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: 10707830
• 负责人: Bill Thelin
• 金额: $ 91.37万
• 依托单位: ALTIS BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707830
• 关键词: 3-Dimensional; Address; Adopted; Adverse event; Age; Animal Model; Animals; Apical; Area; Benchmarking; Biological Assay; Biological Models; Body mass index; Cell Differentiation process; Cell Survival; Cell model; Cell physiology; Cells; Cellular Structures; Cessation of life; Clinical; Clinical Data; Clinical Trials; Colon; Competence; Complement; Culture Media; Data; Data Analyses; Data Set; Development; Diarrhea; Disease; Dose; Dose Limiting; Drug Evaluation; Drug Exposure; Drug Kinetics; Drug Screening; Drug Targeting; Engineering; Epithelium; Excision; Exposure to; Fee-for-Service Plans; Feedback; Female; Gastrointestinal tract structure; Goals; Human; Image; Impairment; In Vitro; Industry; Inflammation; Intestines; Life Cycle Stages; Manufacturer; Marketing; Measures; Microinjections; Modeling; Oncology; Organoids; Outcome; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiology; Plasma; Population; Proliferating; Property; Protocols documentation; Publishing; Rattus; Rho-associated kinase; Risk; Running; Safety; Small Intestines; Source; Technology; Testing; Therapeutic; Therapeutic Effect; Tissue Donors; Tissues; Toxic effect; Toxicology; Transformed Cell Line; Transverse colon; Tumor Cell Line; Tumor-Derived; Ulcer; Validation; Work; adverse event risk; cell type; clinical translation; clinically relevant; commercial launch; cost effective; demographics; detection assay; drug mechanism; ethnic diversity; experimental study; gastrointestinal; gastrointestinal epithelium; gastrointestinal system; improved; in vitro Model; in vivo; interest; intestinal barrier; intestinal epithelium; jejunum; kinase inhibitor; lead optimization; male; meter; monolayer; novel; novel therapeutics; phase 1 study; phase 2 study; phase III trial; preclinical safety; preclinical study; regenerative cell; repaired; research clinical testing; risk mitigation; screening; screening services; self-renewal; side effect; stem; stem cell population; stem cell proliferation; stem cells; success; therapeutic candidate; timeline

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 included assays related to stem cell proliferation, self- renewal, and differentiation competency. In our Phase 1 studies, we developed a 5 day, 96-well, plate-based screening assay (termed StemTox) and tested a panel of pharmaceutical agents known to trigger GI side effects. The results revealed a highly sensitive and dose-dependent inhibition of stem cell proliferation and impairment of intestinal barrier formation. Furthermore, our assay detected “adverse” concentrations for compounds that closely paralleled the clinical plasma Cmax associated with human clinical outcomes. In this Phase 2 application, we propose to complete optimization of the StemTox screen, generate a commercial validation data set (using multiple tissue donors and cell lots of transverse colon), and expand the model to integrate cell differentiation and long-term drug exposures. Collectively, this work will result in the commercial launch of the StemTox assay, as well as, enable several follow-on experimental paradigms to more comprehensively assess off-target drug mechanisms and repeat- dosing experiments (to parallel current IND-enabling nonclinical study requirements).

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。其中包括与干细胞增殖有关的测定 更新和差异能力。 在我们的第一阶段研究中，我们开发了5天的96孔基于板的筛选测定法（称为stesttox） 并测试了一组已知触发GI副作用的药物。结果表明了高度 敏感和剂量依赖性抑制干细胞增殖和肠屏障形成的损害。 此外，我们的测定检测到与临床紧密平行的化合物的“不利”浓度 血浆CMAX与人类临床结果相关。在此阶段2应用程序中，我们建议完成 优化StemTox屏幕，生成商业验证数据集（使用多个组织供体和 细胞大量的横向结肠），并扩展模型以整合细胞分化和长期药物暴露。 总的来说，这项工作将导致StemTox分析的商业发布，并启用几个 后续实验范式更全面地评估靶向脱靶药物机制和重复 给药实验（达到平行的目录的非临床研究要求）。