MASTER’S THESIS REPORT

Thesis name:

ASSOCIATION OF FTO GENE VARIANTS
WITH OSTEOPOROTIC VERTEBRAL FRACTURES

IN VIETNAMESE WOMEN
Master’s student:
Huỳnh Chí Thiện
Supervisor:
PhD. Dr. Nguyễn Thị Thanh Hương
PhD. Dr. Lê Đình Tùng

HCMC, 12-2017


Table of Contents
1

Review of the literature

2

Contents of research

3

Materials and methods

4

Results and discussion

5

Conclusions


1. REVIEW OF THE LITERATURE
1.2. Osteoporosis

Normal

Osteoporosis

 Definition: Bone strength
 Classification: primary and secondary osteoporosis
Primary osteoporosis: postmenopausal and senile


1. REVIEW OF THE LITERATURE

(World Health Organization, 1994)


1. REVIEW OF THE LITERATURE
1.3. Vertebral fracture assessment by radiograph
1. Genant’s
2. Quantitative morphometry (QM)


(IFO-VFI, part 2)

Figure 1.8. QM with 6-point placements on radiographs

Genant’s grading


1. REVIEW OF THE LITERATURE
1.4. FTO gene

• 410 kb (410,507 bp)
• Catalyzing the demethylation
of nucleic acid


1. REVIEW OF THE LITERATURE
CẬP NHẬT NGHIÊN CỨU
Intron 1

Bich Tran et al (2013)
SNP rs 1121980, rs 9939609
rs17817449, rs 1558902
rs 9930506, rs 1421085

Gaurav Garg et al (2014)
SNP rs 1121980, rs 9939609
Jianmin Xu (2016)
SNP rs 7206790, rs 9939609

Intron 8


Yan Guo et al (2011)
SNP rs 16952955
rs 2540766
rs 2540784
rs 16952951
rs 12447427
rs 2689247


2. CONTENTS OF RESEARCH
1. Choosing study subjects and SNPs

2. Preparing samples
3. Designing primers & Optimizing the methods

4. Investigating allele frequencies
5. Determining the correlation


3. MATERIALS & METHODS
 Location: 108, Bạch Mai, Việt Xô hospital
 Study subjects: Vietnamese menopausal women ≥ 40 years old
 Patient group: having osteoporosis vertebral fractures
 Control group: not having
Exclusion criterion: secondary osteoporosis
 Sample size:


3. MATERIALS & METHODS

3.1. CAPS (rs1121980)

Figure 1.11. SNP cleavage with restriction enzyme (S. Kim, 2007)


3. MATERIALS & METHODS
3.2. 4ARMS (rs16952955)
(OF)

(IF)

(IR)

(OR)

Figure 1.12. Schematic presentation of the 4ARMS method (Shu Ye, 2001)


3. MATERIALS & METHODS
3.1. 3P (rs9939609)


4. RESULTS & DISCUSSION


4. RESULTS & DISCUSSION


4. RESULTS & DISCUSSION



4. RESULTS & DISCUSSION


4. RESULTS & DISCUSSION


4. RESULTS & DISCUSSION


4. RESULTS & DISCUSSION


5. CONCLUSIONS
- Providing the initial data about allele frequency of SNPs: FTO
Asean osteoporosis SNP-rs16952955, popular SNP rs9939609, and
rs1121980 in Northern Vietnamese women.
- Recomending two effective genotyping methods: 4ARMS and 3P as
well as optimization.
- 4ARMS and 3P are more cost-efficient and labor-intensive than
CAPS, especially time-consuming for laboratory.
- Building the basis for studies on the correlation between SNP and
osteoporosis in general, and osteoporotic fracture in particular.


