Diabetologia

, Volume 55, Issue 10, pp 2845–2847 | Cite as

GATA6 inactivating mutations are associated with heart defects and, inconsistently, with pancreatic agenesis and diabetes

  • A. Bonnefond
  • O. Sand
  • B. Guerin
  • E. Durand
  • F. De Graeve
  • M. Huyvaert
  • L. Rachdi
  • J. Kerr-Conte
  • F. Pattou
  • M. Vaxillaire
  • M. Polak
  • R. Scharfmann
  • P. Czernichow
  • P. Froguel
Research Letter

Keywords

Congenital heart disease GATA6 Neonatal diabetes mellitus Pancreatic agenesis Whole-exome sequencing 

Abbreviations

GATA6

GATA-binding protein 6

NDM

Neonatal diabetes mellitus

To the Editor: Pancreatic agenesis is an extremely rare cause of permanent neonatal diabetes mellitus (NDM) in humans. It can be associated with severe intrauterine growth retardation as well as a plethora of abnormalities or malformations in the heart, biliary tract, gut, thyroid or brain [1, 2, 3]. To date, mutations in three genes have been shown to cause pancreatic agenesis, namely, PDX1 [1], PTF1A [2], and most recently GATA6 [3], all three of which encode transcription factors. The GATA6 study reported heterozygous coding mutations in a dozen individuals with permanent NDM presumed to be due to pancreatic agenesis [3]. The genetic evidence supporting the pathogenicity of these mutations seemed quite strong as the reported mutations had arisen de novo and none of these mutations was present in the 1,000 Genome Project database [3]. The authors concluded that GATA6 may have a key role in human pancreatic development, with strong implications for beta cell regenerative medicine in diabetes [3]. In the present study, we aimed to qualify this conclusion by reporting a case study of a non-consanguineous family.

We analysed the whole exome of two sisters from a French non-consanguineous family (electronic supplementary material [ESM] Methods). The youngest sister presented with severe intra-uterine growth retardation, pancreatic agenesis (with permanent NDM and exocrine pancreatic insufficiency requiring insulin treatment and enzyme replacement therapy), ventricular septal defects requiring surgery during the neonatal period, gall bladder agenesis, bicornuate uterus and neonatal pancytopenia (Table 1). Her older sister presented with patent ductus arteriosus requiring surgery when she was 1 year old, gall bladder agenesis and proctorrhagia (Table 1). Contrary to her sister, she did not present with NDM. At 13 years, she showed normal fasting plasma glucose level (5.4 mmol/l) but she had high glucose values 2 h after an oral glucose challenge (14.2 mmol/l). MRI examination revealed a small pancreas, suggesting pancreatic hypoplasia, but no enzyme replacement therapy was needed. Of note, another pregnancy of the mother was terminated at 22 weeks of amenorrhea, as the male fetus presented with severe hypoplasia of the left heart. The mother has mild valvular abnormalities whereas the father is healthy (Table 1). We had previously searched for mutations in several genes known to be involved in NDM in the two sisters (ESM Methods). By whole-exome sequencing, we identified in both sisters a novel heterozygous deletion of two nucleotides in the fifth exon of GATA6 (c.1504_1505del; p.Lys502Aspfs*5), which leads to a premature termination codon (ESM Fig. 1). The presence of the mutation was investigated in parents, the mother's parents and 378 French adults with normal fasting glucose (ESM Methods). The mutation was not present in the controls. Interestingly, the mother carries the frameshift mutation (ESM Fig. 1), however neither the father nor the mother's parents are carriers, implying that the mutation had arisen de novo in the mother, who is not diabetic. Indeed, since we started to study the family in 1998 (she is now 49), her metabolic profile has always been normal. Therefore, in contrast to the study by Allen et al [3], we demonstrate that GATA6 mutations do not always occur de novo in affected children. Furthermore, the penetrance of the p.Lys502Aspfs*5 frameshift mutation for diabetes is incomplete, whereas it seems complete for heart anomalies (Table 1). In fact, there is a spectrum of phenotypes in the three carriers of the family. This statement is supported by several studies that have reported loss-of-function mutations in GATA6 causing human congenital heart diseases (e.g. persistent or patent truncus arteriosus, atrial septal defects, pulmonary valve stenosis, tetralogy of Fallot) [4, 5]. Importantly, diabetes was never reported in the analysed carriers [4, 5]. It shows that when patients with GATA6 mutations have been primarily selected for congenital cardiac diseases but not for NDM, they are not usually diabetic. In the study by Allen et al it is noteworthy that 14 out of 15 carriers of a GATA6 mutation presented with severe cardiac malformations [3]. The only patient without a heart anomaly carried a non-coding splice-site mutation, of which deleterious functionality was not proven [3].
Table 1

