introductory of bioinformatics? · asthma nkx restless leg syndrome gallstone disease multiple...
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Introductory of Bioinformatics?
Dr . Amr El Kelish
Lecture of Plant Physiology, Faculty Of Science, Suez Canal University
PhD. Technical University Of Munich, Germany
16-08-2015
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Confirmed genetic contributors
to common human diseases (April 2007)
2000 2006 2007 2001
CF3 GCKR FTO
CDKN2A 8q24 #2 8q24 #3 8q24 #4 8q24 #5 8q24 #6
ATG16L1 IRGM 5p13 3p21 10q21 NKX2
PTPN2 IL12B
CDKN2A IGF2BP2 CDKAL1
HHEX SLC30A8
Cholesterol
Obesity
Coronary Dz
KCNJ11
2002 2003 2004 2005
PPAR
Type 2 Diabetes
IBD5 NOD2
Crohn’s Disease
CFH
Age Related Macular Degeneration
CFB/C2 LOC387715
8q24 IL23R
TCF7L2
Prostate cancer
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Cholesterol Obesity Coronary Disease QT interval Atrial Fibrillation Type 2 Diabetes Prostate cancer Breast cancer Colon cancer
KCNJ11
2003 2000
PPAR
2001
IBD5 NOD2
2005 2006 2002
CTLA4
2004
PTPN22
Age Related Macular Degeneration Crohn’s Disease Type 1 Diabetes Systemic Lupus Erythematosus Asthma Restless leg syndrome Gallstone disease Multiple sclerosis Rheumatoid arthritis Glaucoma
CD25 IRF5
PCSK9 CFH
2007
NOS1AP IFIH1
PCSK9 CFB/C2
LOC387715 8q24 IL23R
TCF7L2
CDKN2A 8q24 #2 8q24 #3 8q24 #4 8q24 #5 8q24 #6
ATG16L1 5p13
10q21 IRGM
NKX2-3 IL12B 3p21 1q24
PTPN2 TCF2
CDKN2B/A
IGF2BP2 CDKAL1
HHEX SLC30A8
MEIS1
LBXCOR
1
BTBD9 C3
8q24 ORMDL3
4q25 TCF2 GCKR FTO
C12orf0 ERBB3
KIAA030 CD226 16p13 PTPN2 SH2B3 FGFR2 TNRC9
MAP3K1 LSP1 8q24
LOXL1 IL7R TRAF1 STAT4 ABCG8 GALNT2 PSRC1 NCAN TBL2 TRIB1 KCTD10 ANGLPT3 GRIN3A
Confirmed genetic contributors
to common human diseases (Sept 2007)
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Omic’ Technologies
• Holistic view of the molecules that make up a cell, tissue or
organism.
• They include the universal detection of genes (genomics),
mRNA (transcriptomics), proteins (proteomics) and
metabolites (metabolomics) in a specific biological sample in
a non-targeted and non-biased manner.
• This can also be referred to as high-dimensional biology; the
integration of these techniques is called Systems Biology
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Central Dogma of life
Metabolomics
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Complexity level
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Genomics
• It is the systematic study of an organism's genome.
• The genome is the total DNA of a cell or organism.
• The human genome contains 3.2 billion bases
• They can reveal abnormalities such as chromosomal insertions
and deletions or abnormal chromosomal numbers in a process
called comparative genomic hybridisation.
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Research area of Genomics
• Functional genomics
Describe gene (and protein) functions and interactions)
• Structural genomics
Describe the 3-dimensional structure of every protein encoded by a given genome).
• Epigenomics
Chemical changes to DNA and histone proteins of an organism that can be passed down to an offspring
• Metagenomics Study of genetic material recovered directly from environment samples
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Importance of Genomics for Human
• All humans have 99.9% identical genetic makeup.
• The remaining 0.1% may provide useful information about diseases.
• Goal :
To show why some people get sick from certain infections and environmental changes while others do not.
Genetic Disorders
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Transcriptomics
• It is the total mRNA in a cell or organism and the template for
protein synthesis.
• Reflects the genes that are actively expressed at any given
moment.
