Pancreatic agenesis caused by GATA6 gene mutation: a case report and literature review / 中国新生儿科杂志

Objective:To explore the genotype and clinical phenotype characteristics of patients with pancreatic agenesis caused by GATA6 gene mutations and to improve the clinical understanding of pancreatic agenesis.Methods:The clinical data of a newborn with pancreatic agenesis admitted to the Second Hospital of Shandong University were retrospectively analyzed. Relevant literature published until October 31, 2022, were retrieved from China National Knowledge Infrastructure, Wanfang Database, VIP Database, Chinese Medical Journal Full Text Database, PubMed, Embase and SCI Database with the terms of "pancreatic agenesis", "GATA6", "pancreatic agenesis/hypoplasia" and "GATA6 Translation Factor". The characteristics of gene variants and clinical manifestations of patients diagnosed with pancreatic agenesis caused by GATA6 gene mutation were retrieved and summarized.Results:This case was a full-term male infant who developed insulin dependent hyperglycemia and fatty diarrhea 2 d after birth, accompanied by intrauterine growth restriction, congenital heart disease, and cryptorchidism. Genetic testing showed a novel heterozygous mutation of GATA6 (c.1366C>T) which was consistent with the autosomal dominant inheritance pattern. The phenotype and genotype between the proband and his parents were consistent with the cosegregation. The ACMG mutation was rated as pathogenic variant. Intravenous infusion of insulin, subcutaneous injection of insulin, or long-acting insulin were not effective. After continuous subcutaneous pumping of aspartic insulin combined with oral pancreatic enzyme replacement therapy, the infant's condition was improved and discharged. Follow up to age of 15 months, the patient still relied on continuously subcutaneous pump to control blood glucose, pancreatic exocrine function was back to normal, and the development was generally normal. A total of 59 cases were reported in 22 articles, with the case from our hospital, there were 60 patients in total. Among them, 47 were probands and 13 were family members, about 61.7% (29/47) of which were de novo mutations. There were 39 variants, of which 28.2% (11/39) were missense mutations and 71.8% (28/39) were functional deletion variations. Mutations of GATA6 gene had a broad phenotype spectrum. The phenotypes mainly included neonatal diabetes mellitus ( n=39) and pancreatic exocrine insufficiency ( n=39). Other extra-pancreatic features included different types of congenital heart disease ( n=54), congenital biliary abnormalities ( n=23), intestinal developmental disorders ( n=16), neurocognitive disorders ( n=18) and endocrine abnormalities ( n=15). Conclusions:The heterozygous variations of GATA6 gene lead to pancreatic hypoplasia and a broad phenotype spectrum. The pancreatic phenotypes mainly include neonatal diabetes mellitus and pancreatic exocrine insufficiency, and extra-pancreatic phenotypes include congenital heart disease and other developmental abnormalities.

Long non-coding RNA GATA6-AS1 inhibits proliferation,apoptosis,and metastasis of 5-fluorouracil resistant gastric cancer cells SGC7901/5-FU by regulating GATA6 / 肿瘤

