YIPF5 mutations cause neonatal diabetes and microcephaly through endoplasmic reticulum stress
Elisa De Franco, Maria Lytrivi, Hazem Ibrahim, Hossam Montaser, Matthew N. Wakeling, Federica Fantuzzi, Kashyap Patel, Céline Demarez, Ying Cai, Mariana Igoillo-Esteve, Cristina Cosentino, Väinö Lithovius, Helena Vihinen, Eija Jokitalo, Thomas W. Laver, Matthew B. Johnson, Toshiaki Sawatani, Hadis Shakeri, Nathalie Pachera, Belma Haliloglu, Mehmet Nuri Ozbek, Edip Unal, Ruken Yıldırım, Tushar Godbole, Melek Yildiz, Banu Aydin, Angeline Bilheu, Ikuo Suzuki, Sarah E. Flanagan, Pierre Vanderhaeghen, Valérie Senée, Cécile Julier, Piero Marchetti, Decio L. Eizirik, Sian Ellard, Jonna Saarimäki-Vire, Timo Otonkoski, Miriam Cnop, Andrew T. Hattersley
Elisa De Franco, Maria Lytrivi, Hazem Ibrahim, Hossam Montaser, Matthew N. Wakeling, Federica Fantuzzi, Kashyap Patel, Céline Demarez, Ying Cai, Mariana Igoillo-Esteve, Cristina Cosentino, Väinö Lithovius, Helena Vihinen, Eija Jokitalo, Thomas W. Laver, Matthew B. Johnson, Toshiaki Sawatani, Hadis Shakeri, Nathalie Pachera, Belma Haliloglu, Mehmet Nuri Ozbek, Edip Unal, Ruken Yıldırım, Tushar Godbole, Melek Yildiz, Banu Aydin, Angeline Bilheu, Ikuo Suzuki, Sarah E. Flanagan, Pierre Vanderhaeghen, Valérie Senée, Cécile Julier, Piero Marchetti, Decio L. Eizirik, Sian Ellard, Jonna Saarimäki-Vire, Timo Otonkoski, Miriam Cnop, Andrew T. Hattersley
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Research Article Cell biology Genetics

YIPF5 mutations cause neonatal diabetes and microcephaly through endoplasmic reticulum stress

Abstract

Neonatal diabetes is caused by single gene mutations reducing pancreatic β cell number or impairing β cell function. Understanding the genetic basis of rare diabetes subtypes highlights fundamental biological processes in β cells. We identified 6 patients from 5 families with homozygous mutations in the YIPF5 gene, which is involved in trafficking between the endoplasmic reticulum (ER) and the Golgi. All patients had neonatal/early-onset diabetes, severe microcephaly, and epilepsy. YIPF5 is expressed during human brain development, in adult brain and pancreatic islets. We used 3 human β cell models (YIPF5 silencing in EndoC-βH1 cells, YIPF5 knockout and mutation knockin in embryonic stem cells, and patient-derived induced pluripotent stem cells) to investigate the mechanism through which YIPF5 loss of function affects β cells. Loss of YIPF5 function in stem cell–derived islet cells resulted in proinsulin retention in the ER, marked ER stress, and β cell failure. Partial YIPF5 silencing in EndoC-βH1 cells and a patient mutation in stem cells increased the β cell sensitivity to ER stress–induced apoptosis. We report recessive YIPF5 mutations as the genetic cause of a congenital syndrome of microcephaly, epilepsy, and neonatal/early-onset diabetes, highlighting a critical role of YIPF5 in β cells and neurons. We believe this is the first report of mutations disrupting the ER-to-Golgi trafficking, resulting in diabetes.

Authors

Elisa De Franco, Maria Lytrivi, Hazem Ibrahim, Hossam Montaser, Matthew N. Wakeling, Federica Fantuzzi, Kashyap Patel, Céline Demarez, Ying Cai, Mariana Igoillo-Esteve, Cristina Cosentino, Väinö Lithovius, Helena Vihinen, Eija Jokitalo, Thomas W. Laver, Matthew B. Johnson, Toshiaki Sawatani, Hadis Shakeri, Nathalie Pachera, Belma Haliloglu, Mehmet Nuri Ozbek, Edip Unal, Ruken Yıldırım, Tushar Godbole, Melek Yildiz, Banu Aydin, Angeline Bilheu, Ikuo Suzuki, Sarah E. Flanagan, Pierre Vanderhaeghen, Valérie Senée, Cécile Julier, Piero Marchetti, Decio L. Eizirik, Sian Ellard, Jonna Saarimäki-Vire, Timo Otonkoski, Miriam Cnop, Andrew T. Hattersley

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Figure 6

iPSCs from patients IIIa and IIIb differentiated into β cells are sensitive to ER stress–induced apoptosis.

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iPSCs from patients IIIa and IIIb differentiated into β cells are sensit...
(A) Representative immunostaining of dispersed stage 7 aggregates stained for insulin (INS, green) and glucagon (GCG, red). Nuclei were visualized with DAPI (blue). (B) Quantification of immunostained cells (expressed as percent of total cells) in dispersed stage 7 control (n = 25) and patient cells (n = 11). Blue squares represent patient cells (2 patients, 2 iPSC lines for each); black circles and squares represent healthy control (1 iPSC line) and corrected patient cells (2 iPSC lines from 1 patient), respectively. (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). (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. mRNA expression was normalized to the geometric mean of reference genes β-actin and GAPDH. The median is shown by a horizontal line in the box plots; 25th and 75th percentiles are at the bottom and top of the boxes; whiskers represent minimum and maximum values, and data points independent experiments. Comparisons were done by multiple t test followed by Bonferroni’s correction for multiple comparisons (B), ANOVA followed by Bonferroni’s correction for multiple comparisons (C and D), and paired-ratio t test (E). *P < 0.05, **P < 0.01, ***P < 0.001 treatment vs. DMSO; #P < 0.05, ##P < 0.01 vs. control and corrected cells as indicated.