Robustness of the Intestinal Stem Cell Niche

肠道干细胞生态位的稳健性

• 批准号: 9262951
• 负责人: Xiling Shen
• 金额: $ 30.61万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-06 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9262951
• 关键词: 3D Print; Abdomen; Ablation; Address; Algorithms; Alleles; Animals; Area; Behavior; Biological Assay; Biological Models; Cell Differentiation process; Cell Lineage; Cells; Chronic; Clinical; Colon Carcinoma; Columnar Cell; Development; Disease; Engraftment; Environment; Epithelium; Event; Feedback; Genetic Engineering; Homeostasis; Image; Imaging Techniques; Implant; Individual; Injectable; Injection of therapeutic agent; Intestines; LGR5 gene; Label; Lasers; Left; Ligands; Link; Microscopy; Modeling; Monitor; Movement; Mus; Mutation; Natural regeneration; Nature; Operative Surgical Procedures; Organoids; Paneth Cells; Pathway interactions; Physiologic pulse; Plasticizers; Recovery; Reporter; Resolution; Role; Signal Pathway; Signal Transduction; Small Intestines; Specific qualifier value; Stem Cell Research; Stem cell transplant; Stem cells; System; Systems Analysis; Systems Biology; Techniques; Technology; Testing; Tissues; Transgenic Mice; Validation; adult stem cell; animal imaging; base; blastocyst; cell type; computer framework; daughter cell; dynamic system; improved; in vivo; in vivo Model; innovation; insight; interdisciplinary approach; intestinal crypt; intestinal epithelium; multi-scale modeling; multiphoton imaging; notch protein; novel; public health relevance; regenerative; resilience; spatiotemporal; stem cell biology; stem cell niche; stem cell population; theories; time use; tool; transcriptome

 DESCRIPTION (provided by applicant): The intestinal epithelium is one of the fastest regenerative tissues in the body. Regeneration is sustained by a group of stem cells, termed crypt base columnar cells (CBCs), at the bottom of the intestinal crypt. In the past 5 years, studies of CBCs have helped transform the dogma of stem cell biology. Rather than being largely quiescent, undergoing asymmetric division, and following the unidirectional differentiation hierarchy, CBCs are highly proliferative, capable of symmetric division, and replaceable by more differentiated cell types. Since then, similar stem cell populations and regulatory principles have been identified in many other tissues. However, all these discoveries converge on a central question: how does the stem cell niche control plasticity in order to keep a constant number of CBCs? Since proliferative CBCs have been linked to colon cancer, and stem cell transplants are being used clinically for treating a variety of diseases, it is importantto understand the underlying control. Cellular dynamics Innovative multiphoton imaging and laser ablation technologies will be used to investigate how cells divide, move and recover loss inside the stem cell niche, focusing on four fundamental questions: (1) are CBCs capable of both symmetric and asymmetric division, (2) do individual CBCs in the same niche have equal proliferative potential, (3) how does the niche recover from cell loss, and (4) how do cells outside the niche dedifferentiate and reenter the niche? Signaling dynamics Using the 3D intestinal organoid assay, a systematic study will be carried out to search for feedback and crosstalk mechanisms among major signaling pathways. Dynamical systems analysis will be performed to understand their impact on the niche control circuitry. Integration and Validation To integrate experimental findings and computational insights, a stochastic, multiscale model will be built that captures cellular division, movement and signaling events in the stem cell niche. This model will be used to test hypotheses on the niche control circuitry. A novel engraftment assay through injection of genetically engineered stem cells into blastocysts has been developed for in vivo validation. Innovation Novel in vivo GI imaging techniques have been developed by using openable abdominal window, 3-D printed intestinal support, labeled vasculature roadmap, and tracking algorithms. Through the window, a special laser can ablate a single cell inside the niche without damaging the surrounding tissue. A computational framework will integrate dynamical systems analysis and multiscale modeling to study the regulatory circuitry that controls cellular and signaling dynamics inside the niche. Predictions will be validated in novel chimeric mice with intestinal crypts derived from blastocyst-injected cells. Preliminary findings The stem cell niche undergoes extensive reorganization after loss of one CBC, attesting to its dynamic nature. Various feedback and crosstalk mechanisms have been identified to improve robustness.