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Transcriptomics
Specific gene expression Genes and signatures determined by particular genetic, epigenetic
regulatory factors, environmental exposures
Exploratory approaches E.g global gene expression in tumors versus healthy tissues,
differential responses to distinct environmental exposures
Disease etiology and classification Patterns/signatures rather than single markers can improve
knowledge about etiology and diagnosis
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Medicine
Disease-associated expression patterns (diagnosis)
Cell-cycle monitoring (cancer research)
Treatment-induced expression pattern (drug development and
response)
Biology
Development and Morphology (juveniles vs adults)
Interactions between organisms (antagonistic, mutualistic)
Interactions between organisms and their environments
(temperature, radiation, draught, toxins and heavy metals)
Evolution (within- and between species variation)
Functional analyses (wild type vs mutant)
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Proteomics
• Set of all expressed proteins in a cell, tissue or organism.
• Proteomics aims to characterize information flow within the cell
and the organism, through protein pathways and networks.
• The proteome is a dynamic reflection of both genes and the
environment and is thought to hold special promise for
biomarker discovery.
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Tumor markers
• Prognostic markers: malignant potential, disease recurrence
• Positioning markers: positional information (e.g. for surgery)
• Predictive markers: response to different therapies
• Diagnostic markers: detection of malignant disease
Sensitivity = detected positives / real positives
Specificity = detected negatives / real negatives
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Metabolomics
• Study of global metabolite profiles in a system (cell, tissue or
organism) under a given set of conditions.
• Additionally, as the downstream product, the metabolome is
closest to the phenotype of the biological system studied.
• Metabolom is more physically and chemically complex than
the other ‘omes’.
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Systems Biology
• Biological research focusing on the systematic
study of complex interactions in biological
systems using integration models.
• The ultimate aim is to understand whole
systems, e.g. complex cellular pathways, by
studying the effect of altered external factors
on the genome, transcriptome, proteome and
metabolome simultaneously
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Why Omics ?
• The basic aspect of these approaches is that a
complex system can be understood more thoroughly
if considered as a whole.
• Omic technology can be applied not only for the
greater understanding of normal physiological
processes but also in disease processes where they
play a role in screening, diagnosis and prognosis as
well as aiding our understanding of the aetiology of
diseases.
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Omics impact
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Human genetic variation
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Results in Paradigm shift in Life sciences
• Past experiments where hypothesis driven
– Evaluate hypothesis
– Complement existing knowledge
• Present experiments are data driven
– Discover knowledge from large amounts of data
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Life sciences research: from gene to function
Gene DNA
NH2
COOH
Protein
Genome-wide micro-array analysis
“High-throughput” protein-analysis
mRNA AAAAAAAAA
function-2 function-1 function-n
Whole-genome sequence projects
Protein function: -prediction by bioinformatics
-proof by laboratory research
cell nucleus
Gene expression by
RNA synthesis
mRNA translation by protein synthesis
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New types of data Genomes
DNA & RNA sequence
Gene expression
Protein sequence
Protein families, motifs and domains
Protein structure
Protein interactions
Chemical entities
Pathways
Systems
Literature and ontologies
36 EBI Overview
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37 EBI Overview
Genomes Ensembl
Ensembl Genomes EGA
Nucleotide sequence EMBL-Bank
Microarray & gene expression data
ArrayExpress
Proteomes UniProt, PRIDE
Protein families, motifs and domains
InterPro
Protein structure PDBe
Protein interactions IntAct
Chemical entities ChEBI
Pathways Reactome
Systems BioModels
Literature and ontologies CiteXplore, GO
37
Databases: molecules to systems
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Developments towards Bio-informatics & e-Science
• Experiments become increasingly more complex
• Driven by increase of detector developments
• Results in an increase in amount and complexity of data
• Something has to be done to harness this development
– Bio-informatics to translate data into useful biological, medical, pharmaceutical & agricultural knowledge
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Role of bioinformatics
cell
Dat
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g Genomics
Transcriptomics
Proteomics
Metabolomics
Integrative/System Biology
RNA
protein
metabolites
DNA
methodology Bioinformatics
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Genetic Research Deals
with Inherited Traits
DNA isolation.
Use bioinformatics to research differences in DNA
sequences.
Genetic researchers study inherited traits by analyzing DNA sequences.
How are we similar? How are we different?
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The Practices of Scientific Research
Hypothesis and Prediction
Data Collection and Analysis
Interpretation and Evaluation
The Scientist
Observations of Nature
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What is Bioinformatics?
