The contributions of inflammatory cocktail and stromal cell origin on a scaffold-free 3D biofabricated SSuPer tissue of endometriosis and normal endometrium

炎症混合物和基质细胞起源对子宫内膜异位症和正常子宫内膜无支架 3D 生物制造 SSuPer 组织的贡献

• 批准号: 10218664
• 负责人: Shannon Michelle Hawkins
• 金额: $ 23.78万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218664
• 关键词: 3-Dimensional; 3D Print; Address; Affect; Biological; Biological Markers; Bioreactors; Cell Communication; Cell Culture Techniques; Cell Differentiation process; Cell Separation; Cell physiology; Cell secretion; Cells; Cellular Morphology; Cellular Spheroids; Clinical; Culture Media; Data; Decidual Cell Reactions; Defect; Disease; Endometrial; Endometrial Stromal Cell; Endometriomas; Endometrium; Epithelial; Epithelial Cells; Extracellular Matrix; Fibroblasts; Foundations; Functional disorder; Growth; Gynecologic; Gynecology; Histologic; Human; Hysterectomy; Immunohistochemistry; In Vitro; Infertility; Inflammation Mediators; Inflammatory; Intervention; Lesion; Mediator of activation protein; Methods; Modeling; Molecular; Morbidity - disease rate; National Institute of Child Health and Human Development; Operative Surgical Procedures; Pathogenesis; Perfusion; Peritoneal Fluid; Phenotype; Play; Pregnancy; Pregnancy Outcome; Pregnancy loss; Preparation; Process; Progesterone; Progesterone Receptor Status; Recurrence; Reproducibility; Reproductive Health; Research; Resistance; Risk; Role; Signal Transduction; Specimen; Strategic Planning; Stromal Cells; Testing; Tissue Model; Tissues; Uterine cavity; Uterus; Validation; Woman; base; biofabrication; bioprinting; cytokine; design; endometriosis; eutopic endometrium; healthy pregnancy; high reward; high risk; human disease; implantation; in vitro Model; inflammatory marker; innovation; intercellular communication; new therapeutic target; novel; novel therapeutics; personalized medicine; pregnant; reconstitution; response; scaffold; steroid hormone; three dimensional structure; three-dimensional modeling; transcriptomics; validation studies

Endometriosis is a debilitating disease that affects up to 10% of U.S. women. While treatments exist, more than 50% of women with endometriosis will have disease recurrence. The lack of useful models continues to impede the discovery of aspects of pathophysiology that are amenable to novel interventions. Therefore, there is an urgent need to develop novel models of endometriosis to advance the field significantly. This application proposes to create 3D biofabricated SSuPer (Self-Supporting Perfused) tissues that recapitulate either endometriosis or normal endometrium using the Kenzan method. The Kenzan method uses the Regenova Bio 3D Printer to place multicellular spheroids onto a microneedle array, called a Kenzan. On Kenzan, the spheroids fuse through cell-cell interaction and secretion of their native extracellular matrix, allowing more intrinsic intercellular communication. Intercellular communication between epithelial and stromal cells is paramount in endometrial function. Preliminary data shows that the Kenzan method effectively biofabricates the first scaffold-free 3D bioprinted constructs comprised of both endometriotic epithelial and endometrial stromal cells. Further, SSuPer tissues contain microchannels that can be perfused in a biologically inert FABRICA bioreactor. Because human endometrial stromal fibroblasts are critical to uterine function, stromal fibroblasts from women with endometriosis can be used to model the abnormal cellular function, aberrant molecular signaling, and pro-inflammatory transcriptomic profiles that occur in endometriosis. Additionally, pro- inflammatory mediators are elevated in the peritoneal fluid of women with endometriosis and likely play a role in progesterone resistance. This application's objective is to biofabricate 3D models of both endometriosis and normal endometrium, which recapitulate the inflammatory, transcriptomic, and steroid hormone response faithfully. The central hypothesis is that both inflammatory cytokines and stromal cell origin (i.e., endometriosis or not) independently affect the endometriotic or endometrial phenotype of each SSuPer tissue. SSuPer tissues, comprised of epithelial and stromal cells from either endometriosis or normal endometrium, will be biofabricated and perfused with either media or media with an inflammatory cocktail. SSuPer tissues will be determined to be either endometriosis or normal endometrium by inflammatory marker secretion, expression of endometriosis biomarkers, and global transcriptomic changes. Secondary validation studies will focus on steroid hormone response. The research proposed in this application is innovative because it will generate novel scaffold-free 3D biofabricated SSuPer tissues that maintain high biological fidelity to endometriosis or normal endometrium. Rigorously validated in vitro models will provide useful models for investigating the pathophysiology of endometriosis and other uterine dysfunction diseases such as infertility and recurrent pregnancy loss. Highly reproducible models are necessary for testing novel therapies.

子宫内膜异位症是一种使人衰弱的疾病，影响着多达 10% 的美国女性。虽然存在治疗方法，但超过 50%患有子宫内膜异位症的女性会出现疾病复发。缺乏有用的模型继续阻碍 发现适合新干预措施的病理生理学方面。因此，有一个 迫切需要开发新的子宫内膜异位症模型来显着推进该领域的发展。这个应用程序 提议创建 3D 生物制造 SSuPer（自支撑灌注）组织，该组织可概括 子宫内膜异位症或正常子宫内膜使用 Kenzan 方法。 Kenzan 方法使用 Regenova Bio 3D 打印机将多细胞球体放置到称为 Kenzan 的微针阵列上。 Kenzan 上的球体 通过细胞间相互作用和天然细胞外基质的分泌进行融合，从而实现更多内在的 细胞间通讯。上皮细胞和基质细胞之间的细胞间通讯对于 子宫内膜功能。初步数据表明 Kenzan 方法有效地生物制造了第一个 由子宫内膜异位上皮和子宫内膜组成的无支架 3D 生物打印结构 基质细胞。此外，SSuPer 组织含有微通道，可以在生物惰性的环境中进行灌注。 FABRICA 生物反应器。由于人子宫内膜基质成纤维细胞对于子宫功能至关重要，因此基质 来自患有子宫内膜异位症的女性的成纤维细胞可用于模拟异常的细胞功能 子宫内膜异位症中发生的分子信号传导和促炎转录组谱。此外，亲 患有子宫内膜异位症的女性腹腔液中炎症介质升高，可能在 黄体酮抵抗。该应用程序的目标是生物制造子宫内膜异位症和子宫内膜异位症的 3D 模型 正常子宫内膜，概括炎症、转录组和类固醇激素反应 忠实。中心假设是炎症细胞因子和基质细胞起源（即子宫内膜异位症） 或不）独立影响每个 SSuPer 组织的子宫内膜异位或子宫内膜表型。 SSuPer 组织， 由来自子宫内膜异位症或正常子宫内膜的上皮细胞和基质细胞组成，将被生物制造 并用培养基或含有炎症混合物的培养基进行灌注。 SSuPer 组织将被确定为 子宫内膜异位症或正常子宫内膜通过炎症标志物分泌、子宫内膜异位症的表达 生物标志物和整体转录组变化。二次验证研究将集中于类固醇激素 回复。本申请中提出的研究具有创新性，因为它将生成新颖的无支架 3D 生物制造的 SSuPer 组织，对子宫内膜异位症或正常子宫内膜保持高生物保真度。 严格验证的体外模型将为研究病理生理学提供有用的模型 子宫内膜异位症和其他子宫功能障碍性疾病，如不孕症和反复流产。高度 可重复的模型对于测试新疗法是必要的。