Objective:To investigate the effects of regulation of GATA6 expression by long non-coding RNA(lncRNA)GATA6 antisense RNA 1(GATA6-AS1)on the proliferation,apoptosis,and metastasis of 5-fluorouracil(5-FU)resistant gastric cancer(GC)cells SGC7901/5-FU,and to explore the underlying mechanisms. Methods:Parental SGC7901 cells and SGC7901/5-FU cells were used as controls(treated with blank culture medium).Empty vector pcDNA3.1(control for GATA6-AS1 overexpression),recombinant vector pcDNA3.1-GATA6-AS1(for GATA6-AS1 overexpression),shNC(negative control for GATA6-AS1 silencing)or shGATA6-AS1(for GATA6-AS1 silencing)were transfected into SGC7901/5-FU cells by Lipofectamine 2000 to overexpress or silent GATA-AS1.The mRNA expression of GATA6-AS1 and GATA6 in cells of different treatment groups were examined by real-time fluorescence quantitative PCR.After 5-FU treatment,the proliferation,migration,invasion and apoptosis were analyzed by CCK-8 assay,wound-healing assay,Transwell assay,and flow cytometry(FCM)assay,respectively.The protein expression levels of Bax,Bcl-2,Caspase 3 and GATA6 in different treatment groups were examined by Western blotting.The stability of GATA6 mRNA was evaluated after α-amanitin treatment.The relationship between GATA6-AS1 and GATA6 was studied by RNA pull-down experiment.Furthermore,the SGC7901 cell transplantation tumor model was established using Balb/c nude mice,and the grouping was the same as in vitro experiments with 12 mice in each group.The tumor growth was recored and the tumor inhibition rate was calculated after 5-FU treatment.The histopathological changes of tumor tissue in each group was assessed by HE staining.The protein expression of proliferating cell nuclear antigen(PCNA)in tumor tissues of nude mice in each group was detected by Western blotting. Results:Compared with parental SGC7901 cells(control group),the expression of GATA6-AS1 and GATA6 mRNA was significantly downregulated in SGC7901/5-FU cells(P＜0.05);the proliferation,migration and invasion of SGC7901/5-FU cells was significantly decreased while the apoptosis of SGC7901/5-FU cells was significantly decreased(P＜0.05);the protein expression levels of GATA6,Bax and Caspase 3 were significantly decreased while that of Bcl-2 was significantly increased(P＜0.05).Compared with SGC7901/5-FU-pcDNA group,the above changes showed opposite trends in SGC7901/5-FU cells overexpressing GATA6-AS1.Compared with SGC7901/5-FU-shNC group,the expression of GATA6-AS1 and GATA6 mRNA was significantly downregulated in GATA6-AS1-silencing SGC7901/5-FU cells(P＜0.05);the proliferation,migration and invasion of GATA6-AS1-silencing SGC7901/5-FU cells was significantly decreased while apoptosis of GATA6-AS1-silencing SGC7901/5-FU cells was significantly decreased(P＜0.05);the protein expression levels of GATA6,Bax and Caspase 3 were significantly decreased while that of Bcl-2 was significantly increased(P＜0.05).GATA6-AS1 can increase the stability of GATA6 mRNA and positively regulate the expression of GATA6.In vivo characterization showed that,compared with mice transplanted with parental SGC7901 cells,the tumor volume and PCNA protein expression in tumor tissues were significantly increased in mice transplanted with drug resistant SGC7901/5-FU cells after 5-FU treatment(P＜0.05).Mice in SGC7901/5-FU-GATA6-AS1 group showed significantly reduced tumor volume and PCNA protein expression compared with the SGC7901/5-FU-pcDNA group(P＜0.05),and pathological analysis of the tumor tissues revealed milder development of GC in SGC7901/5-FU-GATA6-AS1 group.Whereas,mice in SGC7901/5-FU-shGATA6-AS1 group showed significantly increased tumor volume and PCNA protein expression compared with the SGC7901/5-FU-shNC group(P＜0.05),and histopathological analysis of the tumor tissues revealed severer development of GC in SGC7901/5-FU-shGATA6-ASl group. Conclusion:Overexpression of GATA6-AS1 can promote the expression of GATA6,inhibit the growth of GC cells,and thus reverse the resistance of SGC7901/5-FU cells to 5-FU.

Clinical characteristics and genetic analysis of a child with specific type of diabetes mellitus caused by missense mutation of GATA6 gene / 浙江大学学报·医学版

A 2-year-old boy was admitted to Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine in Nov 30th, 2018, due to polydipsia, polyphagia, polyuria accompanied with increased glucose levels for more than 2 weeks. He presented with symmetrical short stature [height 81 cm (－2.2 SD), weight 9.8 kg (－2.1 SD), body mass index 14.94 kg/m2 (P10-P15)], and with no special facial or physical features. Laboratory results showed that the glycated hemoglobin A1c was 14%, the fasting C-peptide was 0.3 ng/mL, and the islet autoantibodies were all negative. Oral glucose tolerance test showed significant increases in both fasting and postprandial glucose, but partial islet functions remained (post-load C-peptide increased 1.43 times compared to baseline). A heterozygous variant c.1366C>T (p.R456C) was detected in GATA6 gene, thereby the boy was diagnosed with a specific type of diabetes mellitus. The boy had congenital heart disease and suffered from transient hyperosmolar hyperglycemia after a patent ductus arteriosus surgery at 11 months of age. Insulin replacement therapy was prescribed, but without regular follow-up thereafter. The latest follow-up was about 3.5 years after the diagnosis of diabetes when the child was 5 years and 11 months old, with the fasting blood glucose of 6.0-10.0 mmol/L, and the 2 h postprandial glucose of 17.0-20.0 mmol/L.

