Microorganism & Human Cancer

Riyani Wikaningrum Bag. Mikrobiologi FK Universitas YARSI

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Viruses and Human Cancer

Virus acts as a cofactor

Viruses act as initiators of neoplastic process various mechanisms

Interaction Tumor Virus - Host

1. Persistent infection 2. Host immune response 3. Mechanisms of action by Human Cancer Virus 4. Cell susceptibility to viral infection 5. Retention of tumor virus nucleic acid in host cell
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Genes unrestrained cell growth

genes that are stimulatory for growth and which cause cancer when hyperactive oncogenes genes that inhibit cell growth and which cause cancer when they are turned off antioncogenes or tumor-suppressor genes
Viruses cancers virus carry a copy of one of these genes virus alter expression of the these genes

Malignant transformation
Changes Details

Morphology
Growth, contact

Loss of shape; rounding

Decreased adhesion to surface Loss of contact inhibition of growth and movement Increased ability to grow from a single cell Increased ability to grow in suspension Capacity for continued growth (immortalization)

Cellular properties

DNA synthesis induced

Chromosomal changes
Appearance of new antigens (viral or cellular in origin) Biochemical properties Loss of fibronectin Reduced cAMP

Viruses
Epstein-Barr virus

Cancer
Burkitt's lymphoma Nasopharyngeal carcinoma Hodgkin's disease

Strength of association
++ ++ ++

Viral genome in cancer cells

+ + +

Cofactor
Malaria Nitrosamine s ? cigarettes ? HSV2

Human papillomavirus

Cervical cancer

Skin cancer Hepatitis B virus Hepatitis C virus HTLV1 HSV2 Liver cancer Liver cancer

++ ++

+ + -

? UV light ? Aflatoxin ? Hepatocyte regeneration -

T-cell leukemia Cervical cancer

++

Tumor Viruses
For most viruses: Replication Lysis Progeny virions

Genome

all viral proteins

Lytic Life Cycle

Non-structural and structural proteins made
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Tumor Viruses
Virus

Latent Life Cycle

Cell

Integration (often)

Transformation

Some virus-specific proteins expressed (early functions) - No mature virus Viral structural proteins are not expressed

Changes in the properties of host cell - TRANSFORMATION

Sometimes latency may terminate cell must be infected by 8 complete virus

Tumor Viruses
Transformation:
Loss of growth control
Reduced adhesion

Motility
Invasion Ability to form tumors - viral genes interfere with control of cell replication and other aspects of the cell phenotype

Transformed cells frequently exhibit chromosomal aberrations

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Tumor Viruses
TRANSFORMATION
VIRAL TRANSFORMATION
The changes in the biological functions of a cell that result from
REGULATION

of the cells metabolism by viral genes and that confer on the infected cell certain properties characteristic of
NEOPLASIA
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Tumor Viruses
Both DNA and RNA tumor viruses can
transform cells Integration of viral genome into the host chromosomes often occurs

Similar mechanisms of transformation by each type of tumor virus

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Two Major Classes of Tumor Viruses

DNA Tumor Viruses
DNA viral genome
DNA-dependent DNA polymerase (Host or viral)

Host RNA polymerase

Viral mRNA Similar to host cell!

Viral protein
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RNA Tumor Viruses

Viral RNA genome
Reverse transcriptase (Virus-encoded)

Viral DNA genome (integrated)

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

Viral genomic RNA

Splicing (Host splicing enzymes)

messenger RNA

viral protein

Important: Use HOST RNA polymerase to make its genome An enzyme that normally makes mRNA

Virus

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DNA Tumor Viruses

DNA genome
Host RNA polymerase II

mRNA
Host enzymes

protein

virus

OR TRANSFORMATION In transformation usually only EARLY functions are expressed

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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
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DNA Tumor Viruses In Human Cancer

Papilloma Viruses

Epidermodysplasia verruciformis
wart malignant skin squamous cell carcinoma

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DNA Tumor Viruses In Human Cancer

