Molecular Diagnosis of Neonatal Diabetes Mellitus Using Next-Generation Sequencing of the Whole Exome

Background: Accurate molecular diagnosis of monogenic non-autoimmune neonatal diabetes mellitus (NDM) is critical for patient care, as patients carrying a mutation in KCNJ11 or ABCC8 can be treated by oral sulfonylurea drugs instead of insulin therapy. This diagnosis is currently based on Sanger sequencing of at least 42 PCR fragments from the KCNJ11, ABCC8, and INS genes. Here, we assessed the feasibility of using the next-generation whole exome sequencing (WES) for the NDM molecular diagnosis. Methodology/Principal Findings: We carried out WES for a patient presenting with permanent NDM, for whom mutations in KCNJ11, ABCC8 and INS and abnormalities in chromosome 6q24 had been previously excluded. A solution hybridization selection was performed to generate WES in 76 bp paired-end reads, by using two channels of the sequencing instrument. WES quality was assessed using a high-resolution oligonucleotide whole-genome genotyping array. From our WES with high-quality reads, we identified a novel non-synonymous mutation in ABCC8 (c.1455G.C/p.Q485H), despite a previous negative sequencing of this gene. This mutation, confirmed by Sanger sequencing, was not present in 348 controls and in the patient’s mother, father and young brother, all of whom are normoglycemic. Conclusions/Significance: WES identified a novel de novo ABCC8 mutation in a NDM patient. Compared to the current Sanger protocol, WES is a comprehensive, cost-efficient and rapid method to identify mutations in NDM patients. W

New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk.

Levels of circulating glucose are tightly regulated. To identify new loci influencing glycemic traits, we performed meta-analyses of 21 genome-wide association studies informative for fasting glucose, fasting insulin and indices of beta-cell function (HOMA-B) and insulin resistance (HOMA-IR) in up to 46,186 nondiabetic participants. Follow-up of 25 loci in up to 76,558 additional subjects identified 16 loci associated with fasting glucose and HOMA-B and two loci associated with fasting insulin and HOMA-IR. These include nine loci newly associated with fasting glucose (in or near ADCY5, MADD, ADRA2A, CRY2, FADS1, GLIS3, SLC2A2, PROX1 and C2CD4B) and one influencing fasting insulin and HOMA-IR (near IGF1). We also demonstrated association of ADCY5, PROX1, GCK, GCKR and DGKB-TMEM195 with type 2 diabetes. Within these loci, likely biological candidate genes influence signal transduction, cell proliferation, development, glucose-sensing and circadian regulation. Our results demonstrate that genetic studies of glycemic traits can identify type 2 diabetes risk loci, as well as loci containing gene variants that are associated with a modest elevation in glucose levels but are not associated with overt diabetes

New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk

Levels of circulating glucose are tightly regulated. To identify new loci influencing glycemic traits, we performed meta-analyses of 21 genome-wide association studies informative for fasting glucose, fasting insulin and indices of beta-cell function (HOMA-B) and insulin resistance (HOMA-IR) in up to 46,186 nondiabetic participants. Follow-up of 25 loci in up to 76,558 additional subjects identified 16 loci associated with fasting glucose and HOMA-B and two loci associated with fasting insulin and HOMA-IR. These include nine loci newly associated with fasting glucose (in or near ADCY5, MADD, ADRA2A, CRY2, FADS1, GLIS3, SLC2A2, PROX1 and C2CD4B) and one influencing fasting insulin and HOMA-IR (near IGF1). We also demonstrated association of ADCY5, PROX1, GCK, GCKR and DGKB-TMEM195 with type 2 diabetes. Within these loci, likely biological candidate genes influence signal transduction, cell proliferation, development, glucose-sensing and circadian regulation. Our results demonstrate that genetic studies of glycemic traits can identify type 2 diabetes risk loci, as well as loci containing gene variants that are associated with a modest elevation in glucose levels but are not associated with overt diabetes

A genome-wide association search for type 2 diabetes genes in African Americans.

