Engineering the ovarian microenvironment and deciphering folliculogenesis in a biomimetic matrix

工程卵巢微环境并破译仿生基质中的卵泡发生

• 批准号: 10116957
• 负责人: Claire Elizabeth Tomaszewski
• 金额: $ 3.99万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2022-12-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10116957
• 关键词: 3-Dimensional; Address; Adipose tissue; Affect; Aftercare; Autologous Transplantation; Basement membrane; Biological; Biomimetics; Cancer Survivor; Cell physiology; Cells; Chemistry; Chemotherapy and/or radiation; Childhood; Complex; Cryopreservation; Cytokine Network Pathway; Deposition; Development; Endocrine; Engineering; Environment; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Fertility; Fertilization in Vitro; Gene Expression Regulation; Genetic Transcription; Goals; Gonadotropins; Growing Follicle; Growth; Growth Factor; Hematologic Neoplasms; Hormones; Human; Hydrogels; In Vitro; Infertility; Lead; Live Birth; Mus; Nature; Oocytes; Outcome; Ovarian; Ovarian Follicle; Ovarian Tissue; Ovary; Paracrine Communication; Patients; Peptides; Population; Primordial Follicle; Process; Property; Quality of life; Risk; Role; Signal Transduction; Standardization; Structure; Supporting Cell; System; Systems Biology; Systems Development; Time; Translating; Transplantation; Woman; Work; autocrine; cancer cell; cancer therapy; cytokine; egg; ethylene glycol; fertility preservation; folliculogenesis; girls; granulosa cell; innovation; insight; novel; oocyte maturation; oocyte retrieval; ovarian reserve; paracrine; prepuberty; primary ovarian insufficiency; reproductive function; restoration; scaffold; self assembly; side effect; stem cells; success; theca cell; transcription factor

The long-term goal of this work is to establish a broad fertility preservation option for women facing infertility as a result of gonadotoxic treatments. The overall objective of this proposal in working towards the long term goal and mitigating the risks associated with autotransplantation is to create a biomimetic environment that promotes in vitro growth of immature follicles. The low success rates of follicle development, or folliculogenesis, are attributed to the complex and poorly understood paracrine, autocrine and endocrine signaling between the cells in a follicle, neighboring follicles and their microenvironment. The central hypothesis is that recreating the ovarian microenvironment through co-encapsulation of follicles with adipose-derived stem cells (ADSCs) in a hydrogel which retains cell-secreted extracellular matrix (ECM) will supply the necessary support for primary follicle growth in vitro. The rationale for the proposed work is that by recapitulating the natural ovarian microenvironment and deciphering transcription factor and cytokine networks, the culture system can be further developed to promote folliculogenesis of human follicles. In the first aim, ECM-sequestering peptides will be incorporated in a biomimetic poly(ethylene glycol) (PEG) hydrogel system using Michael-type addition chemistry to facilitate deposition of ECM components and mimic the native ovarian tissue. The follicle basement membrane is composed of ECM proteins and it functions as a structural support for the cells, a selective barrier for materials entering the follicle, and a scaffold for retaining growth factors. It is continuously remodeled as the follicle grows, but cell-secreted ECM molecules are unable to adhere to unmodified PEG for self-assembly. By integrating ECM-sequestering peptides in the PEG hydrogels, the structural and biological roles of ECM can be restored for in vitro folliculogenesis. In the second aim, Transcriptional Activity Cellular Array (TRACER) will be used to determine the dynamic transcription factor (TF) activity in granulosa cells of growing follicles when cultured in a biomimetic hydrogel which includes paracrine and ECM support. This information will give insight to the internal cell processes which lead to follicle growth and survival. The contribution of this work is expected to be a novel in vitro follicle culture that supports primary follicle growth, and a better understanding of the underlying mechanisms which drive folliculogenesis. The contribution of this work will be significant because it will guide the development of a standardized in vitro culture for maturation of human follicles and a safe fertility preservation option for patients unable to produce mature eggs as a result of gonadotoxic treatments. The proposed work is innovative in that it will be the first instance of follicle culture in a synthetic ECM-sequestering matrix, and the first time TRACER will be used to study follicles in PEG.

这项工作的长期目标是为促性腺毒性治疗而为面临不育症的妇女建立广泛的生育能力。该提案的总体目标是朝着长期目标努力并减轻与自动移植相关的风险是创造一个仿生环境，从而促进未成熟卵泡的体外生长。卵泡发育或卵泡生成的成功率低归因于卵泡，相邻卵泡及其微环境之间的复杂且知之甚少的旁分泌，自分泌和内分泌信号。中心假设是，通过与脂肪衍生的干细胞（ADSC）共囊化来重现卵巢微环境，该水凝胶保留了保留细胞分泌的细胞外基质（ECM）的水凝胶，将为体外体外的原发性卵泡生长提供必要的支撑。拟议工作的理由是，通过概括天然卵巢微环境和解密转录因子和细胞因子网络，可以进一步开发培养系统来促进人类卵泡的卵泡发生。在第一个目的中，ECM序列的肽将掺入使用迈克尔型添加化学的仿生聚（PEG）水凝胶系统中，以促进ECM成分的沉积并模仿天然卵巢组织。卵泡地下膜由ECM蛋白组成，它是对细胞的结构支持，进入卵泡的材料的选择性屏障以及保留生长因子的脚手架。随着卵泡的生长，它不断重塑，但是细胞分泌的ECM分子无法粘附在未修改的钉子上以进行自组装。通过在PEG水凝胶中整合ECM序列的肽，可以恢复ECM的结构和生物学作用以进行体外卵泡发生。在第二个目标中，转录活性细胞阵列（示踪剂）将用于确定在培养在A中的卵泡颗粒细胞中的动态转录因子（TF）活性 仿生水凝胶，包括旁分泌和ECM载体。该信息将深入了解导致卵泡生长和生存的内部细胞过程。这项工作的贡献预计将是一种新型的体外卵泡培养物，支持原发性卵泡生长，并更好地理解驱动卵泡发生的基本机制。这项工作的贡献将是重要的，因为它将指导标准化体外培养的人类卵泡成熟，并为由于性腺毒性治疗而无法产生成熟卵的患者的安全生育能力保存选择。拟议的工作具有创新性，因为它将是合成ECM序列矩阵中卵泡培养的第一个实例，并且首次使用示踪剂来研究PEG中的卵泡。