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

我们建议利用血吸虫血吸虫来源的IPSE(白介素4诱导)进行治疗 曼氏血吸虫卵的精华成分)来缓解膀胱过敏症。膀胱癌的发病率 某些患者的过敏症(即间质性膀胱炎/膀胱痛综合征和膀胱过度活动) 人口超过40%，现有的治疗方法对许多人无效。因此，新的治疗方法是 需要的。曼氏链球菌IPSE同源物(M-IPSE)通过三种不同的机制改变宿主生物学：1) 移位到靶细胞的核中改变基因转录；2)与嗜碱性粒细胞和肥大细胞上的IgE结合以 触发IL-4的分泌；以及3)隔离趋化因子。我们已经证明了苏云金杆菌的同源物 IPSE(H-IPSE)转位到尿路上皮细胞核，结合嗜碱性粒细胞上的IgE。我们的电子数据显示H- IPSE绑定CCL2。综上所述，这些发现说明H-IPSE和M-IPSE共享许多宿主调控 功能，但它们的区别尚不清楚。免疫和非免疫对膀胱的调节可能是一种新的 膀胱过敏症的处理方法。阻断膀胱神经CCL2-CCR2的相互作用 背根神经节减轻啮齿动物模型的疼痛。IL-4减轻异环磷酰胺诱导的小鼠膀胱炎 减少膀胱疼痛和过度活动。然而，静脉注射IL-4对患者有不良反应。我们有 发现单次静脉注射H-IPSE优于MESNA和重组IL-4 抑制异环磷酰胺引起的膀胱出血，减少尿频和疼痛行为 通过IL-4和核定位序列依赖的机制。总而言之，单剂量的H- IPSE通过与已知途径一致的途径减轻两种小鼠模型的膀胱过敏反应 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 通过定义和利用其核定位信号-、IL-4-和IL-4治疗膀胱超敏反应 趋化因子结合依赖机制。为了检验这一假设，我们将在目标1中定义和优化 异丙肾上腺素治疗膀胱超敏反应的机制研究 工程突变体具有：1)更强的核转位能力，将抑制Pain-2的转录- 相关基因；2)诱导IL-4产生的能力增加；以及3)加强趋化因子的隔离。 在目标2中，我们将确定IPSE的慢性毒性潜力。我们假设H-IPSE和M-IPSE 核上诱导肠上皮细胞或树突状细胞凋亡的突变体和突变体 如果长期服用，易位将导致毒性。最后，我们推论IPSE变体/突变体 它们不会在核转位到尿路上皮细胞时触发癌症基因相关转录 导致膀胱癌。我们的工作的独特之处在于它证明了一种尿路病原体衍生的宿主调节 分子可用于治疗膀胱疾病模型。这些研究的完成将产生 更好地了解膀胱过敏症和膀胱炎的多重免疫和非免疫方面 有可能为这一系列症状确定治疗候选药物。

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.