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：
10671614
负责人：
Andrea Jones
金额：
$ 2.63万
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 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10671614
关键词：
Age Angiopoietins Apoptotic Atlases Autologous Transplantation Automobile Driving Basement membrane Bioinformatics Biological Biomimetics Cancer Survivor Cell Proliferation Cell secretion Cells Characteristics Chemistry Clinic Clinical Complex Computational Biology Cortex of ovary Coupled Cryopreservation Data Set Deposition Development Diameter Dissociation Embryo Encapsulated 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 Recurrent Malignant Neoplasm Reproducibility Research Risk Role Signal Transduction Somatic Cell Sorting Stains Standardization Stimulus Stromal Cells Supplementation Support System Suspensions System Tissue Engineering Tissues Translating Translations Vascular Endothelial Growth Factors Woman Work aged angiogenesis anticancer treatment autocrine 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 meter next generation sequencing novel oocyte cryopreservation oocyte retrieval paracrine prepuberty primary ovarian insufficiency prospective reproductive risk mitigation scaffold self assembly single cell sequencing single-cell RNA sequencing standard of care success three dimensional structure transcription factor transcriptome transcriptomics translational potential

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孕症的女性建立广泛的生育力保留选择。该提案努力实现所提出的目标并减轻与自体移植相关的风险的总体目标是创建一个新的方法。促进人类卵泡从原始阶段体外发育的仿生环境小卵泡培养的成功率较低，很大程度上归因于复杂且知之甚少的旁分泌、自分泌和内分泌。核心假设是，人类卵泡的转录谱将通过设计保留细胞分泌的细胞外基质（ECM）的仿生水凝胶来揭示驱动发育和重建卵巢微环境的机制。这项工作的基本原理是破译驱动卵泡激活和早期发育的机制，并重现自然卵巢。在 ECM 隔离水凝胶中的微环境中，未来的培养系统可以应用于临床，用于冷冻保存的卵泡的成熟以及随后的受精和妊娠。在第一个目标中，单细胞 RNA 测序将用于分析人类卵泡和支持性基质细胞。第二个目标是，利用迈克尔型加成化学将 ECM 螯合肽纳入仿生聚乙二醇 (PEG) 水凝胶系统中，以促进卵泡的基底膜由 ECM 蛋白组成，其功能是卵泡细胞的结构支持、分子进入卵泡的选择性屏障以及保留可溶性生长因子和细胞因子的支架。它在卵泡发育过程中不断重塑，但细胞分泌的 ECM 分子无法粘附到未修饰的 PEG 上进行自组装。通过水凝胶，可以在体外卵泡发生中恢复 ECM 的结构和生物学作用，这项工作的贡献将是生殖年龄卵巢的单细胞图谱，突出显示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，以及一种新颖的方法。支持人类卵泡发育的体外卵泡培养系统这项工作的贡献将是重大的，因为它将指导人类卵泡成熟的标准化体外培养和安全的生育力保存选择的开发。这项工作的创新之处在于，它将是第一个来自生殖健康老年女性的单细胞数据集，也是第一个在合成 ECM 隔离基质中进行人类卵泡培养的实例。