Gene related to aging, obesity and myocardial infarction, Fragile X Syndrome, Hirschsprung disease, Hearing impairment.

1. Gene related to Aging, Obesity, and
Myocardial Infarction
Fragile X Syndrome, Hirschsprung Disease,
Hearing Impairment
MAY SOE THU
5836362 MTMT/M
1

2. Outline
Aging and related genes
Obesity and related genes
Myocardial Infarction and related genes
Fragile X syndrome
Hirschsprung disease
Hearing Impairment
Update Article
2

3. Aging
The process during which structural and functional changes accumulate in an organism
as a result of the passage of time
Deterioration results in a high susceptibility to environmental challenges, leading to
age-associated pathologies that ultimately cause death
3

4. GenAge Model Organisms of Aging-Related Genes
mouse (Mus musculus)
fruit fly (Drosophila melanogaster)
roundworm (Caenorhabditis elegans)
baker's yeast (Saccharomyces cerevisiae)
http://genomics.senescence.info/genes/ 4

5. http://www.nature.com/nrg/journal/v6/n11/box/nrg1706_BX1.html
LongevityPathway
5

6. Gene related with Human Aging
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3295054/pdf/ad-2-3-186.pdf 6

7. LMNA
A protein encoded by LMNA gene
Encodes both lamin A and C, two components of the lamina,
a layer of the inner nuclear membrane that may interact
with chromatin
Involved in nuclear stability, chromatin structure and gene
expression
http://www.premierexhibitions.com/exhibitions
Progeria
Mutation  Hutchinson-Gilford progeria syndrome, Emery-Dreifuss muscular
dystrophy, familial partial lipodystrophy, limb girdle muscular dystrophy, dilated
cardiomyopathy, etc.
7

8. WRN
An enzyme encoded by WRN gene
A member of the RecQ Helicase family
Also known as Werner syndrome ATP-dependent
helicase or DNA helicase or RecQ-like type 3
An only RecQ Helicase that contains 3' to
5' exonuclease activity
Important in repairing of double stranded breaks, non-
homologous end joining, and base excision repair
Essential in telomere maintenance and replication,
especially the replication of the G-rich sequences
Werner Syndrome
http://depts.washington.edu/monnatws/projects.php 8

9. ERCC8
A protein encoded by ERCC gene
Involved in transcription and maybe in DNA repair
Mutation  Hereditary Cockayne syndrome
CS cell : abnormally sensitive to ultraviolet radiation
defective in the repair of transcriptionally active genes
https://www.emaze.com 9https://ghr.nlm.nih.gov/gene/ERCC8

10. Obesity
A condition in which excess fat accumulates in the body
It can have an adverse effect on health
Obesity : BMI > 30 kg/m2
10http://drpaulwizman.com/about-obesity/obesity

11. Obesity Trends Among U.S. Adults, 2014
11

12. http://www.ijem.in/text.asp?2014/18/7/17/145049 12

13. Obesity related Genes
Odd Ratio
http://www.nature.com/nrg/journal/v10/n7/fig_tab/nrg2594_F2.html 13

14. FTO gene
Fat mass and obesity associated protein (alpha-ketoglutarate-dependent dioxygenase)
Chromosome 16
http://www.cnschronicle.com/2013/12/ 14

15. CTNNBL1 gene
Beta-catenin-like protein 1
On chromosome 20q11.2
A protein involved in Wnt/β-catenin signaling pathway which is a central
pathway in adipogenesis
Biological process – Apoptosis, Gene expression, mRNA splicing, somatic
diversification of immunoglobulins
15

16. Myocardial Infarction
Also called ‘Heart Attack’
Irreversible necrosis of heart muscle secondary to prolonged ischaemia.
Due to an imbalance in oxygen supply and demand, which is most often caused
by plaque rupture with thrombus formation in a coronary vessel, resulting in an
acute reduction of blood supply to a portion of the myocardium
16http://ereidmiller.com/myocardial-infarction-symptoms

17. Differentiation between MI types 1 and 2
according to the condition of the coronary arteries
17
http://www.nature.com/nrcardio/journal

18. MI related Genes
Fig: Common genetic variations in genes involved in LDL-cholesterol metabolism that have an effect
on the risk of myocardial infarction
Jeanette Erdmann, et al. Genetic cause of Myocardial Infarction, Dtsch Arztebl Int 2010; 107(40): 694–9 18

19. MI related Genes
Jeanette Erdmann, et al. Genetic cause of Myocardial Infarction, Dtsch Arztebl Int 2010; 107(40): 694–9 19

