目的分析一对确诊为二尖瓣脱垂并重度关闭不全的罕见双卵双生的双胞胎病例的基因突变,探讨基因突变与二尖瓣脱垂的关系。方法术中取腱索及血液标本送检,采用全外显子测序和基因拷贝数变异(copy number variations,CNV)分析。结果超声检...目的分析一对确诊为二尖瓣脱垂并重度关闭不全的罕见双卵双生的双胞胎病例的基因突变,探讨基因突变与二尖瓣脱垂的关系。方法术中取腱索及血液标本送检,采用全外显子测序和基因拷贝数变异(copy number variations,CNV)分析。结果超声检查结果显示,二尖瓣病变部位和病变形态在二者之间一致,均为二尖瓣前后叶稍冗长,腱索纤细,收缩期脱向左房。基因检测结果显示,均存在骨形成蛋白2型受体(Bone morphogenetic protein receptor type-2,BMPR2)基因突变。结论BMPR2基因外显子出现突变可能与二尖瓣脱垂相关,如能证实这一基因突变的作用,可为以后的临床诊断和治疗提供帮助。展开更多
Objective： Bone morphogenetic protein receptor 2（BMPR2） and hypoxia-inducible factor 1-α（HIF1-α） existed abnormal expression in several types of cancer. However, their expressions and related roles in osteosarc...Objective： Bone morphogenetic protein receptor 2（BMPR2） and hypoxia-inducible factor 1-α（HIF1-α） existed abnormal expression in several types of cancer. However, their expressions and related roles in osteosarcoma are largely unknown.Methods：To investigate the clinical significance of BMPR2 and HIF1-αin osteosarcoma,we analyzed their expression levels in 103 osteosarcoma specimens by immunochemistry.Meanwhile,we conducted a follow-up to examine the metastatic behavior and overall survival（OS）of osteosarcoma patients.Results：Among 103 tissues,61 cases had BMPR2-positive expression and 57 cases had HIF1-αpositive expression.A significant correlation was noticed between BMPR2 and HIF1-αexpression in osteosarcoma specimens（P=0.035）.Receiver-operating characteristic（ROC）curves were calculated to investigate the predictive value of the two markers in tumor metastasis.By means of univariate and multivariate analysis,BMPR2 and HIF1-αexpression,as well as higher tumor grade,were identified as significant risk factors for OS in patients with osteosarcoma.Kaplan-Meier survival analysis revealed that the patients with BMPR2 and HIF1-αpositive expression had worse OS compared with patients with BMPR2-negative or HIF1-α-negative staining.Conclusions：It can be concluded that BMPR2 and HIF1-αexpression is highly correlated with metastatic behavior in patients with osteosarcoma and can serve as predictive markers for metastasis and OS of these patients.展开更多
Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor,...Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II(BMPR2), E1 A binding protein p300(EP300), transforming growth factor-β2(TGFβ2), and tumor necrosis factor, and alpha-induced protein 3(TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcriptpolymerase chain reaction(qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference. Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control(P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control(P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group(P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3(r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control(P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines(P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells(P<0.05).Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.展开更多
In this study, RLM-RACE was used to identify the transcriptional start site 387 bp upstream of the translational start. Evolutionarily conserved transcription factor binding sites were identified, and a series of luci...In this study, RLM-RACE was used to identify the transcriptional start site 387 bp upstream of the translational start. Evolutionarily conserved transcription factor binding sites were identified, and a series of luciferase reporter constructs driven by BMPR2 promoter elements used to determine their functional relevance. We found the promoter area from 983 bp to 90 bp upstream of the transcriptional start gave maximal activity, greater than longer constructs, with an area between 570 bp and 290 bp upstream of the transcriptional start containing an important repressor element. To characterize this repressor, we used a combination of EMSA, mutation of the EGR1 binding site, transfection with EGR1 and NAB1 constructs, and mutation of the NAB1 binding site within the EGR1 protein. From this we conclude that EGR1 is essential to BMPR2 transcription, but that NAB1 binding to EGR1 causes it to act as a repressor.展开更多
