discovering the genes controlling response to trypanosoma congolense infection harry noyes...
Post on 21-Dec-2015
212 views
TRANSCRIPT
![Page 1: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/1.jpg)
Discovering the genes controlling response to Trypanosoma
congolense infectionHarry Noyes
University of Liverpool
![Page 2: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/2.jpg)
Participants• ILRI, Nairobi• Morris Agaba• John Gibson• Fuad Iraqi• Steve Kemp• Hassan Musa• Joel Mwakya• Daniel Mwanga• Jan Naessens• Joseph Nganga• Moises Ogugu• John Wambugu
• University of Manchester
• Andy Brass• Helen Hulme• Leo Zeef• Leanne Wardlesworth
• University of Liverpool
• Anthea Broadhead• Derek Daly• Kate Goodheart• Harry Noyes• Katie Rennie
Roslin InstituteAlan ArchibaldSusan AndersonLaurence Hall
FundingWellcome Trust
![Page 3: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/3.jpg)
Trypanosomosis
Is a fatal disease of livestock.
The livestock equivalent of sleeping sickness in humans
T brucei rhodesiense T gambiense
T. congolense, T. vivax
![Page 4: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/4.jpg)
Bovins
Bovins et GlossinesGlossines
CattleTsetseCattle and tsetse
Distribution of Trypanosomiasis
N’DamaBoran
![Page 5: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/5.jpg)
Trypanosoma brucei
![Page 6: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/6.jpg)
QuickTime™ and aNone decompressor
are needed to see this picture.
![Page 7: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/7.jpg)
Mouse models of trypanotolerance.
0
10
20
30
40
50
60
70
80
90
100
1 21 41 61 81 101 121
Days Post Challenge
% S
urv
ival
F6
AJ
C57BL
Survival of F6 and parental mice
![Page 8: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/8.jpg)
X =
C57BL6 resistant
AJ and Balb/c susceptible
Creating mapping populations
![Page 9: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/9.jpg)
Trypanosoma infection response (Tir) loci
C57/BL6 x AJ and C57/BL6 x BALB/C
Iraqi et al Mammalian Genome 2000 11:645-648 Kemp et al. Nature Genetics 1997 16:194-196
![Page 10: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/10.jpg)
D17
MIT
408
D17
MIT
029
DI1
7MIT
234 D
17M
IT17
7
D17
MIT
091
D17
MIT
072
5CM
40
35
30
25
20
15
10
5
0
LO
D S
CO
RE
D17MIT16
Tir1 locus in the F6
![Page 11: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/11.jpg)
Contribution of 10 genes from Boranand N’Dama
cattle to reduction in degree of trypanosomosisBoran (relatively susceptible)
The N’Dama and Boran each contribute trypanotolerance alleles at 5 of the 10 most significant QTL, indicating that a synthetic breed could
have even higher tolerance than the N’Dama.
