A Novel IL-35 Expressing Probiotic Platform for Inducing Allergen Specific Tolerance

项目摘要 我们的目标是开发VTC-L35作为治疗哮喘的新的耐受诱导益生菌平台。 根据美国疾病控制与预防中心的数据，美国有超过2500万人患有哮喘。在全球，近3.5亿 人们患有哮喘，每年有30多万人死于这种疾病(CDC)。 由于哮喘从根本上说是一种免疫失调疾病，目前许多哮喘治疗方法都是 免疫抑制药，包括类固醇。虽然这些在控制哮喘症状方面是有效的 许多患者的效果是暂时的，有相当大的副作用，需要持续给药。 哮喘“发作”是由各种环境刺激引发的，其中最常见的是 空气过敏原。脱敏方案是存在的，但考虑到大多数 哮喘患者有多种过敏原，通常是不明的。因此，迫切需要新的疗法来 让哮喘患者耐受他们自己的“个性化”过敏原。 最近，我们项目组开发了一种新型的乳酸乳球菌(L.lactis)研究菌株，该菌株表达IL-35 (职训局-L35)口服VTC-L35可有效降低糖尿病的发病率和疾病严重程度。 胶原性关节炎小鼠模型中预防和治疗方案中的炎症 模特(CIA)。VTC-L35诱导CIA小鼠CCR6+和CCR6CD39+−+Treg细胞。对他们的调查 诱导发现CCR6+和CCR6CDFoxp3+/或CD39+−+T细胞都是−的来源。 VTC-L35诱导的IL-10。 我们相信，在个人环境过敏原存在的情况下口服VTC-L35将 诱导抗原特异性调节细胞，恢复对个人过敏原的耐受性，逆转哮喘 症状。为了验证我们的方法，我们计划演示口服VTC-L35在 低剂量屋尘螨(HDM)抗原的存在可预防过敏和呼吸道症状 老鼠模型。随后，我们将开发和验证基因组整合的临床菌株。高层 本次快速跟踪SBIR应用的目的是：1)展示我们的VTC-L35研究菌株诱导 无可检测到短期毒性的小鼠HDM过敏模型的耐受性；2)建立和表征一种 从基因组整合操纵子表达人IL-35的临床候选VTC-L35；以及3)开发非 GLP和GLP临床前数据集，以支持VTC-L35 IND批准。 该方案的完成将建立一种新型的表达IL-35的益生菌平台，可应用于 广泛治疗哮喘、过敏，甚至一系列自身免疫性疾病。

Project Summary Our goal is to develop VTC-L35 as a novel tolerance-inducing probiotic platform for the treatment of asthma. According to the CDC, more than 25 million people in America suffer from asthma. Globally, nearly 350 million people suffer from asthma, with more than 300,000 annual deaths attributed to the disease (CDC). Because asthma is fundamentally a disease of immune dysregulation, many current asthma treatments are immunosuppressive agents, including steroids. While these are effective at managing asthma symptoms for many patients, the effects are temporary, have considerable side-effects, and require constant administration. Asthma “attacks” are triggered by a variety of environmental stimuli, the most common of which are aeroallergens. Desensitization protocols exist but are intensive and impractical given the fact that most asthmatics have multiple, often unidentified, allergens. As a result, new therapies are desperately needed to tolerize asthmatics against their own “personalized” allergens. Recently, our project team developed a novel Lactococcus lactis (L. lactis) research strain expressing IL-35 (VTC-L35)26. Oral administration of VTC-L35 effectively reduced the incidence and disease severity of inflammation in both prophylactic and treatment protocols in the mouse model of collagen-induced arthritis model (CIA). VTC-L35 induced CCR6+ and CCR6− CD39+ CD4+ Treg cells in CIA mice. Inquiry into their induction revealed that both CCR6+ and CCR6− Foxp3+/or− CD39+ CD4+ T cells act as the source of the IL-10 induced by VTC-L35. We believe that oral administration of VTC-L35 in the presence of an individual’s environmental allergens will induce antigen-specific regulatory cells and restore tolerance to an individual’s allergens, reversing asthma symptoms. To validate our approach, we plan to demonstrate that oral administration of VTC-L35 in the presence of low amounts of house dust mite (HDM) antigens prevents allergic and airway symptoms in a mouse model. Subsequently, we will develop and validate a genome-integrated clinical strain. The high-level aims of this Fast-track SBIR application are to 1) demonstrate that our VTC-L35 research strain induces tolerance in a mouse HDM allergy model with no detectable short-term toxicity; 2) develop and characterize a VTC-L35 clinical candidate expressing human IL-35 from a genome-integrated operon; and 3) develop non- GLP and GLP preclinical datasets to support VTC-L35 IND approval. Completion of this proposal will establish a novel IL-35-expressing probiotic platform that can be applied to treat asthma, allergies broadly and even an array of autoimmune diseases.