African Americans are disproportionately affected by type 2 diabetes (T2DM) yet few studies have examined T2DM using genome-wide association approaches in this ethnicity. The aim of this study was to identify genes associated with T2DM in the African American population. We performed a Genome Wide Association Study (GWAS) using the Affymetrix 6.0 array in 965 African-American cases with T2DM and end-stage renal disease (T2DM-ESRD) and 1029 population-based controls. The most significant SNPs (n = 550 independent loci) were genotyped in a replication cohort and 122 SNPs (n = 98 independent loci) were further tested through genotyping three additional validation cohorts followed by meta-analysis in all five cohorts totaling 3,132 cases and 3,317 controls. Twelve SNPs had evidence of association in the GWAS (P<0.0071), were directionally consistent in the Replication cohort and were associated with T2DM in subjects without nephropathy (P<0.05). Meta-analysis in all cases and controls revealed a single SNP reaching genome-wide significance (P<2.5×10(-8)). SNP rs7560163 (P = 7.0×10(-9), OR (95% CI) = 0.75 (0.67-0.84)) is located intergenically between RND3 and RBM43. Four additional loci (rs7542900, rs4659485, rs2722769 and rs7107217) were associated with T2DM (P<0.05) and reached more nominal levels of significance (P<2.5×10(-5)) in the overall analysis and may represent novel loci that contribute to T2DM. We have identified novel T2DM-susceptibility variants in the African-American population. Notably, T2DM risk was associated with the major allele and implies an interesting genetic architecture in this population. These results suggest that multiple loci underlie T2DM susceptibility in the African-American population and that these loci are distinct from those identified in other ethnic populations

Response letter to “Latent class analysis of 216 patients with adult-onset Still’s disease” by Sugiyama et al.

Abstract Sugiyama et al. recently described in “Latent class analysis of 216 patients with adult-onset Still’s disease,” baseline characteristics, laboratory tests, treatment, relapse, and death of adult-onset Still’s disease (AOSD) patients from a Japanese hospital. They identified two subgroups: Class 1 (n=155) with a younger age and typical symptoms of AOSD and Class 2 (n=61) with older patients and fewer typical symptoms of AOSD. In 2022, VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) syndrome, an established X-linked disease associated with a somatic mutation in UBA1, is considered as a differential diagnosis for AOSD particularly in elderly. These patients from Class 2 could benefit from more explorations for mild myelodysplasia and VEXAS

Le lymphome de la zone marginale ganglionnaire, un diagnostic d'exclusion (à propos de 24 cas)

Le lymphome de la zone marginale ganglionnaire est une pathologie rare, de présentation hétérogène, pour laquelle il n existe pas encore de marqueurs immunohistochimiques ou moléculaires spécifiques. Le diagnostic différentiel avec d autres lymphomes à petites cellules B peut s avérer parfois extrêmement difficile. La mise en place d une stratégie dans la réalisation des examens complémentaires est donc indispensable. Nous avons caractérisé sur le plan morphologique, immunohistochimique et moléculaire 24 cas de lymphome de la zone marginale ganglionnaire. L étude immunohistochimique a été réalisée avec les anticorps suivants : CD20, CD79a, CD3, CD5, CD21, CD23, CD43, CD10, Bcl6, Bcl2, cyclineD1, Sox11et Ki67. Une recherche de réarrangement de BCL1, BCL2, MALT1 pour l ensemble des cas, et de BCL6 pour 3 cas, a été effectuée. L exon 5 de MYD88 a été étudié pour la totalité des cas. Quatre types d architecture ont été observés : diffuse (37.5%), nodulaire (4.5%), nodulaire et diffuse (41.5%) et splénique (16.5%). Le polymorphisme cellulaire était prédominant (67%). La différenciation lymphoplasmocytaire était la plus fréquente (67%), tandis que le contingent monocytoïde pur était peu rencontré (4.5%). Une composante minoritaire était cependant souvent retrouvée (33%). L immunophénotype classique était observé dans 41.5% des cas. Les cellules tumorales de 5 cas étaient positives avec l anticorps anti-CD5. Cinq autre cas présentaient un marquage avec l anticorps anti-CD23 et un cas avec les anticorps anti-CD5 et anti-CD23. Les cellules tumorales de 3 cas étaient partiellement positives avec l anticorps anti-Bcl6. Trois cas ont été exclus au cours de l étude. Les cellules tumorales de l un d entre eux étaient positives avec les anticorps anti-cycline D1 et anti-Sox11. L étude FISH avait mis en évidence une t(11;14). Les deux autres cas présentaient un marquage partiel des cellules tumorales par l anticorps anti-Bcl6, ainsi qu une amplification du gène BCL6 en FISH. La mutation L265P de MYD88 a été retrouvée dans un cas dont la morphologie, le phénotype et l étude moléculaire étaient en faveur d un lymphome de la zone marginale ganglionnaire. Devant l importante variabilité de présentation morphologique et immunophénotypique des lymphomes de la zone marginale ganglionnaire, nous mettons en avant l intérêt d une étude immunohistochimique globale face à un lymphome à petites cellules B, notamment le bénéfice de l usage de différents marqueurs du centre germinatif et de la cycline D1. La FISH ainsi que la recherche de la mutation L265P de MYD88 présentent également un intérêt dans ce diagnostic.LILLE2-BU Santé-Recherche (593502101) / SudocSudocFranceF

