1.Association of the polymorphism of platelet membrane glycoprotein I a gene with myocardial infarction.
Yong-hui ZHAO ; Yan-ni WANG ; Jia-qing ZHU ; Ai-qun MA ; Chang-zong CUI ; Qing-bin ZHAO
Chinese Journal of Medical Genetics 2003;20(5):417-420
OBJECTIVEPlatelet membrane glycoprotein (GP) Ia/IIa complex is the major collagen receptor on platelets. Platelet activation by GP Ia/ IIa dependent adhesion leads to cellular events that catalyze prothrombin conversion and fibrin clot formation. Correlation between the polymorphism of platelet membrane GP Ia gene and myocardial infarction (MI) was explored.
METHODSA total of 137 patient s with myocardial infarction and 175 controls with no history of coronary heart disease, thrombogenic and hemorrhagenic diseases were studied by case-control. Platelet GP I a gene 807 C/T polymorphisms were checked by polymerase chain reaction-sequence specific primers.
RESULTSThere were significant differences in the distribution of T and C alleles between MI and control groups (T:42.70% vs 32.00%, C:57.30% vs 68.00%, P<0.001). No matter among all subjects or among subjects aged CONCLUSIONThe above data suggest that there is a strong association between the presence of GP Ia T allele and MI. T allele ca n be a marker of genetic susceptibility to MI. These need to be substantiated by a large scale and prospective study.
Aged
;
Alleles
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Female
;
Gene Frequency
;
Genotype
;
Humans
;
Integrin alpha2
;
genetics
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Male
;
Middle Aged
;
Myocardial Infarction
;
genetics
;
pathology
;
Polymorphism, Genetic
2.Identification of compound heterozygous mutations in the ITGA2B gene in a Chinese patient with Glanzmann thrombasthenia.
Jia-yong ZHENG ; Yan-hui JIN ; Yong-lin ZHU ; Pei-pei JIN ; De-ting ZHANG ; Zi-bing JIN
Chinese Medical Journal 2010;123(11):1397-1401
BACKGROUNDGlanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by the tendency to hemorrhage and the inability of platelets to aggregate in response to agonists. GT is caused by a defect of the platelet glycoprotein IIb/IIIa complex. The objective of this study was to describe the clinical features and the genetic cause of GT in a 6-year-old girl from south China.
METHODSA three-generation family was studied. The proband patient aged 6 years and her parents undertook examinations of platelet counts, blood film, bleeding time, platelet aggregation, and flow cytometry. All coding exons of the ITGA2B and ITGB3 genes were amplified by polymerase chain reaction (PCR), and direct sequencing was performed for mutational screening on the patient and normal controls consisted of 52 healthy blood donors. Reverse transcription PCR was conducted to test for exon skipping.
RESULTSThe proposita patient showed dispersing platelets, prolonged bleeding time, and severely reduced platelet aggregation in response to the physiological agonists adenosine diphosphate (ADP), epinephrine, collagen, and ristocetin. Flow cytometric measurements showed that the contents of alphaIIb and beta3 were significantly decreased. Sequencing results demonstrated two different types of heterozygous mutations existed in the alphaIIb gene (c.2930delG and IVS15-1delG). The compound mutations were also confirmed in the patient's mother and father separately.
CONCLUSIONSThe alphaIIbbeta3 deficiency of the proband was caused by two compound ITGA2B mutations, which were first reported in Chinese GT patients. The IVS15-1delG was first confirmed to cause an exon skipping.
Asian Continental Ancestry Group ; Child ; Female ; Flow Cytometry ; Heterozygote ; Humans ; Integrin alpha2 ; genetics ; Integrin beta3 ; genetics ; Mutation ; Pedigree ; Reverse Transcriptase Polymerase Chain Reaction ; Thrombasthenia ; genetics ; metabolism ; pathology
3.Association of the platelet membrane glycoprotein I a C807T gene polymorphism with aspirin resistance.
