Targeting the E1 Control Point of Protein Ubiquitination in Cancer

靶向癌症中蛋白质泛素化的 E1 控制点

基本信息

批准号：
10535648
负责人：
Christina Marie Zeina
金额：
$ 3.89万
依托单位：
HARVARD MEDICAL SCHOOL
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：
2022-08-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10535648
关键词：
Affinity Antibodies Antineoplastic Agents Apical Apoptosis Binding Binding Sites Biochemical Biology Bortezomib Cell Proliferation Cell Survival Cells Charge Chemicals Chemoresistance Complex Crystallization Dana-Farber Cancer Institute Degradation Pathway Deuterium Development Drug resistance Enzymes Genetic Goals Homeostasis Human Hydrogen In Vitro Laboratories Lead Leukemic Cell Libraries Malignant Neoplasms Mass Spectrum Analysis Measures Molecular Molecular Conformation Multiple Myeloma Mutagenesis Mutation N-terminal Oncology Organism Pathologic Pathway interactions Peptide Hydrolases Peptides Permeability Pharmacology Physicians Polyubiquitin Proteasome Inhibition Proteins Proteome Quality Control Reporting Research Resistance Scientist Series Shapes Source Stress Structure Surface System Technology Testing Therapeutic Training Programs Ubiquitin Ubiquitination Validation X-Ray Crystallography adduct alpha helix anti-cancer base cancer care cancer cell cancer therapy design drug development drug discovery efficacy testing in vitro activity in vivo individualized medicine inhibitor innovation knock-down leukemia medical schools mouse model multicatalytic endopeptidase complex multidisciplinary mutation screening novel novel therapeutic intervention phase I trial pointed protein protein aminoacid sequence proteotoxicity prototype small molecule stapled peptide thioester treatment strategy tumor uptake

项目摘要

PROJECT SUMMARY The ubiquitin-proteasome system (UPS) is a homeostatic enzymatic cascade that tags defective or unwanted cellular proteins with ubiquitin, resulting in their efficient proteasomal degradation. Protein quality control is critical to cancer cell survival due to the heightened demands of cell proliferation and the damage sustained to the cancer cell proteome from a host of stresses. Multiple myeloma is a classic example of a human cancer exquisitely dependent on the proteasome due to massive overproduction of antibody proteins and is thus susceptible to proteasome inhibition by bortezomib. FDA approval of bortezomib heralded the development of multiple approaches to modulate the UPS for therapeutic benefit by targeting the E1, E2, and E3 ubiquitin transfer enzymes and deubiquitinases. UBE1 is the most apical enzyme of the ubiquitination cascade and is singularly responsible for 99% of ubiquitin charging of E2 proteins. The small molecule TAK-243 was developed to target the ATP binding pocket of UBE1 and forms a covalent adduct with the C-terminus of ubiquitin, resulting in arrest of enzymatic activity. TAK-243 demonstrated anti-tumor activity in vitro, in mouse models, and in a small phase 1 trial, underscoring the potential of blocking UBE1 as an anti-cancer strategy. However, point mutagenesis at the TAK-243 binding site can cause resistance, mandating alternative approaches to targeting UBE1 in cancer. Examination of the structures of E1/E2 complexes revealed a surface groove on E1 bound by the N-terminal a- 1 helix of E2 (E2h1), suggesting that an a-helical mimic of E2h1 could be developed as a potential UBE1 inhibitor. Natural motifs such as E2h1 often unfold when taken out of context of the native protein, resulting in loss of bioactive shape and rapid proteolytic degradation. Peptide stapling reinforces the natural a-helical shape of bioactive peptides and confers stabilized structure, protease resistance in vivo, enhanced target binding affinity, and favorable pharmacology. Applying this technology to UBE1 targeting, the Walensky lab recently generated a prototype inhibitor using the E2h1 sequence of UBE2A. The stapled peptide bound to the UBE1 surface groove and blocked ubiquitin transfer to E2 proteins, resulting in suppression of protein ubiquitination in vitro. Here, I propose to combine our stapled peptide approach to drug development and our recent discovery of a targetable helix-in-groove interaction on UBE1 to advance a unique treatment strategy for human leukemias. Specifically, I aim to (1) design, synthesize, and characterize novel stapled peptide inhibitors of E1 (SPIEs) that target a helix- in-groove interface between E1 and E2 proteins; (2) define the conformational consequences of E1 targeting by SPIEs and the structure of a lead SPIE/E1 complex; and (3) advance lead SPIEs to testing in UPS-dependent leukemia cells to evaluate mechanism of action and anti-cancer efficacy. I am excited to pursue the proposed training program for my chemical biology graduate studies in the laboratory of Dr. Loren Walensky at the Dana- Farber Cancer Institute and Harvard Medical School, and look forward to developing as a creative, independent, and innovative physician-scientist at the forefront of cancer drug discovery and cancer care.

