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1.
Front Genet ; 15: 1414451, 2024.
Article in English | MEDLINE | ID: mdl-38978877

ABSTRACT

Background: Rabson-Mendenhall syndrome (RMS), a rare disorder characterized by severe insulin resistance due to biallelic loss-of-function variants of the insulin receptor gene (INSR), presents therapeutic challenges (OMIM: 262190). This case study explores the efficacy of adjunctive therapy with sodium-glucose cotransporter 2 inhibitors (SGLT2is) in the management of RMS in an 11-year-old male patient with compound heterozygous pathogenic variants of INSR. Methods: Despite initial efforts to regulate glycemia with insulin therapy followed by metformin treatment, achieving stable glycemic control presented a critical challenge, characterized by persistent hyperinsulinism and variable fluctuations in glucose levels. Upon the addition of empagliflozin to metformin, notable improvements in glycated hemoglobin (HbA1c) and time in range (TIR) were observed over a 10-month period. Results: After 10 months of treatment, empagliflozin therapy led to a clinically meaningful reduction in HbA1c levels, decreasing from 8.5% to 7.1%, along with an improvement in TIR from 47% to 74%. Furthermore, regular monitoring effectively averted normoglycemic ketoacidosis, a rare complication associated with SGLT2 inhibitor therapy. Conclusion: This case highlights the potential of SGLT2i as adjunctive therapy in RMS management, particularly in stabilizing glycemic variability. However, further research is warranted to elucidate the long-term efficacy and safety of this therapeutic approach in RMS and similar insulin resistance syndromes.

2.
Diabetologia ; 54(7): 1693-701, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21544516

ABSTRACT

AIMS/HYPOTHESIS: The aim of this study was to investigate the genetic aetiology of permanent diabetes mellitus with onset in the first 12 months of age. METHODS: We studied 46 probands with permanent, insulin-requiring diabetes with onset within the first 6 months of life (permanent neonatal diabetes mellitus [PNDM]/monogenic diabetes of infancy [MDI]) (group 1) and eight participants with diabetes diagnosed between 7 and 12 months of age (group 2). KCNJ11, INS and ABCC8 genes were sequentially sequenced in all patients. For those who were negative in the initial screening, we examined ERN1, CHGA, CHGB and NKX6-1 genes and, in selected probands, CACNA1C, GCK, FOXP3, NEUROG3 and CDK4. The incidence rate for PNDM/MDI was calculated using a database of Italian patients collected from 1995 to 2009. RESULTS: In group 1 we found mutations in KCNJ11, INS and ABCC8 genes in 23 (50%), 9 (19.5%) and 4 (8.6%) patients respectively, and a single homozygous mutation in GCK (2.1%). In group 2, we identified one incidence of a KCNJ11 mutation. No genetic defects were detected in other loci. The incidence rate of PNDM/MDI in Italy is estimated to be 1:210,287. CONCLUSIONS/INTERPRETATION: Genetic mutations were identified in ~75% of non-consanguineous probands with PNDM/MDI, using sequential screening of KCNJ11, INS and ABCC8 genes in infants diagnosed within the first 6 months of age. This percentage decreased to 12% in those with diabetes diagnosed between 7 and 12 months. Patients belonging to the latter group may either carry mutations in genes different from those commonly found in PNDM/MDI or have developed an early-onset form of autoimmune diabetes.


Subject(s)
Diabetes Mellitus/etiology , Diabetes Mellitus/genetics , ATP-Binding Cassette Transporters/genetics , Diabetes Mellitus/epidemiology , Female , Genetic Predisposition to Disease , Germinal Center Kinases , Humans , Infant , Infant, Newborn , Insulin/genetics , Male , Mutation , Potassium Channels, Inwardly Rectifying/genetics , Protein Serine-Threonine Kinases/genetics , Receptors, Drug/genetics , Sulfonylurea Receptors
3.
J Endocrinol Invest ; 34(9): e275-80, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21666412

