High-throughput identification of molecular targets responsible for drug-induced peripheral neuropathies.

高通量鉴定导致药物引起的周围神经病变的分子靶标。

基本信息

批准号：
10371819
负责人：
Alex Nechiporuk
金额：
$ 39.6万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
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：
2022-09-22 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10371819
关键词：
Acute Address Adult Afferent Neurons Aftercare Animals Antineoplastic Agents Axon Back CRISPR screen CRISPR/Cas technology Cell Proliferation Cell Survival Chronic Clinic Clinical Confocal Microscopy Cutaneous Cytotoxic Chemotherapy Cytotoxic agent Data Dermal Distal Dose Drug Targeting Embryo FDA approved Functional disorder Future Image In Vitro Intervention Knowledge Larva Lead Libraries Ligands Maintenance Malignant Neoplasms Mediating Molecular Molecular Target Morphology Mus Natural regeneration Nervous system structure Neurons Neuropathy Pain Pathway interactions Patients Peripheral Peripheral Nervous System Peripheral Nervous System Diseases Pharmaceutical Preparations Pharmacology Pharmacotherapy Phenotype Phosphotransferases Process Proto-Oncogene Protein c-kit Quality of life Rapid screening Reagent Receptor Inhibition Receptor Protein-Tyrosine Kinases Regimen Reporting Role Sensory Severities Signal Pathway Signal Transduction Skin Spinal Ganglia System Testing Therapeutic Time Tissue Sample Toxic effect Transgenic Organisms Tyrosine Kinase Receptor Inhibition Work Zebrafish axon growth base cancer cell cancer therapy chemotherapy density dosage effective therapy genetic approach human tissue imaging platform in vivo inhibitor inhibitor therapy knock-down loss of function molecular drug target mouse model mutant neurotoxic neurotoxicity new therapeutic target novel anticancer drug novel therapeutic intervention painful neuropathy precision drugs prevent receptor response screening side effect somatosensory therapeutic target time use translational study

项目摘要

Summary Precision drug therapies have emerged as an effective and increasingly common form of cancer treatment. These therapies frequently employ multi-kinase inhibitor drugs (MKIs) that each target multiple receptor tyrosine kinases, sometime in combination with “conventional” cytotoxic chemotherapy drugs. One common side effect of many cancer drug treatments is damage to the patient's peripheral nervous system, termed drug- induced peripheral neuropathies (DIPNs). Most of these DIPNs are caused by the “dying back” of distal sensory axons that innervate the skin, leading to sensory pain and dysfunction. Several commonly used MKIs induce peripheral neuropathies, however, specific targets responsible for these painful DIPNs are unknown. To address this knowledge gap, we established a high-content screening approach that allows rapid identification of neurotoxic compounds in zebrafish. Using this approach, we showed that three MKIs known to produce DIPNs in patients led to the reduced density of distal cutaneous somatosensory axons in zebrafish. Live imaging demonstrated that axon retraction is the cellular basis for this reduced dermal axon density, consistent with a “dying back” pathophysiology. Furthermore, these results were replicated in mouse dorsal root ganglia neurons. Initial screening for MKI targets underlying this neurotoxic effect found that loss of the receptor tyrosine kinase c-Kit, but not other shared kinase targets, led to reduced cutaneous axon density. c-Kit is expressed in a subset of vertebrate sensory neurons in embryos and adults and its ligand SCF is expressed in the skin, but the specific role of this ligand-receptor in axon maintenance or DIPNs has not been defined. Importantly, application of one of these MKIs in c-kit mutants did not exacerbate axon density loss, indicating that Kit is a major target for an MKI in the peripheral nervous system. Based on preliminary data we propose in Aim 1 to: 1) implement a high-content screening approach in zebrafish to identify MKIs that induce distal sensory axon toxicity in vivo and then identify their molecular targets; and 2) validate these results in mammalian DRG culture. Aim 2 will characterize the downstream mechanisms underlying the neurotoxicity of c-Kit receptor loss-of-function described in our preliminary data. Our work will identify and characterize the molecular targets and cellular bases of painful MKI-induced peripheral neurotoxicity. This, in turn, will provide new pathways and points of potential intervention to study in the context of a major, but unaddressed, clinical problem in cancer drug therapies. In addition, we will also establish a workflow and reagents for future study of candidate targets of DIPNs.

概括精准药物疗法已成为一种有效且日益常见的癌症治疗形式。这些疗法经常采用多激酶抑制剂药物 (MKI)，每种药物都针对多个受体酪氨酸激酶，有时与“传统”细胞毒性化疗药物联合使用。许多癌症药物治疗的副作用是损害患者的周围神经系统，称为药物- 诱发周围神经病 (DIPN) 大多数 DIPN 是由远端“死亡”引起的。支配皮肤的感觉轴突，导致感觉疼痛和功能障碍。几种常用的 MKI。诱发周围神经病变，然而，造成这些痛苦的 DIPN 的具体靶点尚不清楚。为了解决这一知识差距，我们建立了一种高内涵筛选方法，可以快速识别使用这种方法，我们发现了三种已知会产生的 MKI。患者体内的 DIPN 导致斑马鱼远端皮肤体感轴突密度降低。成像表明轴突回缩是真皮轴突密度降低的细胞基础，一致此外，这些结果在小鼠背根神经节中得到了复制。对导致这种神经毒性作用的 MKI 靶点的初步筛选发现受体的丢失。酪氨酸激酶 c-Kit，而不是其他共享激酶靶点，导致皮肤轴突密度降低。在胚胎和成体的脊椎动物感觉神经元亚群中表达，其配体 SCF 在皮肤，但这种配体受体在轴突维持或 DIPN 中的具体作用尚未确定。重要的是，在 c-kit 突变体中应用这些 MKI 之一并没有加剧轴突密度损失，表明根据我们在中提出的初步数据，该套件是周围神经系统中 MKI 的主要目标。目标 1 是：1) 在斑马鱼中实施高内涵筛选方法，以识别诱导远端体内感觉轴突毒性，然后确定其分子靶标；2) 验证这些结果；目标 2 将表征哺乳动物 DRG 神经毒性的下游机制。我们的初步数据中描述的 c-Kit 受体功能丧失将被识别并表征。反过来，这将提供痛苦的 MKI 诱导的周围神经毒性的分子靶点和细胞基础。在重大但尚未解决的临床背景下进行研究的新途径和潜在干预点此外，我们还将为未来的研究建立工作流程和试剂。 DIPNs 的候选目标。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Single-Cell Analysis of Rohon-Beard Neurons Implicates Fgf Signaling in Axon Maintenance and Cell Survival.

Rohon-Beard 神经元的单细胞分析表明 Fgf 信号传导参与轴突维持和细胞存活。

DOI：
发表时间：
2024-04-17
期刊：
影响因子：
0
作者：
Tuttle, Adam M;Miller, Lauren N;Royer, Lindsey J;Wen, Hua;Kelly, Jimmy J;Calistri, Nicholas L;Heiser, Laura M;Nechiporuk, Alex V
通讯作者：
Nechiporuk, Alex V