Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells
<p><strong>Poster by Abdoulaye Diane, Razik Bin Abdul Mu-u-min, Asma Allouch, and Heba H. Al-Siddiqi (Hamad Bin Khalifa University)</strong></p> <p>Background: Impaired insulin secretion contributes to the pathogenesis of diabetes mellitus type 1 (T1DM) through autoimmu...
محفوظ في:
| المؤلف الرئيسي: | |
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| مؤلفون آخرون: | , , |
| منشور في: |
2023
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| الموضوعات: | |
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| _version_ | 1864513564697427968 |
|---|---|
| author | Abdoulaye Diane (14152749) |
| author2 | Razik Bin Abdul Mu-u-min (15430008) Asma Allouch (15430010) Heba H Al-Siddiqi (12204889) |
| author2_role | author author author |
| author_facet | Abdoulaye Diane (14152749) Razik Bin Abdul Mu-u-min (15430008) Asma Allouch (15430010) Heba H Al-Siddiqi (12204889) |
| author_role | author |
| dc.creator.none.fl_str_mv | Abdoulaye Diane (14152749) Razik Bin Abdul Mu-u-min (15430008) Asma Allouch (15430010) Heba H Al-Siddiqi (12204889) |
| dc.date.none.fl_str_mv | 2023-05-17T11:39:29Z |
| dc.identifier.none.fl_str_mv | 10.57945/manara.22785233.v1 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/poster/Exendin-4_promotes_GSIS_without_increasing_-cell_number_through_upregulation_of_glucose-sensing_apparatus_and_mitochondrial_oxidative_phosphorylation_machinery_genes_in_hPSC-derived_-cells/22785233 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biomedical and clinical sciences Medical biotechnology Diabetes Exendin-4 Mitochondria hPSC-derived β-cells Differentiation |
| dc.title.none.fl_str_mv | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| dc.type.none.fl_str_mv | Image Poster info:eu-repo/semantics/publishedVersion image |
| description | <p><strong>Poster by Abdoulaye Diane, Razik Bin Abdul Mu-u-min, Asma Allouch, and Heba H. Al-Siddiqi (Hamad Bin Khalifa University)</strong></p> <p>Background: Impaired insulin secretion contributes to the pathogenesis of diabetes mellitus type 1 (T1DM) through autoimmune destruction of pancreatic β-cells and the pathogenesis of diabetes mellitus type 2 (T2DM) through β-cell dedifferentiation and other mechanisms. Emerging β-cell replacement with human pluripotent stem cell (hPSC)–derived β-cells may provide remedial cell therapy. Most in vitro differentiation protocols have generated hPSC-derived β-cells with either immature phenotype such as impaired or weakened glucose-stimulated insulin secretion (GSIS) relative to primary β-cells. Evidence has shown that β-cell mitochondria play a central role in coupling glucose metabolism to insulin exocytosis. Therefore, the impairment of GSIS in hPSC-derived β-cells may be attributed to impaired glucose sensing and/or mitochondrial dysfunction. </p> <p>Objective: The aim of this study is to investigate the effect of Exendin-4 (GLP-1 receptor analog), known to promote mitochondrial function, on enhancing maturation and functionality of hPSC-derived β-cells.</p> <p>Methods: Differentiation of hPSC into β-cells was carried out in a stepwise 3D differentiation protocol using 30 ml spinner flask adapted from the protocol of Veres et al. (2019). This consists of 6 stages: S1, definitive endoderm; S2, gut tube endoderm; S3, pancreatic progenitors 1; S4, pancreatic progenitors 2; S5, endocrine precursors; and S6, β-like cells. To determine the differentiation efficiency, relevant stage-specific marker expression was assessed at each stage using flow cytometry. Finally, to further test the effect of GLP-1 receptor analog on the functionality of hPSC-derived β-cells, 50 nM exendin-4 was added to the suspension culture during the last three days of differentiation and GSIS was performed. Gene expression in cell clusters was determined by RT-PCR. Data are mean ± SEM (n=6)</p> <p>Results: Flow cytometry data for relevant stage-specific markers showed 96 % OCT4 positive, 89% SOX17 and 83% PDX1 positive cells, indicating good pluripotency, high definite endoderm and pancreatic progenitor induction, respectively. As for β-cell markers, we found 41.4% NKX6.1/insulin double positive cells at final stage 6, indicating a generation of β-cells. Expression profiling during differentiation confirmed the generation of insulin-expressing β-cells. However, GSIS data showed no difference in c-peptide secretion between low (2.8mM) and high (20 mM) glucose but high response to direct cellular depolarization-mediated c-peptide by KCl; suggesting a lack of functional β-cells. Interestingly, addition of Exendin-4 (50 nM) during the last 3 days of the differentiation significantly enhanced GSIS associated with increased expression of glucose-sensing apparatus genes (Glut2, G6P2C, GCK) and genes encoding mitochondrial oxidative phosphorylation machinery.</p> <p>Conclusion: Our data demonstrated for the first time in 3D differentiation of hPSC-derived β-cells that addition of Exendin-4 promotes GSIS through upregulation of both glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes without increasing β-cell number.</p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_27ef46b5b674b7860c50926e98dcf78f |