Objective:Pulmonary atresia(PA)is a rare type of complex cyanotic congenital heart defect characterized primarily by an undeveloped pulmonary valve or pulmonary artery.Therefore,defining a disease-causing gene mutatio...Objective:Pulmonary atresia(PA)is a rare type of complex cyanotic congenital heart defect characterized primarily by an undeveloped pulmonary valve or pulmonary artery.Therefore,defining a disease-causing gene mutation in a pulmonary atresia family is a possible method of genetic counseling,future prenatal diagnosis,and therapeutic approaches for pulmonary atresia.Methods:Blood samples were collected from six PA family members,and genomic DNA was extracted using the QIAamp DNA Blood Mini Kit.Gene detection was performed using a second-generation sequencing gene panel.Results:Genetic testing results indicated that a heterozygous mutation originating from maternal inheritance was detected in the BMPR2 gene of the proband’s genomic DNA.The pathogenic gene was c.2804C>T(p.A935V).The mutation was also detected in the genomic DNA of the proband’s elder brother(III-1),but not in other family members.Conclusion:To the best of our knowledge,this is the first study to report the BMPR2 variant responsible for pulmonary atresia.The frequency of the c.2804C>T(p.A935V)mutation detected in this family is extremely low in the normal population(1/246048).The mutation was highly conserved among different species.Sorting intolerant from tolerant(SIFT)predicts it to be a harmful mutation.展开更多
Disrupted bone morphogenetic protein type 2 receptor(BMPR2)signaling in endothelial cells drives pulmonary arterial hypertension(PAH).However,targeted recovery of this signaling pathway by lipid nanoparticles(LNPs)has...Disrupted bone morphogenetic protein type 2 receptor(BMPR2)signaling in endothelial cells drives pulmonary arterial hypertension(PAH).However,targeted recovery of this signaling pathway by lipid nanoparticles(LNPs)has not been explored as a therapy.Here,we employed Design of Experiments to optimize the delivery efficiency of LNPs targeting pulmonary endothelial cells developed by our laboratory,resulting in a remarkable 35-fold increase in a simplified three-component formulation without helper lipids.Administration of BMPR2 mRNA LNPs effectively reversed established PAH in two experimental rat models(monocrotaline or SU5416-hypoxia)by reversing pulmonary vascular remodeling.Specifically,BMPR2 mRNA LNPs replenished the expression of BMPR2 protein and subsequently activated downstream pathways,as confirmed by elevated levels of p-SMAD1/5/9 and ID1 proteins.The relief of pulmonary arterial occlusion was demonstrated by thinned pulmonary arterial media and decreased proportion of full muscularized vessels.Alleviation of right ventricular hypertrophy was indicated by declined Fulton index,the cross-sectional area of right ventricular cardiomyocytes as well as collagen deposition.Effective recovery of right ventricular function was evidenced by increased pulmonary artery flow acceleration time/pulmonary artery flow ejection time ratio.These findings underscore the potential of restoring BMPR2 signaling through pulmonary endothelial cell-specific LNPs for treating PAH.展开更多
Background Familial pulmonary arterial hypertension （FPAH） is an autosomal dominant disorder characterized by plexiform lesions of endothelial cells in pulmonary arterioles which leads to elevated pulmonary arterial...Background Familial pulmonary arterial hypertension （FPAH） is an autosomal dominant disorder characterized by plexiform lesions of endothelial cells in pulmonary arterioles which leads to elevated pulmonary arterial pressure, right-sided heart failure and death. Heterozygous mutations in the bone morphogenetic protein type II receptor gene （BMPR2） have been found to underlie a majority of FPAH cases. More than 140 distinct mutations have been identified in FPAH cases and in idiopathic pulmonary arterial hypertension （IPAH） cases, but only one mutation has been reported in Chinese patients. Methods A three-generation pedigree of FPAH and another 10 patients with IPAH were collected. In the family, two of the 9 surviving and one deceased family member were diagnosed as FPAH. The entire protein-coding region and intron/exon boundaries of the BMPR2 gene were amplified by PCR using DNA samples from affected individuals. Direct sequencing of PCR products was performed on both the sense and antisense strands. To confirm the segregation of the mutation within the family and exclude the presence of the mutation in normal subjects, the relevant exon was amplified by PCR, followed by mutation-specific RPLP analysis. Results In the Chinese pedigree with FPAH an A-to-T transition at position 1157 in exon 9 of the BMPR2 gene was identified which resulted in a Glu386Val mutation. We confirmed the segregation of the mutation within the family and excluded the presence of the mutation in a panel of 200 chromosomes from normal subjects. No mutation was detected in BMPR2 in the other 10 patients with IPAH. Conclusions This amino acid substitution occurs at a glutamic acid that is highly conserved in all type Ⅱ TGF-β receptors. The nearly invariant Glu forms an ion pair with an invariant Arg at position 491 thereby helping to stabilize the large lobe. Substitution of Arg at position 491 is the most frequently observed missense mutation in FPAH, but until now no mutations at position 386 have been found in FPAH. The predicted functional impact of the Glu386Val mutation and its absence in healthy controls support the mutation as the cause of FPAH.展开更多