Epidermodysplasia verruciformis Papilloma virus

2008 Dermatology Online Journal

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DNA Tumor Viruses In Human Cancer

Papilloma Viruses urogenital cancer wart malignant squamous cell carcinoma

Squamous cell carcinoma: Larynx Esophagus Lung

All histologically similar

10% of human cancers may be HPV-linked

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DNA Tumor Viruses In Human Cancer

Papilloma Viruses
>100 types identified - most common are types 6 and 11 Most cervical, vulvar and penile cancers are ASSOCIATED with types 16 and 18 (70% of penile cancers) EPIDEMIOLOGIAL STUDIES BUT: HPV 16 and HPV 18 do transform human keratinocytes Effective Vaccine (quadrivalent recombinant HPV 6, 11, 16 and 18 proteins made in yeast - Gardasil) 19

Papilloma Viruses
The important transforming genes in papilloma viruses are: E6 and E7 Early genes - Not encoding structural proteins Oncogenes

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DNA Tumor Viruses In Human Cancer

Polyoma Viruses
Simian virus 40 - juvenile hamster sarcomas, transformation

Polyoma - mouse leukemia, in vitro transformation

Human polyomas (JC and BK) - monkey sarcoma, transformation

Possible association of BK with human prostate cancer

Polyoma virus transforms cells when the genome is incomplete

Early functions are necessary - ONCOGENES

21 JC: PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY (PML)

DNA Tumor Viruses In Human Cancer

Adenoviruses
Highly oncogenic in animals

Only part of virus integrated

Always the same part Early functions E1A region: 2 T antigens E1B region: 1 T antigen E1A and E1B = Oncogenes

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DNA Tumor Viruses In Human Cancer

Common pattern
Early functions (non-structural) proteins are involved in transformation Papilloma: E6 and E7 Polyoma: Large T and small T antigen Adenovirus: E1A and E1B
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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
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DNA Tumor Viruses In Human Cancer

Herpes Viruses
Considerable evidence for role in human cancer
Some very tumorigenic in animals Integrated viral DNA found in small proportion of tumor cells: hit and run

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DNA Tumor Viruses In Human Cancer

Epstein-Barr Virus
Burkitts Lymphoma
Nasopharyngeal cancer Infectious mononucleosis (glandular fever) Transforms human B-lymphocytes in vitro

Burkitts lymphoma: malarial infested regions Nasopharyngeal cancer: China, SE Asia diet?
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DNA Tumor Viruses In Human Cancer

Human herpes virus 8 Kaposis Sarcoma Herpes Virus
Kaposis sarcoma
Hematologic malignancies Primary effusion lymphoma
Multicentric Castleman's disease (MCD) a rare lymphoproliferative disorder (AIDS) MCD-related immunoblastic/plasmablastic lymphoma Various atypical lymphoproliferative disorders
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DNA Tumor Viruses In Human Cancer

Hepatitis B Virus
DNA genome
RNA polymerase II
Host enzyme

RNA Provirus
Reverse transcriptase
Viral enzyme

DNA genome

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Hepatocellular Carcinoma (HCC)

DNA Tumor Viruses In Human Cancer

Hepatitis B continued

Vast public health problem

10% of population in underdeveloped countries are chronic carriers

Long latency

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DNA Tumor Viruses In Human Cancer

Hepatitis B continued

Epidemiology: Strong correlation between HBV and hepatocellular carcinoma China: 500,000 - 1 million new
cases of hepatocellular carcinoma per year Taiwan: Relative risk of getting HCC is 217 x risk of non-carriers
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DNA Tumor Viruses In Human Cancer

Summary

These viruses can transform cells or have lytic life cycle

Often integrate into host genome

In transformation often ONLY early genes (non-structural) are transcribed These are genes that are also necessary for a PRODUCTIVE infection

True viral genes

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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 33

