Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones.
Prader-Willi syndrome (PWS) is a multisystem disorder with neurobehavioral, metabolic, and hormonal phenotypes, caused by loss of expression of a paternally-expressed imprinted gene cluster. Prior evidence from a PWS mouse model identified abnormal pancreatic islet development with retention of aged...
Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023-04-01
|
| Series: | PLoS Genetics |
| Online Access: | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1010710&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849468095231426560 |
|---|---|
| author | Erik A Koppes Marie A Johnson James J Moresco Patrizia Luppi Dale W Lewis Donna B Stolz Jolene K Diedrich John R Yates Ronald C Wek Simon C Watkins Susanne M Gollin Hyun Jung Park Peter Drain Robert D Nicholls |
| author_facet | Erik A Koppes Marie A Johnson James J Moresco Patrizia Luppi Dale W Lewis Donna B Stolz Jolene K Diedrich John R Yates Ronald C Wek Simon C Watkins Susanne M Gollin Hyun Jung Park Peter Drain Robert D Nicholls |
| author_sort | Erik A Koppes |
| collection | DOAJ |
| description | Prader-Willi syndrome (PWS) is a multisystem disorder with neurobehavioral, metabolic, and hormonal phenotypes, caused by loss of expression of a paternally-expressed imprinted gene cluster. Prior evidence from a PWS mouse model identified abnormal pancreatic islet development with retention of aged insulin and deficient insulin secretion. To determine the collective roles of PWS genes in β-cell biology, we used genome-editing to generate isogenic, clonal INS-1 insulinoma lines having 3.16 Mb deletions of the silent, maternal- (control) and active, paternal-allele (PWS). PWS β-cells demonstrated a significant cell autonomous reduction in basal and glucose-stimulated insulin secretion. Further, proteomic analyses revealed reduced levels of cellular and secreted hormones, including all insulin peptides and amylin, concomitant with reduction of at least ten endoplasmic reticulum (ER) chaperones, including GRP78 and GRP94. Critically, differentially expressed genes identified by whole transcriptome studies included reductions in levels of mRNAs encoding these secreted peptides and the group of ER chaperones. In contrast to the dosage compensation previously seen for ER chaperones in Grp78 or Grp94 gene knockouts or knockdown, compensation is precluded by the stress-independent deficiency of ER chaperones in PWS β-cells. Consistent with reduced ER chaperones levels, PWS INS-1 β-cells are more sensitive to ER stress, leading to earlier activation of all three arms of the unfolded protein response. Combined, the findings suggest that a chronic shortage of ER chaperones in PWS β-cells leads to a deficiency of protein folding and/or delay in ER transit of insulin and other cargo. In summary, our results illuminate the pathophysiological basis of pancreatic β-cell hormone deficits in PWS, with evolutionary implications for the multigenic PWS-domain, and indicate that PWS-imprinted genes coordinate concerted regulation of ER chaperone biosynthesis and β-cell secretory pathway function. |
| format | Article |
| id | doaj-art-511e07a3ded143c49933e62d23762e4c |
| institution | Kabale University |
| issn | 1553-7390 1553-7404 |
| language | English |
| publishDate | 2023-04-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Genetics |
| spelling | doaj-art-511e07a3ded143c49933e62d23762e4c2025-08-20T03:25:57ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042023-04-01194e101071010.1371/journal.pgen.1010710Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones.Erik A KoppesMarie A JohnsonJames J MorescoPatrizia LuppiDale W LewisDonna B StolzJolene K DiedrichJohn R YatesRonald C WekSimon C WatkinsSusanne M GollinHyun Jung ParkPeter DrainRobert D NichollsPrader-Willi syndrome (PWS) is a multisystem disorder with neurobehavioral, metabolic, and hormonal phenotypes, caused by loss of expression of a paternally-expressed imprinted gene cluster. Prior evidence from a PWS mouse model identified abnormal pancreatic islet development with retention of aged insulin and deficient insulin secretion. To determine the collective roles of PWS genes in β-cell biology, we used genome-editing to generate isogenic, clonal INS-1 insulinoma lines having 3.16 Mb deletions of the silent, maternal- (control) and active, paternal-allele (PWS). PWS β-cells demonstrated a significant cell autonomous reduction in basal and glucose-stimulated insulin secretion. Further, proteomic analyses revealed reduced levels of cellular and secreted hormones, including all insulin peptides and amylin, concomitant with reduction of at least ten endoplasmic reticulum (ER) chaperones, including GRP78 and GRP94. Critically, differentially expressed genes identified by whole transcriptome studies included reductions in levels of mRNAs encoding these secreted peptides and the group of ER chaperones. In contrast to the dosage compensation previously seen for ER chaperones in Grp78 or Grp94 gene knockouts or knockdown, compensation is precluded by the stress-independent deficiency of ER chaperones in PWS β-cells. Consistent with reduced ER chaperones levels, PWS INS-1 β-cells are more sensitive to ER stress, leading to earlier activation of all three arms of the unfolded protein response. Combined, the findings suggest that a chronic shortage of ER chaperones in PWS β-cells leads to a deficiency of protein folding and/or delay in ER transit of insulin and other cargo. In summary, our results illuminate the pathophysiological basis of pancreatic β-cell hormone deficits in PWS, with evolutionary implications for the multigenic PWS-domain, and indicate that PWS-imprinted genes coordinate concerted regulation of ER chaperone biosynthesis and β-cell secretory pathway function.https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1010710&type=printable |
| spellingShingle | Erik A Koppes Marie A Johnson James J Moresco Patrizia Luppi Dale W Lewis Donna B Stolz Jolene K Diedrich John R Yates Ronald C Wek Simon C Watkins Susanne M Gollin Hyun Jung Park Peter Drain Robert D Nicholls Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones. PLoS Genetics |
| title | Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones. |
| title_full | Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones. |
| title_fullStr | Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones. |
| title_full_unstemmed | Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones. |
| title_short | Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones. |
| title_sort | insulin secretion deficits in a prader willi syndrome β cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones |
| url | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1010710&type=printable |
| work_keys_str_mv | AT erikakoppes insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT marieajohnson insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT jamesjmoresco insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT patrizialuppi insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT dalewlewis insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT donnabstolz insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT jolenekdiedrich insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT johnryates insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT ronaldcwek insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT simoncwatkins insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT susannemgollin insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT hyunjungpark insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT peterdrain insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones AT robertdnicholls insulinsecretiondeficitsinapraderwillisyndromebcellmodelareassociatedwithaconcerteddownregulationofmultipleendoplasmicreticulumchaperones |