Unbiased transcription factor CRISPR screen identifies ZNF800 as master repressor of enteroendocrine differentiation 公告 2023-10-27 14:53:03 Written By Kym Kilbourne Title: Unbiased transcription factor CRISPR screen identifies ZNF800 as master repressor of enteroendocrine differentiation Research Background and Significance Enteroendocrine cells (EECs) are hormone-producing cells located in the epithelium of the digestive tract, playing a crucial role in regulating metabolic activities, including insulin levels, satiety, gastrointestinal secretion, and motility. The generation of EECs involves a finely tuned regulatory network of multiple transcription factors (TFs), yet the composition of this network remains incompletely understood due to the difficulty of efficiently inducing EECs from adult stem cells. Understanding the differentiation mechanisms of EECs is essential for uncovering key regulatory factors in intestinal physiology and pathology. Moreover, research on EECs may provide new strategies for the treatment of related metabolic diseases. Therefore, this study aimed to systematically identify transcription factors regulating EEC differentiation through CRISPR screening technology, in hopes of discovering new regulatory mechanisms and potential therapeutic targets. Research Introduction The research team conducted an unbiased, systematic CRISPR screen using an optimized human small intestine organoid culture system to identify transcription factors that regulate EEC differentiation. They discovered that ZNF800 is a key inhibitor that plays a dominant suppressive role by controlling the endocrine transcription factor network. Research Highlights This study represents the first large-scale CRISPR screening in human small intestine organoids aimed at identifying key transcription factors regulating EEC differentiation. The study reveals ZNF800 as the master inhibitor of EEC differentiation, providing a new perspective for understanding the differentiation mechanisms of EECs. Through single-cell RNA sequencing (scRNA-seq) and ChIP-seq technologies, the study detailed how ZNF800 functions by directly controlling the endocrine TF network, including PAX4. Research Results ZNF800 as the Master Inhibitor of EEC Differentiation: Through CRISPR knockout screening, the study found that ZNF800 is a key transcription factor that plays a dominant inhibitory role in regulating EEC differentiation. After ZNF800 knockout, the number of EECs increased, particularly the number of enterochromaffin (EC) cells, while the number of goblet and Paneth cells decreased. The Impact of ZNF800 on EEC Subtypes: Single-cell RNA sequencing analysis showed that in ZNF800-knockout organoids, the number of early EECs and ECs increased, while the number of other EEC subtypes (such as L cells, D cells, etc.) decreased. This indicates that ZNF800 not only affects the overall differentiation of EECs but may also influence the developmental trajectory of specific EEC subtypes. Direct Downstream Target Genes of ZNF800: Using ChIP-seq technology, the research team discovered that ZNF800 directly binds to the genomic regions of multiple transcription factors, including INSM1, SOX4, and PAX4. These genes are known to play a key role in EEC differentiation. In particular, PAX4, as a direct downstream target gene of ZNF800, its knockout could inhibit the differentiation of EECs induced by ZNF800 knockout, indicating the key role of PAX4 in regulating EC cell differentiation. The Role of PAX4 in EEC Differentiation: The study further found that PAX4 knockout could reduce the number of EECs and redirect the differentiation trajectory of EC cells to other EEC subtypes. This suggests that PAX4 not only plays an inhibitory role in EEC differentiation but may also be involved in determining EEC subtype specificity. Independent Functions of ZNF800 and GFI1: The study also explored the relationship between ZNF800 and GFI1, finding that these two genes independently function to inhibit EEC differentiation. GFI1 knockout did not affect the expression of ZNF800, and the number of EECs in GFI1-knockout organoids did not significantly change. The Impact of ZNF800 on EEC Function: The study assessed the function of EECs by measuring serotonin secretion and found that the secretion of serotonin in ZNF800-knockout organoids decreased, consistent with the increase in the number of EC cells, as EC cells are the main producers of serotonin. Potential Role of ZNF800 in Embryonic Development: The study also mentioned the expression of ZNF800 in human intestinal development, suggesting that it may also play an important role in embryonic development. This finding provides clues for future research on the function of ZNF800 in early development. Research Significance This study not only reveals the important role of ZNF800 in EEC differentiation but also provides new molecular targets for further research on the differentiation mechanisms of EECs. Through the optimized human small intestine organoid model, the study provides a powerful platform for CRISPR-based functional screening, aiding in the future discovery of more factors regulating human intestinal physiology and pathology. These findings offer the possibility for developing new therapeutic strategies, especially in metabolic diseases and related enteroendocrine diseases, by modulating the functions of ZNF800 and PAX4 to affect the differentiation and function of EECs. Paper Link https://pubmed.ncbi.nlm.nih.gov/37883554/ Kym Kilbourne