ABSTRACT

BACKGROUND AND AIM: Metabolic characteristics and rate of progression to overt Type 2 diabetes (T2D) in low-risk European obese children are not well documented. Aim of the study was to investigate differences in insulin sensitivity and secretion in Italian obese children and youngsters with pre-diabetes. METHODS: Ninety-six obese children and youngsters with pre-diabetes, pair-matched with individuals with normal glucose tolerance (NGT) were included in the present study. Participants were screened by oral glucose tolerance. Pre-diabetes was classified as impaired fasting glucose (IFG), impaired glucose tolerance (IGT) and combined IFG-IGT. Homeostasis model assessment of insulin resistance (HOMA-IR), 2-h insulin, insulin sensitivity index (ISI) and disposition index (DI) were calculated to estimate fasting, peripheral and whole body insulin sensitivity and capacity of pancreatic islets to compensate for lower insulin sensitivity, respectively. One-way analysis of variance was used to compare groups. RESULTS: Eleven subjects had IFG (11.5%), 79 IGT (82.3%), 6 combined IFG-IGT (6.3%). Individuals with IFG showed the highest HOMA-IR (p=0.0007), those with IGT the highest 2-h insulin (p<0.0001), those with IFG-IGT the lowest ISI (p<0.0001), with severely reduced DI (p=0.0003). Compared with NGT, DI was 60% lower in those with IFG-IGT. CONCLUSION: IFG is linked primarily to fasting insulin resistance, IGT to peripheral insulin resistance. IFG-IGT is hallmarked by reduced whole body insulin sensitivity and an additional severe defect in DI. Further longitudinal studies are needed to understand whether the different categories of pre-diabetes in European obese adolescents represent real pre-diabetic alterations.


Subject(s)
Obesity/physiopathology , Prediabetic State/physiopathology , Adolescent , Blood Glucose/metabolism , Child , Diabetes Mellitus, Type 2/physiopathology , Disease Progression , Fasting , Female , Glucose Intolerance/physiopathology , Glucose Tolerance Test , Homeostasis , Humans , Insulin Resistance/physiology , Male , Obesity/complications , Prediabetic State/etiology , Risk Factors , Young Adult
4.
Diabetologia ; 53(7): 1354-61, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20393693

ABSTRACT

AIMS/HYPOTHESIS: The results of studies on the genetics of complex traits need to be replicated and to reach robust statistical significance before they can be considered as established. We here tried to replicate the previously reported association between the TRIB3 Q84R polymorphism (rs2295490) and glucose homeostasis. METHODS: Three samples of Europeans with fasting glucose <7.0 mmol/l were studied. In sample 1 (n=791), the association between TRIB3 Q84R and impaired glucose regulation (IGR; defined as impaired fasting glucose and/or impaired glucose tolerance and/or type 2 diabetes by OGTT) and insulin sensitivity (ISI), and its interplay with early-phase insulin secretion (i.e. disposition index [DI]) were analysed. Sample 2 (n=374) and sample 3 (n=394) were used to replicate the association with IGR and insulin sensitivity (by glucose clamp), respectively. Genotyping was performed by TaqMan allele discrimination. RESULTS: R84 carriers were at higher risk of IGR: OR for the additive model 1.54, p=0.004, and 1.63, p=0.027, in samples 1 and 2, respectively. In sample 1, both ISI (p=0.005) and DI (p=0.043) were progressively lower from QQ to QR and RR individuals. A 'triangulation approach' indicated that the association with IGR was mostly mediated by DI rather than by ISI changes (i.e. being the expected ORs 1.51 and 1.25, respectively). In sample 3, glucose disposal was 38.8+/-17.7, 33.8+/-14.4, and 31.6+/-13.3 micromol min(-1)kg(-1), p=0.022, in QQ, QR and RR individuals, respectively. CONCLUSIONS/INTERPRETATION: Our data confirm that the TRIB3 R84 variant affects glucose homeostasis and suggest this effect is due to an alteration of the interplay between insulin sensitivity and secretion.


