functional genomics and proteomics - masterthesissen 2011.pdf · signalling system (ckr‐2&...

Functional Genomics Functional Genomics and Proteomicsand Proteomicsand Proteomicsand Proteomics

Prof. dr. Liliane Schoofs

// / f/ Shttp://bio.kuleuven.be/df/[email protected]

Big ideas from small brains

Conserved mechanisms:

Caenorhabditis elegans

mechanisms: reproduction,

locomotion, aging, digestion, …

Caenorhabditis elegans 302 neuronsHuman brain

~100 billion neurons

Main research topics:A i• Aging

• Cancer• Feeding behavior• Reproduction• Malaria

Characterization of novel neuropeptide-mediated signaling systems in C. elegans

Screening for GPCR ligand

Expression analysis

Neuropeptides are key regulators of animalphysiology (feeding, reproduction) throughbinding of GPCRs.

Behavioral assays

e.g. locomotion RNAi knockdownC. elegans: neurobiological model

Contact:Contact: [email protected]@bio.kuleuven.be

The GnRH receptors of the model organism The GnRH receptors of the model organism C. elegansC. elegans: a : a good target for the development of nematicides?good target for the development of nematicides?good target for the development of nematicides?good target for the development of nematicides?

8 GnRH receptor genes

Techniques:- Molecular biology- Reverse pharmacology

Structure activity studiesConserved GnRH like ligand

Potential

- Structure-activity studies- Phenotyping: egg-laying assays, …- Microfluidics (lab on chip)


nematicide targets ?Lab-on-a chip

The GnRH receptor: a suitable target to b t l i it d t t fli ?combat malaria mosquitoes and tsetse flies?

Major physiological functions in insects regulated by peptide hormones thatMajor physiological functions in insects regulated by peptide hormones that interact with G protein‐coupled receptors

Adipokinetic hormone (AKH) in insectsAdipokinetic hormone (AKH) in insectsevolutionary linked to gonadotropin‐releasing hormone (GnRH)

AKH insect energy metabolism

AKH receptor and other peptide hormone receptors astarget for a new generation of insecticides based onpeptidomimetica

h l l b lTechniques: ‐Molecular biology‐ Reverse pharmacology‐ Structure‐activity studies


mutagenmutagen

Genoom sequencing

Wormsorter Fluorescent reporter protein for reproduction

Genome sequencing


Signalling system (ckr‐2 & nlp‐12) feeding behaviour, energy homeostasis,

Cloning and deorphanizing

Signalling system (ckr 2 & nlp 12) feeding behaviour, energy homeostasis,reproduction, induction of dauer state and parasitism

Parastrongyloides trichosuri ckr‐2 receptor

Structure‐activity studies on parasitic and C. elegans ligandsligands

host

Hairpin RNAi of receptor and ligand & fenotypingAustralian possum


Proteomics and Proteomics and peptidomicspeptidomics of longof long--lived lived C. C. eleganselegans

Many life-extending interventions available in C. elegans→ insulin pathway mutations, dietary restriction,…

Studying long-lived worms to help us combat aging-associated diseases→ interventions that increase lifespan also decrease incidence of cancer, neurodegenerative diseases

Techniques

neurodegenerative diseases,…

PeptidomicsPeptidomicsDifferential labeling of peptides Liquid chromatographyMass spectrometry

HowHow do the do the endocrineendocrinesystemssystems influenceinfluence agingaging??Mass spectrometry

ProteomicsProteomicsGel electrophoresis (2D-DIGE)Mass spectrometry

WhatWhat are the are the effectorseffectorsthatthat controlcontrol agingaging??

Contact:Contact:[email protected]@bio.kuleuven.be

Fatal attraction: food preference in C. elegans

A

Choice between A: E. coli OP50, standard food source

B

B: Aeromonas hydrophila,pathogen

Fatal attraction: Worms display a clear preference towards Aeromonas hydrophila over their normal, non-pathogenic, E. coli OP50 diet

B

y p , p g ,

What are the neuronal circuits in the brain that are controlling this behaviour?

Analyse the choice behaviour of odorant receptor mutants in C. elegans.

Which aphid brain factor determines the switch from asexual Which aphid brain factor determines the switch from asexual to sexual reproduction?to sexual reproduction?

Aphid = agricultural pest, easy to breed, endosymbionts, diverse host-pathogen interactions, reproductive plasticity, viviparous ...

to sexual reproduction?to sexual reproduction?

.g

⇒ Research model

⇒ Genome Sequencing Project⇒ Genome Sequencing Project

⇒ -omics approach for physiological questions

Summertime: asexual LD SDPhotoperiod shortening

F ll it h t d l .Summertime: asexual reproducing females

LD SD Fall: switch towards sexual reproducing females

.destruction of group 1 braincells

determine group 1 content + compare LD with SD

destruction of group 1 braincells ~ sexual reproduction!

. International context: Erasmus when interestedmethods: micro-dissecting, confocal microscopy, proteomics, mass spectrometry, pyrosequencing ...


.

The search for biomarker candidates for colorectal cancer

• Biomarker: try to detect the tumor in an early curable stage• Link between cancer and inflammation• Macrophages and their progenitor cells are an important ‘surrogate’ tissue

• Tumor associated macrophages promote tumorprogression and metastasis

• Phenotypic switch between M1 and M2 macrophages• Easy to isolate from whole blood

• Search for proteins that are differentially expressed between healthy individuals and colorectal patients• Techniques:

• proteomics (gel‐based and gel‐free)• mass spectrometry


functional genomics and proteomics - masterthesissen 2011.pdf · signalling system (ckr‐2&...

Documents