Personalizing Early-Stage Type 1 Diabetes in Children

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Mesenchymal stem cells for the cell-based therapy of type 1 diabetes : an experimental approach

Tese de doutoramento, Medicina (Fisiopatologia), Universidade de Lisboa, Faculdade de Medicina, 2011Human adult mesenchymal stem cells (hMSC) represent a potential source for cellbased therapies in Diabetes Mellitus. Yet, till now, no definitive data exist for clinical application. Aims 1. Evaluation of the pancreatic endocrine differentiation potential of humanmesenchymal stem-cells, immortalized with telomerase (hMSC-TERT); 2. Comparative analysis of the plasticity and molecular signature of human-islet-derived pancreatic cells (hIPC) and bone-marrow-derived hMSC (hBM-MSC). Research strategies: 1. In vitro differentiation of hMSC-TERT into insulin producing cells through: a) Endocrine-promoting culture conditions; b) Ectopic expression of two key regulatory genes of human endocrine pancreas, neurogenin3 (hNGN3) and pancreatic-duodenal homeobox 1 (/hPDX-1) genes in hMSC-TERT. 2. Comparison of the functional signature of hIPC and hBM-MSC by: a) Immunophenotype characterization; b) In vitro mesenchymal and endocrine differentiation of primary hBM-MSC and hIPC; c) Transcriptome analysis for detection of mesenchymal and adult stem-cell related gene markers. Identification of target genes, related to the origin and function of hBM-MSC and hIPC. Results We could demonstrate that hMSC-TERT can be differentiated in vitro into pancreatic endocrine phenotypes with the ability to synthesise and store insulin. Similar to pancreatic development, in hMSC-TERT, NGN3 contributes to the activation of PDX-1 promoter. Insulin secretion in a glucose-sensing manner was not detected in our cell system, which indicates that a mature state of beta-cell was not attained in these phenotypes; hIPC and hBM-MSC display very similar MSC phenotypes. However, our data suggest distinct functions of these populations, which are related to a “mesodermal tissue specification” and “positional memory” of these cells. In hIPC, we confirmed a state of epithelial-mesenchymal transition. Conclusions MSC populations are adult stem cells with distinct levels of plasticity and commitment. Future studies will help to identify those MSC populations with greater potential to replace or regenerate beta cells in type 1 Diabetes.As células estaminais adultas obtidas a partir do estroma de tecidos humanos, representam uma fonte celular potencialmente atractiva para a terapia celular da Diabetes Mellitus. Todavia, não existem ainda resultados definitivos que permitam a sua aplicação clínica. Objectivos 1. Avaliação do potencial de diferenciação endócrina das células estaminais mesenquimatosas humanas, imortalizadas pela telomerase (hMSC-TERT); 2. Análise comparativa da plasticidade e assinatura molecular de células humanas derivadas do ilhéu pancreático (hIPC) e células estaminais mesenquimatosas derivadas da medula óssea (hBM-MSC). Estratégias de investigação 1. Diferenciar in vitro as hMSC-TERT em células produtoras de insulina através de: a) Cultura em meio específico pró-endócrino; b) Expressão ectópica de dois genes reguladores do pâncreas endócrino humano, neurogenina3 (hNGN3) e pancreatic-duodenal homeobox 1 (hPDX-1) nas células hMSC-TERT. 2. Comparar a assinatura molecular funcional das hIPC e hBM-MSC por: a) Caracterização do fenotipo imunológico; b) Diferenciação in vitro das hIPC e hBM-MSC em linhagens mesenquimatosas e endócrinas; c) Análise transcripcional de marcadores do mesênquima e genes associados ao carácter estaminal. Identificação de genes-alvo relacionados com a origem e função das hIPC e hBM-MSC. Resultados As hMSC-TERT são diferenciáveis em fenótipos do pâncreas endócrino com capacidade de síntese de insulina. Tal como ocorre na pancreatogénese, a expressão de NGN3 contribui para a activação do PDX1 nas hMSC-TERT. A ausência de resposta insulínica à glicose indica que as hMSC-TERT modificadas não atingiram a maturidade funcional de células-beta nativas; As hIPC e hBM-MSC apresentam características de MSC, tendo todavia, funções tecidulares distintas que reflectem diferenças moleculares relacionadas com a “especificidade mesodérmica” e “memória posicional” destas células. As hIPC, encontram-se num estado de transição epitelial-mesenquimatosa . Conclusões As populações MSC, são células estaminais adultas com níveis distintos de plasticidade e compromisso tecidular. Estudos futuros permitirão identificar as MSC com maior potencial para substituição ou regeneração de células beta na Diabetes tipo 1

