Aaron Chapla

Maturity onset diabetes of the young in India - a distinctive mutation pattern identified through targeted next-generation sequencing

To establish and utilize a Next‐Generation Sequencing (NGS)‐based strategy to screen for maturity onset diabetes of the young (MODY) gene mutations in subjects with early‐onset diabetes.

Zinc protects against indomethacin-induced damage in the rat small intestine.

The clinical utility of nonsteroidal anti-inflammatory drugs (NSAIDs) is often limited by the adverse effects that they produce in the small intestine. Alterations in the composition and functions of the glycocalyx and brush border membranes of the rat small intestine have been shown to occur in response to indomethacin, an NSAID often used in the study of adverse effects of these drugs. The micronutrient, zinc, has been documented to have cytoprotective effects in the gastrointestinal tract. The aim of this study was to evaluate the potential of zinc to reduce indomethacin-induced small intestinal damage. We pre-treated rats with zinc sulphate (50 mg/kg body weight) 2h before administration of indomethacin (20 mg/kg body weight) and sacrificed the rats 1, 12 or 24h after indomethacin. The extent of small intestinal mucosal damage and the content of lipids and sugars in the mucosa were determined. Bacterial counts in the intestinal lumen and the mucosa were ascertained. Activities of matrix metalloproteinases (MMPs) and levels of metallothionein in the mucosa were also measured. Pre-treatment with zinc sulphate was found to reduce the extent of indomethacin-induced mucosal damage. It also prevented drug-induced changes in the content of lipids and sugars in the mucosa. Drug-induced increases in activities of the MMPs and bacterial counts in the intestine were also attenuated by zinc. Metallothionein levels were significantly higher in animals pre-treated with zinc. We conclude that zinc was effective in protecting against indomethacin-induced small intestinal damage and suggest that it may do so by induction of metallothionein.

Molecular diagnosis of maturity onset diabetes of the young in India

Diabetes is highly prevalent in India and the proportion of younger patients developing diabetes is on the increase. Apart from the more universally known type 1 diabetes and obesity related type 2 diabetes, monogenic forms of diabetes are also suspected to be prevalent in many young diabetic patients. The identification of the genetic basis of the disease not only guides in therapeutic decision making, but also aids in genetic counselling and prognostication. Genetic testing may establish the occurrence and frequency of early diabetes in our population. This review attempts to explore the utilities and horizons of molecular genetics in the field of maturity onset diabetes of the young (MODY), which include the commoner forms of monogenic diabetes.

Comprehensive Maturity Onset Diabetes of the Young (MODY) Gene Screening in Pregnant Women with Diabetes in India.

Pregnant women with diabetes may have underlying beta cell dysfunction due to mutations/rare variants in genes associated with Maturity Onset Diabetes of the Young (MODY). MODY gene screening would reveal those women genetically predisposed and previously unrecognized with a monogenic form of diabetes for further clinical management, family screening and genetic counselling. However, there are minimal data available on MODY gene variants in pregnant women with diabetes from India. In this study, utilizing the Next generation sequencing (NGS) based protocol fifty subjects were screened for variants in a panel of thirteen MODY genes. Of these subjects 18% (9/50) were positive for definite or likely pathogenic or uncertain MODY variants. The majority of these variants was identified in subjects with autosomal dominant family history, of whom five were in women with pre-GDM and four with overt-GDM. The identified variants included one patient with HNF1A Ser3Cys, two PDX1 Glu224Lys, His94Gln, two NEUROD1 Glu59Gln, Phe318Ser, one INS Gly44Arg, one GCK, one ABCC8 Arg620Cys and one BLK Val418Met variants. In addition, three of the seven offspring screened were positive for the identified variant. These identified variants were further confirmed by Sanger sequencing. In conclusion, these findings in pregnant women with diabetes, imply that a proportion of GDM patients with autosomal dominant family history may have MODY. Further NGS based comprehensive studies with larger samples are required to confirm these finding

The H Syndrome: Molecular Diagnosis Using Next-Generation Sequencing

ABSTRACT Objective: H syndrome is a monogenic systemic inherited form of histiocytosis, with characteristic cutaneous findings accompanying systemic manifestations. The major common endocrine manifestations include hypogonadism, short stature, and diabetes mellitus with characteristic genodermatosis and lead to the diagnosis. Here, we report a rare case of H-syndrome, an autosomal recessive non-autoimmune disorder in a 19-year-old woman who presented with short stature, diabetes mellitus, and hypogonadism associated with characteristic hyperpigmentation and hypertrichosis. The molecular diagnosis was established utilizing next-generation sequencing (NGS) technology. Methods: We describe the clinical spectrum of H syndrome with endocrine and non endocrine multisystem involvement. The solute carrier family 29 (nucleoside transporters), member 3 (SLC29A3) gene was screened for molecular diagnosis utilizing NGS based mutational analysis. Results: H syndrome is caused by a mutation in the SLC29A3 gene, which e...