Bilineage embryo-like structure from EPS cells can produce live mice with tetraploid trophectoderm

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.

The role of transcription factor GATA6 in lung development and diseases / 国际儿科学杂志

GATA transcription factors encode a zinc finger DNA binding protein and play an important role in regulating cell proliferation and differentiation.GATA6 is an important member of the GATA transcription factor family,mainly expressed in the tissues and organs derived from endoderm.It is also the most fundamental member of the GATA family located in the airway epithelial cells.It is involved not only in the regulation of normal lung tissue development,but also in the pathogenesis of lung diseases.This review focuses on the role of GATA6 in lung development and some lung diseases.

Screening and functional analysis of GATA-6 mutations in patients with nonsyndromic conotruncal defects / 中华实用儿科临床杂志

Objective@#To explore the molecular and genetic mechanism of transcription factor GATA-6 in nonsyndromic conotruncal defect (CTD) in order to provide evidence for early prevention and inheritance consultation of CTD.@*Methods@#A total of 32 cases of patients with nonsyndromic CTD and 100 healthy individuals were enrolled in the study.A total of 7 exons and bilateral partial intron-exon boundaries of GATA-6 were amplified by means of polymerase chain reaction (PCR). The PCR products were purified and directly sequenced by using an ABI Genetic Analyzer 3100 Automatic DNA sequence equipment.The acquired GATA-6 gene sequence was compared with standard gene sequence published in National Center for Biotechnology Information database, as well as the healthy control group to observe the GATA-6 gene mutations.The mutations were introduced into pcDNA3.1(+ ) by site-directed mutagenesis PCR on the basis of pcDNA3.1(+ )-GATA-6 in order to generate the GATA6-G245R mutant constructs.Wild type GATA-6, GATA-6-G245R and atrial natriuretic factor-luciferase(ANF-luciferase) were cotransfected into HEK 293T cells in vitro, and the CMV-LacZ were cotransfected as internal reference.Luciferase and galactosidase activity were measured by using luminometer 24 h after transfection and detected in the downstream ANF-luciferase reporter gene.@*Results@#A heterozygous missense mutation in the GATA-6 gene was identified in a patient with double outlets of the right ventricle.The mutation was located in Gly245Arg(G245R) in exon 2 of GATA-6.The mutation of pcDNA3.1(+ )-GATA-6 expression vectors were successfully constructed.Through the detection of luciferase reporter gene activity, it was found that GATA-6-G245R and wild-type GATA-6 decreased by 41.3%, and the comparison between them was statistically significant (P<0.001).@*Conclusions@#Transcription factor GATA-6 gene mutation is associated with the occurrence of nonsyndromic CTD.Transcription factor GATA-6 gene may be susceptible gene in human nonsyndromic CTD.

Relationship between the expression level of GATA6 mRNA and the occurrence of congenital heart disease / 中华实用儿科临床杂志

