Investigating the role of apoptosis-resistance and the tumor environment on development and maintenance of sacrococcygeal teratomas

研究细胞凋亡抗性和肿瘤环境对骶尾部畸胎瘤发生和维持的作用

• 批准号: 10749797
• 负责人: Ernesto Javier Rojas
• 金额: $ 3.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749797
• 关键词: Apoptosis; Apoptotic; BAK1 gene; BAX gene; Bioinformatics; Biological Markers; Biological Process; Biology; Carcinogenicity Tests; Career Mobility; Cell Line; Cell Transplantation; Cell model; Cells; Cellular Assay; Chromatin; Clinical; Common Neoplasm; Complex; Development; Developmental Cell Biology; Doctor of Philosophy; Embryo; Environment; Ethics; Exposure to; Failure; Fellowship; Fetus; Future; Gene Expression; Genetic Transcription; Germ Cells; Germ Layers; Germ cell tumor; Goals; Gonadal structure; Growth Factor; Histology; Human; In Vitro; Infrastructure; Inhibition of Apoptosis; Injections; Knowledge; Lead; Maintenance; Mediating; Modeling; Molecular; Morbidity - disease rate; Mus; Names; Newborn Infant; Outcome; Pathway interactions; Patients; Perinatal mortality demographics; Pluripotent Stem Cells; Process; Proliferating; Recurrence; Research Training; Resistance; Risk; Role; Sampling; Scientist; Signal Transduction; Structure of primordial sex cell; Surgeon; Techniques; Technology; Teratoma; Testing; Tissues; Training; Transposase; Tumorigenicity; Work; biomarker development; cell type; clinical training; collaborative environment; experimental study; fetal; human model; improved; in utero; in vitro Model; in vivo; in vivo Model; induced pluripotent stem cell; innovation; insight; migration; molecular targeted therapies; mortality; multiple omics; perinatal health; perinatal morbidity; perinatal period; pluripotency; potential biomarker; sacrum; single-cell RNA sequencing; skills; stem cell biology; stem cells; theories; therapy development; transcriptomics; transplant model; tumor; tumor heterogeneity; tumorigenic

PROJECT SUMMARY/ABSTRACT Sacrococcygeal teratomas (SCTs) are the most common tumor in newborns, have significant perinatal morbidity and mortality, a 35% rate of recurrence, and lack good biomarkers to predict recurrence risk. SCTs are believed to originate from apoptosis-resistant embryonic gamete-precursors known as primordial germ cells (PGCs). In mice, PGCs can revert to pluripotent cells called embryonic germ cells (EGCs) when exposed to a specific cocktail of growth factors, however, establishing an in vitro model of human EGCs has been technically and ethically challenging. The proposed Research Training Plan will leverage human induced pluripotent stem cell derived PGC-like cells (PGC-LCs) and EGC-like cells (EGC-LCs), single cell multi-omic technologies, and an innovative mouse embryonic injection model to study the process by which PGCs lead to teratoma formation. The central hypothesis is that SCTs arise from apoptosis-resistant ectopic PGCs that have been reverted to a pluripotent state resembling PGC to EGC reversion, and the most aggressive SCTs have tumor environment interactions that maintain these pluripotent cells. In Aim 1, the trainee, MD PhD candidate Ernesto Rojas, will define the chromatin and transcriptional landscape of PGC-LC to EGC-LC reversion with and without apoptosis- resistance. Then, using the in utero injection model he developed, he will characterize the differences in tumorigenic potential between PGC-LC and EGC-LCs, with and without inhibition of apoptosis. In Aim 2, the candidate will identify and modulate pathways maintaining pluripotency in SCTs. He will use single-cell transcriptomics to identify pathways that may modulate the tumorigenicity of EGC-LCs, and will functionally validate these pathways using the PGC-LC to EGC-LC reversion model and the transplant model to find those pathways that improve maintenance of EGC-LCs. The results of Aim 1 will provide the first roadmap of the changes in chromatin and gene expression that occur during reversion from germline to pluripotency to teratoma. The results of Aim 2 will provide potential biomarkers and targetable pathways for future clinical use in SCTs. These studies will identify the important developmental trajectories that occur with reversion from PGC to pluripotency, improve insight to aid biomarker development for recurrences of PGC-derived tumors, and provide potential targetable pathways to treat the most aggressive tumors. To successfully complete this project, the candidate will work with Drs. Diana Laird (sponsor) and Tippi Mackenzie (co-sponsor). Dr. Laird is a germ cell expert and Dr. Mackenzie is a fetal surgeon. Together with the candidate, they have established a training plan for him to gain new knowledge and skills in developmental and stem cell biology techniques and bioinformatic analyses, as well as clinical training, and professional development. Through the collaborators named in this proposal, the collaborative environment and excellent infrastructure at UCSF, and with the support provided by this fellowship, the candidate will be well equipped to investigate the role of development on early peri-natal health and advance his career as a surgeon-scientist.