Clinical data of the three carriers of the GATA6 p.Lys502Aspfs*5 mutation

Clinical characteristics

Proband

Older sister

Mother

Pancreas agenesis

x

Diabetes

x (permanent NDM)

x (onset at 13 years)

Cardiac anomaly

x (requiring surgery)

x (requiring surgery)

x

Gall bladder agenesis

x

x

?

Bicornuate uterus

x

Intra-uterine growth retardation

x

Neonatal pancytopenia

x

Proctorrhagia

x

p.Lys502Aspfs*5 heterozygous mutation

x

x

x

The father is healthy and does not carry the mutation

Based on these findings, we needed to question the proposed key role of GATA6 in pancreatic beta cell development using public and in-house expression databases. First, according to public databases, GATA6 expression is unambiguously weak in human whole pancreas (Gene Portal System, http://biogps.org/, last accessed March 2012). In our experiments on isolated human pancreatic islets and beta cells (ESM Methods), this was also the case (ESM Fig. 2a). In a rat model of pancreatic endocrine cell differentiation and proliferation (ESM Methods), Gata6 expression is constantly weak from precursor cells to mature beta cells (ESM Fig. 2b). Moreover, between embryonic days E13.5 and E17, there is no significant change in Gata6 expression in pancreatic epithelium (ESM Fig. 2c). GATA6 expression is also not significantly different between pancreatic epithelium and mesenchyme in rat and human samples, at embryonic day E12 and at ~7–11 weeks of development, respectively (ESM Methods, ESM Fig. 2d, e). Therefore, our results do not suggest a key role of GATA6 in the specific beta cell lineage, but these results would need further investigation as they are limited to gene expression analyses. In mice, in which GATA6 has been further studied, expression-based analyses suggest that GATA6 could play a role in very early pancreas specification [6, 7, 8]. Nevertheless, it has been reported that transgenic mouse embryos expressing a GATA6–engrailed dominant repressor fusion protein in the pancreatic epithelium and in islets revealed two distinct phenotypes, either pancreatic agenesis or a simple reduction in pancreatic tissue [6], implying a putative spectrum of phenotypes, as we noted in our family. Therefore, the role of GATA6 in pancreatic morphogenesis is very variable, which could be due to various epigenetic effects or partial replacement of GATA6 by other transcription factors from the GATA family that are expressed in the pancreas (e.g. GATA4).

Taken together, our results and other genetic studies indicate GATA6 inactivating mutations are associated with heart defects and, inconsistently, with pancreatic agenesis and diabetes. We suggest that in carriers of a GATA6-inactivating mutation, diabetes (when present) is due to a severe multi-tissue developmental defect rather than a specific impairment of beta cell lineage. The present study is limited by a lack of functional analyses of our identified frameshift mutation. Nevertheless, a de novo deletion which leads to a premature termination codon at the 514th amino acid has previously been reported in an NDM patient with pancreatic agenesis [3]. Our deletion, which leads to a termination codon at the 507th amino acid, is likely to have a functional effect similar to that reported by Allen et al [3].

