Download Plataformas de apoyo a la investigación

Plataformas de apoyo a la
investigación:
biobancos y centros de recursos
biológicos
Jornadas:
“La realidad de los biobancos a la luz del nuevo marco
normativo: retos e implicaciones”
Centro de Investigación del Cáncer
Madrid, 16 de diciembre de 2009
SOCIAL REQUIREMENTS FOR SUPPORTING
RESEARCH: THE DELAYING FACTOR
PATIENT
(SOCIAL
ENVIRONMENT)
Requirement for
immediate results
BASIC
BIOMEDICAL
RESEARCH
Long-term
results
TRANSLATIONAL RESEARCH ENVIRONMENT
Hypothesis
Research objectives
CLINICAL
QUESTIONS
CLINICAL
SOLUTIONS
BASIC
BIOMEDICAL
RESEARCH
Research
results
PATIENT
(CLINICAL
PRACTICE)
CRITICAL FACTORS FOR THE SUCCESS OF
TRANSLATIONAL RESEARCH PROGRAMS
-
Adequate legal framework
-
Appropriate institutional organization
-
Physical proximity between basic and clinical
research
-
Translational research groups
-
Critical scientific mass
-
Cultural/educational change
-
Increased research funds
-
Availability of technological platforms
-
Active role of industry with partnership
Biobanks in Medical Research
Fragmentation
Cancer Genome Project struggels
ELSI
with sample shortage
Funding
(Nature Med 2007)
Technical
Follow-up
700,000
Cancer deaths
(North America; 2007)
NCI:
OECD:
WHO/IARC:
ESF:
EU/ESFRI:
BIOBANKING
Biological samples are #1 roadblock
Global Biological Resource Centre Network
Standards for biological resource centres
Science Policy Briefing: Need for integration
Research infrastructure for Biobanks and
Biomolecular Resources (BBMRI)
BIOBANKS & BIOLOGICAL RESOURCE CENTRES
Major role
• To obtain, process and store biological samples and
their associated data.
• To provide DNA samples to research groups.
• To ensure a rational, effective, ethical and legal use of
the available resources.
The real value of a biobank lies on the existence of
“cooperative research projects of excellence”.
Predict usage of samples far in advance: decide on type of sample,
sample conditions and manipulation (sample fractioning, cell & fluid
components of interest…)
“MOVING” BIOMEDICAL RESEARCH TARGETS
Genome
Genes in tissues
Gene expression
profiles (GEP) in
tissues
Tissue
proteomics
“MOVING” BIOMEDICAL RESEARCH TARGETS
Genome
Genes in tissues
Life style
Environment
Gene expression
profiles (GEP) in
tissues
Tissue
proteomics
Biobank
ASSOCIATED DATA
Epidemiologic questionnaire:
Disease questionnaire:
(Specific for each type of disease)
-General Health
(familiar antecedents)
- Nutritional and lifestyle habits
(smoking, alcohol, physical activity…)
- Demographic data
(family, place of living, culture…)
- Genealogic data
(two generations)
- Phenotypic data
(biochemical analysis, virology,
anthropometrical measurements)
Data always included:
- Clinical data
- Diagnostic data
- Outcome data
- Associated diseases
PROTEOMICS: THE WORLD OF PROTEINS
G0/G1 checkpoint
Proteomics 2009
Size exclusion chromatography 0.5-1h
Size
Fractions:
OVERNIGHT
INCUBATION
Bead array
FACS
IP+Western blots: weeks
IP+ Mass-spectrometry: months
ELISA: lack of sandwich pairs
