Supplement Andrea Jones

补充安德里亚·琼斯

• 批准号: 10505465
• 负责人: Andrea Jones
• 金额: $ 0.25万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-19 至 2024-08-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10505465
• 关键词: Age; Atlases; Autologous Transplantation; Automobile Driving; Basement membrane; Bioinformatics; Biological; Biomimetics; Cell secretion; Cells; Chemistry; Clinic; Complex; Cryopreservation; Data Set; Deposition; Development; Endocrine; Environment; Extracellular Matrix; Extracellular Matrix Proteins; Fertility; Fertilization; Future; Gene Expression Profiling; Goals; Growth Factor; Human; Hydrogels; In Vitro; Infertility; Ovarian; Ovarian Follicle; Ovarian Tissue; Ovary; Pathway interactions; Patients; Peptides; Pregnancy; Reproducibility; Research; Role; Signal Transduction; Standardization; Stromal Cells; Support System; System; Translating; Translations; Woman; Work; aged; anticancer treatment; autocrine; cancer therapy; cytokine; design; egg; ethylene glycol; fertility preservation; folliculogenesis; girls; innovation; next generation sequencing; novel; paracrine; primary ovarian insufficiency; reproductive; risk mitigation; scaffold; self assembly; single-cell RNA sequencing; success

PROJECT SUMMARY The long-term goal of the proposed research is to establish a broad fertility preservation option for women undergoing gonadotoxic anticancer treatments and facing infertility. The overall objective of this proposal in working towards the presented goal and mitigating the risks associated with autotransplantation is to create a biomimetic environment that promotes human follicle development from the primordial stage in vitro. The low success rates of small follicle culture are largely attributed to the complex and poorly understood paracrine, autocrine and endocrine signaling between follicular cells, neighboring follicles, and stromal cells. The central hypothesis is that transcriptional profiling of human follicles will reveal mechanisms driving development and recreating the ovarian microenvironment through design of a biomimetic hydrogel which retains cell-secreted extracellular matrix (ECM) will support human follicle development in vitro. The rationale for the proposed work is that by deciphering the mechanisms driving follicle activation and early development, and recapitulating the natural ovarian microenvironment in an ECM-sequestering hydrogel, future culture systems can be translated to the clinic for maturation of cryopreserved ovarian follicles and subsequent fertilization and pregnancy. In the first aim, single cell RNA sequencing will be used to profile human ovarian follicles and supportive stromal cells. In the second aim, ECM-sequestering peptides will be incorporated in a biomimetic poly(ethylene glycol) (PEG) hydrogel system using Michael-type addition chemistry to promote deposition of ECM components and mimic the native ovarian tissue. The follicle’s basement membrane is composed of ECM proteins and it functions as structural support for follicular cells, a selective barrier for molecules entering the follicle, and a scaffold for retaining soluble growth factors and cytokines. It is continuously remodeled during follicle development, 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 follicle development. The contribution of this work will be a single cell atlas of the reproductive-age ovary highlighting mechanisms driving follicle development and stromal cells’ supportive roles in folliculogenesis and a novel in vitro follicle culture system that supports human follicle development. 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 single cell dataset from healthy reproductive aged women and the first instance of human follicle culture in a synthetic ECM-sequestering matrix.

项目摘要 拟议的研究的长期目标是为妇女建立广泛的生育能力 接受性腺毒性抗癌治疗并面临不育症。该提议的总体目标 朝着提出的目标努力并减轻与自动移植相关的风险是创建一个 仿生环境可促进体外原始阶段的人类植物发育。低 小叶培养的成功率很大程度上归因于复杂的旁分泌，旁分泌不良， 卵泡细胞，相邻卵泡和基质细胞之间的自分泌和内分泌信号传导。中央 假设是人卵泡的转录分析将揭示推动发展的机制 通过设计保留细胞分泌的仿生水凝胶的设计来重新创建卵巢微环境 细胞外基质（ECM）将在体外支持人卵泡发育。拟议工作的理由 是通过解密驱动叶利激活和早期发展的机制，并概括了 在ECM序列水凝胶中，天然卵巢微环境可以将未来的培养系统转化为 冷冻保存卵巢福利的成熟以及随后的受精和妊娠的诊所。在第一个 目的，单细胞RNA测序将用于介绍人类卵巢的福利和支持性基质细胞。在 第二个目的，ECM序列的宠物将纳入仿生聚（乙二醇）（PEG）中 水凝胶系统使用迈克尔型添加化学反应来促进ECM组件的沉积和模拟 本地卵巢组织。 Folicle的地下膜由ECM蛋白组成，它起作用 卵泡细胞的结构支持，一种进入卵泡的分子的选择性屏障，以及用于脚手架的脚手架 保留固体生长因子和细胞因子。它在叶片发育过程中不断重塑，但细胞 - 分泌的ECM分子无法遵守未修饰的钉子进行自组装。通过整合ECM- PEG水凝胶中的隔离肽，可以恢复ECM的结构和生物学作用 体外构造的发展。这项工作的贡献将是复制年龄卵巢的单个细胞图集 突出驱动卵泡发育的机制和基质细胞在卵泡发生和 一种支持人卵泡发育的新型体外卵泡培养系统。这项工作的贡献将 要重要，因为它将指导标准化体外培养的发展以成熟的人类 卵泡和安全的生育能力保存选择，适用于由于无法产生成熟卵的患者 性腺毒性治疗。拟议的工作具有创新性，因为它将是健康的第一个单细胞数据集 在合成的ECM序列矩阵中，生殖老化的妇女和人类卵泡培养的第一个实例。