Bioinformatics is the application of computer science and information technology to biology and medicine. Bioinformatics makes it possible to analyze large quantities of complex biological data and can be used to search biological databases, compare sequences, and draw molecular structures.
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Bioinformatics Tools Help Scientists: Organize, Process, and Make Sense of Complex Biological Data Sets
Protein
Bioinformatics Tools:
DNA Sequencing Identify Mutations in
DNA.
DNA RNA
Bioinformatics Tools:
RNA Sequencing Identify tissue specific
gene expression.
Bioinformatics Tools: Protein 3D Structure
visualization.
Question: What kinds of scientific questions can we answer with bioinformatics tools?
Image Source: Wikipedia Commons
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The Practices of Scientific Research
Hypothesis and Prediction
Data Collection and Analysis
Interpretation and Evaluation
The Scientist
Observations of Nature
DNA sequences from tumors
contain genetic mutations.
Genetic differences
contribute to the development of
cancer. Comparisons of genomes from
different cancers identify variants in genes X and Y.
Studies by other scientists
demonstrate that genes X and Y are involved in DNA
repair, which could explain the
increased cancer risk.
Do families with higher rates of cancer also have higher rates of DNA
damage?
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Reference …TTCACCAACATGCCCACA… F T N M P T Patient …TTCACCAACAGGCCCACA… F T N R P T
…TTCACCAACAGGCCCACA…
Extract DNA from Cells.
Sequence DNA.
Compare Patient DNA Sequence to
Reference Sequence.
Search Database to Determine if Patient
Mutation is Associated with Disease.
Patient Sample: Blood or Saliva.
Inside the Gene Machine: How Information from DNA is Acquired and Used for Genetic Testing
Genetic Counselors work with patients to help them decide whether or not to have a genetic test, and help them understand the results of the test.
Lab Technicians work with patient samples in the lab, purifying and sequencing the DNA.
Bioinformatics programmers create computer programs to help biologists analyze data.
Biomedical Researchers perform experiments with patient samples to find different variations of genes that might cause disease. Bioinformatics tools like BLAST and ClustalW are used to compare sequences.
Medical Doctors and Veterinarians use the knowledge gained from genetic testing to care for their patients.
Image Source: Microsoft Clip Art
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How DNA Sequence Data is
Obtained for Genetic Research
Genetic Data
…TTCACCAACAGGCCCACA…
Extract DNA from Cells.
Sequence DNA.
Compare DNA
Sequences to One Another.
Obtain Samples: Blood , Saliva, Hair
Follicles, Feathers, Scales.
TTCAACAACAGGCCCAC TTCACCAACAGGCCCAC TTCATCAACAGGCCCAC
Image Source: Wikipedia Commons
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Which Animals are Most Closely Related to One Another?
Gray wolf
Jack Russell
Toy Poodle
Labradoodle
Coyote
English Shepherd
Cocker Spaniel
Red fox
Image Source: Wikipedia Commons
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Multiple Sequence Alignment
of Canine DNA Sequences
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Color Coding Reveals Differences
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- Retrieving DNA sequences from databases .
- Identifying restriction sites.
- Designing polymerase chain-reaction (PCR) primers.
- Identifying open reading frames (ORFs).
- Predicting elements of DNA/RNA secondary structure .
-Finding repeats.
- Computing the optimal alignment between two or more DNA
sequences
- Finding polymorphic sites in genes (single nucleotide
polymorphisms, SNPs)
- Assembling sequence fragments
- Finding orthologous and paralogous genes.
Application of Bioinformatics
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Restriction sites
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Polymerase chain reaction (PCR)
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Tandem repetitive DNA
Tandem repeats appear in genomic DNA with a wide variety.
A tandem repeat in DNA is two or more similar copies of a DNA
sequence.
Tandem repeats may not have any functional role.
Tandemly repeated DNA sequences are widespread throughout
the human genome.
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The STR loci carry lots of desirable features which makes them
an ideal candidate for diverse applications including:
1- Forensic applications
2- Individual identification
3- True paternity/maternity detection
4- Fine scale genetic mapping
5- Inter and intra group phylogenetic reconstruction.
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Sequence Alignment
It is a way of arranging the sequences of DNA, RNA, or protein to
identify regions of similarity that may be a consequence of
functional, structural, or evolutionary relationships between the
sequences.