| identifier_str_mv | 10.57945/manara.22785233.v1 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/22785233 |
| publishDate | 2023 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cellsAbdoulaye Diane (14152749)Razik Bin Abdul Mu-u-min (15430008)Asma Allouch (15430010)Heba H Al-Siddiqi (12204889)Biomedical and clinical sciencesMedical biotechnologyDiabetesExendin-4MitochondriahPSC-derived β-cellsDifferentiation<p><strong>Poster by Abdoulaye Diane, Razik Bin Abdul Mu-u-min, Asma Allouch, and Heba H. Al-Siddiqi (Hamad Bin Khalifa University)</strong></p> <p>Background: Impaired insulin secretion contributes to the pathogenesis of diabetes mellitus type 1 (T1DM) through autoimmune destruction of pancreatic β-cells and the pathogenesis of diabetes mellitus type 2 (T2DM) through β-cell dedifferentiation and other mechanisms. Emerging β-cell replacement with human pluripotent stem cell (hPSC)–derived β-cells may provide remedial cell therapy. Most in vitro differentiation protocols have generated hPSC-derived β-cells with either immature phenotype such as impaired or weakened glucose-stimulated insulin secretion (GSIS) relative to primary β-cells. Evidence has shown that β-cell mitochondria play a central role in coupling glucose metabolism to insulin exocytosis. Therefore, the impairment of GSIS in hPSC-derived β-cells may be attributed to impaired glucose sensing and/or mitochondrial dysfunction. </p> <p>Objective: The aim of this study is to investigate the effect of Exendin-4 (GLP-1 receptor analog), known to promote mitochondrial function, on enhancing maturation and functionality of hPSC-derived β-cells.</p> <p>Methods: Differentiation of hPSC into β-cells was carried out in a stepwise 3D differentiation protocol using 30 ml spinner flask adapted from the protocol of Veres et al. (2019). This consists of 6 stages: S1, definitive endoderm; S2, gut tube endoderm; S3, pancreatic progenitors 1; S4, pancreatic progenitors 2; S5, endocrine precursors; and S6, β-like cells. To determine the differentiation efficiency, relevant stage-specific marker expression was assessed at each stage using flow cytometry. Finally, to further test the effect of GLP-1 receptor analog on the functionality of hPSC-derived β-cells, 50 nM exendin-4 was added to the suspension culture during the last three days of differentiation and GSIS was performed. Gene expression in cell clusters was determined by RT-PCR. Data are mean ± SEM (n=6)</p> <p>Results: Flow cytometry data for relevant stage-specific markers showed 96 % OCT4 positive, 89% SOX17 and 83% PDX1 positive cells, indicating good pluripotency, high definite endoderm and pancreatic progenitor induction, respectively. As for β-cell markers, we found 41.4% NKX6.1/insulin double positive cells at final stage 6, indicating a generation of β-cells. Expression profiling during differentiation confirmed the generation of insulin-expressing β-cells. However, GSIS data showed no difference in c-peptide secretion between low (2.8mM) and high (20 mM) glucose but high response to direct cellular depolarization-mediated c-peptide by KCl; suggesting a lack of functional β-cells. Interestingly, addition of Exendin-4 (50 nM) during the last 3 days of the differentiation significantly enhanced GSIS associated with increased expression of glucose-sensing apparatus genes (Glut2, G6P2C, GCK) and genes encoding mitochondrial oxidative phosphorylation machinery.</p> <p>Conclusion: Our data demonstrated for the first time in 3D differentiation of hPSC-derived β-cells that addition of Exendin-4 promotes GSIS through upregulation of both glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes without increasing β-cell number.</p>2023-05-17T11:39:29ZImagePosterinfo:eu-repo/semantics/publishedVersionimage10.57945/manara.22785233.v1https://figshare.com/articles/poster/Exendin-4_promotes_GSIS_without_increasing_-cell_number_through_upregulation_of_glucose-sensing_apparatus_and_mitochondrial_oxidative_phosphorylation_machinery_genes_in_hPSC-derived_-cells/22785233CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/227852332023-05-17T11:39:29Z |
| spellingShingle | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells Abdoulaye Diane (14152749) Biomedical and clinical sciences Medical biotechnology Diabetes Exendin-4 Mitochondria hPSC-derived β-cells Differentiation |
| status_str | publishedVersion |
| title | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| title_full | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| title_fullStr | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| title_full_unstemmed | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| title_short | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| title_sort | Exendin-4 promotes GSIS without increasing β-cell number through upregulation of glucose-sensing apparatus and mitochondrial oxidative phosphorylation machinery genes in hPSC-derived β-cells |
| topic | Biomedical and clinical sciences Medical biotechnology Diabetes Exendin-4 Mitochondria hPSC-derived β-cells Differentiation |