文摘目的分析一对确诊为二尖瓣脱垂并重度关闭不全的罕见双卵双生的双胞胎病例的基因突变,探讨基因突变与二尖瓣脱垂的关系。方法术中取腱索及血液标本送检,采用全外显子测序和基因拷贝数变异(copy number variations,CNV)分析。结果超声检查结果显示,二尖瓣病变部位和病变形态在二者之间一致,均为二尖瓣前后叶稍冗长,腱索纤细,收缩期脱向左房。基因检测结果显示,均存在骨形成蛋白2型受体(Bone morphogenetic protein receptor type-2,BMPR2)基因突变。结论BMPR2基因外显子出现突变可能与二尖瓣脱垂相关,如能证实这一基因突变的作用,可为以后的临床诊断和治疗提供帮助。
基金supported by grants from the National Natural Science Foundation of China (No. 81572633)
文摘Objective： Bone morphogenetic protein receptor 2（BMPR2） and hypoxia-inducible factor 1-α（HIF1-α） existed abnormal expression in several types of cancer. However, their expressions and related roles in osteosarcoma are largely unknown.Methods：To investigate the clinical significance of BMPR2 and HIF1-αin osteosarcoma,we analyzed their expression levels in 103 osteosarcoma specimens by immunochemistry.Meanwhile,we conducted a follow-up to examine the metastatic behavior and overall survival（OS）of osteosarcoma patients.Results：Among 103 tissues,61 cases had BMPR2-positive expression and 57 cases had HIF1-αpositive expression.A significant correlation was noticed between BMPR2 and HIF1-αexpression in osteosarcoma specimens（P=0.035）.Receiver-operating characteristic（ROC）curves were calculated to investigate the predictive value of the two markers in tumor metastasis.By means of univariate and multivariate analysis,BMPR2 and HIF1-αexpression,as well as higher tumor grade,were identified as significant risk factors for OS in patients with osteosarcoma.Kaplan-Meier survival analysis revealed that the patients with BMPR2 and HIF1-αpositive expression had worse OS compared with patients with BMPR2-negative or HIF1-α-negative staining.Conclusions：It can be concluded that BMPR2 and HIF1-αexpression is highly correlated with metastatic behavior in patients with osteosarcoma and can serve as predictive markers for metastasis and OS of these patients.
基金supported by the Guangdong Province Key Foundation of Science and Technology Program (Grant No.2009B0507000029)the Guangdong Province Science and Technology Program (Grant No.2012B031800474)a grant from the Overseas Chinese Affairs Office of the State Council Key Discipline Construction Fund (Grant No.51205002)
文摘Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II(BMPR2), E1 A binding protein p300(EP300), transforming growth factor-β2(TGFβ2), and tumor necrosis factor, and alpha-induced protein 3(TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcriptpolymerase chain reaction(qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference. Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control(P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control(P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group(P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3(r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control(P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines(P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells(P<0.05).Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.
文摘In this study, RLM-RACE was used to identify the transcriptional start site 387 bp upstream of the translational start. Evolutionarily conserved transcription factor binding sites were identified, and a series of luciferase reporter constructs driven by BMPR2 promoter elements used to determine their functional relevance. We found the promoter area from 983 bp to 90 bp upstream of the transcriptional start gave maximal activity, greater than longer constructs, with an area between 570 bp and 290 bp upstream of the transcriptional start containing an important repressor element. To characterize this repressor, we used a combination of EMSA, mutation of the EGR1 binding site, transfection with EGR1 and NAB1 constructs, and mutation of the NAB1 binding site within the EGR1 protein. From this we conclude that EGR1 is essential to BMPR2 transcription, but that NAB1 binding to EGR1 causes it to act as a repressor.
