BioMarker

Definition:
Biomarker is a substance used as an indicator of a biologic state
Existence of living organisms or biological process.
A particular disease state
A fragment of DNA sequence
Proteins
Nucleic acids
Carbohydrate
Lipids
Small molecules

Biomarkers
Location of the biomarker:
Location of a particular molecule can also be a marker
Cellular-subcellular locations
Tissue or organ locations
Thymidylate synthase (TS)
nucleus vs the cytoplasm
The higher the level in NE, lower level in the cyto.
Lower survival in colorectal cancer patients
Galectin-3
Galectin-3 is a member of lectin family. It is wildly distributed
In tissue of epithelial, fibroblast, and dendritic cells.
Blood Galectin-3
Predict the outcomes for patients with symptoms of heart failure

Biomarker
Detection of biomarker
Detection of biomarker diagnosis
Self properties, e.g enzymatic activities
Antibodies, IHC, ELISA
Detection of biomarker
Quantitative
a link between quantity of the marker and disease
Qualitative
a link between exist of a marker and disease

Biomarker & Diagnosis

Ideal Marker for diagnosis
Should have great sensitivity, specificity, and accuracy in reflecting total
disease burden. A tumor marker should also be prognostic of outcome and
predictive of tumor recurrence and effectiveness of anti-cancer treatments.
Biomarker for Screening
The marker must be highly specific, minimize false positive and negative
The marker must be able to clearly reflect the early stage of disease
The marker must be easily detected without complicated medical
procedures. The disease markers released to serum and urine are good
targets for application of early screening.
The method for screening should be cost effective.

Samples for biomarker detection

Blood, urine, or other body fluids samples
Tissue samples

Biomarker Discovery and Validation

Correlation: a biomarker vs a disease or status of a disease
Do not need understand functions

Detection: Detection of a particular marker is important

Validation: Build statistical correlation large number of samples
Validation: sensitivity and specificity
Validation: Stand alone vs along with other markers

Common Serum Markers for Cancer

Diagnosis/prognosis
AFP
Lung

CEA

CA15-3

CA19-9

CA125

Pancreas

Kidney

Breast

Ovarian

Cervical

Uterine

x
x

PSA

PSAf

PAP

hTG

HCGb

Ferr

NSE

x
x

x
x

Prostate

x
x

Liver

Gastro
Colon

x
x

Bladder

Brain

x
x

Myeloma

Thyroid
Testicular

x
x

A2M

x
x

Leukemia

B2M

x
x

Common Biomarker for Cancer

AFP (>100 ng/ml)
Alpha fetoprotein (AFP) is typically found in the developing fetus.
Because of the association of the rapid cell growth, this fetal protein is
also used as a tumor marker. non cancerous liver diseases such as
cirrhosis and viral hepatitis can lead to high level AFP

CEA (>10 ng/ml)

Carcinoembryonic antigen (CEA) is produced during fetal development. After
birth, the production of CEA stops and is undetectable. CEA has also been
found elevated in nonmalignant tumors such as pleural effusions. Elevation of
CEA after conventional treatment of neoplasms has been correlated with a
recurrence of cancer

CA19-9 (>37 U/ml)

Carbohydrate Antigen 19-9 (CA 19-9) is present in the fetus in the epithelium
of the fetal stomach. It is primarily used as a marker for pancreatic cancer.
High levels also exist in conditions such as non-malignant liver disease and
other disorders of the gastrointestinal tract.

Common Biomarker for Cancer

hTG (>10 ng/ml )
Thyroglobulin (hTG) is a glycoprotein that is found in thyroid gland. This
protein binds to thyroxine, which controls the rate of metabolic processes
in body.

HCG (>10 mIU/ml )

Human Chorionic Gonadotropin-beta (HCG) is normally produced by the
placenta during pregnancy, an indicator of pregnancy. The protein can be
detected in serum or urine. Non-malignant elevations may be observed in
pregnancy, ulcers, dukes disease, and cirrhosis. Levels of HCG are useful in
monitoring the effectiveness of treatment.

