Elucidating How Primary Cilia Regulate Hedgehog Signaling by Super-Resolution Microscopy

通过超分辨率显微镜阐明初级纤毛如何调节 Hedgehog 信号传导

基本信息

批准号：
10436146
负责人：
Xiaoyu Shi
金额：
$ 24.83万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-18 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10436146
关键词：
3-Dimensional Active Biological Transport Adult Algorithms Basal cell carcinoma Binding Sites Cells Cilia Data Defect Disease Docking Embryo Foundations Frequencies Goals Heterogeneity Homeostasis Image Knock-out Label Malignant Neoplasms Maps Measures Mediating Mediator of activation protein Membrane Microscopy Modeling Molecular Movement Mutation Nature Optics Pattern Prevention Preventive screening Proteins Receptor Signaling Regulation Reporting Resolution Role Signal Transduction Signaling Molecule Techniques Technology Therapeutic Therapeutic Intervention Time Tissues Uncertainty base congenital heart disorder developmental disease insight mutant nanometer resolution novel novel strategies novel therapeutics particle preventive intervention protein protein interaction reconstruction smoothened signaling pathway spatial relationship spatiotemporal structural biology trafficking

项目摘要

Abstract Hedgehog signals are the key regulators of embryonic patterning and adult tissue homeostasis. Consequently, defects in Hedgehog signaling can cause developmental diseases, congenital heart disease, and cancers such as basal cell carcinoma. There is an urgent need to understand the molecular mechanisms that underlie the modulation of Hedgehog signaling pathway for its potential preventative and therapeutic value. It is known ver- tebrate Hedgehog signaling relies on the ciliary trafficking of Hedgehog signaling receptors, among which smoothened (SMO) is the central positive mediator of Hedgehog signaling. If mutations occur in either intrafla- gellar transporters (IFTs), or in the ciliary transition zone, SMO activities can be severely disrupted. However, the molecular mechanism of how IFT particles and transition zone regulate trafficking of SMO is currently un- known. Determination of the molecular regulation mechanisms is the objective of this application. Our prelimi- nary data acquired by Stochastic Optical Reconstruction Microscopy (STORM) showed the colocalization of transition zone proteins with SMO and IFT88, suggesting that transition zone proteins and IFT particles interact with SMO. Based on previous studies and our own primary data, my central hypothesis is that the transition zone serves as a checkpoint for Hedgehog signaling receptors, and IFTs help Hedgehog signaling receptors cross the transition zone. This transition zone checkpoint model represents a novel mechanism for the control of cilium trafficking and the cross-interaction between different ciliary cargos. It could potentially allow new ap- proaches to manipulate Hedgehog signals, and underlie the foundation for treatments of diseases caused by defects in Hedgehog signaling. To approach to the project, I plan to map SMO molecules and IFT particles in the transition zone using multicolor 3D STORM. It will reveal the spatial relationship between SMO molecules and these ciliary components at a resolution of ~15 nm. Algorithms will be developed to reduce the uncertainty of the spatial easements caused by structural heterogeneity and immunostaining, providing a ~ 5nm precision of the distance between investigated proteins, indicating protein-protein interaction. Equally important as the static structural study, I also plan to detect the interactions among SMO molecules, IFT particles, and transition zone proteins dynamically using single-particle tracking and photoconversion imaging. The proposed project will not only offer new insights into the molecular mechanisms of Hedgehog signaling regulation, but also ad- vance a suite of microscopy-based technologies and algorithms that can be broadly applied to the fields of cell signaling and structural biology. Furthermore, the results are expected to have broad impact, because the reg- ulatory components to be identified by this project will provide new mechanisms and new drug screen for pre- ventive and therapeutic interventions. 1

