• Curcumin

• HDACi (Histone deacetylase inhibitor)

• Cantley 2001- suppress bone loss
a. 1179.4b
b. MS-275
c. Sodium butyrate (tissue regeneration)
 Genome-Wide
Association Studies

Causal models of diseases

1. Infinitesimal model (Vischer et el 2008)
▪ common variants are among the major source of genetic
variance or disease susceptibility
▪ Hundreds or thousands of different loci contribute to each
case
2. Rare allele model

▪ Genetic variance is due to highly penetrant variant with allele

frequencies of <1%.

▪ Fewer studies does not support model

3. Straightforward hypothesis

▪ Common variation influences the expression and activity of

genes in the molecular pathway
▪ establishing the background susceptibility that is modified by
rare variants with larger effects.
 •Aggressive Periodontitis
GLT6D1 (glycosyltransferase 6 Schaefer 2010,Howson 2017 (Sudan
domain containing 1) population)

Siglec-5 ( Sialic acid binding IG like Munz 2017 (German,Netherland,Turkey

lectin 5) population)

PF41CXCL5( Platelet Factor 4/Pro platelet Munz 2017

basic protein/CXC Chemokine Ligand)

NeuropeptideY Schaefer 2010

Fereiteg

ANRIL(Antisense non coding RNA Schaefer 2009

In INK4 LOCUS)
 Chronic periodontitis
Defensin alpha 1,alpha 3-Teumer 2013 (germen cases)

Neuropeptide Y Divoris 2013-(europeanamerican)

PF4/CXCL5 Divoris 2013-(european american)

Shimiz u 2015 (Japanese)

Schaefer 2014 (US)

Feng 2014 (US)

Hong 2015 (KORA)

Sonders 2016 (Hispanic)

GENE EXPRESSION
ANALYSIS
DISTINCT CLUSTER OFGENE EXPRESSION

Functional genomes study of gene function

1. Epithelium
2. Area of inflammatory cells in connective tissue
3. Non- inflammatory area in connective tissue

Main steps- probe production

- target production(DNA)
MICROARRAYS

GEO-Gene expression omnibus Hybridization

Less Costly

• Detect DNA or RNA (cDNA)

• Study extent to which certain genes are turned on or off in cell,

tissue

• c DNA is applied to microarray rapid screening , unknown gene

not found
• 21,000 genes –gene chip

• Deposit (more than 10,000) different gene sequence on a small

surface-chip.

• In a row and columns

• Using Northern Blot, RT-PCR

• C DNA labelling with flurodyes

• bind complementary
RNA SEQUENCING
performed on whole biopsies so produce an average of gene
expression of all cells.
SAGE-TRANSCRIPTOMIC
TECHNIQUE

• To produce a snapshot of m RNA in form of small tags.

• Quantitative Analysis.

• Unknown transcripts can be found – does not

require a pre existing clone.

I. Long SAGE

II. RL- SAGE

III. Super- SAGE

GENE THERAPY
a) A normal gene inserted into a non specific location to
replace a non functional gene.

b) An abnormal gene developed for a normal gene.

c) An abnormal gene repaired through selective reverse

mutation.

d) Regulation of gene altered.

Rationale of Gene Therapy
❑ Regeneration of PDL tissues occurs by the appropriate presentation of
selected multiple regulatory signals.

❑ Growth factors topically remain for a limited duration

❑ For long-term exposure of growth factors need Gene therapy

❑ A therapeutic strategy for directed, sustained, regulated protein

expression
Principles

❑ To transfer genetic information to target cells to produce a desired

therapeutic effect.

❑ It can be according to vector:

➢ Viral vectors
➢ Non-viral vectors
Viral Vectors
❑ Known as transduction

❑ DNA or RNA

❑ Prolonged transgene expression

❑ Higher efficiency

DISADVANTAGE:
• mutagenesis
• Cost
Non- Viral Vectors

❑ Known as transfection.

❑ SCID clinical trial complications lead to the importance of non-viral vectors

Classified as:
Physical Chemical

Magnetofection Organic Inorganic

optical transfection
Proelectrolyte CaPO4 ,etc
Mechanism

❖ Positively charged to facilitate entry through the plasma membrane

of target cells.

❖ ADVANTAGES:
• low cost
• Safety
Routes
Ex-vivo In-vivo

▪ Safer ▪ Immediate treatment

▪ Cells screened for ▪ Low transduction
tumorigenicity efficiency
▪ Complex ▪ Possibility of causing
▪ Costly inflammation/immune
response
▪ Difficulty in targeting
cell population
Role in Periodontal Treatment

BMPs
▪ BMP-7 and Noggin gene transfer (Jin et al 2003)

▪ Adenoviral vector (ex-vivo)

▪ Prevent binding of BMP – 2,4,7 to cell surface receptors

▪ Successful PDTE

▪ Similar studies by Chen et al 2008 and Park et al 2015, ex-vivo BMP-2 gene using
MSCs and PDLSCs
 PDGF-A and B

▪ Anusaksathein 2004 and Lin et al 2008

gene delivery using adenovirus

Non- viral vectors in periodontal treatment

▪ Elongoven et al 2013
▪ PDGF- B gene delivery using nano sized CaPO4 particles
Gene Activated Matrix (GAM)

▪ Direct gene transfer using TE and local gene delivery.

▪ GAM in implanted site act as scaffold/ template

▪ Fibroblasts migrate to the template

▪ Take up the gene and PDL regeneration activated

▪ Yang et al 2013
combined Bio-oss with BMSCs transfected BFGF gene
encoding plasmid
Effect of Genetics on Treatment Plan
OUTCOME OF DIFFERENT TREATMENT METHODS
PREDICTED BY GENOTYPE
A Generic multi-causality model for periodontitis

Other factors
(tooth & dention (Epi)genetic factors
related &
stochasticity)

Microbial communities Lifestyle factors

(dental biofilms)

Comorbidities
(systemic diseases)
CONCLUSION

As genetic testing becomes widespread, its our

responsibility to fully understand and counsel the
patients about implications of test result for their
treatment options and their risk of developing various
dental diseases in future
 Questions
01 Implications of genetics in periodontal disease

02 Role of genetics in periodontal disease

03 Genetic markers

Discuss the role of genetic factors in the pathogenesis of

04 periodontal disease

Discuss the role of genetic factors associated with

05 periodontal disease
Thank You