Personalized medicine in the “-omics”

era: how can Nanotechnology help?

Ricardo Franco, REQUIMTE, Portugal
 Personalized Medicine: Definition

“Personalized medicine is the use of diagnostic and

screening methods to better manage the individual
patient’s disease or predisposition toward a disease….

“Personalized medicine will enable risk assessment,

diagnosis, prevention, and therapy specifically tailored to
the unique characteristics of the individual, thus enhancing
the quality of life and public health.”

– NHLBI Strategic Planning, Theme #10

 Addressing the Complexity of
Addressing the complexity of disease
Cardiovascular Disease

Metabolic pathways

Gene expression profile

 A New Biomarker Toolbox:
The output of the “-omics” approach
Shift to Personalized Care

Human Genome Sequence Polymorphisms ~ 10,000,000

(Genomics)

Gene Expression Profiles Microarrays of ~ 25,000 gene

(Transcriptomics) transcripts

Proteome Protein arrays of specific protein

(Proteomics) products ~ 100,000

Metabolome Small molecule metabolites

(Metabolomics) ~ 5000
 Example: Gene Expression
Example: Gene Expression Profiling
Profiling
 Concept of “Biosignature”

Insult or Therapy
Patient A Patient B

Arrays for
genomics,
transcriptomics,
proteomics,
“Biosignatures”
metabolomics

Outcome A Outcome B
 The Ultimate Goal

Personalized
Clinical data
Healthcare
Genomics
Megabytes – Gigabytes
of Data Per Patient
Transcriptomics

The patient has disease X, subclass

Y, which will likely respond to drug Z

Proteomics
 How can Nanotechnology help?

• Producing high throughput diagnostic assays

– laboratory context effective, inexpensive
– at home + easy to use, portable, personalized (“point-of-
care”)

• Creating innovative therapeutic modalities that leverage

the validated systems biology outputs for exquisitely
specific individualized therapy.
 Cheap and reliable health care technologies

The Bill and Melinda Gates Foundation

http://www.grandchallenges.org

“A Point of Care Diagnostic System for the Developing

World”

More
affordable
 Bio-barcode assay for the detection of nucleic acid
and protein targets without PCR
Extraordinarily sensitive, showing high sensitivity for both nucleic acid and protein targets.
Two types of particles are used: (i) a magnetic microparticle with recognition
elements for the target of interest; and (ii) a AuNP with a second recognition agent
(sandwich format) and hundreds of thiolated single-strand oligonucleotide barcodes.

Jwa-Min Nam, et al. Science 301, 1884 (2003)

 Microarray scanometric immunoassay with AuNP
probes and signal enhancement with silver/gold

PSA detection:
• 300 aM (∼9000 copies) in buffer
• 3 fM in 10% serum

Kim, D. et al., Anal. Chem. 81, 9183, (2009)

 Prototype for paper "chip" technology
George Whitesides, Harvard Univ.

Could be used in the developing world to cheaply diagnose deadly diseases

such as HIV, malaria, tuberculosis, hepatitis and gastroenteritis.

"I think it's real. It can be very useful, but it's not the fanciest manifestation of
lab on a chip by any means. If it works and it's cheap, it's good for
everybody.“
Keith Herold, Associate Professor of Bioengineering at the University of Maryland ,

http://www.cnn.com/2010/TECH/02/25/whitesides.chip/index.html
 Non-cross-linking assay – DNA functionalized
gold nanoparticles in a colorimetric kit format

Patent pending: PT 103730

 Assay for Mycobacterium tuberculosis DNA detection
in clinical samples – no PCR needed; inexpensive

P. V. Baptista, et al., Clin. Chem. (2006), 52 (7), 1433; P. V. Baptista, et al., Anal. Bioanal. Chem. (2008), 391, 943
 Genotyping of Single Point Mutations (SNPs) involved
in xenobiotics metabolism and obesity
CYP450 enzymes
90% of xenobiotics are metabolized by:
CYP1A2, CYP2C9, CYP2C19, CYP3A5, CYP3A4 and CYP2D6 enzymes

Genetic variability influences a patient’s response to a particular xenobiotic

CYP2D6
Involved in the metabolism of 20% of prescribed xenobiotics
- such as antiarrhythmics, antidepressants, analgesics, antihistamines, beta-blockers, etc.

“tag” SNP → Haplotype → Genotype → Phenotype

Ultrarapid metabolizer

Extensive metabolizer (i.e. standard)

Poor metabolizer

Human DOR
Polymorphisms in the DOR gene cause obesity and Type 2 diabetes
Armada-Bras J et al (2007) Int J Obesity 31 (Supp. 1s):S20
Baumgartner BG et al (2007) PLoS One, 2:e1183

www.sciencedaily.com
 Genotyping of Single Point Mutations (SNPs) involved
in xenobiotics metabolism and obesity
CYP2D6
detection of *4 haplotype → 1846(G/A) “tag” SNP

Higher

Abs Ratio

Lower
 Genotyping of Single Point Mutations (SNPs) involved
in xenobiotics metabolism and obesity
DOR
detection of DOR1(G/C) SNP

G. Doria, et al. Colloids Surf. B: Biointerfaces (2010), 77, 122

STABVida: our partners in personalized medicine
solutions (genomics; transcriptomics; metabolomics
www.stabvida.com)
 Genotype Significance
Associated with 1.76 cm
Chromossome Locus Gene SNP higher waist perimeter
AA compared to genotype GG
Genotipo Significado
18Genótipo 18q21.32 Significado
MC4R rs12970134
CC Normal CC Normal
Associated with 0.88
DAVID MARISCAL
Orfeu FLORES
cm higer waist perimeter
AG
CG Risco mais elevado em dietas compared to genotype GG
com alto teor em gorduras CG Riesgo en dietas con alto contenido en
grasas

GG Risco mais elevado em dietas GG Riesgo en dietas con alto contenido en

com alto teor em gorduras grasas
GG Normal waist perimeter

Genotipo Significado

CC Normal
DAVID MARISCAL

CG Riesgo en dietas con alto contenido en

grasas

GG Riesgo en dietas con alto contenido en

grasas
Contributors

Eulália Pereira Ricardo Franco

Peter Eaton
Gonçalo Doria (STABVida)

Pedro Baptista
Financing

PTDC/SAU-BEB/66511/2006
PTDC/QUI/64484/2006;
SFRH/BDE/15544/2005
THANK YOU FOR
YOUR ATTENTION