Therapeutic Exploitation of IPSE, a Urogenital Parasite-Derived Host Modulatory Protein, for Bladder Hypersensitivity Syndromes

IPSE（一种泌尿生殖寄生虫衍生的宿主调节蛋白）对膀胱过敏综合征的治疗利用

• 批准号: 9288039
• 负责人: Michael Harrison Hsieh
• 金额: $ 51.59万
• 依托单位: BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-05 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9288039
• 关键词: Adverse effects; Adverse reactions; Affect; Apoptosis; Basophilic Cell; Basophils; Behavior; Binding; Biology; Bladder; Bladder Diseases; Bladder Injury; CCL2 gene; CCL3 gene; Cell Adhesion; Cell Nucleus; Cells; Chinese Hamster Ovary Cell; Chronic; Computer Simulation; Cystitis; Data; Dendritic Cells; Disease model; Dose; Engineering; Enteral; Epithelial Cells; Freezing; Functional disorder; Generations; Genes; Genetic Transcription; Genitourinary system; Hemorrhage; Homologous Gene; Hypersensitivity; IL8 gene; Ifosfamide; IgE; Immune; Immune response; Incidence; Increased frequency of micturition; Interleukin-4; Interstitial Cystitis; Intestines; Intravenous; Lead; Ligands; Link; Malignant neoplasm of urinary bladder; Mesna; Modeling; Mus; Nerve; Nitric Oxide Synthase; Nuclear; Nuclear Localization Signal; Nuclear Translocation; Oncogenes; Overactive Bladder; Pain; Parasites; Pathway interactions; Patients; Peripheral; Production; Property; Proteins; RANTES; Recombinants; Resiniferatoxin; Rodent; Rodent Model; Role; Schistosoma haematobium; Schistosoma mansoni; Spinal Ganglia; Symptoms; Syndrome; TRPV1 gene; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Ulcer; Uropathogen; Urothelial Cell; Variant; Viral; Work; bladder pain; chemokine; cytokine; effective therapy; egg; gastrointestinal; ineffective therapies; intravesical; mast cell; mouse model; mutant; novel therapeutics; pain behavior; pain receptor; patient population; spontaneous pain; standard of care; therapeutic candidate

We propose to therapeutically exploit Schistosoma haematobium-derived IPSE (Interleukin-4 inducing Principle from Schistosoma mansoni Eggs) to mitigate bladder hypersensitivity. The incidence of bladder hypersensitivity (i.e., interstitial cystitis/bladder pain syndromes and overactive bladder) in some patient populations is over 40%, and existing treatments are ineffective for many. Thus, novel therapeutics are needed. The S. mansoni homolog of IPSE (M-IPSE) alters host biology by three distinct mechanisms: 1) translocating into nuclei of target cells to alter gene transcription; 2) binding IgE on basophils and mast cells to trigger IL-4 secretion; and 3) sequestering chemokines. We have shown that the S. haematobium homolog of IPSE (H-IPSE) translocates into urothelial cell nuclei and binds IgE on basophils. Our in silico data show H- IPSE binds CCL2. Collectively, these findings illustrate that H-IPSE and M-IPSE share many host modulatory features, but their differences are unclear. Immune and non-immune modulation of the bladder may be a new management approach for bladder hypersensitivity. Blocking CCL2-CCR2 interactions in bladder nerves and dorsal root ganglia lowers pain in rodent models. IL-4 lessens ifosfamide-induced cystitis in mice and decreases bladder pain and overactivity. Yet, intravenous IL-4 has adverse side effects in patients. We have discovered that a single intravenous dose of H-IPSE is superior to both Mesna and recombinant IL-4 in suppressing ifosfamide-induced bladder hemorrhage, and that it reduces urinary frequency and pain behaviors through IL-4- and nuclear localization sequence-dependent mechanisms. In conclusion, a single dose of H- IPSE lessens bladder hypersensitivity in two mouse models through pathways consistent with the known properties of M-IPSE and H-IPSE. We hypothesize that H-IPSE and M-IPSE variants can be optimized as safe therapeutics for bladder hypersensitivity by defining and exploiting their nuclear localization signal-, IL-4-, and chemokine binding-reliant mechanisms. To test this hypothesis, we will, in Aim 1, define and optimize the mechanisms by which IPSE exerts therapeutic effects in bladder hypersensitivity through the generation of engineered mutants with: 1) greater nuclear translocation ability to which will dampen transcription of pain- related genes; 2) increased capacity to induce IL-4 production; and 3) enhanced sequestration of chemokines. In Aim 2, we will determine the chronic toxicity potential of IPSE. We hypothesize that H-IPSE and M-IPSE variants and mutants that induce apoptosis of intestinal epithelial cells or dendritic cells upon nuclear translocation will cause toxicity when given on a chronic basis. Finally, we theorize that IPSE variants/mutants which do not trigger cancer gene-related transcription upon nuclear translocation into urothelial cells will not cause bladder cancer. Our work is unique in that it demonstrates that a uropathogen-derived host modulatory molecule can be therapeutically exploited in bladder disease models. The completion of these studies will yield a better understanding of the multiple immune and non-immune aspects of bladder hypersensitivity and potentially identify therapeutic candidates for this constellation of symptoms.