Subject(s)
Cell Cycle Proteins/genetics , Glucose/metabolism , Homeostasis/genetics , Insulin Resistance/genetics , Insulin/metabolism , Protein Serine-Threonine Kinases/genetics , Repressor Proteins/genetics , Adolescent , Adult , Aged , Female , Genetic Predisposition to Disease/genetics , Humans , Insulin Secretion , Male , Middle Aged , Polymorphism, Genetic/genetics , Young Adult
5.
Hum Mutat ; 28(11): 1150, 2007 Nov.
Article in English | MEDLINE | ID: mdl-17939176

ABSTRACT

Transglutaminase 2 (TG2 or TGM2) is a multi-functional enzyme which catalyzes transamidation reactions or acts as a G-protein in intracellular signalling. Tgm2-/- Mice lacking TG2 activity are glucose intolerant and show impairment of insulin secretion, suggesting an important physiological role for TG2 in the pancreatic beta cell. We have previously described a TGM2 heterozygous missense mutation ((c.998A>G, p.N333S) in a 14 year-old patient with insulin-treated diabetes and in his diabetic father. The aim of this study was to further investigate the role of TG2 in early-onset type 2 diabetes. We analysed the TGM2 gene in 205 patients with clinically defined Maturity Onset Diabetes of the Young (MODY) or early-onset type 2 diabetes. We found two novel heterozygous mutations (c.989T>G, p.M330R; c.992T>A, p.I331N), which were not detected in 300 normoglycemic controls. All mutations were in residues which are located close to the catalytic site and impaired transamidating activity in vitro. Gene expression of TGM family genes and localization of TG2 in normal human pancreas indicated that TG2 is the only transglutaminase significantly expressed in human pancreatic islet cells. We conclude that reduced TG2 activity can contribute to disorders of glucose metabolism possibly via an impairment of insulin secretion.


Subject(s)
Diabetes Mellitus, Type 2/genetics , GTP-Binding Proteins/genetics , Mutation, Missense , Transglutaminases/genetics , Adolescent , Adult , Age of Onset , Animals , COS Cells , Chlorocebus aethiops , Heterozygote , Humans , Immunohistochemistry , Protein Glutamine gamma Glutamyltransferase 2
8.
Diabetes Res Clin Pract ; 116: 202-4, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27321336

ABSTRACT

We found that in obese youth, plasma glucose above 155mg/dl (8.6mmol/l) at 60min of an OGTT, a predictor of type 2 diabetes (T2D) in adults, was present in 12% with normal glucose tolerance and 57% with impaired glucose tolerance. Performance of elevated 1-h glucose in predicting T2D in overweight youngsters should be tested.


Subject(s)
Blood Glucose/analysis , Diabetes Mellitus, Type 2/epidemiology , Glucose Tolerance Test/methods , Obesity/blood , Adolescent , Adult , Female , Humans , Prevalence
9.
Diabetes ; 41(4): 408-15, 1992 Apr.
Article in English | MEDLINE | ID: mdl-1607067

ABSTRACT

Denaturing gradient gel electrophoresis (DGGE) has been used to screen for mutations in the insulin receptor gene. Each of the 22 exons was amplified by the polymerase chain reaction (PCR). For each exon, one of the two PCR primers contained a guanine-cytosine (GC) clamp at its 5' end. The DNA was analyzed by electrophoresis through a polyacrylamide gel containing a gradient of denaturants. Two geometries for the gels were compared; the gradient of denaturants was oriented either parallel or perpendicular to the electric field. The sensitivity of the technique was evaluated by determining whether DGGE succeeded in detecting known mutations and polymorphisms in the insulin receptor gene. With parallel gels, 12 of 16 sequence variants were detected. The use of perpendicular gels increased the sensitivity of detection so that all 16 sequence variants were successfully detected when DNA was analyzed by a combination of perpendicular and parallel gels. Furthermore, DGGE was used to investigate a patient with leprechaunism whose insulin receptor genes had not previously been studied. Two mutant alleles were identified in this patient. The allele inherited from the father had a mutation substituting alanine for Val-28; in the allele inherited from the mother, arginine was substituted for Gly-366.


