HPV and cervical cancer

林雅雯
分機 18910
Department of Microbiology and Immunology
National Defense Medical Center
 Tumor Viruses

• Retrovirus
• Herpesvirus
• Hepatitis virus
• Human papillomavirus
• ……….
 Tumor viruses-retrovirus
• Retrovirus
– ssRNA
– 1911 Chicken sarcoma
– 1970 Reverse transcriptase
– 1976 Proto-oncogens > 60
– 1980s HTLV-1: T cell lymphoma/leukemia
 Virion Structure
Lipid Envelope Nucleic Acid

Protein
Capsid

Virion
Associated
Spike
Polymerase
Projections
 Virion Components
• Protein
– Structural proteins
– Membrane proteins
– Receptor recognition
– Enzymes
• Genomic nucleic Acid
– DNA
– RNA
• Lipid envelope
– Plasma membrane – Paramyxoviruses
– Nuclear membrane – Herpes viruses
– Golgi membrane - Bunyaviruses
 Virus replication
Maturation
Release

Virion Budding
attachment
to cellular Insertion of virus
receptors proteins into
membrane
Genomic
nucleic acid Virion
Uncoating synthesis Assembly
Newly synthesised
virus proteins

Replication of Protein
Genomic Synthesis
nucleic acid mRNA synthesis
 Baltimore Classification of Viruses
Grou p Gen om e Repl ic atio n Ex am ple
1 dsDNA dsDNA mRNA Herpes simplex
virus

2 ssDNA ssDNA dsDNA mRNA Parvovirus

3 dsRNA dsRNA mRNA Reovirus

4 +ve ssRNA dsRNA +ve ssRNA [Acts as mRNA] Enterovirus

5 -ve ssRNA dsRNA -ve ssRNA mRNA Influenza A

virus

6 ssRNA ssRNA dsDNA mRNA Retrovirus

(e.g. HIV)

7 Nicked dsDNA nicked dsDNA intact dsDNA mRNA Hepatitis B

virus
RNA
 Tumor Viruses
For most viruses:

Replication Lysis Progeny

virions
Lytic Life Cycle

Genome all viral proteins

 Tumor Viruses
Virus

Latent Life Cycle Cell

Integration (usually)

Transformation

Some virus-specific proteins expressed - No mature

virus
Viral structural proteins are not expressed
Sometimes latency may terminate – cell must be infected
by complete virus
Changes in the properties of host cell - TRANSFORMATION
 Tumor Viruses
Transformation:
• Loss of growth control

• Reduced adhesion
• Motility
• Invasion
• Ability to form tumors - viral genes interfere
with control of cell replication
• Transformed cells frequently exhibit
chromosomal aberrations
 TRANSFORMATION
Both DNA and RNA tumor viruses can transform cells
Integration occurs (usually)
Similar mechanisms

VIRAL TRANSFORMATION
The changes in the biological functions of a cell that result from
REGULATION
of the cell’s metabolism by viral genes and that confer on the
infected cell certain properties characteristic of
NEOPLASIA
These changes often result from the integration of the viral
genome into the host cell DNA
Two Major Classes of Tumor Viruses
DNA Tumor Viruses
DNA viral genome
DNA-dependent
Host RNA
DNA polymerase
polymerase
(Host or viral)
Viral mRNA

Viral protein
 RNA Tumor Viruses
Viral RNA genome
Reverse transcriptase (Virus-encoded)

Viral DNA genome (integrated)

DNA-dependent RNA polymerase (Host
IMPORTANT
RNA pol II)

Viral genomic RNA

Splicing (Host splicing enzymes)

messenger RNA
Important: Use HOST
RNA polymerase
to make its genome
viral protein
An enzyme that
normally
Virus makes mRNA
 DNA Tumor Viruses
DNA genome
Host RNA
polymerase II
mRNA
Host enzymes

protein

virus

OR TRANSFORMATION
In transformation usually only EARLY functions are
expressed
 DNA Tumor Viruses In
Human Cancer

Papilloma Viruses
• cause natural cancers in animals
• cause benign warts
• ubiquitous
• epitheliotropic - most human tumors are malignancies of
epithelial cells
 DNA Tumor Viruses In
Human Cancer

Papilloma Viruses

• Epidermodysplasia verruciformis

wart malignant
squamous cell carcinoma
 DNA Tumor Viruses In Human
Cancer

ONCOGENE
A gene that codes for a protein that potentially can
transform a normal cell into a malignant cell
An oncogene may be transmitted by a virus in which
case it is known as a VIRAL ONCOGENE

v-onc
 RNA Tumor Viruses
RNA Genome - Retroviruses
RNA-dependent DNA Polymerase encoded by virus
REVERSE TRANSCRIPTASE
RNA genome
Reverse transcriptase virus

