IRE1a-XBP1 Signaling as a Driver of Chemotherapy-Induced Peripheral Neuropathy

IRE1a-XBP1 信号作为化疗引起的周围神经病变的驱动因素

基本信息

批准号：
10116344
负责人：
Juan R Cubillos-Ruiz
金额：
$ 18.52万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10116344
关键词：
Affect Antineoplastic Agents Behavior Behavior Control Binding Proteins Bioinformatics Biological Assay Biological Markers Biology Blood Cancer Biology Cancer Patient Cells Chemotherapy-induced peripheral neuropathy Clinical Research Complication Cytotoxic agent Development Dinoprostone Endoplasmic Reticulum Enzymes Equilibrium Event Exhibits Flow Cytometry Foundations Genetic Genomics Genotype Hand Homeostasis Host Defense Human Hypersensitivity Immune Immunofluorescence Immunologic Immunologic Surveillance Immunologist Immunology Immunophenotyping In Vitro Inflammatory Inositol Interleukin-1 Internal Medicine Internist Knockout Mice Leg Leukocytes Life Ligation Malignant Neoplasms Mechanics Mediating Mediator of activation protein Modeling Modification Molecular Target Mononuclear Leukocytes Mus Myeloid Cells Nerve Fibers Numbness Oncologist Organelles Paclitaxel Pain Pathway interactions Patient-Focused Outcomes Patients Pattern Peripheral Nerves Pharmaceutical Preparations Pharmacology Phenocopy Phenotype Population Positioning Attribute Process Production Prostaglandins Protein Biosynthesis Proteins Quality of life Reflex action Reporter Research Personnel Role Savings Scientist Signal Transduction Social Impacts Solid Neoplasm Spinal Ganglia Spleen Symptoms TLR4 gene TNF gene Testing Therapeutic Transgenic Mice Translating Tubular formation Tumor Immunity Variant Venus Weight-Bearing state Wild Type Mouse XBP1 gene allodynia anti-cancer base cancer therapy career chemotherapy chronic pain conditional knockout cyclooxygenase 2 cytokine economic impact effective therapy endoplasmic reticulum stress experience experimental study foot improved in vivo inhibitor/antagonist innovation macrophage misfolded protein monocyte multidisciplinary neuroimmunology neuroinflammation new therapeutic target novel pain behavior pain sensitivity painful neuropathy pathogen prevent response sciatic nerve sensor side effect spatiotemporal taxane trafficking transcriptome sequencing transcriptomics translational scientist tumor tumor growth

项目摘要

Summary Chemotherapy-induced peripheral neuropathy (CIPN) occurs in up to 75% of patients who receive cytotoxic agents such as paclitaxel (PTX), and is a major reason to discontinue chemotherapy. These patients experience pain, sensitivity to cold, and imbalance. PTX induces CIPN-related pain by activating Toll-like receptor 4 (TLR4) on monocytes, which induces expression of pro-inflammatory cytokines. Targeting TLR4 may be risky due to the role of TLR4 in immune defense against cancer. Interestingly, TLR4 activation triggers inositol-requiring enzyme 1 alpha-X-Box Binding Protein 1 (IRE1α-XBP1) signaling in immune cells. The inhibition of IRE1α-XBP1 enhances PTX antineoplastic effects by reducing pro-inflammatory factors, suggesting that it could be an attractive target to prevent CIPN and improve the efficacy of PTX for cancer. We have confirmed that PTX causes IRE1α-XBP1 activation and induces a pro-inflammatory phenotype in primary human leukocytes. Notably, leukocytes from mice lacking IRE1α-XBP1 specifically in immune cells (Ern1/Xbp1f/f-Vav1cre) do not display this pro-inflammatory phenotype upon PTX exposure. These conditional knockout (cKO) mice exhibit reduced cold allodynia and hind paw unbalance in a model of neuropathic pain (partial sciatic nerve ligation - PSNL). Additionally, through transcriptomic analyses we found that IRE1α-XBP1 signaling in leukocytes is critically required for the induction of prostanoids and cytokines that have been associated with CIPN. Therefore, we hypothesize that PTX promotes CIPN by activating IRE1α-XBP1 signaling in leukocytes, and that targeting this pathway could be used to prevent CIPN. We will accomplish the following specific aims: 1) Define the role of immune-intrinsic IRE1α-XBP1 in PTX-induced CIPN. We will administer PTX to mice with leukocyte-specific deletion of IRE1α-XBP1 (cKO) as well as their wild type (WT) counterparts. We anticipate that cKO mice will be protected from PTX-induced CIPN related behaviors. Then, we will treat WT mice with selective IRE1α inhibitors (MKC8866 or KIRA8) in order to pharmacologically prevent PTX-induced CIPN behaviors. These studies will define the feasibility and therapeutic potential of targeting IRE1α for CIPN. 2) Establish how PTX influences IRE1α-XBP1 signaling in immune cells to drive CIPN. We will exploit the ER stress-activated indicator (ERAI) transgenic mouse, whose cells express a yellow fluorescent protein variant (Venus) when IRE1α is activated. Immunofluorescence will be used to identify ER-stressed leukocytes in the blood, spleen, sciatic nerves, and dorsal root ganglia during CIPN. Leukocytes will be sorted according to reporter positivity and analyzed via RNA-Seq, immunophenotyping, and functional assays. These experiments will unearth how PTX- induced IRE1α-XBP1 activation influences leukocyte function throughout CIPN progression. Our team of neuro- oncologists, internist clinicians, and basic scientists with expertise in pain biology, neuroimmunology, immunology, cancer biology, IRE1α-XBP1, genomics and bioinformatics is uniquely positioned to test this innovative hypothesis and contribute to the development of novel non-narcotic treatments for chronic pain.