The rare DRB1*04:08-DQ8 haplotype is the main HLA class II genetic driver and discriminative factor of Early-onset Type 1 diabetes in the Portuguese population

IntroductionEarly-onset Type 1 diabetes (EOT1D) is considered a disease subtype with distinctive immunological and clinical features. While both Human Leukocyte Antigen (HLA) and non-HLA variants contribute to age at T1D diagnosis, detailed analyses of EOT1D-specific genetic determinants are still lacking. This study scrutinized the involvement of the HLA class II locus in EOT1D genetic control.MethodsWe conducted genetic association and regularized logistic regression analyses to evaluate genotypic, haplotypic and allelic variants in DRB1, DQA1 and DQB1 genes in children with EOT1D (diagnosed at ≤5 years of age; n=97), individuals with later-onset disease (LaOT1D; diagnosed 8-30 years of age; n=96) and nondiabetic control subjects (n=169), in the Portuguese population. ResultsAllelic association analysis of EOT1D and LaOT1D unrelated patients in comparison with controls, revealed that the rare DRB1*04:08 allele is a distinctive EOT1D susceptibility factor (corrected p-value=7.0x10-7). Conversely, the classical T1D risk allele DRB1*04:05 was absent in EOT1D children while was associated with LaOT1D (corrected p-value=1.4x10-2). In corroboration, HLA class II haplotype analysis showed that the rare DRB1*04:08-DQ8 haplotype is specifically associated with EOT1D (corrected p-value=1.4x10-5) and represents the major HLA class II genetic driver and discriminative factor in the development of early onset disease.DiscussionThis study uncovered that EOT1D holds a distinctive spectrum of HLA class II susceptibility loci, which includes risk factors overlapping with LaOT1D and discriminative genetic configurations. These findings warrant replication studies in larger multicentric settings encompassing other ethnicities and may impact target screening strategies and follow-up of young children with high T1D genetic risk as well as personalized therapeutic approaches

Do GnRH Agonists Really Increase Body Weight Gain? Evaluation of a Multicentric Portuguese Cohort of Patients With Central Precocious Puberty

