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，影响多达10%的美国女性。虽然有治疗方法，但不止是 患有子宫内膜异位症的女性中有50%会复发。缺乏有用的模型继续阻碍着 发现可用于新干预的病理生理学方面。因此，有一个 迫切需要开发新的子宫内膜异位症模型，以显著推进该领域的研究。此应用程序 建议创造3D生物软化SSuPer(自持灌流)组织，它概括了 子宫内膜异位症或正常子宫内膜采用Kenzan法。Kenzan方法使用Regenova Bio 3D打印机将多细胞球体放置在称为Kenzan的微针阵列上。在Kenzan上，球体 通过细胞间的相互作用和分泌天然的细胞外基质进行融合，允许更多的内在 细胞间通信。上皮细胞和基质细胞之间的细胞间通讯在 子宫内膜功能。初步数据显示，Kenzan方法有效地使第一个 无支架3D生物打印构建的子宫内膜异位上皮和子宫内膜 基质细胞。此外，SSuPer组织包含可以在生物惰性中灌流的微通道 法布里卡生物反应器。因为人子宫内膜间质成纤维细胞对子宫功能至关重要， 子宫内膜异位症患者的成纤维细胞可以用来模拟细胞功能的异常、异常 发生在子宫内膜异位症中的分子信号和促炎转录谱。此外，亲- 炎症介质在子宫内膜异位症患者腹腔液中升高，并可能在 黄体酮抵抗。该应用程序的目标是丰富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，它概括了炎症、转录和类固醇激素反应 忠实地。中心假设是炎性细胞因子和间质细胞起源(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症或子宫内膜表型。SSuPer纸巾， 由来自子宫内膜异位症或正常子宫内膜的上皮和间质细胞组成，将被生物软化 并用媒体或带有煽动性鸡尾酒的媒体灌输。SSuPer组织将被确定为 子宫内膜异位症或正常子宫内膜受炎性标志物分泌、表达的影响 生物标志物和全球转录组的变化。二次验证研究将集中在类固醇激素上 回应。本申请中提出的研究具有创新性，因为它将生成新的无脚手架3D 生物软化的SSuPer组织，对子宫内膜异位症或正常子宫内膜保持高度的生物学保真度。 经过严格验证的体外模型将为研究糖尿病的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疾病，如不孕症和反复妊娠丢失。高度 可重复的模型是测试新疗法所必需的。