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; 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 sapococcygeal畸胎瘤（SCTS）是新生儿中最常见的肿瘤，具有明显的围产期发病率 死亡率，复发率为35％，缺乏良好的生物标志物来预测复发风险。相信SCT 起源于抗凋亡的胚胎配子剂，称为原始生殖细胞（PGC）。在 小鼠，PGC可以恢复为多能细胞，称为胚胎生殖细胞（EGC） 然而，生长因素的鸡尾酒，建立人类EGC的体外模型在技术上一直是 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能干细胞 衍生的PGC样细胞（PGC-LCS）和EGC样细胞（EGC-LCS），单细胞多氨基技术以及A 创新的小鼠胚胎注射模型研究PGC导致畸胎瘤形成的过程。 中心假设是SCT是由抗凋亡的异位PGC引起的，这些PGC已恢复为 多能状态类似于PGC与EGC恢复，最具侵略性的SCT具有肿瘤环境 维持这些多能细胞的相互作用。在AIM 1中，学员MD博士候选人Ernesto Rojas Will 将PGC-LC的染色质和转录景观定义为EGC-LC重新恢复，并没有凋亡 - 反抗。然后，使用他开发的子宫内注射模型，他将表征 PGC-LC和EGC-LC之间的致瘤潜力，有或不抑制凋亡。在AIM 2中 候选人将识别并调节维持SCT中多能性的途径。他将使用单细胞 转录组学以识别可能调节EGC-LC的肿瘤性的途径，并在功能上将 使用PGC-LC到EGC-LC归还模型和移植模型验证这些途径，以找到这些途径 改善EGC-LC的维护的途径。 AIM 1的结果将提供第一个路线图 从种系到多能性到畸胎瘤的恢复期间发生的染色质和基因表达的变化。 AIM 2的结果将为SCT中的未来临床使用提供潜在的生物标志物和可靶向途径。 这些研究将确定从PGC重新转换到的重要发展轨迹 多能性，改善洞察力，以帮助生物标志物开发PGC衍生肿瘤的复发，并提供 治疗最具侵略性肿瘤的潜在目标途径。 为了成功完成该项目，候选人将与Drs合作。 Diana Laird（赞助商）和Tippi Mackenzie（共同赞助商）。 Laird博士是生物细胞专家，Mackenzie博士是胎儿外科医生。与 候选人，他们为他制定了培训计划，以获得发展方面的新知识和技能 干细胞生物学技术和生物信息学分析以及临床培训和专业 发展。通过此提案中命名的合作者，协作环境和出色的 UCSF的基础设施以及该奖学金提供的支持，候选人将有足够的能力 调查发展对早期生日健康的作用，并提高他作为外科医生科学家的职业。