Next-Generation Sequencing-Based Genetic Testing For Familial Partial Lipodystrophy

ABSTRACT Objective: Familial partial lipodystrophy (FPL) of the Dunnigan type (FPLD) is an autosomal dominant condition characterized by fat loss in the limbs and trunk, fat accumulation in the head and neck, and early onset diabetes mellitus. Here we describe the establishment and utilization of next-generation sequencing (NGS)-based genetic testing for FPLD. Methods: We describe NGS-based mutational analysis of the lamin A/C (LMNA) gene, followed by confirmation through Sanger sequencing. Results: We report a patient and her mother with accumulation of fat in the neck and face and loss of fat in the limbs and trunk typical of FPLD2, with young onset diabetes mellitus without ketoacidosis. Both subjects had elevated homeostasis model assessment estimated insulin resistance (HOMA-IR) values and serum triglyceride levels, indicating insulin resistance. Dual energy X-ray absorptiometry confirmed typical fat redistribution. NGS-based mutational analysis of the LMNA gene in these patients revealed a hot spot ...

Sirolimus therapy for congenital hyperinsulinism in an infant with a novel homozygous KCNJ11 mutation.

Abstract Background: Congenital hyperinsulinism results in refractory hypoglycemia. If a therapy with diazoxide has been unresponsive this has been treated by subtotal pancreatectomy in the past. This therapeutic option poses an increased risk of developing diabetes at a later stage. There have been a few case reports on the use of sirolimus in such situations in the recent past. Case presentation: Our patient was started on sirolimus very early, on day 29 of life and at the age of 14 months is doing well on sirolimus therapy. His growth and development have been good and he has not had any major complications so far. Genetic testing showed a novel KCNJ11 homozygous mutation on next generation sequencing and the parents were heterozygous carriers. Conclusions: We report the successful use of sirolimus in the management of diazoxide unresponsive congenital hyperinsulinism with diffuse pancreatic involvement. We believe this is the youngest patient to be initiated on sirolimus so far.

Molecular Diagnosis of Wilson's Disease Using Next Generation Sequencing Platform

BackgroundandAim:The coexistence ofWilson’s disease and autoimmune liver disease in a same patients is a rare entity. In this situation, combined treatment with steroid and D-Penicillamine may be effective. Methods:We analysed clinical, histological, laboratory profile for patients with chronic liver disease with aim of finding the etiologyof thedisease after rulingout common causes like alcohol, viruses and drugs. Results: Out of 10 patients 6 were males and 4 were females. Commonest clinical presentation was abdominal distension (80%), abdominal pain (30%), pedal edema (60%), splenomegaly (40%) and upper GI bleed (40%). Laboratory investigation revealed anemia (50%), thrombocytopenia (70%), prothrombin time prolongation in (60%), normal liver function in 60%, abnormal liver function in (40%). Hepatitis A, B, C and E were negative in all the cases. Serum cereloplasmin <20mg% in (30%), normal level in (70%). 24h urinary copper level range of >200 in (40%), 100 to 200 in (40%) and 90 to 100 in (20%). Autoimmune markers revealed ANA strong positivity in (40%), mild positivity in (60%). AMA, ASMA, Anti-LKM-1 were negative in all cases (100%). Liver biopsy showed features of Autoimmune liver disease and PERIPORTAL copper deposition in 80% of cases where 20% showed features of copper deposition in liver. Conclusion: The coexistence of Wilson’s disease and autoimmune liver disease is a rare entity and clinician should have high level of suspicion in diagnosing the problem and medical treatment with steroids and DPencillamine simultaneously tobe started in thesepatients (Figure 1). Figure 1. Liver biopsy.

Monogenic diabetes—diagnostic conundrums

Diabetes mellitus (DM) is a global pandemic [1] that affects nearly 382 million people worldwide [2]. The vast majority of patients (approximately 85 %) are classified into polygenic type 1 diabetes (T1D) or type 2 diabetes (T2D). However, with growing evidence from genomic research, several monogenic causes of diabetes have emerged. Monogenic forms of diabetes include maturity onset of diabetes of young (MODY), neonatal diabetes and rare syndromic forms of diabetes [3].

Carney complex with PRKAR1A mutation: A case report

Carney complex is a multiple endocrine neoplasia syndrome with various features which include myxomas, endocrine tumours and lentigines lesions. We report a case of Carney complex with components of lentigines, ACTH independent adrenal Cushing’s syndrome (with a paradoxical increase in 24 hour urinary cortisol following the high dose (8mg) dexamethasone suppression test – and is likely to be due to primary pigmented nodular adrenal hyperplasia) positive for a protein kinase A type 1A regulatory subunit (PRKAR1A) gene mutation.