Objective To investigate the relationship between the expression of GATA6 mRNA and the occu-rrence of congenital heart disease(CHD). Methods A total of 60 cases of CHD (CHD group)were diagnosed at Obstetrical Department,Rizhao City Maternal and Child Health Hospital,and 60 cases of normal pregnancy women (control group)were collected in March 2013 to May 2017. The expression levels of mRNA of GATA6 in the peripheral blood of pregnant women in 2 groups were detected by using real-time quantitative PCR (qRT-PCR)technique,and the relevant information of 2 groups of mothers before and during pregnancy was collected so as to investigate the rela-tionship between mRNA and GATA6 in CHD. Results There were no significant differences in age,body mass index (BMI),passive smoking,alcohol consumption,pregnancy medication,pregnancy-induced hypertension,high -density lipoproteincholesterol (HDL-C),total cholesterol (TC),triacylglycerol (TG)between pregnant women in the congenital heart disease group and the control group (all P > 0. 05). The CHD group showed a statistical difference significance in the rate of taking folic acid(81. 67％)and gestational diabetes incidence(8. 33％),serum level of LDL-C[(3. 59 ± 0. 46)mmol/ L]compared with the healthy control group [96. 67％,0,(3. 20 ± 0. 44)mmol/ L] (all P < 0. 05). The expression level of GATA6 mRNA in peripheral blood of patients with congenital heart disease (0. 014 ± 0. 005)was significantly lower than that of the control group (0. 129 ± 0. 031),and the difference was statis-tically significant (t = 28. 386,P = 0. 000). Logistic regression analysis showed that the level of in GATA6 mRNA ex-pression increased in the peripheral blood and folic acid intake were protective factors for CHD (all P < 0. 05)during pregnancy;while fetal gestational diabetes,elevated levels of LDL-C are risk factors for congenital heart disease (all P < 0. 05)fetus. Conclusion Down regulation of GATA6 mRNA expression in the maternal peripheral blood is one of the independent risk factors for fetal congenital heart disease.

Effects of GATA6 silencing mediated by lentivirus on apoptosis of human liver cancer Huh-7 cells / 中国病理生理杂志

AIM:To explore the effect of GATA6 gene silencing on apoptosis of hepatocellular carcinoma Huh-7 cells.METHODS:RNA interference vectors of the target gene GATA6 mediated by lentivirus were constructed in vitro to transfect the hepatocellular carcinoma cell line Huh-7.The apoptotic rate of transfected cells was measured by flow cytome-try.The protein expression of GATA6, NF-κB and Bcl-2 in transfected cells was determined by Western blotting.RE-SULTS:The transfection efficiency was 57.4%.The mRNA and protein expression of GATA6 reduced significantly after the carcinoma cell line Huh-7 being transfected by RNA interference vectors mediated by lentivirus.The apoptotic rate of the carcinoma cells with silent GATA6 gene was significantly increased (P<0.05).The protein expression levels of NF-κB and Bcl-2 were also significantly decreased.CONCLUSION: Lentiviral vector-mediated RNA interference of GATA6 has an inhibitory effect on the expression of the gene itself, and promotes the apoptosis of hepatocellular carcinoma cells.Regu-lation of the apoptosis-related protein expression by the NF-κB signaling to influence the proliferation and apoptosis of hepa-tocellular carcinoma cells might be one of the possible mechanisms.

The research progress of GATA6 in heart development and isolated atrial septal defect / 临床儿科杂志

GATA6 transcription factor belongs to the GATA family and contains 2 conserved zinc ifnger DNA binding domains. GATA6 not only presents in embryonic tissues but also found in heart, lung and pancreas and is essential for the maintenance of their function.The present review focuses on the critical roles of GATA6 in heart development and atrial septal defect to provide theoretical basis for diagnosis and treatment of atrial septal defect.

The role of transcription factor GATA-6 in heart development and congenital heart disease / 国际儿科学杂志

Congenital heart disease is caused by abnormal embryonic heart development.According to the research findings of developmental biology,different heart cells must be strictly regulated to ensure proper cell location and development.In the process of heart development,transcription factors play an important role in the regulation of gene expression.Zinc finger transcription factor GATA-6,an early indicator of myocardial cell development,is currently regarded as main candidate gene involved in the pathogenesis of congenital heart disease.GATA-6 gene mutations lead to changes in transcriptional activity of the gene product,which together with other relevant factors may play a role in the pathogenesis of congenital heart disease.

