权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular mechanisms of Hedgehog receptor function

Hedgehog受体功能的分子机制

基本信息

批准号：
10737476
负责人：
PHILIP A BEACHY
金额：
$ 40.45万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10737476
关键词：
Acceleration Address Adult Affinity Anatomy Basal cell carcinoma Binding Binding Proteins Biochemical Biological Biological Assay Bladder Blood - brain barrier anatomy Cations Cause of Death Cell membrane Cells Cholesterol Chronic Chronic Obstructive Pulmonary Disease Cilia Complex Cryoelectron Microscopy Disease Down Syndrome Embryonic Pattern Specification Energy-Generating Resources Erinaceidae Event Family Family member Funding G-Protein-Coupled Receptors Hydrophobicity In Vitro Inflammatory Bowel Diseases Injury Intestines Ligands Link Lipids Lung Maintenance Malignant Neoplasms Measures Mediating Membrane Metals Molecular Molecular Conformation Muscle Natural regeneration Organ Pancreas Pathology Pathway interactions Pattern Pharmaceutical Preparations Play Porifera Postembryonic Primary Neoplasm Proliferating Protein Inhibition Proteins Regulation Resolution Role SHH gene Signal Induction Signal Pathway Signal Transduction Signaling Protein Site Specific qualifier value Sterols Structure Surface Therapeutic Tissue Differentiation Tissues Transducers Work antagonist bone cancer therapy cell type embryo tissue hedgehog signal transduction in vivo inhibitor member morphogens nanobodies neoplastic cell novel novel strategies polypeptide prevent protein complex protein protein interaction protein structure receptor receptor function regenerative repaired resistance mutation smoothened signaling pathway stem cells tissue stem cells transmission process tumor tumor growth tumor progression

项目摘要

Project Summary Hedgehog (Hh) signaling specifies the embryonic tissue pattern of many metazoan organs and maintains this tissue pattern post-embryonically by regulating the expression of proliferation- or differentiation-inducing signals that target adult tissue stem or progenitor cells. Drugs developed to block Hh pathway activity, based on our previous work, have received FDA approval for treatment of ectodermally-derived cancers, such as basal cell carcinoma. In pancreatic, bladder, and other cancers of endodermal origin, Hh pathway activity in tumor- associated stroma presents a barrier to tumor growth and progression, thus suggesting pathway activation rather than inhibition as a therapeutic approach. In addition, pathway activation has a beneficial regenerative role in bone and muscle repair, in reducing pathology associated with inflammatory bowel disease, and in preventing or ameliorating injury and breach of the blood-brain-barrier, among other emerging biological activities. On the other hand, chronic low-level elevation of pathway activity in the lung, as is associated with reduced expression of the Hh pathway inhibitor Hhip (Hh-interacting protein), is genetically linked to chronic obstructive pulmonary disease (COPD), the third leading cause of death worldwide. During the previous funding period for this project we utilized protein structure determination and biochemical and cell biological approaches to establish the molecular mechanism of Hh signaling, in which Hh binding to its receptor Patched1 (Ptch1) activates the pathway by alleviating Ptch1-mediated suppression of the essential transducer and GPCR family member, Smoothened (Smo). We found that cholesterol is the crucial link between Ptch1 and Smo, that cholesterol in the inner leaflet of the membrane is decreased by Ptch1 transport activity, and that Hh binding to Ptch1 blocks this transport activity. These events critically regulate pathway activity, as conformational switching of Smo to its active state requires entry and binding of a sterol from the inner leaflet of the membrane into a central cavity within the Smoothened seven-transmembrane bundle. We also showed how the Dispatched1 (Disp1) transporter, structurally related to Ptch1, uses Na+ flux to power its export and packaging of the dually lipid modified Sonic hedgehog protein signal (ShhNp), enabling it to move through tissues as a soluble morphogen in complex with its carrier Scube2. We propose here to deepen our understanding of Hh signal transduction and pathway regulation by establishing the energy sources and the step-by-step lipid- handling mechanisms of the Ptch1 and Disp1 transporters. We will determine the structure of the ShhNp:Scube2 morphogen, and the mechanism of its release from Disp1. Finally we plan to elucidate the mechanism of Hh signal antagonism by Hhip, using cryo-EM to determine the high-resolution structure and functionally dissect a membrane-associated tent-like Hhip multimeric complex that occludes all receptor-interacting surfaces of the Hh protein. Our findings may provide a basis for new approaches to therapeutic modulation of Hh pathway activity.