项目摘要/摘要 摘要骶尾部畸胎瘤是新生儿最常见的肿瘤，具有显著的围产期发病率。 和死亡率，35%的复发率，并且缺乏良好的生物标志物来预测复发风险。SCT被认为是 起源于抗凋亡的胚胎配子--称为原始生殖细胞(PGCs)的前体。在……里面 小鼠，当暴露于特定的胚胎生殖细胞(EGCs)时，PGCs可以恢复为多能细胞 然而，在生长因子的混合作用下，建立人EGCs的体外模型在技术上和 在道德上具有挑战性。拟议的研究培训计划将利用人类诱导的多潜能干细胞 衍生的PGC样细胞(PGC-LCS)和EGC样细胞(EGC-LCS)、单细胞多组学技术和 创新小鼠胚胎注射模型，研究PGCs导致畸胎瘤形成的过程。 中心假设是SCT起源于抗凋亡的异位PGCs，这些异位PGCs已经恢复到 多能状态类似于PGC到EGC逆转，最具侵袭性的SCT具有肿瘤环境 维持这些多能细胞的相互作用。在目标1中，实习生、医学博士候选人埃内斯托·罗哈斯将 确定PGC-LC到EGC-LC逆转的染色质和转录图谱-有无凋亡- 抵抗。然后，使用他开发的宫内注射模型，他将描述 PGC-LC和EGC-LC之间的致瘤潜力，有和没有抑制细胞凋亡。在目标2中， 候选人将识别和调节维持SCTS多能性的途径。他将使用单细胞 转录学，以确定可能调节EGC-LC致瘤性的途径，并将从功能上 使用PGC-LC到EGC-LC的逆转模型和移植模型验证这些途径，以找到这些途径 改善EGC-LCS维护的途径。目标1的结果将提供第一个路线图 染色体和基因表达的变化，发生在从生殖系到多能畸胎瘤的逆转过程中。 AIM 2的研究结果将为未来SCTS的临床应用提供潜在的生物标志物和靶向途径。 这些研究将确定从PGC恢复到PGC的重要发展轨迹 多能性，改善洞察力，以帮助PGC来源的肿瘤复发的生物标志物的开发，并提供 治疗侵袭性最强的肿瘤的潜在靶向途径。 为了成功地完成这个项目，候选人将与戴安娜·莱尔德博士(赞助商)和蒂皮合作 麦肯齐(共同发起人)。莱尔德博士是生殖细胞专家，麦肯齐博士是胎儿外科医生。与 作为候选人，他们已经为他制定了培训计划，以获得发展和发展方面的新知识和技能 干细胞生物技术和生物信息学分析，以及临床培训和专业人员 发展。通过本提案中点名的协作者，协作环境和优秀 在加州大学旧金山分校的基础设施，并在该奖学金提供的支持下，候选人将完全准备好 研究发育对围产期早期健康的作用，并促进他作为外科科学家的职业生涯。