项目摘要泛素 - 蛋白酶体系统（UPS）是一种稳态的酶促级联，标记有缺陷或不必要的标签含泛素的细胞蛋白，导致其有效的蛋白酶体降解。蛋白质质量控制至关重要由于细胞增殖的需求增强以及对癌症细胞的存活以及对来自许多应力的癌细胞蛋白质组。多发性骨髓瘤是人类癌症的经典例子由于抗体蛋白的大量生产过多而导致的蛋白酶体非常依赖于蛋白酶体，因此容易受到硼替佐米抑制蛋白酶体的影响。 FDA批准Bortezomib宣布了通过针对E1，E2和E3泛素转移来调节UP的多种方法来调节UPS以获得治疗益处酶和去泛素酶。 ube1是泛素化级联的最根尖酶，是单一的负责99％的E2蛋白质泛素充电。小分子tak-243是为了靶向的 UBE1的ATP结合口袋，并与泛素的C末端形成共价加合物，导致逮捕酶活性。 TAK-243在体外表现出抗肿瘤活性，在小鼠模型中和小相中表现出抗肿瘤活性 1个试验，强调阻止UBE1作为抗癌策略的潜力。但是，点诱变 TAK-243结合位点可能会引起抗性，强制靶向癌症中的UBE1的替代方法。对E1/E2复合物的结构的检查表明，N端A-在E1上的表面凹槽 1 E2的螺旋（E2H1），表明可以开发E2H1的A螺旋模拟物作为潜在的UBE1抑制剂。当从天然蛋白的上下文中取出时，诸如E2H1之类的天然主题通常会展开，导致失去生物活性形状和快速蛋白水解降解。肽固定剂增强了自然的A螺旋形状生物活性肽并赋予稳定的结构，体内蛋白酶的抗性，增强靶联接亲和力，和有利的药理学。 Walensky Lab将这项技术应用于UBE1目标，最近生成了使用UBE2A的E2H1序列的原型抑制剂。与UBE1表面凹槽结合的钉肽并阻塞了泛素转移到E2蛋白，从而导致体外抑制蛋白质泛素化。在这里，我建议将我们的固定肽方法结合起来进行药物开发和我们最近发现的目标 UBE1上的螺旋在沟间相互作用，以推进人类白血病的独特治疗策略。具体来说，我的目标是（1）设计，合成和表征靶向螺旋 - 的E1（SPIE）的新型固定肽抑制剂 E1和E2蛋白之间的河流界面；（2）通过间谍和铅SPIE/E1复合物的结构；（3）提前间谍在UPS依赖性中进行测试白血病细胞评估作用机理和抗癌功效。我很高兴追求拟议的我的化学生物学研究生研究培训计划在Dana- Farber Cancer Institute和Harvard Medical School，并期待发展为创意，独立，以及创新的医师科学家在癌症药物发现和癌症护理的最前沿。