Tóm Tắt Luận Án Tiến Sĩ (Tiếng Anh).Pdf

MINSTRY OF EDUCATION MINISTRY OF HEALTH HANOI MEDICAL UNIVERSITY NGUYEN THI HOA STUDY ON POLYMORPHISMS OF SOME GENES ASSOCIATED OSTEOPOROSIS IN MEN Training major Internal Medicine Specialization Inte[.]

MINSTRY OF EDUCATION MINISTRY OF HEALTH

HANOI MEDICAL UNIVERSITY

NGUYEN THI HOA

STUDY ON POLYMORPHISMS OF SOME GENES ASSOCIATED OSTEOPOROSIS IN MEN

Training major : Internal Medicine Specialization : Internal – Osteoarthritis

DOCTORAL THESIS SUMMARY

HANOI – 2023

THIS THESIS WAS PRESENTED AT HANOI MEDICAL

UNIVERSITY

Scientific intructors:

1 Dr Nguyen Thi Thanh Huong

2 Asc Prof Dr Tran Thi Minh Hoa

Reviewer 1: Asc Prof Dr Nguyen Mai Hong

Reviewer 2: Asc Prof Dr Le Thu Ha

Reviewer 3: Prof Dr Tran Huy Thinh

This thesis is to be defended in the university’s council

At: … day … month … year……

This thesis can be found at:

- National library

- Library of Hanoi Medical University

PUBLISHED WORKS RELATED TO THE THESIS

1 Nguyen Thi Hoa, Tran Thi Minh Hoa, Nguyen Thi Thanh Phuong

“Relationship between LRP5 genotype polymorphism at SNP Q89R (rs41494349) with bone density in men” Vietnamese Medical Journal, August 2020 493:263-270

2 Nguyen Thi Hoa, Nguyen Thi Nan, Tran Thi Minh Hoa, Nguyen Thi

tetrahydrofolate reductase (MTHFR) C677T gene polymorphism and osteoporosis in men" Journal of Vietnamese Medicine, May

2022 514(2):221 - 226

3 Nguyen Thi Hoa, Nguyen Thi Nan Tran Thi Minh Hoa, Nguyen Thi Thanh Huong "Relationship between fat mass and obesity gene polymorphism - Associated (FTO) Rs1121980 with osteoporosis in men" Vietnamese Medical Journal November 2022 520(2):161-167

INTRODUCTION

1 The justification of this study

Osteoporosis is a systemic skeletal disease, characterized by a decrease in bone mineral density (BMD) and a deterioration in bone microarchitecture that results in brittle and fragile bones Osteoporotic fractures increase disability, mortality, healthcare cost, and decrease quality of life, especially in the elderly Osteoporosis is more generally considered a disease not only detrimental to women but also prevalent

in men The incidence of osteoporotic fractures in men is about half of that in women Furthermore, the mortality related to hip fractures in elderly men is considerably higher than in women Osteoporosis is a complicated disease affected by many factors, including diet (calcium and protein intake), physical activities, endocrine status, co-morbidities, and genetic factors In particular, genetic factor plays an important role

in osteoporosis pathogenesis Twin- and family-based studies have indicated that 60–85% of the bone mineral density (BMD) variance is determined genetically Up to now, genome-wide association studies (GWASs) and meta-analyses have discovered 518 loci related to BMD, osteoporosis and osteoporotic fractures, but only 20% of the gene polymorphisms in these studies could explain its mechanism impacting bone density and fracture Numerous previous studies showed that three gene polymorphisms LRP5 Q89R, MTHFR C677T and FTO rs1121980 were associated with transformation in bone density and fractures in Caucasians and Asian populations Moreover, In Vietnam, there are no studies conducted on 3 gene polymorphisms LRP5Q89R, MTHFR C677T and FTORs1121980, especially in men

2 Scientific and practical significance of the study

- The first research result defined the characteristics of gene polymorphisms LRP5Q89R, MTHFRC677T and FTORs1121980 in

elderly Vietnamese men

- The study showed that 3 polymorphisms LRP5Q89R, MTHFRC677T and FTOrs1121980 all raise the proportion of osteoporosis in Vietnamese men and were independent of other risk factors: age, BMI, history of fracture, level of physical activities Men having more risk alleles in the 3 gene polymorphisms increase osteoporosis probability Therefore, this information will help clinicians assess osteoporosis risks in young people, especially those with numerous background diseases and drugs affecting bone metabolism, resulting in early diagnosis, early treatment, and minimizing fracture proportion, the fatality after osteoporosis fracture and treatment cost burden for the economy