 描述（由申请人提供）：肠上皮是体内再生最快的组织之一。再生是由一组干细胞维持的，称为隐窝基底柱状细胞（CBCs），在肠隐窝底部。在过去的5年里，CBCs的研究已经帮助改变了干细胞生物学的教条。CBCs不是大部分处于静止状态，经历不对称分裂，并遵循单向分化层次，而是高度增殖，能够对称分裂，并可被更分化的细胞类型取代。从那时起，在许多其他组织中也发现了类似的干细胞群和调节原则。然而，所有这些发现都集中在一个中心问题上：干细胞生态位如何控制可塑性以保持恒定数量的CBC？由于增殖性CBCs与结肠癌有关，并且干细胞移植在临床上被用于治疗各种疾病，因此了解潜在的控制非常重要。细胞动力学创新的多光子成像和激光消融技术将用于研究细胞如何分裂、移动和恢复干细胞生态位内的损失，重点关注四个基本问题：（1）CBCs是否能够对称和不对称分裂，（2）同一小生境中的单个CBCs是否具有相等的增殖潜力，（3）小生境如何从细胞损失中恢复，（4）龛外的细胞如何去分化并重新进入龛内？信号动力学使用3D肠道类器官测定，将进行系统的研究，以寻找主要信号通路之间的反馈和串扰机制。将进行动态系统分析，以了解它们对生态位控制电路的影响。整合和验证为了整合实验发现和计算见解，将建立一个随机的多尺度模型，捕获干细胞生态位中的细胞分裂，运动和信号传导事件。这个模型将被用来测试生态位控制电路的假设。通过将基因工程干细胞注射到胚泡中的新型植入试验已被开发用于体内验证。创新通过使用腹部开窗、3D打印肠道支持、标记的血管路线图和跟踪算法，开发了新型体内GI成像技术。通过窗口，一种特殊的激光可以消融壁龛内的单个细胞，而不会损伤周围的组织。一个计算框架将整合动力系统分析和多尺度建模，以研究控制生态位内细胞和信号动力学的调节电路。预测将在具有源自胚泡注射细胞的肠隐窝的新型嵌合小鼠中验证。初步发现干细胞龛在失去一个CBC后经历了广泛的重组，证明了其动态性质。各种反馈和串扰机制已被确定以提高鲁棒性。

项目成果

Surface Functionalized Graphene Biosensor on Sapphire for Cancer Cell Detection.

蓝宝石上的表面功能化石墨烯生物传感器用于癌细胞检测。

• DOI: 10.1166/jnn.2016.12042
• 发表时间: 2016
• 期刊: Journal of nanoscience and nanotechnology
• 作者: Joe,DanielJ;Hwang,Jeonghyun;Johnson,Christelle;Cha,Ho-Young;Lee,Jo-Won;Shen,Xiling;Spencer,MichaelG;Tiwari,Sandip;Kim,Moonkyung
• 通讯作者: Kim,Moonkyung

Experimental Null Method to Guide the Development of Technical Procedures and to Control False-Positive Discovery in Quantitative Proteomics.

• DOI: 10.1021/acs.jproteome.5b00200
• 发表时间: 2015-10-02
• 期刊: Journal of proteome research
• 影响因子: 4.4
• 作者: Shen X;Hu Q;Li J;Wang J;Qu J
• 通讯作者: Qu J

A miR-34a-Numb Feedforward Loop Triggered by Inflammation Regulates Asymmetric Stem Cell Division in Intestine and Colon Cancer.

• DOI: 10.1016/j.stem.2016.01.006
• 发表时间: 2016-02-04
• 期刊: Cell stem cell
• 影响因子: 23.9
• 作者: Bu P;Wang L;Chen KY;Srinivasan T;Murthy PK;Tung KL;Varanko AK;Chen HJ;Ai Y;King S;Lipkin SM;Shen X
• 通讯作者: Shen X