Notes

Acknowledgements

We are sincerely indebted to the family for participation in the study. We thank P. Gallina (CNRS UMR8199 unit, Lille, France) for clinical data capture. We also thank our collaborators in the French study of neonatal diabetes: K. Busiah (Necker Hospital, Paris, France), H. Cavé (Robert Debré Hospital, Paris, France) and A. Simon (Necker Hospital, Paris, France).

Funding

Our study was supported by the transnational European research grant on Rare Diseases (ERANET-09-RARE-005) and by the French Agence Nationale de la Recherche (ANR-10-LABX-46 and ANR-10-EQPX-07-01).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Contribution statement

AB and PF designed the study, interpreted the data and wrote the manuscript; MV, BG, MP and PC analysed the clinical data and revised the manuscript; LR, JK-C, FP and RS contributed to acquisition of gene expression data (in rat and/or in human) and revised the manuscript; OS and FD performed the bioinformatic analyses and revised the manuscript; ED and MH performed the sequencing and revised the manuscript; PF is the guarantor of the manuscript. All authors of the paper have read and approved the final version submitted.

Supplementary material

125_2012_2645_MOESM1_ESM.pdf (88 kb)
ESM Methods(PDF 88 kb)
125_2012_2645_MOESM2_ESM.pdf (78 kb)
ESM Figure 1Location of the p.Lys502Aspfs*5 frameshift mutation in GATA6. (PDF 77 kb)
125_2012_2645_MOESM3_ESM.pdf (80 kb)
ESM Figure 2Array-based gene expression data analysis of GATA6 in human and rat pancreatic tissue samples, at different stages of development. (a) Expression of GATA6 (and PDX1: positive control) in human pancreatic islets (n = 3) and sorted beta-cells (n = 3). (b) Expression of Gata6 (and Ins1/Ins2: positive controls) in rat pancreatic embryonic samples (n = 6), during different time lapses (E13.5 + 0, 1, 3, 5, and 7 days), mimicking the endocrine cell differentiation and proliferation. (c) Expression of Gata6 (and Ins1/Ins2: positive controls) in rat pancreatic embryonic epithelium (n = 3) (E13.5 and E17). (d) Expression of GATA6 in human pancreatic embryonic epithelium (n = 5) and mesenchyme (n = 5) (between 7–11 weeks of development). (e) Expression of Gata6 (and Ins1/Ins2: positive controls) in rat pancreatic embryonic epithelium (n = 3) and mesenchyme (n = 3) (E12). The dotted lines represent the 75th percentile of expression of all probes present in the microarray. N.S., non-significant. Data are presented as mean ± s.e.m. (PDF 79 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • A. Bonnefond
    • 1
    • 2
  • O. Sand
    • 1
    • 2
  • B. Guerin
    • 3
  • E. Durand
    • 1
    • 2
  • F. De Graeve
    • 1
    • 2
  • M. Huyvaert
    • 1
    • 2
  • L. Rachdi
    • 4
  • J. Kerr-Conte
    • 2
    • 5
  • F. Pattou
    • 2
    • 5
    • 6
  • M. Vaxillaire
    • 1
    • 2
  • M. Polak
    • 4
    • 7
  • R. Scharfmann
    • 4
  • P. Czernichow
    • 7
  • P. Froguel
    • 1
    • 2
    • 8
  1. 1.Genomics and Metabolic Diseases, CNRS UMR8199 – Lille Institute of BiologyLille CedexFrance
  2. 2.Lille Nord de France UniversityLilleFrance
  3. 3.Department of PaediatricsHospital of PauPauFrance
  4. 4.Inserm-U845, Research Center Growth and SignallingParis Descartes University, Sorbonne Paris Cité, Necker HospitalParisFrance
  5. 5.Inserm-U859LilleFrance
  6. 6.Department of Endocrine SurgeryHospital of LilleLilleFrance
  7. 7.Department of Paediatric EndocrinologyNecker Enfants Malades HospitalParisFrance
  8. 8.Department of Genomics of Common Disease, School of Public HealthImperial College LondonLondonUK

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