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.

我们建议通过治疗利用血吸虫血吸虫衍生的IPSE（白介素-4诱导 曼森卵的原理）减轻膀胱超敏反应。膀胱的发生率 某些患者的高敏性（即间质膀胱炎/膀胱疼痛综合征和过度活跃的膀胱） 人口超过40％，现有治疗对许多人的效果无效。因此，新颖的治疗学是 需要。 IPSE的S. Mansoni同源物（M-IPSE）通过三种不同的机制来改变宿主生物学：1） 转移到靶细胞的核中以改变基因转录； 2）在嗜碱性粒细胞和肥大细胞上结合IgE与 触发IL-4分泌； 3）隔离趋化因子。我们已经表明 IPSE（H-IPSE）转移到尿路上皮细胞核中，并在嗜碱性粒细胞上结合IgE。我们的硅数据显示H- IPSE绑定CCL2。总的来说，这些发现表明H-IPSE和M-IPSE共享许多主机调节 特征，但它们的差异尚不清楚。膀胱的免疫和非免疫调节可能是新的 膀胱超敏反应的管理方法。阻止膀胱神经中的CCL2-CCR2相互作用 背根神经节降低了啮齿动物模型的疼痛。 IL-4减少了小鼠IFOSFAMIDE诱导的膀胱炎和 减轻膀胱疼痛和过度活动。然而，静脉注射IL-4在患者中具有不利的副作用。我们有 发现单个静脉剂量的H-IPSE优于Mesna和重组IL-4 抑制IFOSFAMIDE引起的膀胱出血，并降低尿频和疼痛行为 通过IL-4和核定位序列依赖性机制。总之，一剂h- IPSE通过与已知的途径相一致 M-IPSE和H-IPSE的性质。我们假设H-IPSE和M-IPSE变体可以优化为安全 通过定义和利用其核定位信号，IL-4-和 趋化因子结合利益机制。为了检验这一假设，我们将在AIM 1中定义和优化 IPSE通过产生的机制在膀胱超敏反应中发挥治疗作用 具有以下工程的突变体：1）更大的核转运能力会抑制疼痛的转录 - 相关基因； 2）增加诱导IL-4产生的能力； 3）增强趋化因子的隔离。 在AIM 2中，我们将确定IPSE的慢性毒性潜力。我们假设H-IPSE和M-IPSE 诱导肠上皮细胞或树突状细胞凋亡的变体和突变体 慢性化时，易位会导致毒性。最后，我们认为IPSE变体/突变体 在核易位到尿路上皮细胞时不会引发癌症基因相关的转录 引起膀胱癌。我们的工作是独一无二的 分子可以在膀胱疾病模型中进行治疗。这些研究的完成将产生 更好地了解膀胱超敏反应的多种免疫和非免疫方面 有可能确定症状星座的治疗候选者。