基金This work was supported by Shanghai Children’s HospitalChinese National Natural Science Foundation,No.81371499。
文摘Objective:Pulmonary atresia(PA)is a rare type of complex cyanotic congenital heart defect characterized primarily by an undeveloped pulmonary valve or pulmonary artery.Therefore,defining a disease-causing gene mutation in a pulmonary atresia family is a possible method of genetic counseling,future prenatal diagnosis,and therapeutic approaches for pulmonary atresia.Methods:Blood samples were collected from six PA family members,and genomic DNA was extracted using the QIAamp DNA Blood Mini Kit.Gene detection was performed using a second-generation sequencing gene panel.Results:Genetic testing results indicated that a heterozygous mutation originating from maternal inheritance was detected in the BMPR2 gene of the proband’s genomic DNA.The pathogenic gene was c.2804C>T(p.A935V).The mutation was also detected in the genomic DNA of the proband’s elder brother(III-1),but not in other family members.Conclusion:To the best of our knowledge,this is the first study to report the BMPR2 variant responsible for pulmonary atresia.The frequency of the c.2804C>T(p.A935V)mutation detected in this family is extremely low in the normal population(1/246048).The mutation was highly conserved among different species.Sorting intolerant from tolerant(SIFT)predicts it to be a harmful mutation.
基金sponsored by Science and Technology Commission of Shanghai Municipality(YDZX20223100001002,China)the Natural Science Foundation of Shanghai(25ZR1401251)the ShanghaiTech University Startup Grant.
文摘Disrupted bone morphogenetic protein type 2 receptor(BMPR2)signaling in endothelial cells drives pulmonary arterial hypertension(PAH).However,targeted recovery of this signaling pathway by lipid nanoparticles(LNPs)has not been explored as a therapy.Here,we employed Design of Experiments to optimize the delivery efficiency of LNPs targeting pulmonary endothelial cells developed by our laboratory,resulting in a remarkable 35-fold increase in a simplified three-component formulation without helper lipids.Administration of BMPR2 mRNA LNPs effectively reversed established PAH in two experimental rat models(monocrotaline or SU5416-hypoxia)by reversing pulmonary vascular remodeling.Specifically,BMPR2 mRNA LNPs replenished the expression of BMPR2 protein and subsequently activated downstream pathways,as confirmed by elevated levels of p-SMAD1/5/9 and ID1 proteins.The relief of pulmonary arterial occlusion was demonstrated by thinned pulmonary arterial media and decreased proportion of full muscularized vessels.Alleviation of right ventricular hypertrophy was indicated by declined Fulton index,the cross-sectional area of right ventricular cardiomyocytes as well as collagen deposition.Effective recovery of right ventricular function was evidenced by increased pulmonary artery flow acceleration time/pulmonary artery flow ejection time ratio.These findings underscore the potential of restoring BMPR2 signaling through pulmonary endothelial cell-specific LNPs for treating PAH.
文摘Background Familial pulmonary arterial hypertension （FPAH） is an autosomal dominant disorder characterized by plexiform lesions of endothelial cells in pulmonary arterioles which leads to elevated pulmonary arterial pressure, right-sided heart failure and death. Heterozygous mutations in the bone morphogenetic protein type II receptor gene （BMPR2） have been found to underlie a majority of FPAH cases. More than 140 distinct mutations have been identified in FPAH cases and in idiopathic pulmonary arterial hypertension （IPAH） cases, but only one mutation has been reported in Chinese patients. Methods A three-generation pedigree of FPAH and another 10 patients with IPAH were collected. In the family, two of the 9 surviving and one deceased family member were diagnosed as FPAH. The entire protein-coding region and intron/exon boundaries of the BMPR2 gene were amplified by PCR using DNA samples from affected individuals. Direct sequencing of PCR products was performed on both the sense and antisense strands. To confirm the segregation of the mutation within the family and exclude the presence of the mutation in normal subjects, the relevant exon was amplified by PCR, followed by mutation-specific RPLP analysis. Results In the Chinese pedigree with FPAH an A-to-T transition at position 1157 in exon 9 of the BMPR2 gene was identified which resulted in a Glu386Val mutation. We confirmed the segregation of the mutation within the family and excluded the presence of the mutation in a panel of 200 chromosomes from normal subjects. No mutation was detected in BMPR2 in the other 10 patients with IPAH. Conclusions This amino acid substitution occurs at a glutamic acid that is highly conserved in all type Ⅱ TGF-β receptors. The nearly invariant Glu forms an ion pair with an invariant Arg at position 491 thereby helping to stabilize the large lobe. Substitution of Arg at position 491 is the most frequently observed missense mutation in FPAH, but until now no mutations at position 386 have been found in FPAH. The predicted functional impact of the Glu386Val mutation and its absence in healthy controls support the mutation as the cause of FPAH.