The relationship between gene GATA6 defect and congenital heart disease / 国际儿科学杂志

Recent studies have showed GATA6 may be a candidate gene of congenital heart disease (CHD).GATA6 is a member of the GATA transcription factors,and GATA6 is expressed within the precardiac mesoderm from early stages in its specification.Some studies about outflow tract (OFT) development show that GATA6 can directly regulate the migration of cardiac neural crest cells.Researchers find some missense mutation on GATA6 (p.A178V,p.L198V,p.D404Y,p.E460X) by directly sequencing.These mutations may influence transcriptional activity of GATA6 and are closely associated with CHD.Appropriate level of GATA6 in embryos is critical to cardiogenesis,and over-expression GATA6 can disrupt this process.Cardiogenesis needs a cascade of transcription factors includes GATA6 and other factors under fight temporal and spatial control.This paper reviews the latest studies about the relationship between gene GATA6 defect and CHD.

A novel 11;18 translocation in a case of acute myeloid leukemia with maturation.

Acute myeloid leukemia with maturation (AML-M2) is associated with the 8;21 translocation. For the first time in an adult patient with AML-M2, a novel unbalanced translocation involving the short arm of chromosome 11 and long arm of chromosome18 with new breakpoints is presented. CD82 on band 11p11.2 and GATA 6 on 18q11.2 may play a role in the pathogenesis of de novo AML M2. The report with translocation (11;18)(p11.2;q11.2), as the sole cytogenetic abnormality provides more data on the leukemogenesis of de novo AML M2.

Detection and clinical value of novel GATA6 mutations associated with congenital atrial septal defects / 中华检验医学杂志

Objective To identify novel mutations in the GATA6 gene associated with congenital atrial septal defects (ASD).Methods This was a case-control study.A cohort of 220 unrelated Han-race patients with congenital ASD and 200 unrelated ethnically matched healthy individuals used as controls,who were admitted to Tongji University Affiliated Tongji Hospital from January,2007 to October,2011,were recruited.The peripheral venous blood samples from the participants were prepared.All the coding exons and their flanking sequences of the GATA6 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination technique.The acquired sequences were aligned with the sequences derived from GenBank by BLAST to identify the sequence variations.The software ClustalW was used to analyze the conservation of the altered amino acids.Results Three novel heterozygous missense GATA6 mutations,c.250G ＞A (p.A84T),c.649G ＞C (p.G217R) and c.1270A ＞C (p.S424R),were identified in 3 of 220 ASD patients,respectively.None of the three mutations was detected in 200 healthy control individuals.A cross-species alignment of GATA6 encoded protein sequences showed that the mutated amino acids were relatively conserved evolutionarily.Conclusion The identification of novel GATA6 mutations associated with ASD contributes to the reveal of the mechanism involved in the pathogenesis of ASD.

Study on GATA-6 Gene on Inhibition of Endogenous Carbon Monoxide on Pulmonary Vascular Smooth Muscle Cells Proliferation / 实用儿科临床杂志

Objective To investigate the effect of GATA-6 on endogenous carbon monoxide(CO) inhibited pulmonary vascular smooth muscle cells(PVSMCs) proliferation induced by platelet-derived growth factor(PDGF).Methods Tissue mass culture was done to get PVSMCs artery in SD rats.The PVSMCs were stimulated to proliferation by PDGF(20 ?g/L),and the 3 different concentrations of hemin[a substrate and inducer of heme oxygenase-1(HO-1)] add into the cultures to induce CO production.The PVSMC cell proli-feration was detected by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method,DNA synthesis was detected by thymidine incorporation,and GATA-6 mRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR).Results CO inhibited the PVSMCs proliferation induced by PDGF in a dose-dependent and time-dependent manner.Hemin with high concentration markedly inhibited the proliferation and DNA synthesis of PVSMCs.After 2 hours with PDGF,the expression of(GATA-6) mRNA markedly down-regulated,and began returned after 6 hours.However,CO could reversed this down-regulation.Conclusions CO can inhibit the proliferation of PVSMCs induced by PDGF.PDGF can result in the expression of GATA-6 mRNA down-regulated;the down-regulation is reversed by CO.This study suggested that CO maybe inhibit PVSMCs proliferation by regulating expression of GATA-6.