Engineering a biomimetic matrix to promote development of human ovarian follicles in vitro

工程仿生基质促进人类卵泡体外发育

基本信息

批准号：
10311304
负责人：
Andrea Jones
金额：
$ 4.05万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
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/10311304
关键词：
Age Angiopoietins Apoptotic Atlases Autologous Transplantation Automobile Driving Basement membrane Bioinformatics Biological Biomimetics Caliber Cancer Survivor Cell Proliferation Cells Characteristics Chemistry Clinic Clinical Complex Computational Biology Cortex of ovary Coupled Cryopreservation Data Set Deposition Development Dissociation Embryo Endocrine Engineering Environment Extracellular Matrix Extracellular Matrix Proteins Fertility Fertilization Flow Cytometry Fluorescence Future Gene Expression Profile Gene Expression Profiling Germ Cells Goals Growth Growth Factor Hormonal Hormone Responsive Hormones Human Hydrogels In Vitro Individual Infertility Knowledge Life Ligand Binding Malignant Neoplasms Mechanics Methods Molecular Biology Mus Nature Oocytes Ovarian Ovarian Follicle Ovarian Tissue Ovarian tissue cryopreservation Ovary PGF gene Paracrine Communication Pathway interactions Patients Peptide Hydrolases Peptides Platelet-Derived Growth Factor Population Pregnancy Primordial Follicle Procedures Protocols documentation Reproducibility Research Risk Role Savings Signal Transduction Somatic Cell Sorting - Cell Movement Stains Standardization Stimulus Stromal Cells Structure Supplementation Support System Suspensions System Tissue Engineering Translating Translations Vascular Endothelial Growth Factors Woman Work aged angiogenesis anticancer treatment autocrine base cancer cell cancer recurrence cancer risk cancer therapy comparative cytokine cytotoxic design egg ethylene glycol fertility improvement fertility preservation folliculogenesis girls human tissue improved in vivo innovation next generation sequencing novel oocyte cryopreservation oocyte retrieval paracrine prepuberty primary ovarian insufficiency prospective reproductive scaffold self assembly single cell sequencing single-cell RNA sequencing standard of care success three dimensional structure tissue culture transcription factor transcriptome transcriptomics

项目摘要

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.

拟议研究的长期目标是为接受性腺毒性抗癌治疗和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提出的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，从而在体外从原始阶段促进人类植物的发展。小叶培养的成功率低主要归因于卵泡细胞，邻近卵泡和基质细胞之间的复合物，旁分泌，自分泌和内分泌信号传导。中心假设是，人类立法的转录分析将揭示推动发展的机制，并通过设计仿生氢环境（ECM）来促进卵巢微环境，将支持人类立法的体外发展。拟议工作的基本原理是，通过解释驱动毛细血管激活和早期发育的机制，并在ECM序列水凝胶中概括天然的卵巢微环境，可以将未来的培养系统转化为冷冻保存卵巢福利的诊所，以使其成熟，以供冷冻保存的卵巢福利和随后的培养系统。在第一个目的中，单细胞RNA测序将用于介绍人卵巢福利和支持性基质细胞。在第二个目标中，使用迈克尔型添加化学化学，将ECM序列肽掺入仿生聚（PEG）水凝胶系统中，以促进ECM成分的沉积并模仿天然的卵巢组织。 Folicle的地下膜由ECM蛋白组成，它是对卵泡细胞的结构支持，是进入卵泡的分子的选择性屏障，以及保留固体生长因子和细胞因子的支架。在卵泡发育过程中，它不断重塑，但是细胞分泌的ECM分子无法粘附于未修饰的钉子以进行自组装。通过在PEG水凝胶中整合ECM隔离肽，可以恢复ECM的结构和生物学作用以进行体外毛囊发育。这项工作的贡献将是促进卵泡发育的生殖年龄卵巢突出机制的单个细胞地图集，并在卵泡中和基于新颖的体外作用。支持人卵泡发育的卵泡培养系统。这项工作的贡献将是重要的，因为它将指导建立标准化的体外培养物，以使人类卵泡成熟，并为无法产生因性腺毒性治疗而无法产生成熟卵的安全性生育能力保存选择。拟议的工作具有创新性，因为它将是健康生殖老年妇女的第一个单细胞数据集，也是人类卵泡培养物中的第一个单个细胞数据集中，是合成的ECM序列矩阵中的第一个单细胞数据集。