20. PCSK9 & LDLR
http://eurheartj.oxfordjournals.org/content/early/2013/08/15/eurheartj.eht273 20

21. ApoB & ApoE
ApoB - on chromosome 2
- primary apolipoprotein of chylomicrons, VLDL, IDL, and LDL particles
ApoE - on chromosome 19 with apolipoprotein C1 & C2
- an important regulator of cholesterol, fatty acid, and glucose homeostasis
Higher ApoB Higher LDL Plagues
Defects in ApoE Impaired transport HLP
HLP : Hyperlipoproteinaemia
21

22. Fragile X Syndrome
A genetic condition that causes a range of developmental problems including
intellectual disabilities and cognitive impairment
Caused by mutations in the FMR1 gene, which is located on the X chromosome
and whose locus at Xq27.3
Males > Females
Occurrence : approximately 1 in 4000 (M), 1 in 8000 (F)
Physical Characteristics : Protruding ears, Long face, Flat feet,
Soft skin, High-arched palate, etc.
Behavioral Characteristics : Stereotypic movements, Social anxiety.
http://amessageofmeanfrommeghan.com/ 22

23. FMR1 Gene
Bagni C, Tassone F, Neri G, Hagerman R. Fragile X syndrome: causes, diagnosis, mechanisms, and therapeutics. The Journal of clinical investigation. 2012;122(12):4314-22.
23

24. Hirschsprung Disease
Disorder that arise as a consequence of defective
neural crest cell development (Neurocristopathies)
Characterized by a congenital absence of neurons
in a portion of intestinal tract usually distal colon
Occurrence: 1 in 5000 births
http://flipper.diff.org/apprulesitems/items/6712 24

25. Genes related to Hirschsprung Disease
Butler Tjaden NE, Trainor PA. The developmental etiology and pathogenesis of Hirschsprung disease. 2013;162(1):1-15. 25

26. RET and GDNF gene
http://www.nature.com/nrn/journal/v3/n5/fig_tab/nrn812_F1.html
ENCC proliferation & survival, apoptosis, migration and differentiation
26

27. Hearing Impairment
A partial or total inability to hear
Not the same with deafness; no hearing
May occur in one or both ears
In children, it can affect the ability to learn language
In adults, it can cause work-related difficulties
http://www.ncbi.nlm.nih.gov/books/NBK1434/ 27

28. Genetic Association of Hearing Impairment
DFNA: Non-syndromic
deafness, autosomal dominant
DFNB: Non-syndromic
deafness, autosomal recessive
DFNX: Non-syndromic
deafness, X-linked
http://www.cmsact.org/uploads/4/6/7/5/4675465 28

29. http://www.ncbi.nlm.nih.gov/books/NBK1434/ 29

30. GJB2
Gap Junction Beta-2 protein which is encoded by GJB2 gene
Also known as connexin26 (Cx26)
30
GJB2
Non-syndromic
DeafnessMutation
Hinders gap junction
construction

31. 31

32. Obesity
It is characterized by an increase in adipose tissue mass and excess lipid
deposition, and is associated with enlarged adipocytes
Endogenous and exogenous factors such as various stressors, aging, and low
physical activity lead to obesity and may change patterns of gene expression
However the factors responsible for these differences in obesity pathogenesis
between males and females are largely unknown
32

33. Hypothesis
1) To study the expression of the obesity-related genes in young male and
female mice
2) To compare the expression in older males and females fed either a STD of a
high fat diet (HFD) for 35 weeks
3) To compare the expression of these genes and glucose metabolic activity
between males and females
33

34. Gene Selection
At least 400 genes associated with obesity
34
35 genes
Lipid & Glucose
Metabolism
Receptors for
insulin signaling
in WAT

35. Materials and Methods
Statistical Analysis
RNA isolation and RT-PCR
Intraperitoneal glucose tolerance test
Animal Treatment
Animals
35

36. Animal Group
8-week-old MiceMice
129/SvCPJ mice
STD  35 wks
(343kcal/100g,
crude fat 3.8%)
HFD  35 wks
(508kcal/100g,
crude fat 32%)
N = 7 Males + 9 Females
N = 7 Males + 9 Females
36

37. Animal Treatment
8-wk-old Mice 43-wk-old Mice
Fasting  6 hr
Organs and Blood
White Adipose Tissue (WAT)
 Ovaries
 Testes
 Kidney
 Post. Belly Lining
Plasma level
 Glucose
 Insulin
 Leptin
 Adiponectin
37

