Good set of slides for Molecular abd Genetic aspects of Vur and lower Urinary Pathilogy in children

1. A MOLECULAR AND GENETIC
VIEW OF HUMAN
RENAL AND URINARY TRACT
MALFORMATIONS
Adrian S Woolf
Nephro-Urology Unit
Institute of Child Health,
University College London, UK

2. CLINICAL IMPORTANCE OF
KIDNEY MALFORMATIONS
800 children in the UK have
end stage renal failure.
The commonest causes are:
1. Agenesis (absent kidney)
2. Dysplasia (undifferentiated)
3. Hypoplasia (too few nephrons)

3. LOWER URINARY TRACT
MALFORMATIONS
Kidney malformations are
often associated with ureter
and/or bladder malformations:
1. Vesicoureteric reflux
2. Duplicated lower tracts
3. Obstructed lower tracts

4. CAUSES OF KIDNEY AND
LOWER URINARY TRACT
MALFORMATIONS
1. Mutations of genes expressed
during development
2. Gross changes of fetal kidney milieu
e.g. impairment of fetal urine flow
3. Subtle change of fetal kidney milieu
e.g. low protein maternal diet

5. PRIMARY VESICOURETERIC
REFLUX
1. Occurs in 1% babies
2. ‘Reflux nephropathies’ can be
congenital or acquired and account for
10% of end-stage renal failure
2. Excess risk for vesicoureteric reflux
in first degree relatives is x20-50
3. Some families have dominant inheritance
4. 1p13 locus and other undefined loci

6. I:1 I:2
?
II:1 II:2
III:1 III:2 III:3 III:4 III:14 III:5 III:6 III:7 III:15 III:8 III:9 III:10 III:11 III:12 II
IV:1 IV:2
VESICOURETERIC
REFLUX

7. Embryonic urinary bladder Adult urinary bladder
Uroplakin Ib Uroplakin III

8. Vesicoureteric Reflux in the UK
Establishing a DNA collection
www.vur.org.uk

9. DUPLEX KIDNEYS AND
LOWER URINARY TRACTS
1. Occur in 1% of individuals
2. Occurs with dysplastic kidneys and
vesicoureteric reflux
3. Excess risk in first degree relatives
is about x20-30

12. FRASER SYNDROME
1. Autosomal recessive - fused fingers,
membrane across eyes and dysplastic
or absent kidneys
2. FRAS1 gene is mutated - protein
coats the ureteric bud
3. Null mutant mouse metanephros
involutes with excess apoptosis

13. Patient Described by George Fraser

14. Fraser Syndrome vs bl/bl Phenotype

15. Spectrum of Renal Defects in Fras1 targeted
Mutants (b-d)

18. APOPTOTIC
INVOLUTION OF CYSTIC
DYSPLASTIC KIDNEY

19. RENAL CYSTS AND
DIABETES SYNDROME
1. Autosomal dominant or sporadic
2. Renal agenesis, cysts, dysplasia,
hypoplasia, with diabetes (can
require insulin) and uterus
malformations
3. Hepatocyte Nuclear Factor 1β
transcription factor mutations

20. DUK507 Q243fsdelC
I
1 2
II
1 2
Small kidneys
III 2
1 2 3 4 5
NM NM
Bicornuate Single kidney
uterus
IV
1 2
NM NM

21. HNF1β antisense HNF1β sense
m
u

22. ORAL FACIAL DIGITAL
SYNDROME TYPE 1
1. Male embryonic lethal – only
affected females are born
2. X-linked dominant
3. Glomerocystic kidney disease
4. OFD1 gene codes for a
centrosomal/basal body protein

24. OFD1 IMMUNOHISTOCHEMISTRY
ub
m
v
WHOLE KIDNEY
OFD1Ab Preabsorbed OFD1 Ab
EK84 EK75 EK73 EK70
anti ofd1
-120D
OFD1 PROTEIN IS EXPRESSED IN HUMAN
EMBRYONIC KIDNEYpreabsorbed
MESENCHYME
IN VIVO AND IN DERIVED CELL LINES

