Association of hGrb10 Genetic Variations With Type 2 Diabetes in Caucasian Subjects
- Rosa Di Paola, PHD1,
- Ester Ciociola, PHD1,
- Watip Boonyasrisawat, MS2,
- David Nolan, BS2,
- Jill Duffy, BS2,
- Giuseppe Miscio, PHD1,
- Carmela Cisternino, BSC1,
- Grazia Fini, BSC1,
- Vittorio Tassi, MD, PHD1,
- Alessandro Doria, MD, PHD2 and
- Vincenzo Trischitta, MD13
- 1Research Unit of Endocrinology, Scientific Institute “Casa Sollievo della Sofferenza,” San Giovanni Rotondo, Italy
- 2Research Division, Department of Medicine, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts
- 3Department of Clinical Sciences, University “La Sapienza,” Rome, Italy
- Address correspondence to Rosa Di Paola, Research Unit of Endocrinology, Scientific Institute “Casa Sollievo della Sofferenza,”
Viale Padre Pio, 71013 San Giovanni Rotondo, Italy. E-mail: r.dipaola{at}operapadrepio.it; or Vincenzo Trischitta, Department of Clinical Sciences, University “La Sapienza,” Policlinico Umberto I, 00161, Rome, Italy.
E-mail: vincenzo.trischitta{at}uniroma1.it
The genes contributing to type 2 diabetes are mostly unknown (1). Grb10 is an adapter protein that, in target tissues (2,3), interacts with the insulin receptor (3–9), thus affecting downstream signaling (10–12) and insulin action (2,7,10–16). We tested the hypothesis that variants in the Grb10 gene modulate the risk for type 2 diabetes, one of the most frequent
outcomes of insulin resistance.
Resequencing of coding and immediately flanking sequences of hGrb10 (17,18) identified six single nucleotide polymorphisms (SNPs), five of which had a minor allele frequency >5%. Based on their physical
location and their mutual linkage disequilibrium, these five SNPs (i.e., rs1800504, rs2715128, rs2072235, rs4947710, and rs3807550)
could be grouped into two clusters. The rs1800504 and rs4947710 SNPs (each belonging to different clusters) were analyzed
for association with type 2 diabetes in 764 diabetic patients and 323 unrelated control subjects from the east coast of central
Italy (19,20). No significant association with type 2 diabetes was observed for rs1800504 (data not shown). By contrast, the genotype
distributions of rs4947710 (i.e., G/G, G/A, and A/A) were significantly different between case and control subjects (87.2,
12.3, and 0.5% vs. 61.9, 35.8, and 2.3%, respectively, P < 0.0001), with A allele carriers showing a reduced risk of type 2 diabetes (unadjusted odds ratio 0.239 [95% CI 0.17–0.33],
P = 0.0001; age-, sex-, and BMI-adjusted odds ratio 0.235 [95% CI 0.15–0.36], P = 0.0001). A potential biological relevance of rs4947710 was suggested by an in silico analysis (ESEfinder; available at
http://exon.cshl.edu/ESE/), which indicated that the G-to-A substitution of rs4947710 may cause the disruption of a putative consensus motif for the
human Ser/Arg-rich proteins SF2/ASF.
To replicate this association, we studied 731 type 2 diabetic case and 358 nondiabetic control subjects, all being Caucasians
from the Boston area (20). In contrast to what was observed in the Italian population, the genotype distributions of rs4947740 were similar in case
and control subjects (G/G = 83.9%, G/A = 15.2%, and A/A = 1.0% and G/G = 86.0%, G/A = 14.0%, and A/A = 0.0%, respectively,
P = 0.15).
In conclusion, a significant association between the hGrb10 rs4947710 SNP (whose biological function on differential splicing
is suggested by in silico analysis) and type 2 diabetes was found in Caucasian subjects from Italy but not in those from the
U.S. Lack of replication of genotype-phenotype associations is not an uncommon event in the study of complex disorders (21–23) and can arise from the original result being a false-positive because of bias or chance or from the second result being
a false negative because of insufficient power. Such explanations, however, do not seem to account for our conflicting findings.