Guanhua, SU ; Zhaohui, WANG ; Yanping, DING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(6):664-7
To explore the correlation between the C807T polymorphism of platelet membrane glycoprotein I a (GP I a) gene and aspirin resistance in Chinese people, 200 patients with high-risk of atherosclerosis took aspirin (100 mg/d) for 7 days. Platelet aggregation function was detected using adenosine diphosphate (ADP) and arachidonic acid (AA) before and after the administration of aspirin. Then the subjects were divided into three groups according to the results of platelet aggregation function: an aspirin resistant (AR) group, an aspirin semi-responder (ASR) group and an aspirin-sensitive (AS) group. Platelet GP I a gene 807CT polymorphism was examined by means of polymerase chain reaction-sequence specific primers (PCR-SSP). The results showed that T allelic frequency in AR group and ASR group were higher that of AS group (P<0.005), and the prevalence of genotypes (TT+TC) of these two groups was significantly higher than that in AS group (P<0.05). Platelet GP I a T allele was significantly associated with aspirin resistance as revealed by multiple logistic regression (OR=3.76, 95% CI: 2.87-9.58). The results suggest that inherited platelet GP I a variations may have an important impact on aspirin resistance and the presence of GP I a T allele may be a marker of genetic susceptibility to aspirin resistance.
Aspirin/*administration & dosage
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Atherosclerosis/drug therapy
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Atherosclerosis/genetics
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Drug Resistance/*genetics
;
Integrin alpha2/*genetics
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Platelet Aggregation Inhibitors/*administration & dosage
;
Polymorphism, Genetic/*genetics
4.Effects of WWOX on ovarian cancer cell attachment in vitro.
Jie-Qing ZHANG ; Li LI ; Hong-Lin SONG ; Paige ADAM ; Gabra HANI
Chinese Journal of Oncology 2009;31(6):414-417
OBJECTIVETo observe the effects of WWOX on cell attachment in ovarian cancer, and to explore its mechanisms of action.
METHODSAttachment assay was used to assess the adhesion of wwox-transfected PEO1 cells and vector-transfected PEO1 cells that were constructed, as well as PEO1 parent cells. Alpha/beta integrin-mediated cell adhesion assays were designed to identify cells surface integrins in PEO1 clone cells. Integrin function blocking experiments were designed to further determine integrins in PEO1 clone cells according to the integrin that was selected in integrin expression profiling. FACS analysis was used to further detect the level of integrin alpha3 on the cell membrane.
RESULTSAttachment assay showed that adhesion of WWOX-transfected PEO1 cells to fibronectin was significantly slower than that in vector-transfected controls or PEO1 parent cells, cultured on the pre-coated fibronectin for 2 hours (P<0.01). The level of membranous integrins alpha2 and alpha3 in the WWOX-transfected PEO1 cells was significantly decreased, as compared with that in vector-transfected controls (P<0.05), but there was no association with the level of functioning integrins betal or beta2 in clone cells (P>0.05). The attachment assays were repeated after pre-incubating the cells with integrin alpha2 or alpha3 function-blocking antibodies. These results showed that blocking integrin alpha3 significantly reduced the binding to fibronectin of all the PEO1 clonal lines, as compared with cells pre-incubated with a non-specific IgG antibody (P<0.05). In contrast, preincubation with alpha2 blocking antibody had very little effect on fibronectin binding in these cells (P>0.05). FACS analysis showed that membranous integrin alpha3 expression revealed a marked reduction in WWOX-transfected cells than that in vector-transfected cells.
CONCLUSIONWWOX acts as an ovarian tumor suppressor by modulating the interaction between tumor cells and the extracellular matrix, decreasing integrin activity and adhesion of tumor cells to fibronectin. This suggests an important role for loss of WWOX tumor suppressor in promoting attachment and adhesion of ovarian cancer cells on locoregional peritoneum, and further resulting in enhancing locoregional peritoneal tumor spread.
CD18 Antigens ; metabolism ; Cell Adhesion ; Female ; Fibronectins ; metabolism ; Humans ; Integrin alpha2 ; metabolism ; Integrin alpha3 ; metabolism ; Integrin beta1 ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; Oxidoreductases ; genetics ; metabolism ; Protein Binding ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism ; WW Domain-Containing Oxidoreductase
5.Association of the integrin gene polymorphisms with ischemic stroke and plasma lipid levels.
