F31 Mulero Russe

F31 穆莱罗·鲁斯

• 批准号: 10463993
• 负责人: Adriana Mulero-Russe
• 金额: $ 4.68万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10463993
• 关键词: 3-Dimensional; Address; Adhesives; Adult; Affect; American; Architecture; Area; Basement membrane; Biochemical; Biocompatible Materials; Biological Products; Biophysics; Caliber; Cell Therapy; Cells; Chemical Injury; Confocal Microscopy; Cues; Dependence; Development; Disease model; Drug Screening; Elasticity; Encapsulated; Endoderm; Engineering; Engraftment; Epithelial; Epithelial Cells; Extracellular Matrix; Fiber Optics; Formulation; Foundations; Gene Expression; Generations; Growth; Growth Factor; Hindgut; Human; Hydrogels; In Situ; In Vitro; Inflammation; Inflammatory Bowel Diseases; Intestinal permeability; Intestines; Knowledge; Length; Ligands; Maleimides; Measures; Mechanics; Membrane; Mesenchymal; Mus; Organoids; Patients; Polymers; Process; Property; Rattus; Recovery; Reproducibility; Research; Resolution; Small Intestines; Source; Structure; Technology; Therapeutic; Time; Tissues; Tube; Tumor-Derived; Ultraviolet Rays; Work; arm; base; cell type; clinical translation; clinically relevant; clinically translatable; delivery vehicle; density; directed differentiation; epithelial repair; epithelial wound; ethylene glycol; experience; healing; human pluripotent stem cell; in vitro Model; in vivo; inflammatory milieu; injured; innovation; insight; intestinal crypt; intestinal epithelium; intestinal injury; matrigel; microbiota; monolayer; nutrient absorption; polarized cell; process repeatability; programs; repaired; self organization; stem cells; tissue repair; tool; wound; wound healing

PROJECT SUMMARY Inflammatory Bowel Diseases (IBD) currently affect >1.6 million Americans. IBD is characterized by disruption to the intestinal epithelium and a high inflammatory environment. Available treatments target the inflammation through biological agents, however, fewer efforts have focused on epithelium healing and there are no broadly applicable therapies to repair intestinal epithelium. Human intestinal organoids (HIOs) are three-dimensional (3D) multicellular structures, derived from either adult intestinal stem cells or human pluripotent stem cells (hPSCs), that recapitulate human intestinal tissue architecture. HIOs are a promising cell source for intestinal epithelium repair, disease modeling, and drug screening. Previous work has demonstrated that HIOs engraft to the injured intestinal wall in vivo, however, these approaches are significantly limited by the lack of an appropriate delivery vehicle to drive HIO engraftment. HIO generation from hPSCs is multi-stage directed differentiation process comprising three stages: (I) differentiation into a definitive endoderm monolayer, (II) hindgut and primitive tube differentiation into free-floating, self-organized 3D aggregates (human intestinal spheroids, HIS), and (III) intestinal specification into HIOs within a 3D extracellular matrix. This in vitro culture process spans a 2D growth substrate (stage I and II) to a 3D matrix (stage III). Stage III requires culture within Matrigel, a murine tumor- derived basement membrane extract with ill-defined composition, lot-to-lot variability, and limited clinical translation potential. Another roadblock to HIO technologies is the low yield and consistency of HIS differentiation in HIOs. The objectives of this project are to (1) engineer a synthetic hydrogel platform with independent control of the biochemical and biophysical cues guiding the entire in vitro differentiation of hPSCs into HIOs, and (2) deliver HIOs in a synthetic coating to intestinal injuries in vivo. The central hypothesis is that an engineered synthetic matrix with appropriate biophysical and biochemical cues will support the HIO self-organization, growth, and differentiation process and enhance HIO engraftment and healing of intestinal wounds. Aim 1: Engineer a 2D synthetic matrix promoting spheroid generation from hPSCs. Aim 2: Evaluate the maturation of HIOs from the generated spheroids within synthetic niches. Aim 3: Engineer a clinically translatable therapeutic delivery material for HIOs to injured intestinal tissue. The results of this study will increase the clinical relevance the generated HIOs and will provide a scalable and translatable synthetic material for the differentiation and delivery of HIOs.

项目概要 目前，炎症性肠病 (IBD) 影响着超过 160 万美国人。 IBD 的特点是破坏 肠上皮和高炎症环境。可用的治疗方法针对炎症 然而，通过生物制剂，针对上皮愈合的努力较少，并且没有广泛的研究 适用于修复肠上皮的疗法。人类肠道类器官 (HIO) 是三维 (3D) 多细胞结构，源自成体肠道干细胞或人类多能干细胞（hPSC）， 概括了人类肠道组织的结构。 HIO 是一种有前途的肠上皮细胞来源 修复、疾病建模和药物筛选。之前的研究表明，HIO 可以植入受伤者体内 然而，在体内肠壁中，这些方法由于缺乏适当的递送而受到显着限制。 驱动 HIO 植入的车辆。 hPSC 生成 HIO 是多阶段定向分化过程 包括三个阶段：（I）分化为定形内胚层单层，（II）后肠和原管 分化为自由浮动、自组织的 3D 聚集体（人肠球体，HIS），以及 (III) 将肠道规范化为 3D 细胞外基质内的 HIO。这种体外培养过程跨越了二维生长 基底（第一阶段和第二阶段）到 3D 矩阵（第三阶段）。 III 期需要在 Matrigel（一种小鼠​​肿瘤）内进行培养 衍生的基底膜提取物成分不明确、批次间存在差异且临床效果有限 翻译潜力。 HIO 技术的另一个障碍是 HIS 差异化的低产量和一致性 在 HIO 中。该项目的目标是（1）设计一个具有独立控制的合成水凝胶平台 指导 hPSC 整个体外分化为 HIO 的生化和生物物理线索，以及 (2) 将合成涂层中的 HIO 递送至体内肠道损伤处。中心假设是，一个工程化的 具有适当生物物理和生化线索的合成基质将支持 HIO 自组织、生长、 和分化过程并增强 HIO 植入和肠道伤口愈合。目标 1：设计 a 促进 hPSC 生成球状体的 2D 合成基质。目标 2：评估 HIO 的成熟度 合成生态位内生成的球体。目标 3：设计可临床转化的治疗方案 HIO 修复受损肠道组织的材料。这项研究的结果将增加临床相关性 产生 HIO 并将提供可扩展和可翻译的合成材料用于分化和交付 HIO 的数量。