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

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

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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.

概括精确药物疗法已成为一种有效且日益普遍的癌症治疗形式。这些疗法经常采用多激酶抑制剂药物（MKI），每个疗法都靶向多个接收器酪氨酸激酶，与“常规”细胞毒性化学疗法药物结合使用。一个常见许多癌症药物治疗的副作用是对患者外周神经系统的损害，称为药物 - 诱导的周围神经病（DIPNS）。这些DIPN中的大多数是由不同的“死亡”引起的感知皮肤的感觉轴突，导致感觉疼痛和功能障碍。几个常用的MKI 然而，诱导周围神经病，导致这些痛苦DIPN的特定靶标是未知的。到解决了这个知识差距，我们建立了一种高素质筛选方法，可以快速识别斑马鱼中的神经毒性化合物。使用这种方法，我们表明已知三个MKI会产生患者的DIPN导致斑马鱼远端皮肤体感轴突的密度降低。居住成像表明，轴突缩回是这种降低的皮肤轴突密度的细胞基础，一致具有“死亡”病理生理学。此外，这些结果在小鼠背根神经节中复制神经元。对这种神经毒性作用的MKI目标的初步筛选发现接收器的损失酪氨酸激酶C-KIT，但没有其他共同的激酶靶标导致皮肤轴突密度降低。 C-Kit是在胚胎和成人中的脊椎动物感觉神经元子集中表达，其配体SCF在尚未定义皮肤，但是该配体受体在轴突维持或DIPN中的特定作用尚未定义。重要的是，这些MKI中的一个在C-kit突变体中的应用不会加剧轴突密度损失，这表明该套件是周围神经系统中MKI的主要目标。根据初步数据，我们提出目的1至：1）在斑马鱼中实施一种高素质筛查方法，以识别影响不同的MKI 体内感觉轴突毒性，然后确定其分子靶标； 2）验证这些结果哺乳动物DRG文化。 AIM 2将表征神经毒性的下游机制我们的初步数据中描述的c-kit接收器功能丧失。我们的工作将确定并表征疼痛MKI诱导的周围神经毒性的分子靶标和细胞碱基。反过来，这将提供新的途径和潜在干预的点，以研究主要但未解决的临床背景癌症药物疗法的问题。此外，我们还将建立一个工作流和试剂，以将来研究 DIPNS的候选目标。