Ferr (>120 ng/ml )

Ferritin is an iron binding storage found in the liver, spleen, and bone marrow.
Elevated levels observed in non-cancerous conditions include rheumatoid
arthritis and anemia

Common Biomarker for Cancer

NSE (>12 ng/ml )
Neuron Specific Enolase (NSE) is produced by neurons and
neuroendocrine cells of the central and peripheral nervous system.

b2M (high level)

Beta 2-Microglobulin, (B2M), is an 11 kD protein associated with the outer
membrane of many cells including lymphocytes. It is the small subunit of the
MHC class I molecule. B2M is present in small amounts in serum, csf, and urine
of normal people. A non-malignant condition associated with high b2M levels is
pancreatitis

a2M (<500 g/ml )

Alpha 2 Macroglobulin (A2M) is a large protease inhibitor, 720 kD, capable of
irreversibly binding and inhibiting a wide variety of proteases including
plasmin, pepsin, trypsin, chymotrypsin, and cathepsin D.

Ovarian Cancer Diagnosis by CA-125

CA-125, a product of the MUC16 gene, is a mucin made by certain cells in the
body which include those of the uterine tubes, uterus, cervix, and the lining of
the abdominal and chest cavities
CA125 is a membrane glycoprotein that has very short cytoplasmic domain
and a very long extracellular domain. The cellular function of the protein is
still unknown. The protein exist in the cells of normal and cancerous tissues
of ovarian.
The release of proteolytic fragments of CA125 leads to elevation of CA125
level in bloodstream, which is associated with progression of ovarian cancer
and a few other cancer types. The CA125 level can be over 10 folds higher
in ovarian cancer patients compared to the level in normal.
Diagnosis of ovarian cancer is often simple. A pelvic mass is suspicious and
is very often associated with ovarian cancer. However, when the pelvic mass
is detected, the ovarian cancer usually has already advanced to quit
advanced stage. There is no symptom for ovarian cancer, especially during
early stage.

Her-2/neu
The HER (erbB) family member are
Growth factor receptors.
In the HER (erbB) family, only Her-2
did not have identified ligand.
Expression level correlates with growth
of breast cancer cells.

Diagnostic/prognostic applications.
HER2-positive metastatic breast cancer have a more aggressive disease, greater
likelihood of recurrence, poorer prognosis and decreased survival.

Beta-2-microglobulin (b2M)
B2M is the small subunit of the MHC class I
molecule.
How does the protein promotes tumor growth
and metastasis the expression of MHC
class 1 in many tumor cells are repressed.
An hypothesis is that b2M is a signaling
molecule that linked to cell growth and
survival pathway.
What are the target of the signal? Receptors?
Known or unknown pathway?

Biomarker & Targeted Imaging

Imaging probe link to a targeting moiety that target to bio-marker
Basic requirement: the marker must presented on cell surface in high number

Ideally:
The interaction between marker and targeting moiety is strong
The interaction is highly specific
The target internalization high payload
Distribution at neovasculature, minimal tissue penetration

Imaging Targeting PSMA

Antibody is targeting moiety
Antibody conjugated quantum dots

Antibody

Imaging Targeting PSMA

Antibody conjugated to radio-labels

Patient Images
No resolution hardly tissue image
High sensitivity good for detecting metastasis

MR Image Target Her-2

Advantage for both
Her-2 and PSMA imaging
is that the high number
and distribution of the
marker in agiogenesis
tissue
Disadvantage for both
Her-2 and PSMA imaging
is that the ligand for Her2 and PSMA are not
identified. Antibodies are
the only choice

The levels of both Her-2 and PSMA correlate well with aggressiveness of
disease

Therapies Target Biomarkers

Simply target the biomarker by antibody
Therapeutic agent link to a targeting moiety that target to bio-marker
Basic requirement: the marker must presented on cell surface

Ideally:
The interaction is highly specific.
The target internalization, accumulation good # of agent.
Distribution at neovasculature, minimal tissue penetration.
The level of marker correlate with disease progression

Therapy Target Her-2

Herceptin
Herceptin binds to HER2-positive cancer cells and may block them
from dividing and growing.
Herceptin attaches to the HER2-positive cancer cells and may
signal the body's immune system to destroy the cell.
Herceptin can also conjugated with chemotherapy (paclitaxel)
to destroy HER2-positive cancer cells.

Therapy Target PSMA

One important advantage is the quick
endocytosis of the PSMA upon the
antibody binding.
Due to main neovascular distribution,
targeting of PSMA is very easy, less
tissue penetration.
Both Her-2 and PSMA have a good
property is their high level correlate well
with aggressiveness of disease.

Imaging and Therapeutic Agent

The most significance of the double agent is the immediately follow the effectiveness
of treatment. Also follow whether the therapies have been targeted to the sites

Diagnosis by Proteomic
No single marker can
accurately reflecting a
disease. Not even a
panel of markers.

We need analyses a
large number of disease
markers.