概括化学疗法诱导的周围神经病（CIPN）发生在多达75％的接受细胞毒性的患者中诸如紫杉醇（PTX）之类的药物是停止化疗的主要原因。这些患者经历疼痛，对寒冷和失衡的敏感性。 PTX通过激活Toll样受体4（TLR4）引起与CIPN相关的疼痛在单核细胞上，诱导促炎细胞因子的表达。靶向TLR4可能由于 TLR4在针对癌症的免疫防御中的作用。有趣的是，TLR4激活会触发肌醇提高酶 1 Alpha-X-box结合蛋白1（IRE1α-XBP1）在免疫细胞中的信号传导。抑制IRE1α-XBP1 通过减少促炎性因素来增强PTX抗肿瘤作用，表明它可能是有吸引力的目标，可防止CIPN并提高PTX对癌症的效率。我们已经确认PTX原因 IRE1α-XBP1激活并诱导原代人白细胞中的促炎表型。尤其，来自免疫细胞（ERN1/XBP1F/F-VAV1CRE）中缺乏IRE1α-XBP1的小鼠的白细胞不显示此 PTX暴露后的促炎表型。这些有条件的敲除（CKO）小鼠暴露了寒冷神经性疼痛模型（部分坐骨神经结扎-PSNL）模型中的异常性和后爪不平衡。此外，通过转录组分析，我们发现白细胞中的IRE1α-XBP1信号传导至关重要诱导与CIPN相关的前列腺素和细胞因子所必需的。因此，我们假设PTX通过激活白细胞中的IRE1α-XBP1信号来促进CIPN，并且针对此途径可用于防止CIPN。我们将完成以下特定目标：1）定义免疫内膜IRE1α-XBP1在PTX诱导的CIPN中的作用。我们将对小鼠进行PTX IRE1α-XBP1（CKO）及其野生型（WT）对应物的白细胞特异性缺失。我们预料到这一点 CKO小鼠将受到PTX诱导的CIPN相关行为的保护。然后，我们将使用选择性治疗WT小鼠 IRE1α抑制剂（MKC8866或KIRA8），以防止PTX诱导的CIPN行为。这些研究将定义靶向IRE1α对CIPN的可行性和治疗潜力。 2）建立如何 PTX影响免疫细胞中的IRE1α-XBP1信号传导驱动CIPN。我们将利用ER应力激活指示剂（ERAI）转基因小鼠，其细胞在IRE1α时表达黄色荧光蛋白变体（金星）被激活。免疫荧光将用于鉴定血液中的ER应激的白细胞 CIPN期间神经和背根神经节。白细胞将根据记者潜力进行分类，并且通过RNA-SEQ，免疫表型和功能测定法分析。这些实验将发掘PTX- 诱导的IRE1α-XBP1激活会影响整个CIPN进展的白细胞功能。我们的神经团队肿瘤学家，内科医生和具有疼痛生物学专业知识，神经免疫学专业知识的基础科学家，免疫学，癌症生物学，IRE1α-XBP1，基因组学和生物信息学是独特的，可以测试这一点创新的假设，并有助于开发新的非麻醉性治疗方法来促进慢性疼痛。