Mechanisms mediating human enteroendocrine cell differentiation and function

介导人肠内分泌细胞分化和功能的机制

基本信息

批准号：
10739834
负责人：
Daniel Richard Zeve
金额：
$ 16.94万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10739834
关键词：
ATAC-seq Address Agonist Amino Acids Appetite Regulation Attenuated Automobile Driving Award Bioinformatics Boston CNR1 gene CRISPR/Cas technology Cell Differentiation process Cell physiology Cells Childhood Co-Immunoprecipitations Collaborations Cytokine Signaling Data Desire for food Development Plans Developmental Biology Disease Doctor of Philosophy Duodenum Endocrinologist Enhancers Enteroendocrine Cell Enzyme-Linked Immunosorbent Assay Epigenetic Process Exposure to FFAR1 gene FOXO1A gene Flow Cytometry Foundations Future GPR119 receptor Gastrointestinal tract structure Gene Expression Genes Genetic Transcription Glucose Goals Health Homeobox Hormonal Hormone secretion Hormones Human Hydrocortisone Immunofluorescence Immunologic Inflammatory Insulin Interleukin-6 Intestines MAPK8 gene Mediating Medical Mentors Metabolism Methods Modeling Molecular Non-Insulin-Dependent Diabetes Mellitus Nutrient Obesity Obesity Epidemic Organoids Palmitates Pediatric Hospitals Persons Physiological Play Production Proteins Public Health Rectum Regulation Research Resources Role SP600125 Signal Induction Signal Transduction System Testing Time Transcriptional Regulation Twin Multiple Birth United States Up-Regulation Volatile Fatty Acids career development cell growth cell type comparison control cytokine design directed differentiation epigenetic regulation experience hormonal signals improved incretin hormone inhibitor innovation insulin secretion knockout gene new therapeutic target novel therapeutic intervention obesity development rectal response rimonabant single-cell RNA sequencing small molecule stem cells transcription factor treatment strategy

项目摘要

PROJECT SUMMARY The twin epidemics of obesity and type 2 diabetes mellitus (T2DM) continue to worsen, highlighting a growing need to understand the dysregulation of appetite and insulin secretion that contribute to these diseases. Enteroendocrine cells (EECs) are key regulators of both appetite and insulin secretion. Thus, the long-term goal of this project is to understand the transcriptional and epigenetic regulation of EEC differentiation and function, and how this is perturbed in disease states. The overall objective of this application is to identify and evaluate factors that are necessary for EE differentiation and hormone production/secretion. The central hypothesis is that EEC growth and function are controlled by intrinsic and extrinsic factors. Intrinsically, we have shown that FOXO1 inhibition (AS) and, separately, CB1/JNK inhibition (RSP) induce human EE differentiation, but the mechanisms driving this are not known. Extrinsically, nutrients are known to regulate EE hormone production, but it is unclear how other factors, including disease-related cytokines and hormones, alter this response. The rationale for this proposal is that through a better understanding of EEC differentiation and function, unique therapies can be developed to regulate appetite and insulin secretion via EE hormones. The central hypothesis will be evaluated in two specific aims: 1) to identify the role of HES6 and LMX1B in human EE differentiation and to identify the epigenetic and transcriptional changes driven by AS and RSP during early EE differentiation; and 2) to evaluate the impact of cytokines and hormones on nutrient-stimulated EEC function. In Aim 1, we will study gene expression changes (using CRISPR/Cas9, SHARE-seq), epigenetic changes (using SHARE-seq, bulk ATAC-seq), and protein interactions (using co-immunoprecipitation) in human duodenal organoids to investigate the roles of HES6 and LMX1B and to identify new factors involved in early stages of EE differentiation. In Aim 2, we will assess the ability of EECs (derived from duodenal and rectal organoids) to recapitulate the response of native EECs to nutrient stimulation. We will assess hormone production and secretion (using ELISA, qPCR), and how these responses are dysregulated following exposure to obesity/T2DM-associated cytokines and hormones. The candidate for this K08 proposal is Daniel Zeve, MD, PhD, a pediatric endocrinologist with expertise in developmental biology and metabolism. With his mentor, Dr. David Breault, Dr. Zeve has designed a career development plan to achieve scientific independence. This plan will be performed at Boston Children’s Hospital, allowing the applicant access to a multitude of resources throughout the Harvard Medical system. During the award period, the applicant will gain additional experience in SHARE-seq, epigenetics, CRISPR/Cas9, and bioinformatics through multiple avenues, including coursework, seminars, and high-level collaborations. This innovative project, which seeks to unravel the regulation of EEC differentiation and function, will highlight potential novel therapeutic targets for obesity and T2DM, and provide a strong foundation for Dr. Zeve’s scientific independence and future R01 submissions.

项目概要肥胖和 2 型糖尿病 (T2DM) 这两大流行病继续恶化，凸显出日益增长的需要了解导致这些疾病的食欲和胰岛素分泌失调。肠内分泌细胞（EEC）是食欲和胰岛素分泌的关键调节者。该项目的目标是了解 EEC 分化的转录和表观遗传调控功能，以及在疾病状态下它是如何受到干扰的。该应用程序的总体目标是识别和确定。评估 EE 分化和激素产生/分泌所必需的因素。假设 EEC 的生长和功能是由内在和外在因素控制的。研究表明，FOXO1 抑制 (AS) 和 CB1/JNK 抑制 (RSP) 分别可诱导人类 EE 分化，但驱动这种分化的机制尚不清楚。从外部来看，营养素可以调节 EE。激素的产生，但尚不清楚其他因素，包括与疾病相关的细胞因子和激素，如何改变这一回应的理由是通过更好地理解 EEC 的差异。和功能，可以开发独特的疗法来通过 EE 激素调节食欲和胰岛素分泌。中心假设将在两个具体目标上进行评估：1）确定 HES6 和 LMX1B 在人类 EE 分化并识别 AS 和 RSP 驱动的表观遗传和转录变化在早期 EE 分化过程中；2) 评估细胞因子和激素对营养刺激的影响 EEC 功能在目标 1 中，我们将研究基因表达变化（使用 CRISPR/Cas9、SHARE-seq）、表观遗传。变化（使用 SHARE-seq、bulk ATAC-seq）和蛋白质相互作用（使用免疫共沉淀）人十二指肠类器官研究 HES6 和 LMX1B 的作用并确定参与的新因素在目标 2 中，我们将评估 EEC（源自十二指肠和十二指肠）的能力。直肠类器官）来概括天然 EEC 对营养刺激的反应。产生和分泌（使用 ELISA、qPCR），以及暴露后这些反应如何失调该 K08 提案的候选人是医学博士 Daniel Zeve。博士，儿科内分泌学家，具有发育生物学和新陈代谢方面的专业知识。 David Breault，Zeve 博士设计了一个职业发展计划，以实现科学独立。将在波士顿儿童医院进行，使申请人能够获得多种资源在整个哈佛医疗系统中，申请人将获得额外的经验。通过多种途径在 SHARE-seq、表观遗传学、CRISPR/Cas9 和生物信息学领域开展研究，包括这个创新项目旨在揭示这一问题。 EEC 分化和功能的调节，将突出肥胖和肥胖症的潜在新治疗靶点 T2DM，并为 Zeve 博士的科学独立性和未来 R01 提交奠定了坚实的基础。