Subject(s)
Electrophoresis, Polyacrylamide Gel/methods , Mutation/genetics , Receptor, Insulin/genetics , Alanine/analysis , Alleles , Arginine/analysis , Base Sequence , Child , DNA/analysis , DNA/genetics , Exons , Female , Gene Amplification , Genetic Diseases, Inborn/diagnosis , Genetic Diseases, Inborn/genetics , Genetic Testing , Genetic Variation , Humans , Molecular Sequence Data , Nucleic Acid Denaturation , Polymerase Chain Reaction , Polymorphism, Genetic
10.
Diabetes ; 40(1): 118-22, 1991 Jan.
Article in English | MEDLINE | ID: mdl-2015966

ABSTRACT

The nucleotide sequence of the insulin gene was determined in American Pima Indians and Micronesian Nauruans, two populations in whom the prevalence of non-insulin-dependent (type II) diabetes mellitus is the highest in the world. The insulin gene was amplified by the polymerase chain reaction to generate single-stranded DNA suitable for direct sequencing. The nucleotide sequences of the coding and adjacent regions of the insulin gene in six Pima Indians and two Nauruans with type II diabetes were identical to previously published insulin gene sequences of nondiabetic subjects.


Subject(s)
Diabetes Mellitus, Type 2/epidemiology , Ethnicity , Indians, North American , Insulin/genetics , Adult , Arizona , Base Sequence , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/genetics , Exons , Female , Humans , Insulin/blood , Insulin/isolation & purification , Male , Micronesia , Middle Aged , Molecular Sequence Data , Oligonucleotide Probes , Polymerase Chain Reaction/methods , Polymorphism, Genetic , Prevalence , Proinsulin/blood , RNA, Messenger/genetics , RNA, Messenger/isolation & purification
11.
Diabetes ; 47(10): 1625-9, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9753302

ABSTRACT

Leptin, a hormone secreted by adipocytes, decreases food intake and increases energy expenditure. The role of insulin in the regulation of leptin secretion is poorly understood and is still a topic of debate. Insulin increases leptin mRNA synthesis in rodents, but in humans, the available data are discordant. To investigate the role of chronic hyperinsulinemia in the regulation of plasma leptin concentrations, we studied 13 patients with surgically confirmed insulinoma before and after tumor removal, along with 15 healthy control subjects matched for sex, age, and BMI. Immunoreactive plasma leptin levels were measured by radioimmunoassay; leptin mRNA levels were also determined by reverse transcription-competitive polymerase chain reaction in a subgroup of six patients with insulinoma and six control subjects. All determinations were made with subjects in the fasting state. Plasma leptin concentrations correlated positively with leptin mRNA levels (r = 0.880, P < 0.001). Leptin levels, both plasma protein and mRNA, were significantly higher in the insulinoma patients than in the control subjects (plasma protein: 17.5 +/- 3.6 vs. 2.9 +/- 0.4 ng/ml, respectively, P < 0.001; mRNA: 0.98 +/- 0.33 vs. 0.19 +/- 0.064 amol/microg RNA, respectively, P < 0.05), and they correlated positively with fasting plasma insulin levels in the patients with insulinoma (plasma protein: r = 0.686, P < 0.01; mRNA: 0.796, P < 0.05). Finally, removal of the insulin-secreting tumor was followed by the normalization of plasma leptin levels. In summary, in patients with insulinoma, 1) plasma leptin levels and leptin mRNA are elevated; 2) a direct relationship exists between leptin, both circulating protein and mRNA, and insulin concentrations; and 3) plasma leptin returns to normal levels after tumor removal. These data, therefore, support a role for insulin in the chronic regulation of leptin gene expression.


Subject(s)
Gene Expression , Hyperinsulinism/genetics , Obesity/genetics , Proteins/genetics , Proteins/metabolism , Adipose Tissue/chemistry , Adult , Blood Glucose/metabolism , Fasting , Female , Humans , Insulin/blood , Insulinoma/genetics , Insulinoma/surgery , Leptin , Male , Middle Aged , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/surgery , Polymerase Chain Reaction , RNA, Messenger/analysis , RNA-Directed DNA Polymerase
12.
Diabetes ; 48(2): 403-7, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10334321