Aligned sequences of nucleotide or amino acid residues are
typically represented as rows within a matrix.
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Sequence Assembly Aligning and merging fragments of a much longer DNA sequence
in order to reconstruct the original sequence.
In DNA sequencing technology cannot read whole genomes in one
go, but rather reads small pieces of between 20 and 30000 bases,
depending on the technology used.
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Orthologs vs Paralogs
Orthologs
They are genes of common ancestry between species.
Eg: Dog hemoglobin and human hemoglobin, for example are orthologous. Paralogs
They are when genes are duplicated then one of the copies evolves a new function.
Eg: Humans would be myoglobin and hemoglobin; both are oxygen storage proteins derived from the same ancestor gene, but have different functions (one is a transport protein in red blood cells, the other is a storage protein in muscle cells).
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Impact of Genomics in Medicine
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71
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72 EBI Overview
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73 EBI Overview
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Top Reasons To study
Bioinformatics/Computational
Biology
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Computing is the key skill set for 21st century biology
Biology is becoming a more quantitative science.
In the future, new discoveries will require leveraging big
datasets and using advanced analytical methods.
Big data and complex models require computational skills.
There is no way to escape this reality.
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Nobel-prize winning pioneer • Walter Gilbert:
“To use this flood of [sequence] knowledge, which will pour across the computer networks of the world, biologists not only must become computer literate, but also change their approach to the problem of understanding life”
• Sydney Brenner:
“I spent many hours persuading people that computing was not only going to be the essential tool for biological research but would also provide models for analyzing”
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Computational skills are highly transferable
Computational skills transfer across this sector, plus a much
wider market outside of the (bio)science.
Increasing your computational chops will give you a better
chance at landing a job.
You will have a deeper appreciation for how computers
work and more mastery of when you interact with
computers in your daily life
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Computing will help improve your core scientific
skills
Computing forces you to confront and tame the very human
tendency to do science in ad hoc ways and therefore it
naturally develops core scientific skills such as:
• Logically planning experiments
• Collecting data consistently
• Developing reproducible methodology
• Analyzing your data with proper statistical methodology.
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You should use you Ph.D./Post-Doc to develop new
skills
Majority of Biology Ph.D. students have no training in
scientific computing skills beyond using Excel or statistics
package.
So you have to start training in something new, not just
further developing a skill set that you already have.
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You will develop a more unique skill set in Biology
Majority of Biologists have experimental training, but very
few have advanced Computational training.
And because you will be able to get results that many others
cannot, plus the fact that you will have skills that set you
apart from the herd, you will be more competitive on the job
market.
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You will publish more papers
Computational Biology will provide you more than your
fair share of failed experiments.
As a result, you are very likely to publish more papers
per unit time in Computational Biology.
Funding agencies want to see the junior researchers who
have good ideas and can take them to completion.
Publication is the proof that you can finish projects.
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You will have more flexibility in your research
You can move from flexibly from topic to topic more easily
than you can if your skill set is linked to specific
experimental techniques.
This flexibility in scope allows you to satisfy your
intellectual curiosity or chase the latest trend as you wish.
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You will have more flexibility in working
practices
• Unlike being chained to the bench, you can do working
from home.
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Computational research is cost-effective
• Computational Biology research is cheaper than most
experimental work that requires a large consumables
budget.
• Cost-efficiency is also very important when you are
starting your group and for maintaining continuity of
productivity when riding out troughs in funding or group
size.
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A successful scientist ends up in an office
• The truth is that the native habitat for an academic
researchers is sitting in their office in front of their
computer.
Webb Miller (most highly-cited bioinformaticians)
• If you’ve really wanted to do research since you were
young, then ask yourself: why train in skills you will
never ultimately use for the majority of your career, while
somebody else in your lab gets to have fun making all the
discoveries?
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According to Fast Future report
• Computational biology is going to be among the most
popular jobs of the future.
• Besides, most NGS data will sit untouched and unloved
unless there are better methods to collect information
about phenotype.
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Safer and Environment friend
Mason Vail :
“You’re far less likely to spill a pathogen on yourself if you
avoid the wet lab altogether “.
Another reason that came up in recent discussions is that
computational work is more environmentally friendly.
Less plastic waste, less energy spent on autoclaving, etc.
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