Funding Information: We would like to thank all the colleagues who collected data for the National Database: Joana Serra, Unidade de Endocrinologia Pediátrica, Hospital Pediátrico de Coimbra, Portugal; Marcelo Fonseca, Unidade de Endocrinologia Pediátrica, Hospital Pedro Hispano, Matosinhos, Portugal; Maria João Oliveira, Unidade de Endocrinologia Pediátrica, Centro Materno Infantil do Norte, Porto, Portugal; Maria Miguel Gomes, Unidade de Endocrinologia Pediátrica, Hospital de Braga, Portugal; Paula Vieira, Serviço de Pediatria, Hospital São Francisco Xavier, Lisbon, Portugal; Rita Silva, Unidade de Endocrinologia Pediátrica, Hospital de São João, Porto, Portugal; Sónia Gomes, Unidade de Endocrinologia Pediátrica, Centro Hospitalar Universitário de Lisboa Central, Lisbon, Portugal; Susana Figueiredo, Serviço de Pediatria, Hospital de Viana de Castelo, Portugal; and Susana Pacheco, Unidade de Endocrinologia Pediátrica, Hospital Fernando da Fonseca, Amadora, Portugal. We also thank all children, adolescents, and their parents for their cooperation and understanding. This investigator-initiated study was supported by an unconditional research grant from Ipsen. Funding Information: We would like to thank all the colleagues who collected data for the National Database: Joana Serra, Unidade de Endocrinologia Pedi?trica, Hospital Pedi?trico de Coimbra, Portugal; Marcelo Fonseca, Unidade de Endocrinologia Pedi?trica, Hospital Pedro Hispano, Matosinhos, Portugal; Maria Jo?o Oliveira, Unidade de Endocrinologia Pedi?trica, Centro Materno Infantil do Norte, Porto, Portugal; Maria Miguel Gomes, Unidade de Endocrinologia Pedi?trica, Hospital de Braga, Portugal; Paula Vieira, Servi?o de Pediatria, Hospital S?o Francisco Xavier, Lisbon, Portugal; Rita Silva, Unidade de Endocrinologia Pedi?trica, Hospital de S?o Jo?o, Porto, Portugal; S?nia Gomes, Unidade de Endocrinologia Pedi?trica, Centro Hospitalar Universit?rio de Lisboa Central, Lisbon, Portugal; Susana Figueiredo, Servi?o de Pediatria, Hospital de Viana de Castelo, Portugal; and Susana Pacheco, Unidade de Endocrinologia Pedi?trica, Hospital Fernando da Fonseca, Amadora, Portugal. We also thank all children, adolescents, and their parents for their cooperation and understanding. This investigator-initiated study was supported by an unconditional research grant from Ipsen. Publisher Copyright: Copyright © 2022 Leite, Galo, Antunes, Robalo, Amaral, Espada, Castro, Simões Dias and Limbert.Introduction: There are several concerns associated with gonadotropin-releasing hormone agonist (GnRHa) treatment for central precocious puberty (CPP), such as obesity and changes in body mass index (BMI). We aimed to investigate whether any anthropometric differences exist and if they persist over time. Methods: We conducted an observational study of Portuguese children (both sexes) diagnosed with CPP between January 2000 and December 2017, using a digital platform, in order to analyze the influence of GnRHa treatment on BMI-SD score (BMI-SDS). Results: Of the 241 patients diagnosed with CPP, we assessed 92 patients (8% boys) in this study. At baseline, 39% of the patients were overweight. BMI-SDS increased with treatment for girls but then diminished 1 year after stopping GnRHa therapy (p = 0.018). BMI-SDS variation at the end of treatment was negatively correlated with BMI-SDS at baseline (p < 0.001). Boys grew taller and faster during treatment than did girls (p < 0.001), and therefore, their BMI-SDS trajectory might be different. Conclusions: This study showed an increase of body weight gain during GnRHa treatment only in girls, which reversed just 1 year after stopping treatment. The overall gain in BMI-SDS with treatment is associated with baseline BMI-SDS.publishersversionpublishe

Spectrum of molecular alterations detected in the CYP21A2 gene associated with 21-hydroxylase deficiency

A maioria dos doentes com hiperplasia suprarrenal congénita (HSC) apresenta alterações moleculares no gene CYP21A2, o qual codifica a enzima 21-hidroxilase (21-OH). Os doentes com a forma clássica de deficiência em 21-OH (21-OHD) apresentam a síntese de cortisol diminuída no córtex adrenal e, os casos mais graves, também apresentam deficiência de aldosterona. As mulheres com 21-OHD grave apresentam excesso de andrógenos desde a sua vida fetal conduzindo à virilização dos órgãos genitais externos. Tanto homens como mulheres com 21-OHD completa não sintetizam a aldosterona e, consequentemente, logo após o nascimento, podem desenvolver crises de perda de sal se não forem corretamente diagnosticados e tratados. A 21- OHD não clássica é devida à deficiência parcial em 21-OH, os fenótipos clínicos são menos graves, as mulheres não apresentam virilização dos genitais externos ao nascimento, e geralmente os sinais relativos a excesso de androgénios podem surgir durante a infância ou até mais tarde (durante ou após a puberdade). Neste trabalho descrevem-se as alterações e os genótipos mais frequentes encontrados em doentes portugueses não adultos com 21-OHD. As alterações mais frequentes encontradas na forma clássica da HSC são c.293-13C> G, diferentes deleções/quimeras/conversões génicas do gene CYP21A2 e c.518T> A, enquanto na 21-OHD não-clássica a variante c.844G> T é a mais frequente. Estes resultados contribuem para um diagnóstico correto e uma melhor gestão clínica dos doentes, para o seu aconselhamento genético e para oferecer o diagnóstico pré-natal a casais com risco de ter filhos afetados com a forma clássica de 21-OHD.Most of the patients with congenital adrenal hyperplasia (CAH) have molecular alterations in the CYP21A2 gene, which encodes the enzyme 21-hydroxylase (21-OH). Patients with the classic form of 21-OH deficiency (21-OHD) have the synthesis of cor tisol impaired in the adrenal cor tex and, the most severe cases also have aldosterone deficiency. Females with severe 21-OHD, star ting their fetal life have excess of androgens leading to external genitalia virilization at bir th. Both males and females with complete 21-OHD are not able to synthesize aldosterone, consequently soon af ter bir th may develop salt wasting crises if not correctly diagnosed and treated. Non-classic 21-OHD is due to par tial deficiency of 21-OH, the clinical phenotypes are less severe, females don’t present ambiguity of the external genitalia at bir th, usually signs of androgen excess may be present during childhood or even later in life (during or af ter puber ty). We present here the most frequent alterations and genotypes found in non adult Por tuguese patients with 21-OHD. The most frequent alterations found in the classic form of CAH are c.293-13C>G, dif ferent CYP21A2 deletions/quimeras/gene conversions and c.518T>A, while in non-classic 21-OHD the variant c.844G>T is the most frequent. These results contribute to a correct patient diagnosis, to a better clinical care, genetic counseling and to of fer pre-natal diagnosis to couples at risk of having af fected babies with the classic form of 21-OHD.info:eu-repo/semantics/publishedVersio