ABSTRACT

The finding of a reduced insulin-stimulated glucose uptake and glycogen synthesis in the skeletal muscle of glucose-tolerant first-degree relatives of patients with NIDDM, as well as in cultured fibroblasts and skeletal muscle cells isolated from NIDDM patients, has been interpreted as evidence for a genetic involvement in the disease. The mode of inheritance of the common forms of NIDDM is as yet unclear, but the prevailing hypothesis supports a polygenic model. In the present study, we tested the hypothesis that the putative inheritable defects of insulin-stimulated muscle glycogen synthesis might be caused by genetic variability in the genes encoding proteins shown by biochemical evidence to be involved in insulin-stimulated glycogen synthesis in skeletal muscle. In 70 insulin-resistant Danish NIDDM patients, mutational analysis by reverse transcription-polymerase chain reaction-single strand conformation polymorphism-heteroduplex analysis was performed on genomic DNA or skeletal muscle-derived cDNAs encoding glycogenin, protein phosphatase inhibitor-1, phophatase targeting to glycogen, protein kinase B-alpha and -beta, and the phosphoinositide-dependent protein kinase-1. Although a number of silent variants were identified in some of the examined genes, we found no evidence for the hypothesis that the defective insulin-stimulated glycogen synthesis in skeletal muscle in NIDDM is caused by structural changes in the genes encoding the known components of the insulin-sensitive glycogen synthesis pathway of skeletal muscle.


Subject(s)
Carrier Proteins/genetics , DNA Mutational Analysis , Diabetes Mellitus, Type 2/genetics , Endoribonucleases , Glycoproteins/genetics , Intracellular Signaling Peptides and Proteins , Protein Serine-Threonine Kinases/genetics , Proto-Oncogene Proteins/genetics , RNA-Binding Proteins/genetics , 3-Phosphoinositide-Dependent Protein Kinases , Diabetes Mellitus, Type 2/metabolism , Female , Genetic Variation/physiology , Glucosyltransferases , Glycogen/biosynthesis , Humans , Insulin/physiology , Isomerism , Male , Middle Aged , Muscle, Skeletal/metabolism , Phenotype , Phosphoprotein Phosphatases , Proto-Oncogene Proteins c-akt
13.
J Mol Med (Berl) ; 79(5-6): 270-4, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11485019

ABSTRACT

Maturity-onset diabetes of the young (MODY), an autosomal dominant, early-onset form of type-2 diabetes, is caused by mutations in five different genes all leading to defect(s) in the pancreatic beta cell. However, some patients with this form of diabetes do not bear a mutation in any of the known (MODY1-MODY5) loci, a notion prompting the search for new MODY genes. Clinical and genetic data point toward a defect in beta cell function in the majority of patients with MODY, and partners of the glucose-sensing device are reasonable functional candidates. The high-capacity glucose transporter GLUT2 has the ideal kinetic features for performing this task. However, complete GLUT2 deficiency in humans leads to hepato-renal glycogenosis (Fanconi-Bickel syndrome), and heterozygous GLUT2 mutations apparently behave in a recessive manner. Furthermore, in the human beta cell GLUT1 mRNA is predominant when compared to GLUT2 and glucose influx appears to be largely mediated by this low-Km transporter. Thus, we looked for the presence of sequence variants by polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) within the GLUT1 gene in 90 Italian pedigrees negative at the search for mutations in glucokinase (MODY2) and hepatocyte nuclear factor-1alpha (MODY3), the two genes responsible for about 60% of MODY cases in Italian children. We found three already described silent mutations and a new single base deletion in position -173 of the 5' regulatory region. The -173de1A variant, which was detected in the heterozygous or homozygous state in 30.8% of MODY patients examined and is located in a Nuclear Factor Y binding sequence, is not associated with hyperglycemia in affected relatives of MODY probands. In conclusion, it appears from these results that the glucose transporter gene GLUT1 is unlikely to play a major role in the etiology of MODY diabetes.


Subject(s)
Diabetes Mellitus, Type 2/genetics , Monosaccharide Transport Proteins/genetics , Polymorphism, Single-Stranded Conformational , Adolescent , Base Sequence , CCAAT-Binding Factor/metabolism , Child , Child, Preschool , Consensus Sequence/genetics , Exons/genetics , Glucose Transporter Type 1 , Humans , Italy , Mutation/genetics , Polymerase Chain Reaction , Response Elements/genetics , Transcription Factors/metabolism
14.
An Pediatr (Barc) ; 82(1): e30-4, 2015 Jan.
Article in Spanish | MEDLINE | ID: mdl-25027621