Ye-sheng WEI ; Yan LAN ; Yun-guang LIU ; Lan-qing MENG ; Qun-qing XU ; Hai-yuan XIE
Chinese Journal of Medical Genetics 2009;26(2):211-215
OBJECTIVETo study the association of integrin alpha-2 (ITGA2) gene C807T, integrin beta-3 (ITGB3) gene T176C polymorphisms with ischemic stroke and the effect of the polymorphisms on plasma lipid and lipoprotein levels.
METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to detect the integrin genotypes in 265 patients with ischemic stroke and 280 healthy controls. The plasma lipid and lipoprotein levels were measured by routine method.
RESULTSPlasma total cholesterol (TC), triacylglycerol (TG) and low density lipoprotein-cholesterol (LDL-C) in the patients with ischemic stroke were significantly higher than those in the controls (P< 0.05). The distributions of the ITGB3 gene T176C polymorphism were not different between the ischemic stroke group and control group, but the ITGA2 gene C807T polymorphism was significantly different. The relative risk suffering from ischemic stroke of the T allele carrier was 1.455 times as that of the C allele carrier (OR=1.455, 95%CI: 1.134-1.866). The level of plasma lipid in the T allele carriers was significantly higher than that in the C allele carriers (P< 0.05).
CONCLUSIONThe ITGA2 gene C807T polymorphism was associated with ischemic stroke, the 807 T allele may be a genetic risk factor for ischemic stroke. The ITGA2 gene C807T polymorphism may affect ischemic stroke through plasma lipid and lipoprotein levels.
Brain Ischemia ; blood ; genetics ; metabolism ; Cholesterol, LDL ; genetics ; metabolism ; Female ; Genetic Predisposition to Disease ; Humans ; Integrin alpha2 ; genetics ; metabolism ; Integrin beta3 ; genetics ; metabolism ; Lipid Metabolism ; genetics ; Lipids ; blood ; Male ; Middle Aged ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide
6.Effects of the ITGA2B Nonsense Mutation (c.2659C > T, p.Q887X) on Platelet Function in a Mouse Model of Glanzmann's Thrombasthenia Generated with CRISPR/Cas9 Technology.
Fei YANG ; Miao JIANG ; Zeng-Hua LIN ; Zhan-Li XIE ; Zhen-Ni MA ; Li YANG ; Hong LIU ; Zhao-Yue WANG ; Lu ZHOU
Journal of Experimental Hematology 2022;30(2):559-564
OBJECTIVE:
To construct a mouse model of Glanzmann's thrombasthenia (GT) with ITGA2B c.2659 C>T (p.Q887X) nonsense mutation by CRISPR/Cas9 technology, and then further explore the expression and function of glycoprotein αIIbβ3 on the surface of platelet membrane.
METHODS:
The donor oligonucleotide and gRNA vector were designed and synthesized according to the ITGA2B gene sequence. The gRNA and Cas9 mRNA were injected into fertilized eggs with donor oligonucleotide and then sent back to the oviduct of surrogate mouse. Positive F0 mice were confirmed by PCR genotyping and sequence analysis after birth. The F1 generation of heterozygous GT mice were obtained by PCR and sequencing from F0 bred with WT mice, and then homozygous GT mice and WT mice were obtained by mating with each other. The phenotype of the model was then further verified by detecting tail hemorrhage time, saphenous vein bleeding time, platelet aggregation, expression and function of αIIbβ3 on the surface of platelet.
RESULTS:
The bleeding time of GT mice was significantly longer than that of WT mice (P<0.01). Induced by collagen, thrombin, and adenosine diphosphate (ADP), platelet aggregation in GT mice was significantly inhibited (P<0.01, P<0.01, P<0.05). Flow cytometry analysis showed that the expression of αIIbβ3 on the platelet surface of GT mice decreased significantly compared with WT mice (P<0.01), and binding amounts of activated platelets to fibrinogen were significantly reduced after thrombin stimulation (P<0.01). The spreading area of platelet on fibrinogen in GT mice was significantly smaller than that in WT mice (P<0.05).
