Elucidating the role of Fra1 in pancreatic Kras-driven acinar to ductal metaplasia

阐明 Fra1 在胰腺 Kras 驱动的腺泡到导管化生中的作用

基本信息

批准号：
10537870
负责人：
Alina Lin Li
金额：
$ 4.73万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10537870
关键词：
3-Dimensional ATAC-seq Academic Medical Centers Acinar Cell Acute Advisory Committees Affect Agreement Alleles Amino Acid Motifs Attenuated Automobile Driving Award Back Basic Science Binding Sites Biological Assay CSF3 gene Caerulein Cell Culture Techniques Cell Nucleus Cells Characteristics Cholecystokinin Chromatin Colony-Stimulating Factor Receptors Data Analyses Development Doctor of Philosophy Dose Doxycycline Duct (organ) structure Ductal Epithelial Cell Epithelial Epithelial Cells FOSL1 gene Fellowship Fibroblasts Fos-Related Antigens Foundations Functional disorder Goals Homeostasis Individual Inflammation Injury KRAS oncogenesis KRAS2 gene Knockout Mice Knowledge Lesion Link Maintenance Mediator of activation protein Medical center Mentors Metaplasia Metaplastic Cell Modeling Molecular and Cellular Biology Mutation Oncogenic Outcome Pancreas Pancreatic Ductal Adenocarcinoma Pancreatic Injury Pancreatic Intraepithelial Neoplasia Pancreatitis Persons Phenotype Physicians Population Precipitation Process Public Health Regulation Research Personnel Residencies Resources Role Scientist Signal Pathway Signal Transduction Techniques Testing Therapeutic Intervention Tissues Training Translational Research Universities Up-Regulation XCL1 gene acute pancreatitis analog career cell type chromatin remodeling chronic pancreatitis combat effective therapy experience innovation insight mouse model mutant new therapeutic target novel pancreatic metaplasia pre-doctoral progenitor protein complex response to injury single cell sequencing therapeutic target three dimensional cell culture tissue regeneration transcription factor

项目摘要

PROJECT SUMMARY (ABSTRACT) Acute and chronic pancreatitis afflict millions of individuals in the US. Mouse models have revealed that acinar cells can de-differentiate after pancreatic injury to a progenitor-like cell type with ductal characteristics in a process termed acinar-to-ductal metaplasia (ADM). In the absence of oncogenic mutations, ADM lesions can resolve and reform the acinar compartment. However, in the presence of oncogenic Kras mutations, the ADM lesions can continue to de-differentiate to a pre-invasive pancreatic intraepithelial neoplasia (PanIN). The mechanisms that drive PanIN formation in the context of injury and oncogenic mutations are poorly understood, resulting in an absence of targets to combat the persistent ADM. We have identified previously through bulk ATAC-sequencing that the transcription factor Fra1 is differentially active only in the context of mutant Kras and acute inflammation. We have also generated a unique mouse model expressing inducible mutant KRAS (iKras) and Fra1 loxp alleles (Fra1 KO) in a pancreatic epithelial- specific manner. Our preliminary studies demonstrate that Fra1 loss attenuates ADM formation and stromagenesis compared to the FRA1 WT controls. The overarching goal of this proposal is to understand how FRA1 and its interacting partners, which constitute the AP-1 complex, govern ADM progression in the context of mutant Kras and inflammation. My hypothesis is that FRA1 is a central mediator of ADM. I will investigate the cell autonomous and non-cell autonomous effects driving FRA1 activation through the following interrelated Specific Aims: (1) Investigate how Fra1 remodels chromatin in ADM after induction of pancreatitis; (2) Elucidate the role of fibroblast-secreted G-CSF in Fra1 induction during pancreatitis. I will use novel mouse models, ex vivo 3D acinar and ADM cell cultures, and state-of-the-art single-cell sequencing and chromatin precipitation assays to conduct this study. This proposal also encompasses a translational aspect by proposing G-CSFR as a novel therapeutic target for the treatment of acute pancreatitis. Additionally, this project incorporates aspects of pathophysiology, molecular/cellular biology, and quantitative data analysis. This proposal provides me with a robust foundation in both experimental and quantitative analysis and broadens my fundamental knowledge of pancreatic epithelial cell homeostasis and plasticity. With the guidance from experienced mentors (Drs. Rustgi and Sims), my advisory committee and the rich array of resources at Columbia University Irving Medical Center, I will be able to complete my predoctoral PhD training. Completion of this critical milestone will set the stage for my long-term goal as a physician-scientist who conducts basic and translational research in an academic medical center with a focus on tissue inflammation, cellular identity and plasticity, and tissue regeneration.

项目概要（摘要）急性和慢性胰腺炎困扰着美国数百万人。小鼠模型显示腺泡胰腺损伤后，细胞可以去分化为具有导管特征的祖细胞样类型这一过程称为腺泡到导管化生 (ADM)。在没有致癌突变的情况下，ADM 病变可以解决并改造腺泡室。然而，在存在致癌 Kras 突变的情况下，ADM 病变可继续去分化为浸润前胰腺上皮内瘤变（PanIN）。这在损伤和致癌突变的情况下驱动 PanIN 形成的机制知之甚少，导致缺乏对抗持续性 ADM 的目标。我们之前通过批量 ATAC 测序鉴定出转录因子 Fra1 与仅在 Kras 突变和急性炎症的情况下有效。我们还生成了一个独特的鼠标在胰腺上皮细胞中表达诱导突变体 KRAS (iKras) 和 Fra1 loxp 等位基因 (Fra1 KO) 的模型具体方式。我们的初步研究表明 Fra1 缺失会减弱 ADM 的形成与 FRA1 WT 对照相比，基质发生。该提案的总体目标是了解如何 FRA1 及其相互作用的伙伴构成了 AP-1 复合物，在以下情况下控制 ADM 进展：突变的 Kras 和炎症。我的假设是 FRA1 是 ADM 的中心调节因子。我会调查细胞自主和非细胞自主效应通过以下方式驱动 FRA1 激活相互关联的具体目标： (1) 研究 Fra1 在诱导 ADM 后如何重塑染色质胰腺炎; (2)阐明成纤维细胞分泌的G-CSF在胰腺炎期间Fra1诱导中的作用。我将使用新型小鼠模型、离体 3D 腺泡和 ADM 细胞培养物以及最先进的单细胞测序和染色质沉淀测定来进行这项研究。该提案还包括翻译方面提出G-CSFR作为治疗急性胰腺炎的新治疗靶点。另外，这项目结合了病理生理学、分子/细胞生物学和定量数据分析的各个方面。这该提案为我在实验和定量分析方面奠定了坚实的基础，并拓宽了我的视野胰腺上皮细胞稳态和可塑性的基础知识。在经验丰富的导师（Rustgi 博士和 Sims）、我的顾问委员会和丰富的团队的指导下凭借哥伦比亚大学欧文医学中心的资源，我将能够完成我的博士前博士培训。这一重要里程碑的完成将为我作为一名医师科学家的长期目标奠定基础，学术医学中心的基础和转化研究，重点关注组织炎症、细胞身份和可塑性，以及组织再生。