3 Research objectives

1 Determining the genotype of polymorphisms LRP5 Q89R, MTHFR C677T and FTO rs1121980 in male with primary osteoporosis

2 Analyzing the relevance of LRP5 Q89R, MTHFR C677T, FTO

rs1121980 gene polymorphism to bone density and risk factors of males with primary osteoporosis

4 Thesis structure

- The thesis consists of 125 pages, including 4 chapters: 2 pages of introduction, 34 pages of review of literature, 17 pages of subjects and methods, 34 pages of results, 35 pages of discussion, and 3 pages of conclusion and recommendations

- This thesis consists of 34 tables (the results section 30 tables), 9 charts, 15 figures References: a total of 157 references, including 18 articles in Vietnamese, and 139 articles in English

Chapter 1: OVERVIEW

1.1 The epidemiology of osteoporosis

*In the world: The rate of male osteoporosis varies according to

each study, ranging from 2-16% and increases with age The rate of osteoporosis in men is usually 3-4 times lower than in women The NHANES 2005–2006 study was conducted by the US National Centers for Health Statistics on 3157 American adults aged 50 years and older who were measured BMD at 2 sites of the femoral neck and the total hip, the results for found that 49% of elderly women and 30% of older men had decreased bone mass at the femoral neck, the rate of osteoporosis in women was 10% and the rate of osteoporosis in men was 2% In another study in this country conducted on men aged 69 to

74 years, the rate of osteoporosis was 10.2% In Switzerland, the rate of osteoporosis is 6.3% in men aged 50-80 years old and 16.6% in men between the ages of 80-84 In some Asian countries, the rate of osteoporosis is higher in women than in men, similar to whites A study conducted on 7042 people aged 20 and over in 10 centers in different provinces in China (2002-2006) showed that the rate of osteoporosis in women was 31.2% and 10.4% in women men over 50 In Hong Kong, the rate of lumbar spine osteoporosis in men is 7%, in women, it is 37%, the rate of osteoporosis in the femoral neck in men is 6% and in women, it is 16% In Korea, the prevalence of osteoporosis is 35.5% in women and 7.5% in men over the age of 50

*In Vietnam: the studies in the North and the South showed that

the rate of osteoporosis in both men and women was similar to that of some Asian countries and white men According to the data of Nguyen Thi Thanh Huong in the North of Vietnam, a study on 222 men and 612 healthy women aged 13-83, the rate of osteoporosis in the femoral neck

is 9% for men and 17% for women

Research by author Ho Pham Thuc Lan and colleagues on 357 men and 870 women aged 18 - 89 in Ho Chi Minh City showed that the rate

of male osteoporosis is 10%, while the rate of female osteoporosis is 30% Thus, it can be seen that osteoporosis and osteoporotic fractures are really common diseases in Vietnam and need to be studied appropriately

1.2 Genes related osteoporosis

1.2.1 In the world

Reducing BMD is the principal underlying cause of significant morbidity Determination of BMD is Multifactorial and subject to interplay between environmental and Genetic factors Twin and family studies show that genetic factors may account for 50–80% of the interindividual variation of BMD Genetic polymorphisms in a large number of candidate genes have been reported to be associated with the BMD that affect the biochemical, pharmacological and physiological pathways of bone formation and bone resorption The majority of these SNPs that affect BMD and fractures have been shown in many studies with different p-values and differences between different races The most important and widespread SNPs have been identified as involved

in osteoblastogenesis, osteoclastogenesis, osteoblast Wnt signaling, vitamin D, Estrogen, Collagen, Mevalonate as LRP5 genes , LRP6, DKK1, VDR, ColA1, ESR1, ESR2…