The population of the original study was relatively homogeneous, making the possibility of population stratification remote,
and the P value for association with type 2 diabetes was highly significant, making chance an unlikely explanation of the association
finding. The replication study had close to 100% power to detect the odds ratio observed in the original study. Thus, lack
of replication in our study is likely to result from differences in the genetic and/or environmental background of the populations
studied, highlighting the need for large, collaborative studies providing sufficient power to investigate gene-gene and gene-environment
interactions and their differences among populations.
Acknowledgments
This Research was supported by Telethon Grant GGP02423, by Italian Ministry of Health Grant Ricerca Corrente 2005, and National
Institutes of Health Grants DK60837 (Diabetes Genome Anatomy Project) and DK36836 (Genetics Core of the Diabetes and Endocrinology
Research Center at the Joslin Diabetes Center).
References
- ↵
Almind K, Doria A, Kahn CR: Putting the genes for type II diabetes on the map.
Nat Med
7
:
277
–279,2001
- ↵
Morrione A, Valentinis B, Resnicoff M, Xu S, Baserga R: The role of mGrb10alpha in insulin-like growth factor I-mediated growth.
J Biol Chem
272
:
26382
–26387,1997
- ↵
Dey BR, Frick K, Lopaczynski W, Nissley SP, Furlanetto RW: Evidence for the direct interaction of the insulin-like growth
factor I receptor with IRS-1, Shc, and Grb10.
Mol Endocrinol
10
:
631
–641,1996
-
Hansen H, Svensson U, Zhu J, Laviola L, Giorgino F, Wolf G, Smith RJ, Riedel H: Interaction between the Grb10 SH2 domain and
the insulin receptor carboxyl terminus.
J Biol Chem
271
:
8882
–8886,1996
-
Laviola L, Giorgino F, Chow JC, Baquero JA, Hansen H, Ooi J, Zhu J, Riedel H, Smith RJ: The adapter protein Grb10 associates
preferentially with the insulin receptor as compared with the IGF-I receptor in mouse fibroblasts.
J Clin Invest
99
:
830
–837,1997
-
Morrione A, Valentinis B, Li S, Ooi JY, Margolis B, Baserga R: Grb10: a new substrate of the insulin-like growth factor I
receptor.
Cancer Res
56
:
3165
–3167,1996
- ↵
O’Neill TJ, Rose DW, Pillay TS, Hotta K, Olefsky JM, Gustafson TA: Interaction of a GRB-IR splice variant (a human GRB10 homolog)
with the insulin and insulin-like growth factor I receptors: evidence for a role in mitogenic signaling.
J Biol Chem
271
:
22506
–22513,1996
-
Dong LQ, Farris S, Christal J, Liu F: Site-directed mutagenesis and yeast two-hybrid studies of the insulin and insulin-like
growth factor-1 receptors: the Src homology-2 domain-containing protein hGrb10 binds to the autophosphorylated tyrosine residues
in the kinase domain of the insulin receptor.
Mol Endocrinol
11
:
1757
–1765,1997
- ↵
Frantz JD, Giorgetti-Peraldi S, Ottinger EA, Shoelson SE: Human GRB-IRbeta/GRB10: splice variants of an insulin and growth
factor receptor-binding protein with PH and SH2 domains.
J Biol Chem
272
:
2659
–2667,1997
- ↵
Liu F, Roth RA: Grb-IR: a SH2-domain-containing protein that binds to the insulin receptor and inhibits its function.
Proc Natl Acad Sci U S A
92
:
10287
–10291,1995
-
Wick KR, Werner ED, Langlais P, Ramos FJ, Dong LQ, Shoelson SE, Liu F: Grb10 inhibits insulin-stimulated insulin receptor
substrate (IRS)-phosphatidylinositol 3-kinase/Akt signaling pathway by disrupting the association of IRS-1/IRS-2 with the
insulin receptor.