25. C
150
100
75
α-Tubulin OFD1 merge 50
37
25

26. OFD1 PROTEIN - GREEN
ACETYLATED TUBULIN - RED

27. CAUSES OF KIDNEY AND
LOWER URINARY TRACT
MALFORMATIONS
1. Mutations of genes expressed
during development
2. Gross changes of fetal kidney milieu
e.g. impairment of fetal urine flow
3. Subtle change of fetal kidney milieu
e.g. low protein maternal diet

28. ANIMAL MODEL OF POSTERIOR URETHRAL VALVES
AND FETAL NEPHROPATHY AND UROPATHY

29. CAUSES OF KIDNEY AND
LOWER URINARY TRACT
MALFORMATIONS
1. Mutations of genes expressed
during development
2. Gross changes of fetal kidney milieu
e.g. impairment of fetal urine flow
3. Subtle change of fetal kidney milieu
e.g. low protein maternal diet

30. THE EXPERIMENTAL MODEL
Pregnant rats were supplied isocaloric
diets in which the primary variable was
the protein content from the day of
conception to the end of pregnancy
– 18% protein (control)
– 9% protein (mild protein restriction)
– 6% protein (severe protein restriction)
Welham et al Kidney Int 61:1231-1242, 2002

31. Maternal low
protein diet
Rat Increased
embryonic metanephric
day 13 apoptosis Embryonic kidney
Embryonic Decreased cell
day 15 number
Fewer nephron
progenitor cells
Apoptosis
Two weeks
Fewer nephrons
postnatal
Elevated blood Non-renal
Adult
pressure alterations

32. P<0.05
P<0.01
250
200
Series4
Apoptotic
18% Protein
cells/mm2
150
Series5
9% Protein
100
Series10
6% Protein
50
0
Embryonic day 13
Glomerular complement of Systolic blood pressures of offspring
offspring at 2 weeks of age. at 4 weeks and 19 weeks of age.
Systolic blood pressure
20000 200
* * * *
Glomeruli
15000 (mmHg). 150
10000 18% protein
100
9% protein
5000 50
0 0
6% Protein 9% Protein 18% Protein 4 weeks 19 weeks

33. REPRESENTATIONAL
DIFFERENCE ANALYSIS
• Genes ‘upregulated’ in metanephric kidneys
exposed to maternal normal diet:
Cofilin-1, Prox-1, Calmodulin
• Genes ‘upregulated’ in metanephric kidneys
exposed to maternal low protein diet:
Cadherin-11

34. CAUSES OF KIDNEY AND
LOWER URINARY TRACT
MALFORMATIONS
1. Mutations of genes expressed
during development
2. Gross changes of fetal kidney milieu
e.g. impairment of fetal urine flow
3. Subtle change of fetal kidney milieu
e.g. low protein maternal diet

35. ACKNOWLEDGEMENTS FOR STUDIES OF KIDNEY
AND URINARY TRACT MALFORMATIONS
University College London UK
Monica Banerjee, Maria Bitner-Glindzicz, Isky Gordon,
Ambrose Gullett, Peter Cuckow, Chris Fry, Maggie Godley, Monika Hermanns,
Mike Hubank, Maria Kolatsi-Joannou, Dagan Jenkins, Liam McCarthy,
Sue Malcolm, Peter Nyirady, Donald Peebles, Karen Price, Paul Riley,
Leila Romio, Peter Scambler, Naima Smeulders, Niki Thiruchelvam,
Simon Welham, Melissa Whitten, Duncan Wilcox, Paul Winyard, Su-Ping Yang
Elsewhere in UK
Coralie Bingham, Sally Feather, Andrew Fry,
Judith Goodship, Tim Goodship, Andrew Hattersley, Albert Ong,
Jenny Southgate, British Association for Paediatric Nephrology
Elsewhere in the world
TIGM Naples Italy - Brunella Franco and colleagues
NYU New York USA – Tung-Tien Sun and colleagues