InflammaGut: a drug-screenable co-culture system using gut-associated lymphocytes and autologous primary human gut epithelium that reports inflammation

InflammaGut：一种可药物筛选的共培养系统，使用肠道相关淋巴细胞和自体原代人类肠道上皮来报告炎症

• 批准号: 10616555
• 负责人: Christopher Eldridge Sims
• 金额: $ 98.84万
• 依托单位: ALTIS BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10616555
• 关键词: Acute; Affect; Age; Allogenic; American; Apical; Autologous; Bacteria; Biological; Biological Models; Biotechnology; Cell Communication; Cell Compartmentation; Cell Differentiation process; Cell Line; Cell Separation; Cell Survival; Cells; Chronic; Client; Coculture Techniques; Collaborations; Collagen; Colon; Colonic Neoplasms; Complex; Cryopreservation; Detection; Development; Dietary Factors; Disease; Drug Design; Drug Industry; Drug usage; Encapsulated; Engineering; Environment; Environmental Risk Factor; Epithelial Cells; Epithelium; Etiology; Evaluation; Event; Financial Hardship; Gastrointestinal Diseases; Gene Transfer Techniques; General Population; Genes; Genetic; Goals; Health; Human; Hydrogels; Immune; Immune system; Immunocompetent; In Vitro; Individual; Industry Collaboration; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Bowel Diseases; Intervention; Intestines; Laboratories; Lateral; Lymphocyte; Lymphoid Cell; Mediating; Methods; Modeling; Modification; Morbidity - disease rate; Mucous Membrane; NF-kappa B; Organ Donor; Patients; Peripheral Blood Lymphocyte; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological; Pre-Clinical Model; Production; Proteins; Quality of life; Race; Reporter; Reporter Genes; Reporting; Reproducibility; Research; Research Contracts; Science; Scientist; Services; Side; Signal Transduction; Small Business Innovation Research Grant; Small Intestines; Societies; Symptoms; System; Technology; Testing; Therapeutic; Tight Junctions; Time; Transgenic Organisms; Transplantation; Tumor Cell Line; United States; Validation; Withdrawal; cancer cell; cell motility; cohort; commercialization; culture plates; cytokine; demographics; drug candidate; drug market; flexibility; gastrointestinal; gastrointestinal epithelium; gut inflammation; high throughput screening; human model; human stem cells; immunoregulation; improved; innovation; microbiota; monolayer; novel; novel strategies; novel therapeutics; phase 1 study; pre-clinical; programs; screening; self-renewal; sex; stem cells; tissue culture; tumor-immune system interactions

Project Summary A broad range of inflammatory gastrointestinal conditions impact millions of people in the United States and abroad. The etiology of the disease is multifactorial and heterogeneous, but the common underlying feature is an overactive immune system. Genetics, environmental factors, age, sex, race and even the composition of bacteria that reside in the gut lumen are associated with causation of gut inflammation. While some therapeutic advances have been made, a large number of patients do not respond to existing pharmaceutical interventions leaving many to have life-long morbidity. This provides strong rationale to improve existing therapies and discover alternative and novel approaches to deal with inflammatory conditions of the gut. Preclinical models used for drug research and discovery have been historically poor. They are comprised cancer cells that have low physiological relevance and do not possess the immune cell compartment, which is a missing pivotal component to achieve an accurate model of human inflammatory conditions. For these reasons, there is a need in the therapeutics marketplace for an in vitro intestinal platform that accurately recapitulates luminal-epithelial- immune cell axis of the intestines. To meet this need, Altis Biosystems Inc., an early-stage biotechnology company, has collaborated with scientists at academic laboratories to develop a novel, primary-stem cells-based, in vitro intestinal model (termed RepliGut). We have finished an SBIR Phase I program focused on transgenesis and gene editing of intestinal stem cells (ISCs) cultured on RepliGut. We developed reporter-gene ISCs that generate differentiated epithelium and sense and report in real-time key features of gut inflammation, 1) barrier function, and 2) NF-kB activation. Here we expand the scope to generate a first-in-kind co-culture model called, InflammaGutTM. InflammaGut is a tripartite and flexible system that uses fundamental technology of RepliGut and superimposes inflammation reporter genes and co-culture of human epithelium with allogeneic gut-associated lymphocytes (GAL) to recreate the LEI-axis. Four products are envisioned: 1) InflammaGut:ZO1 and InflammaGut:NF-kB reporter gene epithelium, 2) InflammaGut:Co-Culture: an epithelial/gut-associate lymphocyte (GAL) co-culture system, 3) commercializable human gut-associated lymphocytes, and 4) a new contract research service (CRS) using InflammaGut. There is strong engineering and biological method innovation, including, 1) development of Transwell insert enabling scalable culture of RepliGut differentiated human epithelium over a hydrogel encapsulated lymphoid-cell compartment, and 2) isolation, culture, and cryobanking of GAL from human organ donors. InflammaGut will be built, validated, and tested by academic collaborators (founders of the RepliGut technology), Altis Biosystems, and two industry collaborators. 1

项目概要 多种炎症性胃肠道疾病影响着美国和美国的数百万人 国外。该疾病的病因是多因素且异质性的，但共同的基本特征是 过度活跃的免疫系统。遗传、环境因素、年龄、性别、种族甚至成分 肠腔内的细菌与肠道炎症有关。虽然有一些治疗作用 已经取得了进展，大量患者对现有的药物干预措施没有反应 使许多人终生患病。这为改进现有疗法和 发现治疗肠道炎症的替代和新颖方法。临床前模型 用于药物研究和发现的药物历史上一直很少。它们由癌细胞组成，具有低 生理相关性并且不具有免疫细胞区室，这是一个缺失的关键组成部分 以获得人类炎症状况的准确模型。由于这些原因，有必要 体外肠道平台的治疗市场，可准确概括管腔上皮- 肠道的免疫细胞轴。为了满足这一需求，Altis Biosystems Inc.（一家早期生物技术公司） 公司与学术实验室的科学家合作开发了一种新型的、基于原代干细胞的、 体外肠道模型（称为 RepliGut）。我们已经完成了专注于转基因的 SBIR 第一阶段计划 对在 RepliGut 上培养的肠道干细胞 (ISC) 进行基因编辑。我们开发了报告基因 ISC 生成分化的上皮并实时感知和报告肠道炎症的关键特征，1) 屏障 功能，以及 2) NF-kB 激活。在这里，我们扩大了范围，生成了一种名为“首创的共培养模型”， 肠道炎症TM。 InflammaGut 是一个三方灵活的系统，使用 RepliGut 的基础技术和 叠加炎症报告基因并与同种异体肠道相关的人类上皮细胞共培养 淋巴细胞 (GAL) 来重建 LEI 轴。设想了四种产品：1) InflammaGut:ZO1 和 InflammaGut：NF-kB 报告基因上皮，2) InflammaGut：共培养：上皮/肠道相关淋巴细胞 (GAL) 共培养系统，3) 可商业化的人类肠道相关淋巴细胞，以及 4) 新合同 使用 InflammaGut 的研究服务 (CRS)。有较强的工程和生物方法创新， 包括，1) 开发 Transwell 插入物，实现 RepliGut 分化人类的可扩展培养 水凝胶封装的淋巴细胞室上的上皮细胞，以及 2) 分离、培养和冷冻银行 来自人体器官捐献者的 GAL。 InflammaGut 将由学术合作者构建、验证和测试 （RepliGut 技术的创始人）、Altis Biosystems 和两个行业合作者。 1