A Novel Probiotic for the Treatment of Sjogren's Syndrome

项目摘要 我们的目标是开发一种新型的，免疫增强的乳酸乳杆菌益生菌为基础的治疗 干燥综合征(SjS)的治疗。SJS是一种进行性慢性自身免疫性疾病，其特征是 炎性细胞渗入唾液和泪腺，导致腺泡上皮细胞萎缩。 死亡，外分泌功能丧失1-6。至少一半的SjS患者发展为腺外炎症 并有广泛的全身性临床表现，可影响任何器官系统，包括 结缔组织，5-10%的患者患上危及生命的淋巴瘤7、8.SJS是一种衰弱的疾病 在美国，有多达310万人受到影响，其中女性患病的可能性是9倍。 比男人5，10，11更受Sjs的困扰。 SjS的治疗仍然是一个重要的未得到满足的医疗需求。目前的治疗依赖于替代疗法 例如人工唾液和眼部润滑剂或免疫抑制剂12、13。因为多种抗原 参与该疾病过程的有α-fodrin14-17、核糖核蛋白Ro/SSA14、48-50、La/SSB14、48、49和 M3R18，49，51，口服耐受方法变得有问题。因此，刺激调节细胞的能力 与已知该病的抗原特异性无关，它被认为是一种有吸引力的治疗方法。 最初被认为是人类腹泻疫苗，我们发现定居因子抗原I(CFA/I)来自 人肠毒素大肠杆菌(ETEC)在以下情况下可有效诱导自身抗原特异性T调节细胞 口服作为纯化蛋白或通过沙门氏菌或乳酸乳杆菌细菌传递 系统19-21，63。为了避免与大量生产CFA/I蛋白相关的挑战，并改善 CFA/I口服后的粘膜药代动力学(PK)和药效学(PD)特性 我们开发并鉴定了含有乳酸乳杆菌CFA/I的载体表达产物(简称VTC-I)。 CFA)，并在多种自身免疫模型中评估其疗效。事实上，职训局-终审法院在减少 SjS动物模型的临床症状，以及RA和MS22-24的实验模型。在SBIR中 第一阶段，我们在小鼠模型中确定了胃肠(GI)PK和VTC-CFA的最佳有效剂量 SjS的。此外，我们开发了从基因组整合操纵子表达CFA/I的临床候选基因 (简称VTC-CFA-001)，并完成了FDA的IND前会议，以最终确定我们的IND支持计划。 这项申请旨在推动VTC-CFA走向临床。这项建议的主要目的是：1) 优化VTC-CFA-001上游工艺开发以实现一致的制造，2)准备GMP母版 和支持VTC-CFA-001生产的工作细胞库，3)制造VTC-CFA-001以执行剂量 优化和毒理学，4)确定VTC-CFA-001的最佳剂量并识别/表征生物标志物， 和5)完成VTC-CFA-001 GLP毒理学研究，以便进行IND备案。 VTC-CFA-001的成功商业化将为治疗SJS提供深刻的医学进步。

Project Summary Our goal is to develop a novel, immunologically enhanced L. lactis probiotic-based therapeutic for the treatment of Sjögren’s Syndrome (SjS). SjS is a progressive, chronic autoimmune disease characterized by inflammatory cell infiltration of the salivary and lacrimal glands, resulting in acinar epithelial cell atrophy, cell death, and loss of exocrine function1-6. At least half of SjS patients develop extraglandular inflammatory disease and have a wide range of systemic clinical manifestations that can affect any organ system, including connective tissue, and 5-10% of patients develop life-threatening lymphoma7, 8. SjS is a debilitating disease affecting as many as 3.1 million individuals in the U.S.9, 10, with women being nine times more likely to be afflicted with SjS than men5, 10, 11. Treatment of SjS remains a significant unmet medical need. Current treatment relies on replacement therapies such as artificial saliva and eye lubricants or immunosuppressive agents12, 13. Because of the multiple antigens involved in this disease process, i.e., α-fodrin14-17, ribonuclear protein Ro/SSA14, 48-50, La/SSB14, 48, 49, and M3R18, 49, 51, oral tolerance methods become problematic. Thus, the capacity to stimulate regulatory cells independent of knowing the antigen specificity for the disease poses as an attractive therapeutic approach. Originally conceived as a human diarrheal vaccine, we found that colonization factor antigen I (CFA/I) from human enterotoxigenic E. coli (ETEC) is effective at inducing auto-Ag-specific T regulatory cells when administered orally as either a purified protein or delivered via a Salmonella or L. lactis bacterial delivery system19-21, 63. To avoid challenges associated with producing large quantities of CFA/I protein and improve the mucosal pharmacokinetic (PK) and pharmacodynamic (PD) properties of CFA/I following oral administration, we developed and characterized a vector-containing L. lactis-CFA/I expressing product (referred to as VTC- CFA) and evaluated its efficacy in multiple autoimmune models. Indeed, VTC-CFA was effective at reducing clinical symptoms in an SjS animal model, along with experimental models of RA and MS22-24. In the SBIR Phase I, we determined the gastrointestinal (GI) PK and optimal effective dose of VTC-CFA in a murine model of SjS. In addition, we developed our clinical candidate that expresses CFA/I from a genome-integrated operon (referred to as VTC-CFA-001) and completed an FDA pre-IND meeting to finalize our IND-enabling plans. This application is designed to advance VTC-CFA towards the clinic. The key aims of this proposal are: 1) optimize VTC-CFA-001 upstream process development for consistent manufacturing, 2) prepare GMP master and working cell banks to support VTC-CFA-001 production, 3) manufacture VTC-CFA-001 to perform dose optimization and toxicology, 4) Determine optimal dose of VTC-CFA-001 and identify/characterize biomarkers, and 5) complete VTC-CFA-001 GLP toxicology studies to enable IND filing. Successful commercialization of VTC-CFA-001 will provide a profound medical advancement for treating SjS.