ABSTRACT

Insulin resistance syndromes without lipodystrophy are an infrequent and heterogeneous group of disorders with variable clinical phenotypes, associated with hyperglycemia and hyperinsulinemia. The three conditions related to mutations in the insulin receptor gene are leprechaunism or Donohue syndrome, Rabson-Mendenhall syndrome, and Type A syndrome. A case is presented on a patient diagnosed with type A insulin resistance, defined by the triad of extreme insulin resistance, acanthosis nigricans, and hyperandrogenism, carrying a heterozygous mutation in exon 19 of the insulin receptor gene coding for its tyrosine kinase domain that is crucial for the catalytic activity of the receptor. The molecular basis of the syndrome is reviewed, focusing on the structure-function relationships of the insulin receptor, knowing that the criteria for survival are linked to residual insulin receptor function. It is also pointed out that, although type A insulin resistance appears to represent a somewhat less severe condition, these patients have a high morbidity and their treatment is still unsatisfactory.


Subject(s)
Insulin Resistance/genetics , Mutation , Receptor, Insulin/genetics , Child , Diabetes Mellitus , Female , Humans , Insulin/classification , Phenotype , Severity of Illness Index , Syndrome
15.
J Invest Dermatol ; 98(6 Suppl): 77S-81S, 1992 Jun.
Article in English | MEDLINE | ID: mdl-1588128

ABSTRACT

Mutations of the insulin receptor gene have been identified in patients with genetic syndromes of insulin resistance associated with acanthosis nigricans. These mutations impair insulin responses by reducing the number of insulin receptors on the surface of target cells, or by reducing the receptor's ability to bind insulin or to undergo insulin-stimulated autophosphorylation, an important step in insulin action. Studies of mutant receptors expressed in transfection systems have contributed to our understanding of the structure-function relationships of the insulin receptor.


Subject(s)
Acanthosis Nigricans/genetics , Insulin Resistance/genetics , Receptor, Insulin/genetics , Humans , Mutation , Syndrome
16.
J Clin Endocrinol Metab ; 70(5): 1371-4, 1990 May.
Article in English | MEDLINE | ID: mdl-2110574

ABSTRACT

In man, continuous infusion of GH-releasing hormone (GHRH) does not sustain GH secretion, unlike prolonged hypoglycemia. To further evaluate this difference in the stimulation of GH release we measured GH concentrations for 3 h during prolonged insulin-induced hypoglycemia and GHRH-(1-29)NH2 (100 micrograms/h) infusion in normal individuals. We also assessed the GH response to combined and separate administration of insulin and GHRH. Plasma GH levels increased during prolonged hypoglycemia and remained elevated for the third hour (22-24 micrograms/L). GH concentrations increased during GHRH infusion, peaked at 60 min (23.5 micrograms/L), and rapidly declined. Thus, our findings confirmed that prolonged hypoglycemia, unlike GHRH infusion, sustained elevated GH levels and that these high levels did not appear to influence GH secretion from the pituitary. Changes in FFA did not account for the sustained GH secretion. FFA levels initially declined during insulin infusion, but after 3 h of hypoglycemia they returned to near-basal values (basal, 0.1 +/- 0.02 g/L; 180 min, 0.09 +/- 0.02). The maximal GH concentration attained during the combined insulin and GHRH test was significantly higher than that with the insulin tolerance test or GHRH test (insulin plus GHRH, 71.9 +/- 13.5; insulin tolerance test, 34.2 +/- 2.9; P less than 0.025; GHRH test, 27.9 +/- 3.2; P less than 0.02), indicating an additive effect on GH secretion. These data suggest that insulin-induced hypoglycemia stimulates GH secretion through a mechanism partly independent of GHRH. The release from somatostatin inhibition and stimulation through other neuropeptides (e.g. galanin) is suggested as possible causes of hypoglycemia-induced GH secretion.