In recent years with the rapid development of science, the advent

of GWAS (genome-wide association studies) has discovered many new SNPs related to osteoporosis and fracture skeletal Urano et al conducted a meta-analysis of eight-year GWAS studies (2007-2015) that discovered many novel SNPs associated with osteoporosis and fractures in Caucasian and Asian populations Among these SNPs involved in bone biological processes are known: WNT/β-catenin signaling pathway: catenin β1 (CTNNB1), sclerostin (SOST), LRP4, LRP5, GPR177, WNT4, WNT5B, WNT16, dickkopf1 (DKK1),

transmembrane helix protein 4 (sFRP4) fusion gene, Jagged 1 (JAG1), MEF2C and AXIN1 GWAS studies for osteoporosis have also identified three important factors, namely RANK, RANKL and OPG that influence the differentiation of mesenchymal stem cells into osteoblasts However, the authors also found several loci of genes that have not been found to play a role in bone biomechanics (FAM210A, SLC25A13 ) Research by author Kemp et al in 2013 conducted on 14,492 Britons of both sexes found 307 SNPs on 203 loci that affect the

RA at a significant level, of which 153 new loci were discovered compared with that of 203 loci In previous publications, the authors also showed that 12 SNPs were significantly associated with fracture55

In 2021, the author Zhu X and his colleagues 7 conducted a analysis of 512 GWAS studies around the world within 12 years and found 518 loci in the human genome affecting bone loss and fracture

meta-Of these, only 20% of these loci can explain their relationship to bone biomechanics Osteoporosis is a complex multifactorial disease, the influence of genes on osteoporosis, osteoporosis and fractures has been confirmed However, the actual mechanism of dynamic separation in bone remains unclear Therefore, more studies are still needed on subjects of different races, sexes, and ages to elucidate the role of genes

in osteoporosis and fractures

1.2.2 In Vietnam

In Vietnam in 2015, author Ho Pham Thuc Lan and his colleagues from Ton Duc Thang University conducted a study to determine the influence of gene polymorphisms on MS in Vietnamese The study was conducted on 564 healthy people aged 18 years and older with an average age of 47 (180 men and 384 women) living in Ho Chi Minh City, individuals were measured BM by the DXA method at the spinal position lumbar, femoral neck and upper femoral head The study subjects identified 32 gene polymorphisms (of 29 genes: LRP5, SP7, MBL2/DKK1, ESR1, DHCR7, MEF2C, SOST, VDR, RANK,

RANKL, SCL25A13, MEPE ), these gene polymorphisms were identified was found to be associated with MDD in Caucasians and some Asian countries The study results showed that 3 genes SP7 rs2016266, ZBTB40 rs7543680 and MBL2/DKK1 rs1373004 significantly reduced BMD after controlling for age, gender and weight factors These three gene polymorphisms affect from 0.2% to 1.1% of BMD, especially the SP7 rs2016266 gene polymorphism has the highest correlation, the G allele changes 0.02 g/cm2 of BMD in the femoral neck and whole body In, the author Ho Pham Thuc Lan studied 2 polymorphisms rs3736228 and rs599083 of the LRP5 gene and found no significant association of these 2 gene polymorphisms with MDR at all positions

Stemming from the above fact, we conducted a study on the polymorphism of genes related to osteoporosis in men within the framework of the study on the polymorphism of some genes related to osteoporosis and fractures vertebrae The study was conducted on two groups of subjects: postmenopausal women and men Which, we conducted this to determine the association between 3 gene

polymorphisms LRP5 Q89R, MTHFRC677T, and FTORs1121980 with

osteoporosis and vertebral fractures in postmenopausal women and men These three gene polymorphisms are correlated with MS and fracture in specific studies around the world:

CHAPTER 2 OBJECTS AND RESEARCH METHODS

2.1 Location and duration

- Location: patients at Friendship Hospital and Analysis of 3 gene polymorphisms at the Department of Pathophysiology, Hanoi Medical University

- Duration: from March 2016 to December 2021

- Male with age ≥ 50

- The patient has no history of chronic diseases that cause secondary osteoporosis (such as liver disease, chronic obstructive pulmonary disease (COPD), autoimmune disease (rheumatoid arthritis, lupus, psoriasis, .) and kidney disease chronic, cancer, endocrine diseases and disorders related to vitamin D metabolism, bone metabolism such as diabetes, obesity, malabsorption syndrome, hyperthyroidism, Cushing's syndrome, Cushing's disease ) Patients not taking drugs that cause osteoporosis (corticosteroids, hormone replacement, heparin): based on medical records and clinical examination, if symptoms are suspicious, they will be excluded from the study

- Osteoporosis is defined by a T-score ≤ -2.5 in at least 1 of 3 positions of femoral neck, total hip and lumbar spine BMD at the lumbar spine, neck, total hip was measured by dual energy X-ray absorptiometry (DEXA)