RNA genome

RNA Tumor Viruses

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RNA Tumor Viruses

A normal retrovirus has:
3 genes GAG : internal proteins ENV: Envelope glycoproteins POL: Enzymes
Reverse transcriptase RNase H
Integrase Protease
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RNA Tumor Viruses

RNA is:
Diploid Capped and polyadenylated
Positive sense (same as mRNA)

Viral RNA cannot be read as mRNA (even though same sense)

New mRNA must be made Virus must make negative sense DNA before proteins are made

Therefore virus must carry REVERSE TRANSCRIPTASE 36 into the cell

RNA Tumor Viruses

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RNA Tumor Viruses

Groups of Retroviruses
Oncovirinae Lentiviruses
important

Tumor viruses and similar

important

Long latent period Progressive chronic disease Visna HIV

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RNA Tumor Viruses

Retroviruses known to cause human cancer
Human T cell lymphotropic virus -1 (HTLV-1) Adult T cell leukemia, Sezary T-cell leukemia

Africa, Caribbean S. America (Peru, Bolivia)

Some Japanese Islands
Okinawa, Kiyushu, Shikoku (12 - 16% infection rate)

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RNA Tumor Viruses

Human T cell lymphotropic virus -1 (HTLV-1)
UNITED STATES AND OTHER WESTERN COUNTRIES IV DRUG USERS

US rate of infection about one tenth of that of HIV BUT half as prevalent as HIV in IV drug users

Also causes: Tropical spastic paraparesis (affects the gray and white matter of the spinal cord - myelopathy) 1-4% of infected people
Immunosuppression
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RNA Tumor Viruses

Retroviruses known to cause human cancer
Human T cell lymphotropic virus -2 (HTLV-2)

Hairy cell leukemia

Americas, particularly in native American populations New Mexico (Navajo and Pueblo Indians) Florida (Seminole Indians) Seroprevalence in these populations > 20% Women over 50: seroprevalence - up to 50% in some populations

HIV ?

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RNA Tumor Viruses

Retrovirus Life Cycle
Bind to surface receptor Endocytosis

Fusion of membranes

Release of nucleocapsid to cytoplasm

Nucleus

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RNA Tumor Viruses

Parental RNA
Reverse transcriptase

RNA/DNA Hybrid
Reverse transcriptase

Linear DNA/DNA duplex

Circular Duplex DNA

Integrase
Host DNA polymerase

Integration
Host RNA pol II

Replication (DNA genome in cell)

Host splicing enzymes
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Transcription

Viral RNA genome

mRNA

protein

RNA Tumor Viruses

Drawback to this lifestyle Genomic RNA

Reverse transcriptase
DNA

Host RNA pol II

Genomic RNA Pol II is a host enzyme that, in the uninfected cell, makes mRNA
44 When making mRNA, pol II does not copy entire gene to RNA

Problem of using RNA pol II to copy a gene

RT primer

Viral genomicRNA

Reverse transcriptase dsDNA

RNA synthesis initiation site promotor

RNA pol II RNA synthesis termination site

45 Result: New copy of viral RNA is shorter - lacks control sequences

RNA Tumor Viruses

RNA polymerase II will not copy

Upstream sequences from transcription initiation site

Promotors / Enhancers

Down stream sequences from transcription termination site

Enhancers / Poly A site / termination site Perhaps virus could integrate downstream of a promotor etc so that the cell provides sequences OR

Virus provides its own promotors etc

BUT not copied!
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RNA Tumor Viruses

Repeat region

Clue: Difference in the two forms RNA

Repeat region

U5

GAG

POL

ENV

U3

DNA

U3

R LTR

U5

GAG

POL

ENV U3

R LTR

U5
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U5

Viral RNA

U3

R
Reverse transcriptase

U3 promotor
POLII

U5

U3

U5

Long terminal repeats are formed

POLII

RNA initiation site

RNA termination site

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Retroviruses can have only one promotor region

Contained in U3

LTR
POLII

LTR
POLII

RNA initiation site

RNA termination site

Therefore only one long RNA can be made Therefore mRNA requires processing Explains why RNA has to be positive sense
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U5