CONCLUSION
A GT mouse model with ITGA2B c.2659 C>T (p.Q887X) nonsense mutation has been established successfully by CRISPR/Cas9 technology. The aggregation function of platelet in this model is defective, which is consistent with GT performance.
Animals
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CRISPR-Cas Systems
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Codon, Nonsense
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Disease Models, Animal
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Fibrinogen/genetics*
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Humans
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Integrin alpha2/genetics*
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Mice
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Oligonucleotides
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Platelet Glycoprotein GPIIb-IIIa Complex/genetics*
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RNA, Guide
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Thrombasthenia/genetics*
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Thrombin/genetics*
7.Correlation between the polymorphism of glycoprotein Ia gene and acute coronary syndrome.
Yonghui ZHAO ; Yanni WANG ; Jiaqing ZHU
Chinese Medical Sciences Journal 2004;19(1):13-18
OBJECTIVEThe platelet membrane glycoprotein (GP) Ia/IIa plays a major part as a primary collagen receptor in platelet function. Previous studies indicated that variations of GPIa/IIa density and function are associated with the 807 C/T polymorphism of GPIa gene in American and Spanish Caucasian populations. This study investigated the correlation between acute coronary syndrome (ACS) and 807 C/T dimorphism of GPIa gene in Chinese of Han ethnicity.
METHODSA case-control study was carried out, including 75 patients with either acute myocardial infarction (AMI) or unstable angina pectoris (UAP), and 65 controls with no history of coronary heart disease, thrombogenic and hemorrhagenic diseases. Genotypes of GPIa were checked by polymerase chain reaction-sequence specific primers (PCR-SSP) technique.
RESULTSThe frequencies of both homozygotes and heterozygotes for T807 allele (TT+TC) were significantly higher in patients with AMI than in controls (62.16% vs 33.85%, P<0.01; odds ratio 3.21). The prevalence of (TT+TC) genotypes was also markedly higher in patients with UAP than in controls (65.79% vs 33.85%, P < 0.005; odds ratio 3.76). There was significant difference in the distribution of (TT+TC) genotypes not only between all patients and controls (64.00% vs 33.85%, P<0.005; odds ratio 3.47) but also between the two subgroups aged <60 years (70.00% vs 38.24%, P<0.005; odds ratio 3.77). However, there was no significant difference in the distribution of (TT+TC) genotypes between patients with AMI and with UAP. Platelet GPIa T807 allele remained significantly associated with AMI and UAP by multiple logistic regression (odds ratio 4.94).
CONCLUSIONThis study suggests a strong association between presence of GPIa T807 allele and ACS. T807 allele can be a marker of genetic susceptibility to ACS.
Aged ; Alleles ; Angina, Unstable ; genetics ; Asian Continental Ancestry Group ; Case-Control Studies ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Integrin alpha2 ; genetics ; Male ; Middle Aged ; Myocardial Infarction ; genetics ; Odds Ratio ; Polymorphism, Genetic
8.Association of the Gene Polymorphisms of Platelet Glycoprotein Ia and IIb/IIIa with Myocardial Infarction and Extent of Coronary Artery Disease in the Korean Population.
Sungha PARK ; Hyun Young PARK ; Chanmi PARK ; Young Guk KO ; Eun Kyung IM ; Inho JO ; Chol SHIN ; Jong Bok LEE ; Won Heum SHIM ; Seung Yun CHO ; Yangsoo JANG
Yonsei Medical Journal 2004;45(3):428-434
Platelet membrane receptor glycoproteins (GP) are essential for the platelet activation process, and the genetic polymorphisms in the genes that encode platelet glycoproteins have been proposed to influence the risk of acute coronary syndrome and atherosclerosis. In this study, we investigated the role of GPIa, HPA-1 and HPA-3 polymorphisms as putative risk factors for myocardial infarction (MI) and the extent of coronary artery disease. We selected 1, 073 subjects who underwent coronary angiography; 242 had normal or minimal coronary atherosclerosis, and 831 patients had significant coronary artery disease (CAD). The genotype was determined by the methods of single base extension for C807T/G873A polymorphisms of GPIa, and restriction fragment length polymorphism for HPA-1 and HPA-3. The C807T and G873A polymorphisms of GPIa showed complete linkage in the Korean population. For HPA-1 gene polymorphism, only the HPA-1a/a (PlA1/A1) genotype was observed in 192 selected subjects from our study population. The distribution of GPIa (C807T/G873A) and HPA-3 genotypes did not differ significantly between normal subjects and CAD subjects. No significant association between MI and both gene polymorphisms was present. However, for the subgroup analysis of young male patients whose age was less than 56 years, the genotype frequency of HPA-3b/b was significantly lower in patients with MI compared to patients without a history of MI (7.5% vs. 20.0%, p=0.04). The odds ratio for HPA-3 b homozygosity versus the HPA-3a carrier was 0.32 (95% CI, 0.10- 0.99, p=0.04). Conclusively, HPA-3 polymorphism was associated with MI in Korean individuals younger than 56 years of age, but other polymorphisms of GP, which we studied, were not associated with both the extent of coronary atherosclerosis or MI.