J Biol Chem
278
:
8460
–8467,2003
- ↵
Morrione A: Grb10 proteins in insulin-like growth factor and insulin receptor signaling (Review).
Int J Mol Med
5
:
151
–154,2000
-
Morrione A: Grb10 adapter protein as regulator of insulin-like growth factor receptor signaling.
J Cell Physiol
197
:
307
–311,2003
-
Wang J, Dai H, Yousaf N, Moussaif M, Deng Y, Boufelliga A, Swamy OR, Leone ME, Riedel H: Grb10, a positive, stimulatory signaling
adapter in platelet-derived growth factor BB-, insulin-like growth factor I-, and insulin-mediated mitogenesis.
Mol Cell Biol
19
:
6217
–6228,1999
-
Mounier C, Lavoie L, Dumas V, Mohammad-Ali K, Wu J, Nantel A, Bergeron JJ, Thomas DY, Posner BI: Specific inhibition by hGRB10zeta
of insulin-induced glycogen synthase activation: evidence for a novel signaling pathway.
Mol Cell Endocrinol
173
:
15
–27,2001
- ↵
Deng Y, Bhattacharya S, Swamy OR, Tandon R, Wang Y, Janda R, Riedel H: Growth factor receptor-binding protein 10 (Grb10) as
a partner of phosphatidylinositol 3-kinase in metabolic insulin action.
J Biol Chem
278
:
39311
–39322,2003
- ↵
Angrist M, Bolk S, Bentley K, Nallasamy S, Halushka MK, Chakravarti A: Genomic structure of the gene for the SH2 and pleckstrin
homology domain-containing protein GRB10 and evaluation of its role in Hirschsprung disease.
Oncogene
17
:
3065
–3070,1998
- ↵
Blagitko N, Mergenthaler S, Schulz U, Wollmann HA, Craigen W, Eggermann T, Ropers HH, Kalscheuer VM: Human GRB10 is imprinted
and expressed from the paternal and maternal allele in a highly tissue- and isoform-specific fashion.
Hum Mol Genet
9
:
1587
–1595,2000
- ↵
Di Paola R, Frittitta L, Miscio G, Bozzali M, Baratta R, Centra M, Spampinato D, Santagati MG, Ercolino T, Cisternino C, Soccio
T, Mastroianno S, Tassi V, Almgren P, Pizzuti A, Vigneri R, Trischitta V: A variation in 3′ UTR of hPTP1B increases specific
gene expression and associates with insulin resistance.
Am J Hum Genet
70
:
806
–812,2002
- ↵
Bacci S, Ludovico O, Prudente S, Zhang YY, Di Paola R, Mangiacotti D, Rauseo A, Nolan D, Duffy J, Fini G, Salvemini L, Amico
C, Vigna C, Pellegrini F, Menzaghi C, Doria A, Trischitta V: The K121Q polymorphism of the ENPP1/PC-1 gene is associated with
insulin resistance/atherogenic phenotypes, including earlier onset of type 2 diabetes and myocardial infarction.
Diabetes
54
:
3021
–3025,2005
- ↵
Ioannidis JP, Ntzani EE, Trikalinos TA, Contopoulos-Ioannidis DG: Replication validity of genetic association studies.
Nat Genet
29
:
306
–309,2001
-
He W, Rose DW, Olefsky JM, Gustafson TA: Grb10 interacts differentially with the insulin receptor, insulin-like growth factor
I receptor, and epidermal growth factor receptor via the Grb10 Src homology 2 (SH2) domain and a second novel domain located
between the pleckstrin homology and SH2 domains.
J Biol Chem
273
:
6860
–6867,1998
- ↵
Redden DT, Allison DB: Nonreplication in genetic association studies of obesity and diabetes research.
J Nutr
133
:
3323
–3326,2003