38. Intraperitoneal glucose tolerance test (IPGTT)
8-wk-old Mice
(N = 4  Each sex)
21-24-wk-old Mice
(N = 4  Each sex & diet)
Intraperitoneal inoculation of Glucose (2g/10ml/kg)
Collecting Blood samples at 0, 15, 30, 60, 120, 180 min
Area Under Curve (AUC) & Half-life (t1/2)
Fasting  12 hr
38

39. Reverse Transcription-Polymerase Chain Reaction
RNA
Extraction
RT reaction
Primer sets
and PCR
conditions
Electrophoresis
Densitometry
Normalized with
internal control
genes, 36b4
Density ratios of the products  For comparison
39

40. Fundamental parameters of 8-wk-old mice
40

41. Comparison of parameters of 43-week-
old male & female mice
41

42. Comparison of parameters of 43-week-
old male & female mice
STD for 35 weeks did not induce obesity in both mice
42
HFD  Markedly increase in body weight, weight gain, perigonadal, perirenal,
subcutaneous WAT

43. mRNA Expression in Perigonadal WAT
8 week-old mice 43 week-old mice
43

44. 8 week-old mice 43 week-old mice
mRNA Expression in Perigonadal WAT
44

45. 8 week-old mice 43 week-old mice
mRNA Expression in Perigonadal WAT
45

46. 8 week-old mice 43 week-old mice
mRNA Expression in Perigonadal WAT
46
The expression level of 6 genes (Acaca, Fasn, Slc2a1, Slc2a4, Adipoq, Nampt) were
higher in WAT of 43-week-old female mice than in male mice
Females have a greater ability to metabolize glucose in WAT regardless of diet
The expression level of 2 genes (Acaca, & Fasn) were higher in WAT of 8-week-old
female mice than in male mice but the other gene expressions are similar.

47. Glucose Metabolism Capacity
Glucose metabolism capacity is marginally greater in females than in males
47

48. IPTGG for 21-24 week-old 129/Sv mice
after 12 h fasting
oSTD-fed female mice
●STD-fed male mice
□HFD-fed female mice
∎HFD-fed male mice
Half-life  Longer in males
Glucose absorption and metabolism were faster in female mice fed a STD
48

49. Plasma Adiponectin
Plasma adiponectin – Higher in female than males at 8-week old & after 35 weeks
49

50. Lep mRNA and plasma leptin levels
Expression levels of Lep mRNA and plasma leptin levels were upregulated in HFD-fed mice
50

51. Lep mRNA and plasma leptin levels
Expression levels of Lep mRNA and plasma leptin levels were upregulated in HFD-fed mice
Lipid accumulation might mediate this up-regulation in HFD
Levels of plasma glucose, insulin, & adiponectin remain unchanged
despite the consumption of a HFD for 35 weeks
51

52. Discussion (1)
Expression levels of four genes (Slc2a1, Slc2a4, Adipoq, Nampt) were greater in
the WAT of 43-week-old female mice
So females might have a greater ability to metabolize glucose in WAT regardless
of diet fat content
Expression levels of two genes (Acaca & Fasn) were higher in females than males
at both 8 weeks and 43 weeks
So it may be associated with sex, but not aging
52

53. In IPGTTs, the value of half-life was not significantly different between males and
females at 8 weeks but it was lower in females than males fed a STD
It suggests that the capacity to metabolize glucose may change with age in a sex-
dependent manner
Consuming a HFD for 35 weeks significantly increased AUC in male and female
mice compared with mice fed with STD, and AUC & t1/2 was greater in males
It demonstrate that a HFD may decrease the ability of glucose metabolism
Discussion (2)
53

54. Conclusion
54
Sex and aging may cause diet-independent differences in gene
expression levels in female and male mice
Higher expression of these genes in females could contribute to
higher metabolic activity and resistance to obesity

55. 55

56. References
1. S. Rodríguez-Rodero et al. Aging Genetics and Aging. Gene and Aging. June 2011;
Vol.2, Number 3; 186-195.
2. Andrew J. Walley et al. The genetic contribution to non-syndromic human obesity.
Nature Reviews Genetics. July 2009; Vol.10, 431-442.
3. Jeanette Erdmann, et al. Genetic cause of Myocardial Infarction, Dtsch Arztebl Int.
2010;107(40): 694–9.
4. Bagni C, Tassone F, Neri G, Hagerman R. Fragile X syndrome: causes, diagnosis,
mechanisms, and therapeutics. The Journal of clinical investigation. 2012;122(12):4314-22.
5. Butler Tjaden NE, Trainor PA. The developmental etiology and pathogenesis of
Hirschsprung disease. Translational research : the journal of laboratory and clinical
medicine. 2013;162(1):1-15.
56