Aged
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Coronary Arteriosclerosis/epidemiology/*genetics
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Female
;
Gene Frequency
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Genetic Predisposition to Disease/epidemiology
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Genotype
;
Human
;
Integrin alpha2/*genetics
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Integrin beta3/*genetics
;
Korea
;
Male
;
Middle Aged
;
Myocardial Infarction/epidemiology/*genetics
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Platelet Membrane Glycoprotein IIb/*genetics
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*Polymorphism (Genetics)
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Risk Factors
;
Support, Non-U.S. Gov't
9.Role of IL-1alpha in Cisplatin-Induced Acute Renal Failure in Mice.
Jay Wook LEE ; Woo Jin NAM ; Min Jee HAN ; Jung Ho SHIN ; Jin Gun KIM ; Su Hyun KIM ; Hye Ryoun KIM ; Dong Jin OH
The Korean Journal of Internal Medicine 2011;26(2):187-194
BACKGROUND/AIMS: For unknown reasons, caspase-1 -/- mice, protected against cisplatin-induced acute renal failure (ARF), are deficient in interleukin (IL)-1alpha. We thus asked whether IL-1alpha deficiency underlies the mechanism of protection against cisplatin-induced ARF in these mice. METHODS: Cisplatin (30 mg/kg) was injected intraperitoneally into wild-type C57BL/6 mice to produce a cisplatin-induced model of ARF. IL-1alpha was measured in control vehicle- and cisplatin-treated wild-type animals. We also examined whether IL-1alpha -/- mice were similarly protected against cisplatin-induced ARF. Additionally, infiltration of CD11b- and CD49b-positive cells, as markers of macrophages, natural killer, and natural killer T cells (pan-NK cells), was investigated in wild-type and IL-1alpha -/- mice. RESULTS: Compared with vehicle-treated mice, renal IL-1alpha increased in cisplatin-treated wild-type mice beginning on day 1. IL-1alpha -/- mice were shown to be protected against cisplatin-induced ARF. No significant difference in the infiltration of neutrophils or CD11b- and CD49b-positive cells were observed between wild-type and IL-1alpha -/- mice. CONCLUSIONS: Mice deficient in IL-1alpha are protected against cisplatin-induced ARF. The lack of IL-1alpha may explain, at least in part, the protection against cisplatin-induced ARF observed in caspase-1 -/- mice. Investigation of the protective mechanism (s) in IL-1alpha -/- mice in cisplatin-induced ARF merits further study.
Acute Kidney Injury/chemically induced/*immunology/pathology/physiopathology/prevention & control
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Animals
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Antigens, CD11b/analysis
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Apoptosis
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Biological Markers/blood
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Blood Urea Nitrogen
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*Cisplatin
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Creatinine/blood
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Disease Models, Animal
;
Fluorescent Antibody Technique
;
Integrin alpha2/analysis
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Interleukin-1alpha/deficiency/genetics/*metabolism
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Kidney/*immunology/pathology/physiopathology
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Killer Cells, Natural/immunology
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Macrophages/immunology
;
Mice
;
Mice, Inbred C57BL
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Mice, Transgenic
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Natural Killer T-Cells/immunology
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Necrosis
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Neutrophil Infiltration
;
Time Factors