* Exclusion criteria: Patient has been immobilized for 1 month or

more; did not agree to participate in the study

2.2.2 Control group: Select patients into the control group according to

the same criteria as the patient group, but have normal BM with T index

≥ -1 in all 3 positions of the lumbar spine, femoral neck and total hip

2.3 Methods

2.3.1 Study Design: A case-control study, paired by age group

2.3.2 Study sample size

* Study sample size: Sample size in the interaction model between

genes and environment, the sample size was calculated using QUANTO software for paired case-control studies

(http://quanto.software.informer.com) Based on parameters estimated from previous studies in Vietnam and Asian ethnic groups:

- The rate of osteoporosis is 10% after the age of 50

- Number of SNPs included in the survey = 3

- Type I error (α): 0.05 with the two-sided test hypothesis adjusted; sample force is 0.80

- The ratio of alleles of interest (minor alleles) is 0.20-0.40; with the log additive inheritance mode

- Ratio of objects with interactive environmental factors: 0.25-0.4

- The main effect of genetics: 1.25 Main effect of environment: 1.25 The influence of gene-environment interaction: 4.0-6.0

- The ratio of disease control = 1:1

The calculated sample size for each group was 179 During the study, We selected 200 patients with osteoporosis and 200 subjects without osteoporosis as a control with the above inclusion and exclusion criteria

* Sampling method: convenient sample selection, subjects who

come to the clinic will be screened, bone density measured and blood drawn for testing, meeting the criteria of the disease group or the control group will have blood drawn and sent to the University of Medicine Hanoi for genotyping

2.3.3 Research variables and indicators

- Variables in general information of the study subjects: age, occupation, geography, education level were interviewed through a set

of questionnaires

- Clinical epidemiological variables and indicators: height, weight, BMI calculation, smoking, alcohol consumption, medical history, history of fracture, level of physical activity

- Bone mineral density (BMD): measured BMD by double-energy X-ray absorptiometry (DXA) at the lumbar spine and femoral neck, the total hip

- Gene polymorphisms (SNPs): determining single polymorphisms

of 3 genes LRP5 Q89R, MTHFR C677T and FTO rs1121980 of 400

research subjects by 2 methods:

LRP5 Q89R (rs41494349) and FTO rs1121980: determined by

RFLP-PCR (Restriction Fragment Length

Polymorphisms-Polymerase Chain Reaction)

MTHFR C677T determined by ARMS-PCR

Genotyping results were verified for accuracy by gene sequencing

2.3.4 Methods of data analysis

Stata 14.0 analysis software was used for statistical analysis Categorize quantitative and subgroup variables Quantitative variables were tested for normal distribution using the Kolmologov-Smirnov test The frequencies of alleles were tested for distribution according to the Hardy - Weinberg equilibrium using the squared (χ2) test or the Fisher-exact test Compare the mean values of the variables according to the normal distribution by Student T-test and ANOVA test The mean values of non-normally distributed variables were compared using the Kruskall-Wallis test Use OR (Odd ratio) to determine osteoporosis risk

of gene polymorphisms and risk factors, use correlation coefficient r to find the correlation between risk factors and BMD Analysis of the association of genotype and other factors (age, weight, height, BMI and

smoking, alcohol consumption) with osteoporosis by multivariable logistic regression

- Patient-related information is kept confidential, each patient is encrypted with a unique code

- The technique of manipulation on the patient is guaranteed to be professional - This research topic is conducted purely for scientific

purposes and not for any other purpose

CHAPTER 3: RESULTS 3.1 Demographics of all participants

The study involved 400 individuals (200 men with osteoporosis and 200 normal control men) with the criteria for matching by age group with the following characteristics:

*Average age of the osteoporosis group was 74.96±6.73; there was

no significant difference in the average age between the two study groups, in which the oldest is 91 years old and the lowest is 59 years old The age group with the highest percentage is 70-79 (52%); next is the age group over 80 (26%) the lowest is the age group under 70 (22%); The two groups of patients and controls had similar age groups because we took the data according to the matching criteria by age group

*The average height of the disease group (159.02±5.55) was lower than that of the control group (161.7±5.61) statistically significantly (p<0.001); the average weight of the patient group (55.64 ±5.55) was