Correspondence on “Safety of vaccination against SARS-CoV-2 in people with rheumatic and musculoskeletal diseases: results from the EULAR Coronavirus Vaccine (COVAX) physician-reported registry” by Machado et al

We read with great interest the article by Machado et al who describe safety of vaccination against SARS-CoV-2 in people with rheumatic and musculoskeletal disease. 1 The authors observed that vaccine against SARS-CoV-2 is well tolerated with rare report of I-RMD flare and very rare reports of serious adverse events. We observed that the authors included only 27 patients with autoinflammatory diseases. We thus propose to complete their observation with the result of our study about 190 patients with autoinflammatory disease (AID). A web survey assessing adverse effects after COVID-19 vaccination was sent on 7-30 June 2021 to patients with AID followed in the French national adult AID reference centre, and already included in the Juvenile Inflammatory Rheumatism (JIR) cohort. The patients were asked whether they had received a COVID-19 vaccination, the type of vaccine and number of injections. Severe adverse events were defined by the need for hospitalisation. Local reaction, fever, headache, arthralgia, myalgia, allergic reaction, fatigue, nausea, adenopathy, heart disorder, venous thromboembolism and stroke were monitored after the first and the second injection. The survey was sent by email to 445 patients with AID: 225 (50%) patients answered it, 168 aged between 18 and 55 years old and 57 aged above 55 years old. Among the 190 patients who received two doses of COVID-19 vaccines (online supplemental table), most patients had familial Mediterranean fever (FMF) (n=128, 67.4%); other AID were undefined systemic AID (n=20), TNF-α receptor-associated periodic syndrome (n=13), cryopyrin-associated periodic syndrome (n=9), adult-onset still disease (n=9), mevalonate kinase deficiency (n=7) and A20 haploinsufficiency (n=4). Eleven patients declared also having AA amyloidosis (5.7%). Colchicine was the most used treatment (n=138, 72.6%); 37 (19.5%) patients were on biotherapy, mostly interleukin-1 inhibitors (n=33) and 15 patients were not taking any treatment. Forty-six patients had already contracted SARS-CoV-2. Out of the 190 (84.4%) vaccinated patients with AID, BNT162b2 (Pfizer/ BioNTech) (n=157, 82.6%) and ChAdOx1 nCoV-19 (AstraZeneca) (n=22, 11.5%) were the most common vaccines; few patients received CX-024414 (Moderna) (n=11, 5.8%). Eighty-eight patients (46%) developed mild adverse events after the first injection and 70 patients (54%) after the second injection. Among the 153 patients who received BNT162b2, tenderness at the injection site was the most reported event (n=39, 25.5%); others were myalgia (n=28, 18.3%), fever (n=20, 13%) and headache (n=16, 10.5%). Concerning ChAdOx1 nCoV-19, reported events were fever (n=13, 59%), myalgia (n=11, 50%) and intense fatigue (n=2, 9%). Concerning CX-024414, four patients reported fever and myalgia (36%). No severe adverse event requiring hospitalisation has been reported. Twelve patients with FMF (9.3%) reported a mild flare after the first injection, which did not require hospitalisation. No vaccinated patient had developed COVID-19 after the second vaccine injection. Altogether, this study shows that adverse event of COVID-19 vaccination in patients with AID are similar to the expected adverse effects reported in the general population. 2 Especially among patients with FMF on colchicine treatment, the vaccine is very safe and should be highly recommended to patients with risk factors of severe COVID-19, since we previously reported death among such patients with FMF. 3 It also suggests that COVID-19 vaccination does not usually trigger an AID flare; these data were also reported in patients with autoimmune diseases 4 and AID. 5 To our knowledge, this is the largest study describing the effects of COVID-19 vaccination among patients with AID: the vaccine is well tolerated; these data combined with the results from Machado et al 1 could reassure patients displaying immune systemic disorders including AID patients who are still hesitant about COVID-19 vaccination, especially in the actual context of the resurgence of the epidemy

