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的，免疫增强的L。乳酸菌益生菌为基础的治疗， 干燥综合征（Sjogren’s Syndrome，SjS）SjS是一种进行性慢性自身免疫性疾病，其特征在于 炎性细胞浸润的唾液腺和泪腺，导致腺泡上皮细胞萎缩，细胞 死亡和外分泌功能丧失1 - 6。至少有一半的SjS患者发生腺外炎症 疾病，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%的患者发展为危及生命的淋巴瘤7，8。SjS是一种使人衰弱的疾病 在美国，影响多达310万人9，10，其中女性受影响的可能性是女性的9倍。 比男性更容易患上SJS 5，10，11. SjS的治疗仍然是一个重大的未满足的医疗需求。目前的治疗依赖于替代疗法 例如人工唾液和眼睛润滑剂或免疫抑制剂12，13。因为有多种抗原 参与了这一疾病过程，即，α-fodrin 14 - 17，核糖核蛋白Ro/SSA 14，48 - 50，La/SSB 14，48，49和 M3R18，49，51，口服耐受性方法变得有问题。因此，刺激调节细胞的能力 不依赖于已知疾病的抗原特异性，这是一种有吸引力的治疗方法。 最初设想作为一种人大肠杆菌疫苗，我们发现来自大肠杆菌的定植因子抗原I（CFA/I） 人肠毒素E.大肠杆菌（ETEC）在诱导自身抗原特异性T调节细胞时有效， 作为纯化蛋白或通过沙门氏菌或L.乳酸菌传递 system19 - 21，63.为了避免与生产大量CFA/I蛋白相关的挑战并改善CFA/I蛋白的生物学特性， 口服给药后CFA/I的粘膜药代动力学（PK）和药效学（PD）特性， 我们开发并表征了一种含矢量的L.乳酸杆菌-CFA/I表达产物（称为VTC- CFA）并评估其在多种自身免疫模型中的功效。事实上，VTC-CFA有效地减少了 SjS动物模型中的临床症状，沿着RA和MS的实验模型22 - 24。在SBIR中 第一阶段，我们在小鼠模型中确定了VTC-CFA的胃肠道（GI）PK和最佳有效剂量。 关于SJS此外，我们开发了我们的临床候选物，其从基因组整合操纵子表达CFA/I。 （称为VTC-CFA-001），并完成了FDA IND前会议，以最终确定我们的IND启用计划。 该应用旨在将VTC-CFA推向临床。这项建议的主要目的是：1） 优化VTC-CFA-001上游工艺开发，以实现一致的生产，2）准备GMP主文件 和工作细胞库，以支持VTC-CFA-001生产，3）生产VTC-CFA-001，以进行剂量 4）确定VTC-CFA-001的最佳剂量并鉴定/表征生物标志物， 以及5）完成VTC-CFA-001 GLP毒理学研究以实现IND申报。 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.