DNA genome
Integrase virus

Integrates
Host RNA polymerase II host

RNA genome
RNA Tumor Viruses

Viral Oncogene
V-onc

Cellular Proto-oncogene
C-onc
 RNA Tumor Viruses

Proto-oncogene
A cellular (host) gene that is homologous
with a similar gene that is found in a
transforming virus
A cellular oncogene can only induce
transformation after
• mutation
• some other change in the cell’s genome
 RNA Tumor Viruses
The discovery of the acutely transforming
retroviruses that contain
v-oncs explains how cancers may arise as a
result of infection

These viruses cause rapid cancer in animals

in the laboratory
 RNA Tumor Viruses
In contrast:
Chronically transforming retroviruses
cause tumors inefficiently after prolonged period
of time
Avian Leukosis Virus (causes lymphomas)

R U5 GAG POL ENV U3

R
No oncogene! – How does it cause a
tumor?
 RNA Tumor Viruses
ALV can integrate into the host cell genome
at MANY locations
but in tumor it is always at the SAME site
(or restricted number of sites)
Suggests tumor arose from one cell
• Something must be important about this
site for transformation
• Crucial event must be rare
 RNA Tumor Viruses
What is special about this site?
Myelocytoma tumors from several birds all
have the oncogene close to this site

It is close to
C-myc!
Oncogenesis by promotor insertion
 RNA Tumor Viruses

Could C-oncs be involved in NON-VIRAL

cancers?
 RNA Tumor Viruses
What do oncogenes encode?
Proteins that are involved in growth control and
differentiation

Growth factors
Growth factor receptors
Signal transduction proteins
Transcription factors
 DNA Tumor Viruses
Herpes
Genes can be
assigned to
sites on
specific
myb mos
chromosomes
myc

mos and myc :

chromosome 8

fe fes: chromosome
s
15
 Cancers often result from gene
Burkitt’s translocations
Lymphoma
8:14 translocation
Break in
chromosome 14 at
q32

myc

Acute myelocytic
leukemia
7:15
9:18
11:15:17
 Oncogenesis by
rearrangement

Tumor c-onc new promotor

Burkitt’s lymphoma myc (8) Ig heavy (8 to 14)
Ig light (8 to 2)
B-cell chronic lymphocytic bcl-1 Ig heavy (11 to 14)
leukemia bcl-2 Ig heavy (18 to 14)

T cell chronic lymphocytic tcl-1 T cell receptor

leukemia (14 inversion)
T cell chronic lymphocytic myc T cell receptor (8 to 14)
leukemia
 Oncogenes
Mutations in a proto-oncogene are dominant
“gain of function” mutations

Anti-Oncogenes
• Loss of function mutations
• Retinoblastoma
• p53
 Proto-oncogenes
Dominant
Heterozygote mutations Homozygote

Allele 1 Allele 2 Allele 1 Allele

2
Normal Mutant Mutant Mutant

Binds under Mutant Mutant Mutant

special always always always
circumstance binds binds binds
s
Always binds Always binds

Function gained Function gained

 Anti-Oncogenes
Recessive mutations
Mutation growth

Rb Gene Mutant Rb Mutant Rb Mutant Rb

Rb
Rb
protein
Heterozygote Homozygote

Rb Function lost

Binds and controls cell cycle No binding - Growth

Turns off DNA replication continues
 Anti-Oncogenes
Retinoblastoma gene has normal
regulatory function in many cells

Involved in
Retinoblastoma
Lung carcinomas
Breast carcinomas
 Anti-Oncogenes
P53
Inactivated by
• deletion
• point mutation
In a series of colorectal cancers all
showed:
• Allele 1: partial or complete deletion
• Allele 2: Point mutation
 DNA Tumor Viruses
Oncogenes
• Adenovirus E1A region 2
• SV 40 Large T
• Polyoma Large T
• BK virus Large T
• Lymphotropic virus Large T
• Human papilloma Virus-16 E7
All have a sequence in common
Mutations in this region abolish transformation
capacity
 Anti-Oncogenes
Retinoblastoma
Rb Gene Adenovirus E1A