N’Dama (tolerant)
-15-10-505
1015
-15-10-505
1015
![Page 12: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/12.jpg)
From QTL to gene
• Sequence variants– Resequencing
• Structural variants– Agilent tiling arrays
• Expression variation– Affymetrix microarrays
• Congenic mice• Gene networks
![Page 13: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/13.jpg)
![Page 14: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/14.jpg)
![Page 15: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/15.jpg)
![Page 16: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/16.jpg)
Sequence comparisonsChromosome 5, 73-83Mb
![Page 17: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/17.jpg)
From QTL to gene
• Sequence variants– Resequencing
• Structural variants– Agilent tiling arrays
• Expression variation– Affymetrix microarrays
• Congenic mice• Gene networks
![Page 18: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/18.jpg)
Microarray design at each time point
Resistant C57BL/6Susceptible AJ
![Page 19: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/19.jpg)
Harvesting Tissues
![Page 20: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/20.jpg)
![Page 21: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/21.jpg)
![Page 22: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/22.jpg)
From QTL to gene
• Sequence variants– Resequencing
• Structural variants– Agilent tiling arrays
• Expression variation– Affymetrix microarrays
• Congenic mice• Gene networks
![Page 23: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/23.jpg)
Congenic Mice
• Three congenic lines
• One line for each trypanotolerance QTL
• F1 Cross between AJ and C57BL6
• Backcrossed to AJ for six generations selecting for C57 allele at each generation
![Page 24: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/24.jpg)
Development of Congenic mice
C57BL/6 DNA
AJ DNA
QTL
![Page 25: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/25.jpg)
Position of Mmu17 QTL (Tir1)
![Page 26: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/26.jpg)
Position of Mmu5 QTL (Tir2)
![Page 27: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/27.jpg)
Survival of congenic mice
Response of Congenic mice to T. congolense infection
0
20
40
60
80
100
120
21-30 31-40 41-50 51-60 61-70 71-80 81-90 91-100
Days post intection
% survivingTir1AA (n = 20)Tir1CC (n = 25)Tir2AA (n = 60)Tir2CC (n = 120)Tir3AA (n = 20)Tir3CC (n = 90)TirnAA (n = 100)
![Page 28: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/28.jpg)
From QTL to gene
• Sequence variants– Resequencing
• Structural variants– Agilent tiling arrays
• Expression variation– Affymetrix microarrays
• Congenic mice• Gene networks
![Page 29: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/29.jpg)
Cholesterol metabolism
![Page 30: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/30.jpg)
Endogenous cholesterol production increases after
infection
![Page 31: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/31.jpg)
Tir loci C57/BL6 x AJHDL after 6 weeks high fat diet AIL C57BL6 x NZB/BIN
Iraqi et al Mammalian Genome 2000 11:645-648 Wang et al. Genome Research 2003 13:1654-1664Kemp et al. Nature Genetics 1997 16:194-196
Trypanotolerance QTL are the right of each pair of chromosomes HDL QTL are the left hand of each pair
![Page 32: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/32.jpg)
Total Cholesterol levelsCHOLESTEROL
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
0 8 21 4
DAYS POST INFECTION
AJ_HIGH_FAT
AJ_LOW_FAT
BALB_HIGH_FAT
BALB_LOW_FAT
c57_HIGH_FAT
C57_LOW_FAT
![Page 33: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/33.jpg)
Tir2 QTL controls weight
![Page 34: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/34.jpg)
Collaboration with GSF
• Clinical chemistry and cytokines
• InbredC57BL/6, Balb/c, A/J mice
• Congenic Tir1, Tir2, Tir3 mice plus controls
• Serum samples collected pre-infection and at days 3, 9, 17 and 35 post-infection
![Page 35: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/35.jpg)
![Page 36: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/36.jpg)
![Page 37: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/37.jpg)
![Page 38: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/38.jpg)
![Page 39: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/39.jpg)
Inflammatory counter inflammatory switch
C57
AJ/Balb
0 7 93 17
Classically activated macrophages
Alternatively activated macrophagesTh2 signal (IL4, IL10)
![Page 40: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/40.jpg)
AJ and Balb/c produce alternatively activated macrophages early in infection
![Page 41: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/41.jpg)
Intersection of Cholesterol and Inflammatory pathways
Dunn et al Journal of Experimental MedicineVol. 203, No. 2, February 20, 2006 401–412
Th2 bias
Suppression of cholesterol synthesis
![Page 42: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/42.jpg)
NATURE MEDICINE • VOLUME 9 • NUMBER 2 • FEBRUARY 2003
LXR agonsists lower cholesterol and inhibit NFkB mediated inflammatory signals
![Page 43: Discovering the genes controlling response to Trypanosoma congolense infection Harry Noyes University of Liverpool](https://reader038.vdocument.in/reader038/viewer/2022110322/56649d555503460f94a326d7/html5/thumbnails/43.jpg)
Innate immune response controls survival after infection
with T. congolense • Deletion of T cells does not effect survival
time• C57BL/6 (resistant) mice have strong
inflammatory response and high cholesterol• Identification of regulators of both
mechanisms may lead us to the Quantative trait gene (QTG)