Some retroviruses have an extra gene

typical retrovirus

U5

GAG

POL

ENV

U3

Rous Sarcoma Virus

R U5 GAG POL ENV SRC U3 R
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Some retroviruses have an oncogene instead of their regular genes

Avian Myeloblastosis Virus
R U5 GAG POL MYB U3 R

Feline Sarcoma Virus (FSV) R U5

dGAG

FMS

dENV

U3

Avian Myelocytoma Virus (MC29)

R U5
dGAG

MYC

dENV

U3

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RNA Tumor Viruses

Viral Oncogene
V-onc

Cellular Proto-oncogene
C-onc
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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 cells genome
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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

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RNA Tumor Viruses

In contrast:
Chronically transforming retroviruses cause tumors inefficiently after prolonged period of time
Avian Leukosis Virus (causes lymphomas)

U5

GAG

POL

ENV

U3

No oncogene! How does it cause a tumor?

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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
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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

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RNA Tumor Viruses

Could C-oncs be involved in NON-VIRAL cancers?

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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

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DNA Tumor Viruses

How they tumors depended on our knowledge of RNA tumor viruses

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DNA Tumor Viruses Herpes

Genes can be assigned to sites on specific chromosomes

myb

mos myc

mos and myc : chromosome 8

fes

fes: chromosome 15
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Cancers often result from gene translocations Burkitts Lymphoma

8:14 translocation

Break in chromosome 14 at q32

myc

Acute myelocytic leukemia 7:15 9:18 62 11:15:17

Oncogenesis by rearrangement
Tumor
Burkitts lymphoma
B-cell chronic lymphocytic leukemia T cell chronic lymphocytic leukemia T cell chronic lymphocytic leukemia myc

c-onc
myc
bcl-1 bcl-2 tcl-1
(8)

new promotor
Ig heavy (8 to 14)

Ig light (8 to 2)
Ig heavy (11 to 14) Ig heavy (18 to 14) T cell receptor (14 inversion) T cell receptor (8 to 14)
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Oncogenes
Mutations in a proto-oncogene are dominant gain of function mutations However other oncogenic genes show recessive mutations

Anti-Oncogenes
Loss of function mutations Retinoblastoma p53
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Proto-oncogenes
Heterozygote
Allele 1 Allele 2

Dominant mutations

Homozygote
Allele 2

Allele 1

Normal

Mutant

Mutant

Mutant

Binds under special circumstances

Mutant always binds

Mutant always binds

Mutant always binds

Always binds
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Always binds

Function gained

Function gained

Anti-Oncogenes
Recessive mutations

Mutation
Rb Gene Rb
Rb protein

growth
Mutant Rb Mutant Rb

Mutant Rb

Heterozygote

Homozygote Function lost

No binding - Growth continues 66

Rb
Binds and controls cell cycle
Turns off DNA replication

Anti-Oncogenes
Retinoblastoma gene has normal regulatory function in many cells

Involved in

Retinoblastoma
Lung carcinomas Breast carcinomas
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Anti-Oncogenes
P53
Inactivated by

deletion
point mutation

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DNA Tumor Viruses Oncogenes

Adenovirus
SV 40 Polyoma BK virus Lymphotropic virus Human papilloma Virus-16

E1A region 2
Large T Large T Large T Large T E6, E7

All have a sequence in common

Mutations in this region abolish transformation capacity
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Anti-Oncogenes
Retinoblastoma

Rb Gene

Adenovirus E1A

Rb protein

Rb

105kD Rb

Rb Stops replication Cell cycle continues

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

Papilloma proteolysis

P53

DNA
replication replication
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Stops replication

Bacteria and Human Cancer

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Helicobacter pylori
Associated with:
Gastric and duodenal cancer Gastric mucosa-associated lymphoid tissue (MALT) lymphoma Inflammatory reaction to H. pylori chronic atropic gastritis (CAG) metaplasia dysplasia carcinoma Other factor (?) genetic or environmental
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