Carcinoma da tiroide: incidental e não incidental

ResumoIntroduçãoO aumento da incidência do carcinoma da tiroide pode ser consequência de um aumento da deteção de doença subclínica. O objetivo deste estudo foi comparar as características clínico‐patológicas do carcinoma da tiroide diagnosticado incidentalmente, na análise histológica, com o carcinoma com citologia prévia sugestiva.MétodosAnálise retrospetiva dos processos clínicos de 102 doentes com carcinoma da tiroide. O grupo I, constituído por 69 doentes diagnosticados de forma não incidental, foi comparado com o grupo II, constituído por 33 doentes diagnosticados de forma incidental.ResultadosA idade média foi 51,5 e 56,6 anos nos grupos I e II, respetivamente (p=0,10). Houve maior prevalência do sexo feminino, sendo do sexo masculino 18,8% doentes do grupo I e 12,1% do grupo II (p=0,39). A tiroidite de Hashimoto foi diagnosticada em 21,7 e 15,2% de doentes nos grupos I e II, respetivamente (p=0,53). O carcinoma papilar foi o mais frequente (87,0% no grupo I e 97,0% no grupo II). Na classificação TNM, ambos os grupos apresentaram percentagens semelhantes no estádio III (I 21,7% e II 21,2%). Não existiu diferença estatisticamente significativa na dimensão média do tumor (p=0,05), focalidade (p=0,72), invasão capsular (p=0,07) ou linfovascular (p=0,33). O grupo II não apresentou envolvimento ganglionar em comparação ao grupo I em que ocorreu em 11,6% (p=0,05). O tratamento com iodo radioativo foi mais frequente no grupo I (68,1 vs. 45,5%) p=0,03.ConclusõesNão se verificou diferença estatisticamente significativa, entre os grupos, relativamente à idade, sexo, presença de tiroidite, dimensão do tumor, focalidade, invasão capsular ou linfovascular. Em ambos os grupos registou‐se uma percentagem semelhante de doentes no estádio III. Contudo, os carcinomas incidentais parecem ser biologicamente menos agressivos que os não incidentais.AbstractIntroductionIncreasing incidence of thyroid cancer might be a consequence of an increase detection of subclinical disease. The aim of this study was to compare the clinical and pathologic characteristics of nonincidentally discovered (NID) thyroid cancer with incidentally discovered (ID) on postoperative pathology.MethodsA retrospective medical record review of 102 patients with thyroid cancer was performed. A group I of 69 patients with NID thyroid cancer was compared with a group II of 33 patients with ID thyroid cancer.ResultsAt diagnosis the mean age was 51.5 years for the group I and 56.6 years for the group II (p=0.10). The rate of male was 18.8% in the group I and 12.1% in the group II (p=0.39). Hashimoto's thyroiditis was present in 21.7% and 15.2% of patients in groups I and II, respectively (p=0.53). Papillary tumor was the most frequent in both groups (87.0% in group I and 97.0% in group II). At TNM, stage III was present at a similar rate in both groups (I 21.7% and II 21.2%). There was no statistical difference in the mean size of tumor (p=0.05), focality (p=0.72), capsular involvement (p=.07) and lymphovascular invasion (p=0.33). There weren’t lymph nodes metastasis in group II compared with a rate of 11.6% in group I (p=0.05). Radioactive iodine treatment was most frequent in group I (68.1% vs 45.5%) p=0.03.ConclusionsThere weren’t significant differences in the age, sex, presence of thyroiditis, size of tumor, focality, capsular or lymphovascular invasion between the groups. There was a similar rate of stage III in both groups, nevertheless the ID thyroid cancer seems to be biologically less aggressive than NID