项目摘要 Hedgehog（Hh）信号指定了许多后生动物器官的胚胎组织模式，并维持这种模式。通过调节增殖或分化诱导信号的表达，针对成人组织干细胞或祖细胞的药物根据我们的研究，以前的工作，已获得FDA批准用于治疗外胚层来源的癌症，如基底细胞癌， carcinoma.在胰腺癌、膀胱癌和其他内胚层起源的癌症中，肿瘤中的Hh通路活性，相关间质对肿瘤生长和进展提供了屏障，因此表明通路活化，而不是而不是抑制作为治疗方法。此外，通路激活在细胞凋亡中具有有益的再生作用。骨和肌肉修复，减少与炎症性肠病相关的病理，以及预防或改善血脑屏障的损伤和破坏，以及其它新兴的生物活性。上另一方面，肺中通路活性的慢性低水平升高，与表达减少相关， Hh通路抑制剂Hhip（Hh相互作用蛋白）的基因与慢性阻塞性肺疾病有关，慢性阻塞性肺病（COPD）是全球第三大死亡原因。在该项目的前一个资助期内，我们利用蛋白质结构测定和生物化学方法，和细胞生物学方法来建立Hh信号转导的分子机制，其中Hh结合其受体Patched1（Ptch1）通过减轻Ptch1介导的对必需的传感器和GPCR家族成员Smoothened（Smo）。我们发现胆固醇是 Ptch 1和Smo，即膜内小叶中的胆固醇因Ptch 1转运活性而减少，而Hh与Ptch 1的结合阻断了这种转运活性。这些事件严格地调节通路活性， Smo向其活性状态的构象转换需要来自Smo的内小叶的固醇的进入和结合。将膜插入平滑七跨膜束内的中心腔中。我们还展示了 Dispatched1（Disp1）转运蛋白在结构上与Ptch1相关，它利用Na+流为其输出和包装提供动力双重脂质修饰的Sonic hedgehog蛋白信号（ShhNp），使其能够在组织中移动，可溶性形态发生蛋白与其载体Scube2复合。我们建议在这里加深我们对Hh的理解通过建立能量来源和分步脂质来调节信号传导和途径调节 Ptch 1和Disp 1转运蛋白的处理机制。我们将确定ShhNp：Scube 2的结构形态素，以及其从Disp 1释放的机制。最后，我们计划阐明Hh的机制信号拮抗作用，使用冷冻电镜确定高分辨率结构和功能解剖，一种膜相关的帐篷样Hip多聚体复合物，封闭Hh的所有受体相互作用表面蛋白我们的研究结果可能为Hh通路活性的治疗性调节提供新方法的基础。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Itraconazole inhibits enterovirus replication by targeting the oxysterol-binding protein.

DOI：
10.1016/j.celrep.2014.12.054
发表时间：
2015-02-03
期刊：
Cell reports
影响因子：
8.8
作者：
Strating JR;van der Linden L;Albulescu L;Bigay J;Arita M;Delang L;Leyssen P;van der Schaar HM;Lanke KH;Thibaut HJ;Ulferts R;Drin G;Schlinck N;Wubbolts RW;Sever N;Head SA;Liu JO;Beachy PA;De Matteis MA;Shair MD;Olkkonen VM;Neyts J;van Kuppeveld FJ
通讯作者：
van Kuppeveld FJ

Simultaneous measurement of smoothened entry into and exit from the primary cilium.

同时测量初级纤毛的平滑进入和退出。

DOI：
10.1371/journal.pone.0104070
发表时间：
2014
期刊：
PloS one
影响因子：
3.7
作者：
Kim,Jynho;Hsia,ElaineYC;Kim,James;Sever,Navdar;Beachy,PhilipA;Zheng,Xiaoyan
通讯作者：
Zheng,Xiaoyan

Hedgehog pathway modulation by multiple lipid binding sites on the smoothened effector of signal response.

Hedgehog 通路通过信号响应平滑效应器上的多个脂质结合位点进行调节。

DOI：
10.1016/j.devcel.2013.07.015
发表时间：
2013-08-26
期刊：
DEVELOPMENTAL CELL
影响因子：
11.8
作者：
Myers, Benjamin R.;Sever, Navdar;Chong, Yong Chun;Kim, James;Belani, Jitendra D.;Rychnovsky, Scott;Bazan, J. Fernando;Beachy, Philip A.
通讯作者：
Beachy, Philip A.

The role of ciliary trafficking in Hedgehog receptor signaling.