MINUTES
ARRAYS OF BEADS COUPLED TO DIFFERENT
ANTIBODIES CAPTURING MOLECULES OF INTEREST
Anti-CD3
Anti-CD4
Anti-CD8
Anti-bcr
Anti-JNK
Anti-ERK
Anti-Akt
Anti-Abl
Anti-STAT5
Lund-Johansson et al, Cell Mol Proteomics, 2008
G1
(singlets)
Alexa-488
Pacific Blue
mIgG PE/Cy7
Forward Scatter
Alexa-647
Side Scatter
BEAD ARRAY (n=1300 proteins)
Pacific Orange
mIgG PE
Resolution of protein complexes with dual specificity
BETA 2 MICROGLOBULIN /MHC CLASS I
Go/G1 Cell cycle checkpoint
QUIESCENT CELLS
100000
Protein/bead (PE MFI)
Protein/bead (PE MFI)
10000
Cdk6 p19/INK4d Control Ig
1000
10000
B2M-01
B2M-02
p16
100
Cdk6
10
v 70
6
0
44
8
15
1000
PROTEIN SIZE
2m/HLAI/X 2m/HLAI
2m
100
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22
PROTEIN SIZE
75
43
Resolution of protein complexes with dual specificity
Lund-Johansson et al, Cell Mol Proteomics, 2008
Complexity of multiplexing protein
measurements
- Presence of specific proteases (i.e. decreased or undetected
protein levels)
Cocktail of protease inhibitors (i.e. small molecules)
- Protein localization: membrane, cytoplasmic and nuclear (i.e.
membrane or DNA binding with difficult access for Abs).
DNA digestion
- Protein status (i.e. phosphorylated vs non-phosphorylated monomeres
vs multimeres and protein complexes with varying access to antibodies)
Inhibition of phosphatases
One condition per tube for detection of all proteins
STORAGE OF PB SAMPLES FROM BREAST CANCER
Purify cells of
interest*
*Abs against cell surface structural/non-functional proteins
Immunophenotypic identification of PB
B-cells with a CLLCLL-like phenotype
Clonal B
cells
Normal B
cells
*0.35% of all B-cells & O.O3% of all leucocytes
Frequency of PB CLL
CLL-like BB-cell clones in
healthy adults according to age
Found in 94/637 cases (14.8%)
100
90
75%
80
26%
Percentage of cases
70
60
21.3%
50
18.5%
40
30
5.9%
20
5.7%
10
0
40
45
50
55
60
65
70
Age (years)
75
80
85
90
95
Nieto et al, Blood, 2009
“MOVING” BIOMEDICAL RESEARCH TARGETS
Genome
Genes in tissues
Life style
Environment
Gene expression
profiles (GEP) in
tissues
Tissue
proteomics
Single cell
proteomics
Tissue
homeostasis
Regulation of
GEP in single
cells in a tissue
FROM BIOBANKS TO BIOLOGICAL
RESOURCE CENTRES
ADDITIONAL TASKS
EXPANDED
SERVICES
NEW ACTIVITIES
Biomol.Resources:
- Genomics
- Proteomics
- Lipidomics
Biobanking Research:
-New
-New
-New
-New
technologies
collaborations
services
nodes
Biobanking Education:
-
Pre-analytics
Cell purification
Cryobiology
Nanotechnology
- EQC program
- Master degree
CENTRO DE CARACTERIZACIÓN DE RECURSOS BIOLÓGICOS
INCORPORACIÓN DE INVESTIGACIÓN EN PROCEDIMIENTOS Y
TECNOLOGÍAS DE ÚLTIMA GENERACIÓN
FASE PREANALÍTICA
FRACCIONAMIENTO Y
PURIFICACIÓN CELULAR
OBTENCIÓN
DE LÍNEAS
CELULARES
NANOTECNOLOGÍA
MOLECULAR
CRIOBIOLOGÍA
BBMRI: A EUROPEAN HUB STRUCTURE
THE DISTRIBUTED HUB STRUCTURE OF BBMRI
Secretariat
(statutory seat)
National members
Biobanks
Biomolecular resources
Technology centers
Associated partners:
Hospitals
Universities
Service providers …
MUCHAS GRACIAS
www.bancoadn.org