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 拟议研究的长期目标是为妇女建立一个广泛的生育保留选择 正在接受性腺毒性抗癌治疗并面临不育。本提案的总体目标是 努力实现所提出的目标并减轻与自体移植相关的风险是创建一个 仿生环境，促进人类卵泡从体外原始阶段发育。低 小卵泡培养的成功率很大程度上归因于复杂且知之甚少的旁分泌， 滤泡细胞、相邻滤泡和基质细胞之间的自分泌和内分泌信号传导。中央 假设人类卵泡的转录谱将揭示驱动发育的机制， 通过设计一种仿生水凝胶重建卵巢微环境， 细胞外基质（ECM）将在体外支持人卵泡发育。拟议工作的理由 通过破译驱动卵泡激活和早期发育的机制， 天然卵巢微环境中的ECM螯合水凝胶，未来的培养系统可以转化为 冷冻保存的卵泡成熟以及随后的受精和妊娠的临床。上 目的：单细胞RNA测序将用于分析人类卵泡和支持性基质细胞。在 第二个目标是将ECM螯合肽掺入仿生聚乙二醇（PEG）中， 使用迈克尔型加成化学促进ECM组分和模拟物沉积水凝胶系统 天然卵巢组织卵泡的基底膜由ECM蛋白组成，其功能是 毛囊细胞的结构支持，分子进入毛囊的选择性屏障，以及 保留可溶性生长因子和细胞因子。它在卵泡发育过程中不断重塑，但细胞- 分泌的ECM分子不能粘附到未修饰的PEG上进行自组装。通过整合ECM- 在PEG水凝胶中螯合肽，ECM的结构和生物学作用可以恢复， 体外卵泡发育这项工作的贡献将是生殖年龄卵巢的单细胞图谱 强调驱动卵泡发育的机制和基质细胞在卵泡发生中的支持作用， 一种支持人类卵泡发育的新型体外卵泡培养系统。这项工作的贡献将 因为它将指导标准化的体外培养的发展，用于人胚胎的成熟， 卵泡和一个安全的生育能力保存选择的患者无法产生成熟的卵子，由于 性腺毒性治疗。拟议的工作是创新的，因为它将是第一个来自健康的单细胞数据集。 生殖年龄妇女和人类卵泡培养的第一个实例在合成ECM螯合矩阵。