Unbiased transcription factor CRISPR screen identifies ZNF800 as master repressor of enteroendocrine differentiation 公告 2023-10-27 14:53:03 Written By Kym Kilbourne Title: Unbiased transcription factor CRISPR screen identifies ZNF800 as master repressor of enteroendocrine differentiation Research Background and Significance Enteroendocrine cells (EECs) are hormone-producing cells located in the epithelium of the digestive tract, playing a crucial role in regulating metabolic activities, including insulin levels, satiety, gastrointestinal secretion, and motility. The generation of EECs involves a finely tuned regulatory network of multiple transcription factors (TFs), yet the composition of this network remains incompletely understood due to the difficulty of efficiently inducing EECs from adult stem cells. Understanding the differentiation mechanisms of EECs is essential for uncovering key regulatory factors in intestinal physiology and pathology. Moreover, research on EECs may provide new strategies for the treatment of related metabolic diseases. Therefore, this study aimed to systematically identify transcription factors regulating EEC differentiation through CRISPR screening technology, in hopes of discovering new regulatory mechanisms and potential therapeutic targets. Research Introduction The research team conducted an unbiased, systematic CRISPR screen using an optimized human small intestine organoid culture system to identify transcription factors that regulate EEC differentiation. They discovered that ZNF800 is a key inhibitor that plays a dominant suppressive role by controlling the endocrine transcription factor network. Research Highlights This study represents the first large-scale CRISPR screening in human small intestine organoids aimed at identifying key transcription factors regulating EEC differentiation. The study reveals ZNF800 as the master inhibitor of EEC differentiation, providing a new perspective for understanding the differentiation mechanisms of EECs. Through single-cell RNA sequencing (scRNA-seq) and ChIP-seq technologies, the study detailed how ZNF800 functions by directly controlling the endocrine TF network, including PAX4. Research Results ZNF800 as the Master Inhibitor of EEC Differentiation: Through CRISPR knockout screening, the study found that ZNF800 is a key transcription factor that plays a dominant inhibitory role in regulating EEC differentiation. After ZNF800 knockout, the number of EECs increased, particularly the number of enterochromaffin (EC) cells, while the number of goblet and Paneth cells decreased. The Impact of ZNF800 on EEC Subtypes: Single-cell RNA sequencing analysis showed that in ZNF800-knockout organoids, the number of early EECs and ECs increased, while the number of other EEC subtypes (such as L cells, D cells, etc.) decreased. This indicates that ZNF800 not only affects the overall differentiation of EECs but may also influence the developmental trajectory of specific EEC subtypes. Direct Downstream Target Genes of ZNF800: Using ChIP-seq technology, the research team discovered that ZNF800 directly binds to the genomic regions of multiple transcription factors, including INSM1, SOX4, and PAX4. These genes are known to play a key role in EEC differentiation. In particular, PAX4, as a direct downstream target gene of ZNF800, its knockout could inhibit the differentiation of EECs induced by ZNF800 knockout, indicating the key role of PAX4 in regulating EC cell differentiation. The Role of PAX4 in EEC Differentiation: The study further found that PAX4 knockout could reduce the number of EECs and redirect the differentiation trajectory of EC cells to other EEC subtypes. This suggests that PAX4 not only plays an inhibitory role in EEC differentiation but may also be involved in determining EEC subtype specificity. Independent Functions of ZNF800 and GFI1: The study also explored the relationship between ZNF800 and GFI1, finding that these two genes independently function to inhibit EEC differentiation. GFI1 knockout did not affect the expression of ZNF800, and the number of EECs in GFI1-knockout organoids did not significantly change. The Impact of ZNF800 on EEC Function: The study assessed the function of EECs by measuring serotonin secretion and found that the secretion of serotonin in ZNF800-knockout organoids decreased, consistent with the increase in the number of EC cells, as EC cells are the main producers of serotonin. Potential Role of ZNF800 in Embryonic Development: The study also mentioned the expression of ZNF800 in human intestinal development, suggesting that it may also play an important role in embryonic development. This finding provides clues for future research on the function of ZNF800 in early development. Research Significance This study not only reveals the important role of ZNF800 in EEC differentiation but also provides new molecular targets for further research on the differentiation mechanisms of EECs. Through the optimized human small intestine organoid model, the study provides a powerful platform for CRISPR-based functional screening, aiding in the future discovery of more factors regulating human intestinal physiology and pathology. These findings offer the possibility for developing new therapeutic strategies, especially in metabolic diseases and related enteroendocrine diseases, by modulating the functions of ZNF800 and PAX4 to affect the differentiation and function of EECs. Paper Link https://pubmed.ncbi.nlm.nih.gov/37883554/ Kym Kilbourne