我们建议治疗性地利用血吸虫血吸虫衍生的IPSE（白细胞介素-4诱导 原理来自曼氏血吸虫卵），以减轻膀胱超敏反应。膀胱癌的发病率 超敏反应（即，间质性膀胱炎/膀胱疼痛综合征和膀胱过度活动症 人口超过40%，现有的治疗方法对许多人无效。因此， needed.色葡萄IPSE的mansoni同源物（M-IPSE）通过三种不同的机制改变宿主生物学：1） 转移到靶细胞的细胞核中以改变基因转录; 2）结合嗜碱性粒细胞和肥大细胞上的IgE， 触发IL-4分泌;和3）隔离趋化因子。我们证明了S.血吸虫同系物 IPSE（H-IPSE）易位到尿路上皮细胞核中并结合嗜碱性粒细胞上的IgE。我们的电脑数据显示- IPSE结合CCL2。总的来说，这些发现说明H-IPSE和M-IPSE共享许多宿主调节因子， 他们的特点，但他们的区别还不清楚。膀胱的免疫和非免疫调节可能是一种新的 膀胱超敏反应的管理方法。阻断膀胱神经中的CCL2-CCR2相互作用， 背根神经节减轻啮齿动物模型的疼痛。IL-4减轻小鼠中异环磷酰胺诱导的膀胱炎， 减少膀胱疼痛和过度活动。然而，静脉注射IL-4对患者有不良副作用。我们有 发现单次静脉注射H-IPSE优于美司钠和重组IL-4， 抑制异环磷酰胺诱导的膀胱出血，并减少尿频和疼痛行为 通过IL-4和核定位序列依赖性机制。总之，单剂量的H- IPSE在两种小鼠模型中减轻膀胱超敏反应，其途径与已知的 M-IPSE和H-IPSE的性质。我们假设H-IPSE和M-IPSE变体可以被优化为安全的 通过定义和利用它们的核定位信号-IL-4-和 趋化因子结合依赖机制。为了验证这一假设，我们将在目标1中定义并优化 IPSE通过产生 工程突变体具有：1）更大的核转位能力，其将抑制疼痛的转录， 相关基因; 2）增加诱导IL-4产生的能力;和3）增强趋化因子的螯合。 在目标2中，我们将确定IPSE的潜在慢性毒性。我们假设H-IPSE和M-IPSE 诱导肠上皮细胞或树突细胞凋亡的变体和突变体， 当长期给予时，易位将导致毒性。最后，我们推测IPSE变体/突变体 其在核转位进入尿路上皮细胞时不触发癌基因相关转录， 会导致膀胱癌。我们的工作是独特的，因为它表明，尿路病原体来源的宿主调节 分子可以在膀胱疾病模型中进行治疗性开发。完成这些研究后， 更好地了解膀胱超敏反应的多重免疫和非免疫方面， 潜在地识别针对该症状群的治疗候选物。