Association of diabetic ketoacidosis and HbA1c at onset with year-three HbA1c in children and adolescents with type 1 diabetes: Data from the International SWEET Registry

Piccini, Barbara/0000-0002-2684-8638; Kim, Jae Hyun/0000-0002-0203-7443; Jefferies, Craig/0000-0002-0541-6094; Schwandt, Anke/0000-0003-3041-0759WOS: 000503343500001PubMed: 31797499Objective To establish whether diabetic ketoacidosis (DKA) or HbA1c at onset is associated with year-three HbA1c in children with type 1 diabetes (T1D). Methods Children with T1D from the SWEET registry, diagnosed = 12%]). To adjust for demographics, linear regression was applied with interaction terms for DKA and HbA1c at onset groups (adjusted means with 95% CI). Association between year-three HbA1c and both HbA1c and presentation at onset was analyzed (Vuong test). Results Among 1420 children (54% males; median age at onset 9.1 years [Q1;Q3: 5.8;12.2]), 6% of children experienced DKA with coma, 37% DKA without coma, and 57% no DKA. Year-three HbA1c was lower in the low compared to high HbA1c at onset group, both in the DKA without coma (7.1% [6.8;7.4] vs 7.6% [7.5;7.8], P = .03) and in the no DKA group (7.4% [7.2;7.5] vs 7.8% [7.6;7.9], P = .01), without differences between low and medium HbA1c at onset groups. Year-three HbA1c did not differ among HbA1c at onset groups in the DKA with coma group. HbA1c at onset as an explanatory variable was more closely associated with year-three HbA1c compared to presentation at onset groups (P = .02). Conclusions Year-three HbA1c is more closely related to HbA1c than to DKA at onset; earlier hyperglycemia detection might be crucial to improving year-three HbA1c.Insulet; Sanofi; Medtronic EuropeMedtronic; Lilly Diabetes Excellence Centre; DexCom Inc.; AbbottAbbott Laboratories; Boehringer-IngelheimBoehringer IngelheimInsulet; Sanofi; Medtronic Europe; Lilly Diabetes Excellence Centre; DexCom Inc.; Boehringer-Ingelheim; Abbot

Telemonitoring, Telemedicine and Time in Range During the Pandemic:Paradigm Change for Diabetes Risk Management in the Post-COVID Future

People with diabetes are at greater risk for negative outcomes from COVID-19. Though this risk is multifactorial, poor glycaemic control before and during admission to hospital for COVID-19 is likely to contribute to the increased risk. The COVID-19 pandemic and restrictions on mobility and interaction can also be expected to impact on daily glucose management of people with diabetes. Telemonitoring of glucose metrics has been widely used during the pandemic in people with diabetes, including adults and children with T1D, allowing an exploration of the impact of COVID-19 inside and outside the hospital setting on glycaemic control. To date, 27 studies including 69,294 individuals with T1D have reported the effect of glycaemic control during the COVID-19 pandemic. Despite restricted access to diabetes clinics, glycaemic control has not deteriorated for 25/27 cohorts and improved in 23/27 study groups. Significantly, time in range (TIR) 70–180 mg/dL (3.9–10 mmol/L) increased across 19/27 cohorts with a median 3.3% (− 6.0% to 11.2%) change. Thirty per cent of the cohorts with TIR data reported an average clinically significant TIR improvement of 5% or more, possibly as a consequence of more accurate glucose monitoring and improved connectivity through telemedicine. Periodic consultations using telemedicine enables care of people with diabetes while limiting the need for in-person attendance at diabetes clinics. Reports that sustained hyperglycaemia and early-stage diabetic ketoacidosis may go untreated because of the lockdown and concerns about potential exposure to the risk of infection argue for wider access to glucose telemonitoring. Therefore, in this paper we have critically reviewed reports concerning use of telemonitoring in the acute hospitalized setting as well as during daily diabetes management. Furthermore, we discuss the indications and implications of adopting telemonitoring and telemedicine in the present challenging time, as well as their potential for the future