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

项目摘要 广泛的炎症性胃肠道疾病会影响美国数以百万计的人和 国外。该疾病的病因是多因素和异质性的，但共同的潜在特征是 过度活动的免疫系统。遗传学，环境因素，年龄，性别，种族，甚至 居住在肠道内的细菌与肠道炎症的因果关系有关。而有些治疗 已经取得了进步，许多患者对现有药物干预措施没有反应 留下许多人有终身发病率。这为改善现有疗法和 发现替代性和新颖的方法来应对肠道的炎症状况。临床前模型 用于药物研究和发现历史上一直很差。它们由癌细胞组成 生理相关性，不具备免疫细胞室，这是缺失的关键成分 实现人类炎症条件的准确模型。由于这些原因， 体外肠平台的治疗市场 肠的免疫细胞轴。为了满足这种需求，Altis Biosystems Inc.是一种早期生物技术 公司与学术实验室的科学家合作，开发了一种基于新型的，基于单元的小细胞， 体外肠模型（称为复制）。我们已经完成了针对转基因的SBIR I期计划 以及在复制中培养的肠道干细胞（ISC）的基因编辑。我们开发了记者 - 基因的ISC 在肠道炎症的实时关键特征中产生差异化的上皮以及感官和报告，1）障碍 功能和2）NF-KB激活。在这里，我们扩展范围，以生成一种称为“ 炎症。炎症是一种三方且灵活的系统，它使用了Repligut和 叠加炎症记者基因和人类上皮共培养与同种异体肠相关 淋巴细胞（GAL）重新创建lei轴。设想了四种产品：1）炎症：ZO1和 炎症：NF-KB记者基因上皮，2）炎症：共培养：上皮/肠道/肠道促进淋巴细胞 （GAL）共培养系统，3）可商业化的人类肠道相关淋巴细胞和4）新合同 研究服务（CRS）使用炎症。有强大的工程和生物学方法创新， 包括，1）Transwell的发展插入启用可扩展的文化，以补充分化的人类 水凝胶封装的淋巴细胞室上的上皮，以及2）分离，培养和冷冻鸟 来自人体器官捐献者的GAL。炎症将由学术合作者建立，验证和测试 （Repligut Technology的创始人），Altis Biosystems和两个行业合作者。 1