Exenatide induces frataxin expression and improves mitochondrial function in Friedreich ataxia. (Ile98Ser)-β cells under basal condition (Figure 6E). Reiling JH, et al. Amyotrophic lateral sclerosis (ALS): also known as Lou Gehrig's disease in Canada and the United States, as motor neurone disease (MND) in Australia, Ireland, New Zealand, South Africa, and the United Kingdom, and Charcot disease in francophone countries; is a neurodegenerative neuromuscular disease that results in the progressive loss of motor neurons that control voluntary muscles. |, Find articles by | PUMA expression was also induced by YIPF5 silencing (Supplemental Figure 2G), while BIM expression was not altered. PubMed Nakagawa H, Hazama K, Ishida K, Komori M, Nishimura K, Matsuo S. Inhibition of PLD1 activity causes ER stress via regulation of COPII vesicle formation. PubMed | |, Find articles by The insulin-positive KO cells sustained high BiP expression in vivo, consistent with persistent ER stress. Electronic publication, Version JCI The mature β cells in the KO and YIPF5Ile98Ser grafts showed a 3.3- and 3-fold reduction, respectively, in the percentage of cytoplasmic insulin-positive area, and a 5.7- and 5.4-fold increase, respectively, in the proinsulin area (Figure 5, D and G). (E) mRNA expression of CHOP, BiP, sXBP1, DP5, and PUMA assessed by qPCR in stage 7 aggregates from control and corrected (n = 4–8, black) and patient cells (n = 5–7, blue) exposed for 48 hours to vehicle (DMSO), thapsigargin, or tunicamycin. absence - nezvestnosť - absencia - roztržitosť - nedostatok - neprítomnosť - neúčasť - nepozornosť - zamyslenosť - chýbanie niečoho - náhla a krátka strata pamäti - nedostávanie sa niečoho . (D) Percentage of INS+BiPhi cells per total number of INS+ cells (n = 4–8). Time course experiments in EndoC-βH1 cells exposed to thapsigargin showed that YIPF5 knockdown induced ER stress markers (Figure 3G and Supplemental Figure 2, C–F). PubMed Taniguchi M, et al. YC reprogrammed patient PBMCs into iPSCs. | iPSCs from patients IIIa and IIIb differentiated into β cells are sensitive to ER stress–induced apoptosis. Transient transfection was performed using 30 nM siRNA and Lipofectamine RNAiMAX lipid reagent (Invitrogen/Life Technologies) as previously described (54). SavvyVcfHomozygosity was used to identify large (>3 Mb) homozygous regions in the genome sequencing data (https://github.com/rdemolgen/SavvySuite). Statistical significance was assessed by 2-way ANOVA test with Bonferroni correction in A, by multiple t test with Bonferroni correction in B, and by 1-way ANOVA test with Bonferroni correction in C–E. Kranjc T, Dempsey E, Cagney G, Nakamura N, Shields DC, Simpson JC. **P < 0.01, ***P < 0.001, ****P < 0.0001. Aggregates were exposed to 1.6 mM glucose, 16.7 mM glucose, or 16.7 mM glucose plus 10 μM forskolin for 1 hour. Mutations in 5 genes that dysregulate signaling in the PERK branch of the ER stress response cause β cell dysfunction and apoptosis by perturbing translational control. iPSCs from patients IIIa and IIIb differentiated into β cells are sensitive to ER stress–induced apoptosis. mRNA expression was normalized to the geometric mean of reference genes β-actin and GAPDH. All compounds were from Sigma-Aldrich. | (E) Percentage of INS+BiPhi cells per total number of INS+ cells (n = 3–6). in: All these mutations are predicted to be deleterious; however, it is possible that function of the YIPF5 protein is not completely lost. VL planned and conducted morphometric analyses. Scale bars: 25 μm. The differentiation into β cells was done as previously described (62, 63). | The disease severity was variable between families, with 5 affected individuals in family II dying in early infancy (the homozygous YIPF5 mutation could be confirmed only in the proband, as DNA was not available from siblings), while patient I is reported to have normal neuromotor development at the age of 5 years and his epilepsy resolved at the age of 2 years. Medium was changed every second day for 1 week. 20], accessed May 18, 2020), and both affect residues that are highly conserved across species (up to Saccharomyces cerevisiae). La nouvelle plateforme HorseID vous donne la possibilité d’encoder des mutations directement dans la base