Algorithmes diagnostiques et thérapeutiques des maladies auto-inflammatoires monogéniques présentant des fièvres récurrentes chez l'adulte

International audienceAutoinflammatory diseases (AID) are defined as disorders of innate immunity. They were initially defined in contrast to autoimmune diseases because of the lack of involvement of the adaptive immune system and circulating autoantibodies. The 4 monogenic AIDs first described are called the "historical" AIDs and include: Familial Mediterranean Fever (associated with MEFV mutations), cryopyrinopathies (associated with NLRP3 mutations), Tumor Necrosis Factor Receptor-associated Periodic Syndrome (associated with TNFRSF1A mutations) and Mevalonate Kinase Deficiency (associated with MVK mutations). In the last 10 years, more than 50 new monogenic AIDs have been discovered due to genetic advances. The most important discovery for adult patients is VEXAS syndrome associated with somatic UBA1 mutations leading to an autoinflammatory disease affecting mostly elderly men. Diagnosis of monogenic AIDs is based on personal and family history and detailed analysis of symptoms associated with febrile attacks, in the context of elevated peripheral inflammatory markers. This review proposes a practical approach of the diagnosis of the main monogenic AIDs among adult patients to guide the clinician

Long-Term Outcomes after Pelvic Organ Prolapse Repair in Young Women

The aim of the study was to describe the long-term outcomes of Pelvis Organ Prolapse (POP) repair in women under 40 years old. A retrospective chart review of all POP repairs performed in women ≤40 years old between January 1997 and December 2015 in the Gynecologic Surgery Department of Lille University Hospital was performed. Inclusion criteria were all women ≤40 years old who underwent a POP repair with a stage ≥2 POP according to the Baden and Walker classification. The study population was separated into three groups: a sacrohysteropexy group, a vaginal native tissue repair (NTR) group, and a transvaginal mesh surgery (VMS) group. The primary outcome was reoperation procedures for a symptomatic recurrent POP. Secondary outcomes were other complications. During the study period, 43 women ≤ 40 years old who underwent a POP repair were included and separated into three groups: 28 patients (68%), 8 patients (19%), and 7 patients (16%) in the sacrohysteropexy, VMS, and NTR groups respectively. The mean followup time was 83 ± 52 months. POP recurrence, reoperated or not, was essentially diagnosed in the VMS group (87.5%) and the NTR group (50%). POP recurrence repairs were performed for nine patients (21%): 7%, 62.5%, and 25% in the sacrohysteropexy, VMS, and NTR groups, respectively. Global reoperation concerned 10 patients (23%) whatever the type of POP surgery, mainly patients from the VMS group (75%) and from the NTR group (25%). It occurred in only 7% of patients from the sacrohysteropexy group. Two patients (4%) presented a vaginal exposure of the mesh (in the VMS group). De novo stress urinary incontinence was encountered by nine patients (21%): 29% and 12.5% in the sacrohysteropexy and NTR groups, respectively. Despite the risk of recurrence, POP repair should be proposed to young women in order to restore their quality of life. Vaginal native tissue repair or sacrohysteropexy should be performed after explaining to women the advantages and disadvantages of each procedure

Long-Term Assessment of Pelvic Organ Prolapse Reoperation Risk in Obese Women: Vaginal and Laparoscopic Approaches

The aim of this study was to compare reoperation risks after pelvic organ prolapse repair at 5-year follow-up between obese, overweight, and normal-weight women and to assess these risks accounting for the surgical procedure. We performed a retrospective chart review of all the women who underwent POP repair by transvaginal mesh surgery between January 2005 and January 2009 or laparoscopic sacrocolpopexy between January 2003 and December 2013 at the Gynecologic Surgery Department of the Lille University Hospital. During the study period, 744 women who underwent POP repair were divided into three groups: 382 (51%), 240 (32%), and 122 (16%) in the nonobese group (BMI p = 0.80), nor among the women who underwent transvaginal mesh surgery or laparoscopic sacrocolpopexy. The risks of reoperation for POP recurrence, stress urinary incontinence, or mesh-related complications did not significantly differ between the three BMI groups in the overall population nor accounting for the surgical procedure. In conclusion, obesity does not seem to be a risk factor of reoperation for POP recurrence, SUI, or mesh-related complications in the long term regardless of the surgical approach