Crohn Disease : from gene to therapy

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Crohn Disease from gene to therapy

• Prof Jean-Pierre Hugot
• Hopital Robert Debré
• Paris, France
• Jean-pierre.hugot_at_rdb.aphp.fr

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Gordon Oppenheimer, Burill B Crohn and Leon
Ginzburg, 1969
From  BB Crohn, Life and Work , Falk Foundation
2000
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Treatments for Crohn Disease

• Antibiotics
• Probiotics
• Nutritional support
• 5 amino-salicylates
• Steroids

• Immunosupressive drugs
• Antibodies against TNF and other chemiokines
  (IL12, ICAM, a4integrins)
• Pig parasites

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Bouma G et al. Nature Rev Immunol 2003
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IBD loci
DLG5
IBD3
IBD7
IBD2
SLC22A4/5
1 2 3 4 5 6
7 8 9 10 11 12
IBD8
IBD6
IBD4
CARD15
13 14 15 16 17 18
19 20 21 22 Y X
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Ibd1-Nod2-Card15
N ter
C ter
NF-kB Activation
Oligomerisation
Bacterial recognition
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Why to discover genes?

• To make a diagnosis
• Before disease occurrence (if prevention
  possible) or to avoid invasive procedures
  (sensibiility, sensitivity).

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Proportion of mutated patients (all non
conservative variations )
except P268S
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Why to discover genes?

• To make a diagnosis
• Before disease occurrence (if prevention
  possible) or to avoid invasive procedures
  (sensibiility, sensitivity).
• To classify the disease (definition of disease
  subgoups)

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NOD2/CARD15 mutations are associated with

• A younger age of onset
• A more frequent ileal involvement
• An higher complication rate
• Lesage et al. Am J Hum Genet 2002
• Vermeire et al. Am J Hum Genet 2002
• Cuthbert et al. Gastroenterology 2002
• Hampe et al. Lancet 2002
• Amhad et al. Gastroenterology 2002
• Abreu et al. Gastroenterology 2002
• Louis E et al. Gut 2003

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Why to discover genes?

• To make a diagnosis
• Before disease occurrence (if prevention
  possible) or to avoid invasive procedures
  (sensibiility, sensitivity).
• To classify the disease (definition of disease
  subgoups)
• To develop a treatment
• Gene therapy (in the future?)
• Pharmacogenetics

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Genotype/phenotype relationship response to
Infliximab.
Vermeire S et al. Gastroenterology in press
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Why to discover genes?

• To make a diagnosis
• Before disease occurrence (if prevention
  possible) or to avoid invasive procedures
  (sensibiility, sensitivity).
• To classify the disease (definition of disease
  subgoups)
• To develop a treatment
• Gene therapy (in the future?)
• Pharmacogenetics
• Drug discovery (disease mechanisms)

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From Ogura et al. Nature 2001
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How a defect in a proinflammatory molecule
..can induce an inflammation?
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CARD15 -/- mouse
/
-/-
Pauleau AL et al. Mol Cel Biol 2003
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Interaction between TLR2 and NOD2
Watanabe T Nat Immunol 2004
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How CD mutations induce the inflammation?

• A defect in the innate immunity induces an excess
  of response by the adaptive immunity which is
  deleterious for the mucosa.
• CD mutations are gain of fonction mutations
  (Maeda S Science 2005)
• CD mutations do not inhibit pro-inflammatoy
  pathways (Watanabe T Nature Immunol 2004 Chen CM
  J Biol Chem 2004)
• CD mutations no more activate anti-inflammatory
  pathways (IL10) (Netea MG Eur J Immunol 2004)

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Unpublished 
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CARD15 immunohistochemistry
normal colon
Crohn
Berrebi D et al. Gut 2003
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Card15 immunohistochemistry
Berrebi D et al. Gut 2003
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NOD2 and Paneth cells
Crohn normal ileum
Card15
Lysozyme
Ogura Y et al. Gut 2003
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Screening for Card15/Nod2 interacting small
molecules

• Loss or gain of function?
• Which cell line?
• Role of MDP?

Toward a specific curative thérapy?
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Why to discover genes?

• To make a diagnosis
• Before disease occurrence (if prevention
  possible) or to avoid invasive procedures
  (sensibiility, sensitivity).
• To classify the disease (definition of disease
  subgoups)
• To develop a treatment
• Gene therapy (in the future?)
• Pharmacogenetics
• Drug discovery (disease mechanisms)
• Prevention (environmental factors)

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From Rose J et al. Gut 1988
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ENVIRONMENTAL FACTORS FOR CD

• Tobacco
• Refined sugars
• Fast-food and Cola
• Microparticles
• Tooth paste
• Chewing-Gum
• Margarin
• Fibres
• Backer yeast
• Alcohol
• Caffe
• Corn-flakes
• Curry

• Hot water
• Refrigeration
• Perinatal Infections
• Infections in childhood
• Antibiotics
• Adenoïdectomy
• Breast feeding
• Life events
• Oral contraceptives

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Commensal flora.
Or specific pathogenic bacteria ?
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Most popular infectious agents for CD

• Mycobacterium paratuberculosis
• E coli,
• Y enterocolitica
• L monocytogenes
• Pseudomonas species

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way of life
Flora?
Dlg5?
Specific bacteria?
Octn 1/2?
Card15
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