de données. Human β cell and islet culture and treatment. cause … All authors reviewed and improved the manuscript. ATH, BH, MNO, EU, RY, TG, MY, KP, and BA analyzed the clinical data. PubMed PubMed A membrane protein enriched in endoplasmic reticulum exit sites interacts with COPII. PubMed Schwarz JM, Cooper DN, Schuelke M, Seelow D. MutationTaster2: mutation prediction for the deep-sequencing age. 2007-11-13 22:16:19 - Dans la maladie et de la santé J'ai récemment conduit par une entreprise qui avait un panneau situé à l'avant, qui suit: « Fermé pour cause de maladie. JCI | Consistent with this, ER morphology was severely affected in YIPF5-KO hESC-β cells, but we did not observe the whorl-like structures described in YIPF5-deficient HeLa cells (34), suggesting that phenotypic specificity may be due to YIPF5 cell-specific functions. JCI | Nous partageons également des informations sur l'utilisation de notre site avec nos partenaires de médias sociaux, de publicité et d'analyse, qui peuvent combiner celles-ci avec d'autres informations que vous leur avez fournies ou qu'ils ont collectées lors de votre utilisation de leurs services. Cells were then exposed to Krebs containing 0 mM or 20 mM glucose or 20 mM glucose plus 10 μM forskolin for 40 minutes. Aggregates were lysed by sonication and acid-ethanol for determination of insulin and DNA contents. In EndoC-βH1 cells, YIPF5 expression was strongly upregulated by brefeldin A, which blocks ER-to-Golgi transport, suggesting that transcriptional induction of YIPF5 could be part of a compensatory mechanism to overcome the inhibition of trafficking. in: Loss of β cell function after in vivo implantation. | | Senée, V. This pathway plays a vital role in ER homeostasis and β cell survival, with inhibition of Sar1, which initiates COPII assembly, causing alterations in ER morphology and severe ER stress in β cells (46). CCTOP: a Consensus Constrained TOPology prediction web server. However, ESCp. In order to examine whether the induction of CHOP, a proapoptotic transcription factor in the PERK branch of the ER stress response, sensitizes YIPF5-deficient β cells to apoptosis, we double-knocked-down CHOP and YIPF5 (Figure 3I and Supplemental Figure 2H). The YIPF5 expression pattern we observed in human embryonic brain, showing high expression in progenitor and neuronal compartments of the developing cortex in addition to the choroid plexus within the cerebral ventricles, is consistent with YIPF5 playing a role in neural progenitors and/or neurons during development of the cerebral cortex, which is mostly affected by primary microcephaly. Email or Phone: Password: Forgot account? All had diabetes, with the age at diagnosis ranging from the neonatal period to early infancy. In addition to the KO, we generated an isogenic YIPF5Ile98Ser mutation using CRISPR/Cpf1–mediated homology-directed repair (HDR) (Supplemental Figure 5). | Les cookies nous permettent de personnaliser le contenu et les annonces, d'offrir des fonctionnalités relatives aux médias sociaux et d'analyser notre trafic. YIPF5 knockdown did not significantly affect basal β cell survival, but YIPF5-depleted β cells were markedly sensitized to thapsigargin (Figure 3D). Thapsigargin is presented by solid lines and nontreated cells by dashed lines. in: |, Find articles by PubMed Google Scholar Human β cell and islet culture and treatment. Taken together, our results support YIPF5’s role in ER-to-Golgi transport and point toward YIPF5 loss sensitizing β cells to ER stress–induced apoptosis. Human islets from nondiabetic organ donors (n = 4, 2 female and 2 male donors; age 62 ± 9 years; BMI 27 ± 3 kg/m2; cause of death: 3 cerebral hemorrhage, 1 cardiovascular disease) were isolated by collagenase digestion and density gradient purification, and cultured as previously described (53). Yoshida Y, et al. 1Institute of Biomedical and Clinical Science, University of Exeter Medical School, Exeter, United Kingdom. In secretion experiments, data points represent the average of biological duplicates. (A) Partial pedigrees and summary of clinical features of the 6 patients with homozygous YIPF5 mutations. These data strongly suggest that while β cells are highly dependent on YIPF5 function, α cells appear to be