Molecular Biology

for
Clinicians
Prof Ernesto Gil Deza MD
Instituto Oncologico Henry Moore
Universidad de Salvador
 Objectives

Definition

Pathogenesis

Clinical applications
Molecular Diagnostic Tools

DNA RNA
Citogenetics Transc. Profiling
FISH and CISH Northern blot
CGH RT-PCR
ISH, FISH & CISH
CGH arrays
SNPs
PCR sequencing Proteins
Western blot
Southern blot
2D electrophoresis
Microsatellite MALDI-TOF
instability
SELDI-TOF
Microarray IHC
Molecular Biology
Some problems...
 Sample tissue

Gross Tumor Specimen

Fine Needle Aspiration

Microdissection of Pure Tumor Cells

Paraffin block

Frozen sample

Fresh tissue
 Quantitative
Human Genome

Nuclear Mitocondrial
3000 Mb 16.6 kb
~30.000 genes 37 genes

Genes Extragenetic
30 % 70%

80%
20%

Low number Higher number

10% 90% of copies of copies

Encoded Non encoded

 Statistics
20.000 - 50.000 genetic features
• for only few outcomes:

Response/No responce

Recurrence/No recurrence

Survival/death
• Greater chances of false positive results
Pathogenesis
Oncogenesis

Hannahan and Weinberg Cell 2000

Genetic instability

Preinvasive

Invasive
Clinical applications
Clinical applications
Risk prediction

Cancer diagnosis

Differential diagnosis

Tumor subtypification

Pharmacogenetics

Prediction of clinical outcomes

Novel targeted therapeutics

Modified Workman & Johnsto JCO October 10th 2005
 Risk prediction

Low penetrance high risk

BRCA1

BRCA2

High penetrance low risk

Ethical issues
 Cancer diagnosis

Differencial diagnosis ADH - DCIS - CDI

DNA microarray of ductal lavage

Nanoparticles and blood products

 Tumor
Subtypifications
Disagreement in ICH
Molecular Breast Cancer Subtype Dendrogram

Carey - ASCO 2005

Classification requirements

Texto
Prognosis
Prognostic and natural history

2000 1870 500 BC

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 “Due to the nature of the genes included in the
21 gene assay, which are mainly proliferation genes
and ER-related genes, it could be argued that the
assay is an expensive replacement for ER and Ki67 inmunohistochemical
assays”
Paik and Kim, page 21
 Molecular predictive
factors

ER

PR

Her2
• All are good NEGATIVE predictors.
Expectations and reality
•We can...
Identify hallmarks in tumorigenesis.
Classify tumors according to molecular subtypes
Predict high risk in a few patients
Avoid toxicity in some patients
Design more sophisticated predictive tools

•But...

•Can we save lives, cure more or take care of our

patients better than we do actually?
 Conclusion

“I have loved no darkness,

Sophisticated no truth,
Nursed no delusion,
Allowed no fear”

Aequanimitas

Sir William Osler

1880
Thank you very much
for your attention