F31 Mulero Russe

F31 穆莱罗·鲁斯

• 批准号: 10651644
• 负责人: Adriana Mulero-Russe
• 金额: $ 4.77万
• 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/10651644
• 关键词: 3-Dimensional; Address; Adhesives; Adult; Affect; American; Architecture; Area; Basement membrane; Biochemical; Biocompatible Materials; Biological Products; Biophysics; Cell Therapy; Cells; Cellular Structures; Chemical Injury; Confocal Microscopy; Cues; Dependence; Development; Diameter; Disease model; Drug Screening; Elasticity; Encapsulated; Endoderm; Engineering; Engraftment; Epithelial Cells; Epithelium; Extracellular Matrix; Fiber Optics; Formulation; Foundations; Gene Expression; Generations; Growth; Growth Factor; Hindgut; Human; Hydrogels; Immunologic Deficiency Syndromes; In Situ; In Vitro; Inflammation; Inflammatory Bowel Diseases; Intestinal permeability; Intestines; Knowledge; Length; Ligands; Maleimides; Measures; Mechanics; Membrane; Mesenchymal; Mus; Organoids; Patients; Polymers; Process; Proliferating; Property; Rattus; Recovery; Reproducibility; Research; Resolution; Small Intestines; Source; Specific qualifier value; Structure; Technology; Therapeutic; Time; Tissues; Tube; Tumor-Derived; Ultraviolet Rays; Work; arm; cell type; clinical translation; clinically relevant; delivery vehicle; density; directed differentiation; epithelial repair; epithelial wound; ethylene glycol; experience; healing; human pluripotent stem cell; human stem cells; in vitro Model; in vivo; inflammatory milieu; injured; innovation; insight; intestinal crypt; intestinal epithelium; intestinal injury; matrigel; microbiota; monolayer; nutrient absorption; optical fiber; polarized cell; process repeatability; programs; repaired; self organization; stem cells; tissue repair; tool; translational potential; 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.

项目摘要 炎症性肠道疾病（IBD）目前影响> 160万美国人。 IBD的特征是中断 到肠上皮和高炎症环境。可用治疗目标针对炎症 但是，通过生物学剂，更少的努力集中在上皮愈合上，没有广泛的努力 适用修复肠上皮的疗法。人肠癌（HIO）是三维（3D） 多细胞结构，源自成年肠干细胞或人多能干细胞（HPSC）， 概括了人类肠组织结构。 HIO是肠上皮的有前途的细胞来源 修复，疾病建模和药物筛查。以前的工作表明，Hios植入了受伤的 但是，体内肠壁，这些方法由于缺乏适当的递送而受到明显的限制 驾驶HIO植入的车辆。 HPSC的HIO生成是多阶段的定向分化过程 包括三个阶段：（i）分化为确定的内胚层单层，（ii）后肠和原始管 区分自由浮动，自组织的3D骨料（人肠球体，他的）和（iii） 3D细胞外基质内的HIO中的肠道规范。这种体外培养过程跨越了2D增长 底物（I和II期）至3D矩阵（第三阶段）。第三阶段需要在Matrigel内培养，鼠肿瘤 - 衍生的地下膜提取物，具有不确定的成分，批次到lot的可变性和有限的临床 翻译潜力。 Hio Technologies的另一个障碍是他的分化的低收益和一致性 在hios。该项目的目标是（1）工程师一个具有独立控制的合成水凝胶平台 将HPSC的整个体外分化引导到HIO的生化和生物物理提示，以及（2） 将HIO在体内递送到体内肠道损伤中。中心假设是设计 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植入和肠道伤口的愈合。目标1：工程师A 2D合成基质从HPSC促进球体产生。目标2：评估HIO的成熟 合成生态位中产生的球体。目标3：工程师临床上可翻译的治疗交付 HIO的材料受伤的肠组织。这项研究的结果将增加临床相关性 生成的HIO，并将提供可扩展和可翻译的合成材料，用于分化和交付 hios。