able to survive without YIPF5. We believe that utilizing registries and biobanks to reveal other monogenic atypical forms of diabetes is an important approach to gaining insight and suggest that an insulin sensitizer may alleviate ER stress associated with YIPF5 disruption by decreasing the demand for insulin secretion. Pathogenic variants in 11 genes (ABCC8, KCNJ11, CNOT1, EIF2AK3, SLC19A2, IER3IP1, PTF1A, NEUROD1, MNX1, WFS1, and NKX2-2) are known to cause neonatal diabetes with neurological features, ranging from developmental delay to structural abnormalities such as microcephaly (3, 4). JCI The nature and position of the variants are consistent with at least a partial loss of protein function. (D) Percentage of cytoplasmic area covered by proinsulin or insulin in insulin-positive cells (n = 3–4). Kalender Scale bars: 25 μm. | Eizirik, D. | Written informed consent was given by the parents. (Ile98Ser)-β cells, supporting the previously suggested notion that whorl formation is uncoupled from other YIPF5 functions (35) and our hypothesis that the mutations identified in our patients do not completely abolish YIPF5 function. The cerebrospinal fluid provides a proliferative niche for neural progenitor cells. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.00001. | Log In. Grafts of YIPF5Ile98Ser-mutant aggregates also showed clear signs of increased ER stress when harvested at 3 months after implantation (Figure 5, E and H). In addition, the human YIPF5 gene appears to be intolerant to loss-of-function variants based on gnomAD gene constraints (pLI = 0.99), further supporting that loss-of-function variants are likely strongly deleterious in humans. Neither variant was listed in the gnomAD database (>120,000 individuals [ref. The assignment of the authorship order for the first 6 authors is mainly chronological and reflects the equal contributions from the 3 research groups: EDF identified the causative gene in patients with neonatal diabetes and microcephaly, ML performed the first functional studies suggesting that ER stress may be the causative mechanism, HI and HM generated the genetically modified models and performed their functional studies, HI also generated the mutation correction in patient-derived iPSCs, MNW developed the genome sequencing and homozygosity mapping pipelines that allowed identification of the causative gene, and FF performed studies in patient-derived iPSCs. Patel, K. | PubMed (C and D) Apoptosis was assessed by staining with DNA-binding dyes in vehicle- (DMSO-)treated, thapsigargin-treated, and tunicamycin-treated control and corrected (n = 10) and patient (n = 6–7) stage 7 aggregates (C) or by luminescence produced by annexin V binding in time course experiments (means ± SEM; n = 10 control and corrected lines and n = 5 patient lines) (D). (35) identified 5 YIPF5 residues (3 cytoplasmic and 2 in the transmembrane domain) that, when mutated, resulted in ER whorl formation in HeLa cells. Proinsulin accumulation, increased ER stress signaling, and reduced insulin content in YIPF5-knockout stem cell–derived β cells. 12Kanuni Sultan Suleyman Training and Research Hospital, Department of Pediatric Endocrinology, Istanbul, Turkey. JCI Special thanks of gratitude to Jarkko Ustinov and Solja Eurola (Biomedicum Stem Cell Center, University of Helsinki) for excellent technical and experimental support. Google Scholar, Find articles by Diagramme à utiliser pour dessiner les châtaignes (si moins de 3 épis ou autres marques) B. Formulaire de mutation. iPSC culturing and differentiation into β cells. 7Yeditepe University Hospital, Istanbul, Turkey. Written informed consent for inclusion of patient photographs in scientific publications was collected by the referring clinicians from one parent of each patient. Parents were first cousins (I-1, I-2) and had four children sharing three different pathologies (Fig. Hormone content was normalized for total protein content. Having two C677T variants and elevated homocysteine levels may cause a slightly higher risk for blood clots. We thank the families for participating in the study. YIPF5 is expressed in human pancreatic tissue and brain. In the WT and YIPF5Ile98Ser grafts, 55% and 42% of the cells were insulin-positive, respectively.