Planar culture of gastrointestinal stem cells for screening pharmaceuticals for adverse event risk

胃肠道干细胞平面培养用于筛选药物不良事件风险

• 批准号: 10482465
• 负责人: Christopher Eldridge Sims
• 金额: $ 25.66万
• 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: 2022-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10482465
• 关键词: 3-Dimensional; Address; Adherent Culture; Adverse event; Animal Model; Animals; Apical; Area; Ascending colon; Benchmarking; Biological Assay; Biomimetics; Cell Culture Techniques; Cell Proliferation; Cell Survival; Cell physiology; Cells; Cellular Structures; Cessation of life; Characteristics; Clinical; Clinical Trials; Collaborations; Collection; Colon; Data; Descending colon; Detection; Development; Diarrhea; Disease; Dose; Dose-Limiting; Drug Evaluation; Drug Screening; Duodenum; Electrical Resistance; Engineering; Epithelial; Exposure to; Gastrointestinal Physiology; Gastrointestinal tract structure; Goals; Human; In Vitro; Industry; Inflammation; Intestines; Laboratories; Lead; Literature; Manufacturer Name; Measurement; Measures; Membrane; Microinjections; Modality; Modeling; Monitor; Oncology; Organ Donor; Organ Transplantation; Organoids; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase; Phenotype; Physiology; Population; Property; Rattus; Reproducibility; Risk; S phase; Sampling; Screening procedure; Side; Small Intestines; Source; Stains; Surface; Technology; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Tissue Donors; Tissues; Toxic effect; Transformed Cell Line; Transverse colon; Tumor Cell Line; Tumor-Derived; Ulcer; Variant; adverse event risk; base; biobank; cell type; cost effective; drug testing; gastrointestinal; gastrointestinal epithelium; gastrointestinal system; high risk; ileum; in vitro Assay; in vitro Model; in vivo; indexing; intestinal epithelium; jejunum; manufacturing process; monolayer; novel; novel therapeutics; phase III trial; preclinical safety; preclinical study; prevent; programs; regenerative cell; repaired; research clinical testing; scaffold; screening; side effect; stem; stem cell differentiation; stem cell population; stem cell proliferation; stem cells; stemness; success; therapeutic candidate; transcriptomics

Project Summary GI side effects, such as ulcers and diarrhea, represent the most common source of adverse events for pharmaceuticals. GI stem cells are responsible for repairing and replenishing GI epithelium, and pharmaceutical inhibition of these functions likely contribute to adverse event risk. Currently there are no high-throughput and cost-effective means of screening candidate therapeutics for effects on GI stem cells. Animal models are fraught with confounds (e.g., rats generally do not exhibit diarrhea until death is imminent) and current in vitro models like the Caco-2 tumor cell line do not include a normal stem cell population. While 3D-organoid cultures have a stem cell component, access to the apical aspect of the monolayer for compound exposure is not possible without low-throughput microinjection. Altis Biosystems, Inc. has developed a proprietary culture platform, RepliGut® Planar, enabling primary human GI cells to form an epithelium for drug screening. In preliminary efforts, we initiated development of a GI stem cell-specific platform called RepliGut® StemScreen to address the unmet need for high-throughput, cost-effective GI stem cell screening, including a range of assays to measure properties that might lead to adverse events. These include assays for proliferation using S-phase staining and assessing downstream differentiation and ability to establish barrier function via non-destructive transepithelial electric resistance (TEER) analysis. A panel of pharmaceutical agents associated with clinically adverse GI side effects, tested in a classic EdU assay using StemScreen planar cultures, demonstrated dose-dependent inhibition of stem cell proliferation and prevented stem cell differentiation into a mature epithelium with barrier function. During Phase I we will: 1) characterize monolayer proliferation properties of donor cells in the Altis biobank(initially transverse colon stem cells will be evaluated, with other regions tested in Phase II) from three donors and 2) test a selection of therapeutic agents with known GI adverse event risks for effects on stem cell proliferation, impacting differentiation and the formation of a functional epithelial barrier (indexed as TEER). Demonstrating that the StemScreen platform provides an in vitro screening tool capable of predicting a drug’s risk for GI adverse events in Phase I will enable deeper characterization of stemness, stem cells from other GI regions (e.g., ascending and descending colon; small intestine regions including jejunum, etc.), and stem cell phenotypes. In Phase 2, we plan externally validate the platform in collaboration with leading industry laboratories prior to commercial launch.

项目摘要 GI副作用，例如溃疡和腹泻，代表了广告事件的最常见来源 药品。 GI干细胞负责修复和复制GI上皮和药物 对这些功能的抑制可能导致不良事件风险。目前没有高通量 筛查候选疗法对GI干细胞的影响的成本效益手段。动物模型是 混杂（例如，在死亡即将死亡之前，大鼠通常不存在腹泻）和当前的体外模型 像Caco-2肿瘤细胞系一样，不包括正常的干细胞群体。而3D-核文化具有 干细胞组件，无法访问单层的顶端进行复合曝光 没有低通量显微注射。 Altis Biosystems，Inc。开发了一个专有的文化平台， Repligut®平面，使原代人GI细胞形成用于药物筛查的上皮。在初步 努力，我们启动了一个称为repligut®stemscreen的GI干细胞特异性平台，以解决 未满足的高通量，具有成本效益的GI干细胞筛选，包括一系列测量 可能导致广告事件的属性。这些包括使用S期染色和 评估下游的分化和通过非破坏性换皮建立屏障功能的能力 电阻（TEER）分析。 与临床上不良GI副作用相关的一组药物面板，在经典中进行了测试 使用干屏平面培养物的EDU测定，证明了剂量依赖性抑制干细胞增殖 并防止干细胞分化为具有屏障功能的成熟上皮。在第一阶段我们将：1） 表征供体细胞在Altis Bioband（最初是横向结肠茎）中的单层增生特性 将评估细胞的其他区域，在II阶段中测试了三个捐赠者，2）测试选择的 具有已知GI不良事件的治疗剂对干细胞增殖的影响风险，影响 分化和功能性上皮屏障的形成（索引为TEER）。证明 STEM屏幕平台提供了一种体外筛查工具，能够预测药物发生GI不良事件的风险 在第一阶段，将使其他胃肠道区域的干细胞更深入地表征（例如，上升 并下降结肠；小肠区域，包括空肠等）和干细胞表型。在第2阶段， 我们计划在商业之前与领先的行业实验室合作验证平台 发射。