TÀI LIỆU THAM KHẢO
1. Michaelsson K., Melhus H., Ferm H., Ahlbom A. & Pedersen N. L. "Genetic liability to fractures in the elderly”. Arch Intern
Med. 165(16), 1825-30 (2005)
2. Svanhild Waterloo, Luai A Ahmed, Jacqueline R Center, John A Eisman, Bente Morseth, Nguyen D Nguyen, Tuan
Nguyen, Anne J Sogaard and Nina Emaus. “Prevalence of vertebral fractures in women and men in the population-based Tromsø

Study”. BMC Musculoskeletal Disorders. 13:3 (2012)
3. Dana Bliuc, Nguyen D. Nguyen, Vivienne E. Milch, Tuan V. Nguyen, John A. Eisman, Jacqueline R. Center. “Mortality Risk
Associated With Low-Trauma Osteoporotic Fracture and Subsequent Fracture in Men and Women”. JAMA.301(5):513-521
(2009)
4. J B Richards, F Rivadeneira, M Inouye, T M Pastinen, N Soranzo, S G Wilson, T Andrew, M Falchi, R Gwilliam, K R Ahmadi,
A M Valdes, P Arp, P Whittaker, D J Verlaan, M Jhamai, V Kumanduri, M Moorhouse, J B van Meurs, A Hofman, H A P Pols, D
Hart, G Zhai, B S Kato, B H Mullin, F Zhang, P Deloukas, A G Uitterlinden, T D Spector. “Bone mineral density, osteoporosis,
and osteoporotic fractures: a genome-wide association study”. Lancet.371(9623):1505-12 (2008)
5. Gao X, Shin YH, Li M, Wang F, Tong Q, et al. “The fat mass and obesity associated gene FTO functions in the brain to regulate
postnatal growth in mice”.PLoS One.5:e14005 (2010)
6. Bich Tran, Nguyen D. Nguyen, Jacqueline R. Center, John A. Eisman, and Tuan V.Nguyen. “Association between fat-massand-obesity-associated (FTO) gene and hip fracture susceptibility”. Clin Endocrinol (Oxf).81(2):210-7 (2014)
7. Yan Guo, Hui Liu, Tie-Lin Yang, Siyang M. Li, Siyuan K. Li, Qing Tian, Yong-Jun Liu, Hong-Wen Deng. “The fat mass and
obesity associated gene, FTO, is also associated with osteoporosis phenotypes.” PLoS One.6(11):e27312 (2011)
8. Gaurav Garg, Jitender Kumar, Fiona E. McGuigan, Martin Ridderstrale, Paul Gerdhem, Holger Luthman, Kristina Akesson.
“Variation in the MC4R Gene Is Associated with Bone Phenotypes in Elderly Swedish Women”. PLoS One.9(2):e88565 (2014)
9. H. T. T. Nguyen, B. von Schoultz, D. M. T. Pham, D. B. Nguyen, Q. H. Le, D. V. Nguyen, A. L. Hirschberg, and T. V. Nguyen.
“Peak bone mineral density in Vietnamese women”. Arch Osteoporos. 4(1-2): 9–15 (2009)
10. Jane A. Cauley DrPH. “Defining ethnic and racial differences in osteoporosis and fragility fractures”. Clin Orthop Relat
Res.469:1891–1899 (2011)
11. Chun Wang, Zeng Zhang, Hao Zhang, Jin-Wei He, Jie-Mei Gu, Wei-Wei Hu, Yun-Qiu Hu, Miao Li,Yu-Juan Liu , Wen-Zhen
Fu, Hua Yue, Yao-Hua Ke, and Zhen-Lin Zhang. Susceptibility genes for osteoporotic fracture in postmenopausal Chinese
women. J Bone Miner Res.27(12):2582-91 (2012)
12. Frayling TM, Timpson NJ, Weedon MN, Zeggini E, Freathy RM, Lindgren CM, et al. A common variant in the FTO gene is
associated with body mass index and predisposes to childhood and adult obesity. Science, 316: 889-894 (2007)
13. Dina C, Meyre D, Gallina S, Durand E, Korner A, Jacobson P, et al. Variation in FTO contributes to childhood obesity and
severe adult obesity. Nature genetics, 39: 724-726 (2007)
14. Gerken T, Girard CA, Tung YC, Webby CJ, Saudek V, Hewitson KS et al. The obesity-associated FTO gene encodes a 2oxoglutarate-dependent nucleic acid demethylase. Science, 318: 1469-1472 (2007)
…