345 The Role of TCF7L2 in Hepatic Metabolic Zonation
OBJECTIVES/GOALS: Single nucleotide polymorphisms in the transcription factor 7-like 2 (TCF7L2) gene are associated with Type 2 Diabetes (T2D) and nonalcoholic fatty liver disease (NAFLD). The metabolic function of TCF7L2 in the liver remains to be fully elucidated, but we hypothesized that TCF7L2 c...
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2023-04-01
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| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866123003898/type/journal_article |
| _version_ | 1797840420056596480 |
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| author | Iriscilla I. Ayala Skanda K Hebbale Chris E. Shannon Ivan Valdez Marcel Fourcaudot Terry M. Bakewell Madelaine Sholto Thomas Vallim Sami Heikkinen |
| author_facet | Iriscilla I. Ayala Skanda K Hebbale Chris E. Shannon Ivan Valdez Marcel Fourcaudot Terry M. Bakewell Madelaine Sholto Thomas Vallim Sami Heikkinen |
| author_sort | Iriscilla I. Ayala |
| collection | DOAJ |
| description | OBJECTIVES/GOALS: Single nucleotide polymorphisms in the transcription factor 7-like 2 (TCF7L2) gene are associated with Type 2 Diabetes (T2D) and nonalcoholic fatty liver disease (NAFLD). The metabolic function of TCF7L2 in the liver remains to be fully elucidated, but we hypothesized that TCF7L2 contributes to NAFLD through regulation of zonal metabolic pathways. METHODS/STUDY POPULATION: Using single nuclei RNA sequencing, we examined Tcf7l2 expression in periportal (PP) hepatocytes around the portal triad and pericentral (PC) hepatocytes surrounding the central vein of the liver. To visualize TCF7L2 transcriptional activity we used a TCF reporter mice, which expresses an H2B-eGFP fusion protein downstream of the conserved TCF DNA binding site. We disrupted Tcf7l2 transcriptional activity in mouse liver by breeding mice with a floxed Tcf7l2 exon 11, which encodes part of the DNA binding domain (DBD), to albumin-Cre mice (Hep-TCF7L2ΔDBD). Eight-week-old mice were fed a choline-deficient amino acid-defined high fat (CDAHFD) diet for 8 weeks. In liver samples harvested from these mice, we examined disruption to several key zonated metabolic pathways, and quantified the development of fibrosis. RESULTS/ANTICIPATED RESULTS: Single nuclei analysis revealed that Tcf7l2 mRNA was expressed primarily in parenchymal cells of the liver but was ubiquitous across the liver lobule. However, in immunofluorescence analysis of TCF reporter mice, the transcriptional activity of TCF7L2 was highly restricted to PC hepatocytes. Classic PC hepatocyte markers, including glutamine synthetase (Glul), were absent in Hep-TCF7L2ΔDBD mice. Following the CDAHFD, Hep-TCF7L2ΔDBD mice developed more severe fibrosis in histological analysis, and expressed elevated levels of genes involved in fibrogenesis, collagen synthesis and TGFβsignaling. Hep-TCF7L2ΔDBD mice also displayed hepatic cholesterol accumulation following the CDAHFD, which was likely the result of impaired pericentral bile acid synthesis. DISCUSSION/SIGNIFICANCE: Our results suggest that TCF7L2 plays an important role in the regulation of zonated metabolic pathways, which may contribute to the development of fibrosis. Ongoing analyses are exploring the mechanisms regulating the zonal transcriptional activity of TCF7L2. |
| first_indexed | 2024-04-09T16:14:49Z |
| format | Article |
| id | doaj.art-8e32245c74074c31875e6eb756175b22 |
| institution | Directory Open Access Journal |
| issn | 2059-8661 |
| language | English |
| last_indexed | 2024-04-09T16:14:49Z |