Rb Rb
protein 105kD

Rb

Rb

Stops replication Cell cycle continues

 Anti-Oncogenes
p53
P53 gene P53 gene P53 gene
Hepatitis C Papilloma

P53 P53 P53

Papilloma
proteolysis
P53 DNA

Stops replication replication replication

 Tumor virus-Human
papillomaviruses

Circular dsDNA
~8k bp
~100 types
8 genes

High risk HPV 31

 HPV and Cervical cancer
• 1974~1976
– postulating the role of HPV in cervical cancer
• 1976
– The role of HPV in Pap smear and mild dysplasia
• 1983
– The first HPV isolated from cervical cancer biopsy (HPV 16)
• 1984
– Isolation of HPV 18 from cervical cancer biopsy
• 1985
– Active E6/E7 in cervical cancer
• 1989
– The transforming properties of E6 and E7
 HPV and Cervical cancer
• 1987
– First epidemiologic study
– High rate of infection in young women
• 1992
– A large-scale epidemiological study in Columbia and Spain that
provides convincing evidence that high risk HPVs are the main
risk factor for CC.
• 1995
– Prevalence of HPV in cervical cancer: a worldwide perspective
• 1995
– HPV 16 and 18 were defined as causative agents for cervical
cancer by IARC
 HPV and Cervical cancer
• 1998
– The natural history of HPV infection
– 14% per year, 44% in 3 years
– Average clearance time 8 months
• 1999~2001
– Type-specific persistent infection is the major risk
factor for cervical cancer
• 2000
– Viral load as a risk factor
• 2002
– The causal role of HPV in cervical cancer is beyond
reasonable questioning
 HPV pathogenesis

Oncogene (2003) 22, 5201–5207

 The spectrum of cervical neoplasia

‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧

Normal LSIL HSIL SCC

HPV pathogenesis- viral oncoproteins

• E6-P53
• E7-Rb
• E5-transmembrane
proteins
• E4 (?)
• E2-+/- regulation of
URR
• L1 and L2
Cell cycle and P53/Rb
High-risk HPV E6 promotes p53 degradation via
the ubiquitin pathway
Model for the concerted action of the HPV
oncoproteins in virus-induced cellular
transformation
E6/E7 binding proteins
E6/E7 functions
Functions of the E6 and E7 oncoproteins, and their interaction
with each other in steps that lead to cell immortalization
Nat Rev Cancer. 2002 May;2(5):342-50. Review.
Systemic and host-cell controls that interfere with HPV-induced
progression towards malignant proliferation
HPV pathogenesis-Immune evasion
The biology of HPV infection
How the adaptive immune system ‘sees’ and responds to tumour
(or other foreign) antigens inside cells
 Immune Evasion Mecahnisms
• Subversiion of IFN responses
– E7 blocks IFN-alpha inducible genes
– E7 inhibits IFN-beta promoter
• Oncoprotein escape
• E7 is tolerogenic
• Others
– E7: similar to some human proteins
– E6/E7 reduce IFN-alpha production in NK cell
– E6 can down regulate IL-18
– E5 affect Ag processing and presentation in APCs
– ………….
Protective specific immunity to E7 epithelial tumour antigen by
administration of an inflammatory stimulus
E7 in peripheral epithelium tolerizes the CTL response.
Natural history of HPV infection

30% 3yr
HPV
14%
annually
60% 3 yr 5-10% 3yr
Normal LSIL HSIL
70% 1st yr 30% 1st yr
91% 2nd yr 9% 2nd yr ~10% 2 yr <5% 2 yr
8 mo
HPV (-) HPV (+)
HPV and non-HPV factors that contribute to HPV-induced
malignant progression
 HPV is necessary , but not
sufficient

‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧

Co-Factors: Environmental? Genetic?

Normal LSIL HSIL SCC

HPV-16 類病毒顆粒疫苗
• 以基因工程技術於酵母菌體內大量合成 HPV
病毒 L1 鞘膜蛋白，經活體結合為類病毒顆粒
(virus like particle, VLP) 後純化為疫苗。
• 可充分激發人體免疫系統，產生高效價之抗病
毒抗體。
• 不帶任何病毒基因，無致病危險性。

人類乳突病毒顆粒 類病毒顆粒
 Perspectives
• Mechanism exploration
• Clinical applications
– Diagnosis
– Treatment
– Vaccine
N Engl J Med 2003;349:2042-54.
NATURE REVIEW/CANCER VOLUME4, DECEMBER
2004
Interaction between DNA Methylation and Histone Methylation
Signaling Networks in Cancer

Hanahan and Weinberg, 2000
Cell 100:57
 The End
Thanks for your attention