Subject(s)
Growth Hormone/metabolism , Hypoglycemia/metabolism , Adult , Dose-Response Relationship, Drug , Female , Growth Hormone/administration & dosage , Growth Hormone-Releasing Hormone/administration & dosage , Growth Hormone-Releasing Hormone/physiology , Humans , Hypoglycemia/chemically induced , Infusions, Intravenous , Insulin/administration & dosage , Insulin Infusion Systems , Male , Pituitary Gland/drug effects , Pituitary Gland/physiology
17.
J Clin Endocrinol Metab ; 78(5): 1153-8, 1994 May.
Article in English | MEDLINE | ID: mdl-8175972

ABSTRACT

Molecular scanning techniques, such as denaturing gradient gel electrophoresis (DGGE), greatly facilitate screening candidate genes for mutations. We have used DGGE to screen for mutations in the insulin receptor gene in a family in which four of five daughters were affected by type A insulin resistance in association with acanthosis nigricans and hyperandrogenism. DGGE did not detect mutations in any of the 22 exons of the insulin receptor gene. Nevertheless, Southern blot analysis suggested that there was a deletion of exon 3 in the other paternal allele of the insulin receptor gene. Analysis of the father's cDNA confirmed that exon 3 was deleted from mRNA molecules derived from one of his two alleles of the insulin receptor gene. Furthermore, the father was found to be hemizygous for a polymorphic sequence (GACAsp at codon 234) in exon 3 that was not inherited by any of the five daughters. Instead, all five daughters inherited the paternal allele with the deletion mutation. We did not detect mutations in the mother's insulin receptor gene. Furthermore, the clinical syndrome did not segregate with either of the mother's two alleles of the insulin receptor gene. Although the youngest daughter inherited the mutant allele from her father, she was not clinically affected. The explanation for the incomplete penetrance is not known. These results emphasize the importance of specifically searching for deletion mutations when screening candidate genes for mutations. Furthermore, the existence of apparently asymptomatic carriers of mutations in the insulin receptor gene, such as the father in the present study, suggests that the prevalence of mutations in the insulin receptor gene may be higher than would be predicted on the basis of the observed prevalence of patients with extreme insulin resistance.


Subject(s)
Gene Deletion , Insulin Resistance/genetics , Receptor, Insulin/genetics , Adolescent , Adult , Alleles , Amino Acid Sequence , Base Sequence , Blotting, Southern , Child , Exons , Humans , Molecular Sequence Data , Mutation
18.
J Clin Endocrinol Metab ; 85(3): 1323-6, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10720084

ABSTRACT

Increasing evidence suggests that defects in genes encoding transcription factors that are expressed in the pancreatic beta-cells may be important contributors to the genetic basis of type 2 diabetes mellitus. Maturity-onset diabetes of the young (MODY) now exists in five subtypes (MODY1-5), four of which are caused by mutations in transcription factors hepatocyte nuclear factor-4alpha (HNF-4alpha), HNF-1alpha, insulin promoter factor-1 (IPF-1), and HNF-1beta (MODY1, -3, -4, and -5). Recent evidence from the British population even suggested that IPF-1 may be a predisposing gene for type 2 diabetes. Thus, highlighting the potential role of this transcription factor in the genetic basis of Danish and Italian MODY as well as in Danish patients with late-onset type 2 diabetes mellitus, we have examined the human IPF-1 gene for mutations by single strand conformation polymorphism and heteroduplex analysis in 200 Danish patients with late-onset type 2 diabetes and in 44 Danish and Italian MODY patients. In the patients with late-onset type 2 diabetes we identified a noncoding G insertion/deletion polymorphism at nucleotide -108, a silent G54G, and a rare missense D76N variant. Moreover, a Danish MODY patient was carrier of an A140T variant. Neither the D76N nor the A140T segregated with diabetes, and their transcriptional activation of the human insulin promoter expressed in vitro was indistinguishable from that of the wild type (115 +/- 21% and 84 +/- 12% vs. 100%). We conclude that variants in IPF-1 are not a common cause of MODY or late-onset type 2 diabetes in the Caucasian population, and that in terms of insulin transcription both the N76 and the T140 mutations are likely to represent functionally normal IPF-1 variants with no direct role in the pathogenesis of MODY or late-onset type 2 diabetes mellitus.


