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

• 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
• 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

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.

总结 化疗诱导的周围神经病变（CIPN）发生在高达75%的接受细胞毒性药物治疗的患者中。 药物如紫杉醇（PTX），并且是停止化疗的主要原因。这些患者经历 疼痛，对寒冷的敏感，以及失衡PTX通过激活Toll样受体4（TLR 4）诱导CIPN相关疼痛 单核细胞上，其诱导促炎细胞因子的表达。靶向TLR 4可能是有风险的， TLR 4在癌症免疫防御中的作用有趣的是，TLR 4激活触发肌醇需要酶， 1 α-X-盒结合蛋白1（IRE 1 α-XBP 1）在免疫细胞中的信号传导。IRE 1 α-XBP 1的抑制作用 通过减少促炎因子来增强PTX的抑制作用，这表明它可能是一种 有吸引力的靶点，以防止CIPN和提高PTX对癌症的疗效。我们已经证实PTX会导致 IRE 1 α-XBP 1激活并诱导原代人白细胞中的促炎表型。值得注意的是， 来自免疫细胞中特异性缺乏IRE 1 α-XBP 1的小鼠的白细胞（Ern 1/Xbp 1f/f-Vav 1cre）不显示这一点 促炎表型。这些条件性基因敲除（cKO）小鼠表现出降低的寒冷 在神经性疼痛模型（部分坐骨神经结扎- PSNL）中的异常性疼痛和后爪失衡。 此外，通过转录组学分析，我们发现白细胞中的IRE 1 α-XBP 1信号传导是关键的， 所需的前列腺素类和细胞因子的诱导已与CIPN。所以我们 假设PTX通过激活白细胞中的IRE 1 α-XBP 1信号来促进CIPN， 靶向该途径可用于预防CIPN。我们将实现以下具体目标：1） 明确免疫内源性IRE 1 α-XBP 1在PTX诱导的CIPN中的作用。我们将给小鼠注射PTX， IRE 1 α-XBP 1（cKO）及其野生型（WT）对应物的白细胞特异性缺失。我们预计 将保护cKO小鼠免于PTX诱导的CIPN相关行为。然后，我们将用选择性的 IRE 1 α抑制剂（MKC 8866或KIRA 8），以预防PTX诱导的CIPN行为。 这些研究将确定靶向IRE 1 α治疗CIPN的可行性和治疗潜力。2)确定如何 PTX影响免疫细胞中的IRE 1 α-XBP 1信号传导以驱动CIPN。我们将利用内质网应激激活 指示剂（ERAI）转基因小鼠，当IRE 1 α 被激活了免疫荧光将用于鉴定血液、脾脏、坐骨神经和骨髓中的ER应激白细胞。 神经和背根神经节。将根据报告基因阳性对白细胞进行分类， 通过RNA-Seq、免疫表型分析和功能测定进行分析。这些实验将揭示PTX- 诱导的IRE 1 α-XBP 1活化在整个CIPN进展中影响白细胞功能。我们的神经- 肿瘤学家，内科临床医生，以及在疼痛生物学，神经免疫学， 免疫学、癌症生物学、IRE 1 α-XBP 1、基因组学和生物信息学是检验这一点的唯一方法 创新的假设，并有助于开发新的非麻醉性治疗慢性疼痛。