| publishDate | 2023-04-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj.art-8e32245c74074c31875e6eb756175b222023-04-24T05:55:57ZengCambridge University PressJournal of Clinical and Translational Science2059-86612023-04-01710310310.1017/cts.2023.389345 The Role of TCF7L2 in Hepatic Metabolic ZonationIriscilla I. Ayala0Skanda K Hebbale1Chris E. Shannon2Ivan Valdez3Marcel Fourcaudot4Terry M. Bakewell5Madelaine Sholto6Thomas Vallim7Sami Heikkinen8University of Texas Health Science Center at San AntonioDiabetes Division, University of Texas Health, San Antonio, TX, USADiabetes Division, University of Texas Health, San Antonio, TX, USADiabetes Division, University of Texas Health, San Antonio, TX, USADiabetes Division, University of Texas Health, San Antonio, TX, USADiabetes Division, University of Texas Health, San Antonio, TX, USADepartment of Medicine, Division of Cardiology, University of California, Los Angeles (UCLA), Los Angeles, CA, USADepartment of Medicine, Division of Cardiology, University of California, Los Angeles (UCLA), Los Angeles, CA, USADepartment of Medicine, University of Eastern Finland, Kuopio, Finland Luke Norton, Diabetes Division, University of Texas Health, San Antonio, TX, USAOBJECTIVES/GOALS: Single nucleotide polymorphisms in the transcription factor 7-like 2 (TCF7L2) gene are associated with Type 2 Diabetes (T2D) and nonalcoholic fatty liver disease (NAFLD). The metabolic function of TCF7L2 in the liver remains to be fully elucidated, but we hypothesized that TCF7L2 contributes to NAFLD through regulation of zonal metabolic pathways. METHODS/STUDY POPULATION: Using single nuclei RNA sequencing, we examined Tcf7l2 expression in periportal (PP) hepatocytes around the portal triad and pericentral (PC) hepatocytes surrounding the central vein of the liver. To visualize TCF7L2 transcriptional activity we used a TCF reporter mice, which expresses an H2B-eGFP fusion protein downstream of the conserved TCF DNA binding site. We disrupted Tcf7l2 transcriptional activity in mouse liver by breeding mice with a floxed Tcf7l2 exon 11, which encodes part of the DNA binding domain (DBD), to albumin-Cre mice (Hep-TCF7L2ΔDBD). Eight-week-old mice were fed a choline-deficient amino acid-defined high fat (CDAHFD) diet for 8 weeks. In liver samples harvested from these mice, we examined disruption to several key zonated metabolic pathways, and quantified the development of fibrosis. RESULTS/ANTICIPATED RESULTS: Single nuclei analysis revealed that Tcf7l2 mRNA was expressed primarily in parenchymal cells of the liver but was ubiquitous across the liver lobule. However, in immunofluorescence analysis of TCF reporter mice, the transcriptional activity of TCF7L2 was highly restricted to PC hepatocytes. Classic PC hepatocyte markers, including glutamine synthetase (Glul), were absent in Hep-TCF7L2ΔDBD mice. Following the CDAHFD, Hep-TCF7L2ΔDBD mice developed more severe fibrosis in histological analysis, and expressed elevated levels of genes involved in fibrogenesis, collagen synthesis and TGFβsignaling. Hep-TCF7L2ΔDBD mice also displayed hepatic cholesterol accumulation following the CDAHFD, which was likely the result of impaired pericentral bile acid synthesis. DISCUSSION/SIGNIFICANCE: Our results suggest that TCF7L2 plays an important role in the regulation of zonated metabolic pathways, which may contribute to the development of fibrosis. Ongoing analyses are exploring the mechanisms regulating the zonal transcriptional activity of TCF7L2.https://www.cambridge.org/core/product/identifier/S2059866123003898/type/journal_article |
| spellingShingle | Iriscilla I. Ayala Skanda K Hebbale Chris E. Shannon Ivan Valdez Marcel Fourcaudot Terry M. Bakewell Madelaine Sholto Thomas Vallim Sami Heikkinen 345 The Role of TCF7L2 in Hepatic Metabolic Zonation Journal of Clinical and Translational Science |
| title | 345 The Role of TCF7L2 in Hepatic Metabolic Zonation |
| title_full | 345 The Role of TCF7L2 in Hepatic Metabolic Zonation |
| title_fullStr | 345 The Role of TCF7L2 in Hepatic Metabolic Zonation |
| title_full_unstemmed | 345 The Role of TCF7L2 in Hepatic Metabolic Zonation |
| title_short | 345 The Role of TCF7L2 in Hepatic Metabolic Zonation |
| title_sort | 345 the role of tcf7l2 in hepatic metabolic zonation |
| url | https://www.cambridge.org/core/product/identifier/S2059866123003898/type/journal_article |
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