抽象的刺猬信号是胚胎图案和成人组织稳态的关键调节剂。最后，刺猬信号的缺陷会导致发育疾病，先天性心脏病和癌症作为基底细胞癌。迫切需要了解基于的分子机制刺猬信号通路的调节，以预防和治疗价值。这是已知的 Tebrate Hedgehog信号依赖于刺猬信号受体的睫毛贩运，其中平滑（SMO）是刺猬信号传导的中心阳性介体。如果在两种情况内发生突变 Gellar转运蛋白（IFTS）或在睫状过渡区中，SMO活动可能会严重中断。然而， IFT颗粒和过渡区如何调节SMO运输的分子机制目前不存在已知。确定分子调节机制是该应用的目的。我们的预赛随机光学重建显微镜（Storm）获得的NARY数据显示带有SMO和IFT88的过渡区蛋白质，表明过渡区蛋白和IFT颗粒相互作用与Smo。根据以前的研究和我们自己的主要数据，我的中心假设是过渡区域充当刺猬信号受体的检查点，IFTS有助于刺猬信号受体越过过渡区。此过渡区检查点模型代表了控制的新机制纤毛贩运和不同睫状炼油之间的跨界。它可能允许新的ap- 操纵刺猬信号的规定，并为疾病疗法的基础是刺猬信号的缺陷。要处理该项目，我计划映射SMO分子和IFT颗粒使用多色3D风暴的过渡区。它将揭示SMO分子之间的空间关系这些睫状成分的分辨率约为15 nm。将开发算法以减少不确定性由结构异质性和免疫染色引起的空间地役权，提供约5nm的精度研究的蛋白质之间的距离，表明蛋白质 - 蛋白质相互作用。同样重要静态结构研究，我还计划检测SMO分子，IFT颗粒和过渡之间的相互作用区域蛋白质使用单粒子跟踪和光转换成像动态。拟议的项目不仅将提供有关刺猬信号调节的分子机制的新见解，而且还将 Vance一套基于显微镜的技术和算法，可以广泛应用于细胞领域信号传导和结构生物学。此外，预计结果将产生广泛的影响，因为该项目将要确定的胰腺成分将提供新的机制和新药物筛查，以预先现象和治疗干预措施。 1

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Local enrichment of HP1alpha at telomeres alters their structure and regulation of telomere protection.

DOI：
10.1038/s41467-018-05840-y
发表时间：
2018-09-04
期刊：
Nature communications
影响因子：
16.6
作者：
Chow TT;Shi X;Wei JH;Guan J;Stadler G;Huang B;Blackburn EH
通讯作者：
Blackburn EH

Nanotopography Enhances Dynamic Remodeling of Tight Junction Proteins through Cytosolic Liquid Complexes.

DOI：
10.1021/acsnano.0c04866
发表时间：
2020-10-27
期刊：
ACS nano
影响因子：
17.1
作者：
Huang X;Shi X;Hansen ME;Setiady I;Nemeth CL;Celli A;Huang B;Mauro T;Koval M;Desai TA
通讯作者：
Desai TA

Expansion Microscopy of Ciliary Proteins.

纤毛蛋白的放大显微镜。

DOI：
10.1007/978-1-0716-3507-0_4
发表时间：
2024
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Park,Sohyeon;Shi,Xiaoyu
通讯作者：
Shi,Xiaoyu

Cationic peptides erase memories by removing synaptic AMPA receptors through endophilin-mediated endocytosis.

阳离子肽通过内皮素介导的内吞作用去除突触 AMPA 受体，从而消除记忆。

DOI：
10.21203/rs.3.rs-3559525/v1
发表时间：
2023
期刊：
Research square
影响因子：
0
作者：
Stokes,Eric;Zhuang,Yinyin;Toledano,Michael;Vasquez,Jose;Azouz,Ghalia;Hui,May;Tyler,Isabella;Shi,Xiaoyu;Aoto,Jason;Beier,KevinT
通讯作者：
Beier,KevinT

Polarized endosome dynamics engage cytoplasmic Par-3 that recruits dynein during asymmetric cell division.

DOI：
10.1126/sciadv.abg1244
发表时间：
2021-06
期刊：
Science advances
影响因子：
13.6
作者：
Zhao X;Garcia JQ;Tong K;Chen X;Yang B;Li Q;Dai Z;Shi X;Seiple IB;Huang B;Guo S
通讯作者：
Guo S