Subject(s)
Diabetes Mellitus, Type 2/genetics , Homeodomain Proteins , Mutation, Missense/genetics , Trans-Activators/genetics , 3T3 Cells , Adult , Age of Onset , Aged , Aged, 80 and over , Animals , DNA/genetics , DNA Mutational Analysis , Denmark , Female , Heterozygote , Humans , Male , Mice , Middle Aged , Mutagenesis , Polymorphism, Single-Stranded Conformational , Transcriptional Activation/genetics , White People
19.
J Clin Endocrinol Metab ; 71(1): 164-9, 1990 Jul.
Article in English | MEDLINE | ID: mdl-2196279

ABSTRACT

Mutations in the insulin gene can impair the bioactivity of the insulin molecule. Previously, two classes of mutations have been identified: 1) those that impair posttranslational processing of proinsulin to insulin, and 2) those that alter the structure of the insulin molecule, thereby reducing the affinity of the molecule for the insulin receptor. We have investigated two apparently unrelated patients, both of which have mutations that inhibit the conversion of proinsulin to insulin. By directly sequencing genomic DNA amplified by polymerase chain reaction, we have demonstrated that both patients are heterozygous for the same point mutation converting codon 65 from an arginine (CGT) to a histidine (CAT) codon. Because Arg65 is one of the two dibasic amino acids at the site of proteolytic cleavage between the insulin A-chain and C-peptide, this mutation explains the impairment in the cleavage of proinsulin to insulin. Interestingly, the same His65 mutation has been identified in the insulin gene of a Japanese kindred with familial hyperproinsulinemia. Thus, this mutation has occurred in three apparently unrelated kindreds from two different racial groups. This observation is consistent with the hypothesis that the dinucleotide sequence CpG, the first two nucleotides in the arginine (CGT) codon, is a "hot spot" for mutations.


Subject(s)
Arginine , DNA/analysis , Histidine , Insulin/genetics , Proinsulin/blood , Proinsulin/genetics , Alleles , Amino Acid Sequence , Base Sequence , Blood Glucose/analysis , Codon , Exons , Gene Amplification , Genes , Humans , Hyperinsulinism/genetics , Hyperinsulinism/metabolism , Insulin/biosynthesis , Male , Middle Aged , Molecular Sequence Data , Mutation , Nucleic Acid Hybridization , Peptide Hydrolases , Polymerase Chain Reaction , Proinsulin/metabolism
20.
Endocrinol Metab Clin North Am ; 18(1): 123-43, 1989 Mar.
Article in English | MEDLINE | ID: mdl-2645123

ABSTRACT

Humoral autoimmunity plays an important role in the pathogenesis of two forms of hypoglycemia. In one syndrome, antireceptor autoantibodies bind to the insulin receptor, mimic insulin action, and cause fasting hypoglycemia. In most patients with autoantibodies to the insulin receptor, there is other evidence of autoimmune disease as well. Interpretation of the standard tests used in evaluation of hypoglycemia may be confusing in these patients. For example, antireceptor antibodies may inhibit insulin binding, thereby inhibiting insulin clearance and elevating levels of plasma insulin. Nevertheless, because hypoglycemia suppresses beta-cell secretion, C-peptide levels are usually low. This constellation of data is consistent with surreptitious insulin injection. The most important laboratory test in the differential diagnosis is a direct assay for the presence of antibodies directed against the insulin receptor. Therapy with prednisone appears to alleviate the hypoglycemia rapidly, usually within 24 hours. This effect of prednisone appears to result from antagonism of the effects of antireceptor antibodies without actually lowering their titer. The natural history of this syndrome is that the antireceptor antibodies disappear and the syndrome resolves over a time course of several months to several years. In North America, the presence of anti-insulin antibodies in a hypoglycemic patient most commonly suggests that the patient has been immunized with exogenous insulin. However, some patients--especially in Japan--develop spontaneous autoantibodies directed against insulin. These antibodies can cause hypoglycemia, which is generally reactive in that it occurs several hours after a meal or a glucose challenge rather than in a fasting state. The most effective therapy is frequent small feedings and avoidance of large meals.


Subject(s)
Autoantibodies/immunology , Autoimmune Diseases/immunology , Hypoglycemia/immunology , Insulin Antibodies/immunology , Receptor, Insulin/immunology , Autoantibodies/analysis , Autoimmune Diseases/diagnosis , Autoimmune Diseases/therapy , Humans , Hypoglycemia/etiology , Hypoglycemia/therapy , Immunologic Tests , Insulin Antibodies/analysis , Syndrome
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