Mechanisms of human appendicular and cardiovascular comorbidities: An analysis of heterogeneity and lineage trajectories of the lateral plate mesoderm

人类四肢和心血管合并症的机制：侧板中胚层的异质性和谱系轨迹分析

• 批准号: 10642590
• 负责人: Robert L Lalonde
• 金额: $ 9.97万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10642590
• 关键词: ALDH1A2 gene; Adult; Advisory Committees; Affect; Bar Codes; Birth; Blood Vessels; Branchial arch structure; CRISPR/Cas technology; Cardiac; Cardiovascular system; Cell Lineage; Cells; Clinical Management; Collaborations; Communication; Complex; Congenital Abnormality; Data; Defect; Development; Development Plans; Developmental Biology; Diagnosis; Diagnostic; Disease; Educational process of instructing; Embryo; Experimental Designs; Fibroblast Growth Factor; Forelimb; GATA4 gene; Genes; Goals; Health; Heart; Heart Abnormalities; Heterogeneity; Human; Image; Individual; Institution; Joints; Label; Lateral; Light; Limb Development; Limb structure; Link; Lung; Medical; Mentors; Mentorship; Mesoderm; Microscopy; Modeling; Mutagenesis; Mutation; Nature; Operative Surgical Procedures; Organ; Patients; Pattern; Pectoral; Phase; Phenotype; Population; Research; Research Personnel; Research Proposals; Resources; Severities; Signal Pathway; Signal Transduction; Skeleton; Specific qualifier value; Structure; Symptoms; Testing; Tissues; Training; Transgenic Organisms; Work; Writing; Zebrafish; body system; cardiogenesis; career; career development; cell type; comorbidity; disease phenotype; experimental study; fundamental research; genetic approach; genome sequencing; human disease; improved; insight; lung development; model organism; pediatric department; progenitor; single-cell RNA sequencing; stem cells; success; supportive environment; therapeutic development; transcriptome; transcriptomics; treatment strategy; whole genome

Human diseases are seldom confined to individual tissues or organs, but instead present with a spectrum of comorbidities. A common example includes a group of congenital diseases with linked heart, limb, and lung defects. Using zebrafish as model, the goal of my research is to uncover how individual causative genes drive multi-organ comorbidities in the heart, limb, and lung. Understanding the underlying mechanisms will contribute to the development of therapeutics across disease groups and improve predictive diagnostics of patients with complex multi-organ diseases. In the adult body, the heart, limb, and lung vasculature seem unrelated at the structural level, yet share a close developmental origin in the embryo. These structures all derive from uncommitted lateral plate mesoderm that harbors progenitor cells for numerous organ systems. My overarching hypothesis posits that the lineages of the heart, limb, and lung blood vessels (cardiopharyngeal vasculature) derive from a common progenitor population within the lateral plate mesoderm, and form in an interconnected manner through the action of TBX-FGF and lineage-specific signaling inputs. In Aim 1, I will test the lineage connection between heart, pectoral fin, and cardiopharyngeal vasculature using transgenic lineage labeling and CRISPR-Cas9-based barcoding experiments. My work will directly test my hypothesis that heart, limb, and lung defects are linked by a shared developmental origin of the affected cell types. In Aim 2, I will functionally test the contribution of TBX-FGF signaling pathways, and lineage-specific inputs on joint heart, limb, and cardiopharyngeal vasculature lineage patterning. These experiments will test the impact of shared and lineage-specific signaling perturbation on co-occurrence and severity of heart, pectoral fin, and cardiopharygneal vasculature defects. Upon completion of my aims I will have tested how shared lineage origins, and perturbation of shared signaling pathways and lineage-specific inputs, contribute to human diseases with linked heart, limb, and lung defects. Altogether, my proposed research aims, my technical training, and my career development plan will provide the basis for my career goal of becoming an independent investigator at a leading research institution. During the K99 phase, I will receive mentorship from my mentor Dr. Christian Mosimann and my advisory team, and technical training from my collaborative team consisting of light sheet microscopy, scRNA-seq experimental design, and cardiovascular phenotypic analysis. My career development plan will further my training in intellectual development and collaboration, scientific writing and communication, and mentorship and teaching, facilitating my transition to an independent investigator during the R00 phase. The section of Developmental Biology within the Department of Pediatrics at the CU Anschutz Medical Campus is an exceptionally supportive environment, providing extensive resources that will contribute to my continued academic success and professional development, facilitating the accomplishment of my career goals.

人类疾病很少局限于单个组织或器官，而是存在 合并症。一个常见的例子包括一组链接心脏，肢体和肺的先天性疾病 缺陷。使用斑马鱼作为模型，我的研究的目的是发现单个因果基因如何驱动 心脏，肢体和肺中的多器官合并症。了解基本机制将有助于 跨疾病组开发治疗剂，并改善患者的预测诊断 复杂的多器官疾病。在成人身体中，心脏，肢体和肺脉管系统似乎无关 结构水平，但在胚胎中具有密切的发育起源。这些结构都来自 未承诺的侧板中胚层可为众多器官系统携带祖细胞。我的总体 假设认为心脏，肢体和肺血管的谱系（心脏咽喉血管） 源自侧板中胚层内的公共祖细胞种群，形成在相互连接的 通过TBX-FGF和谱系特异性信号输入的作用。 在AIM 1中，我将测试心脏，胸鳍和心脏咽喉血管之间的谱系连接 使用转基因谱系标记和基于CRISPR-CAS9的条形码实验。我的工作将直接测试我的 假设心脏，肢体和肺部缺陷与受影响细胞的共同发育起源联系起来 类型。在AIM 2中，我将在功能上测试TBX-FGF信号通路和谱系特异性的贡献 关节心脏，肢体和心脏咽管谱系模式的输入。这些实验将测试 共享和谱系特异性信号扰动对心脏同时发生和严重程度的影响，胸鳍， 和心咽血管缺陷。完成目标后，我将测试如何共享谱系 起源和共享信号通路和谱系特异性输入的扰动有助于人类疾病 有连接的心脏，肢体和肺部缺陷。 我拟议的研究目的，我的技术培训以及我的职业发展计划总共将提供 我职业生涯目标的基础是成为领先的研究机构的独立研究者。期间 在K99阶段，我将获得我的导师克里斯蒂安·莫斯曼（Christian Mosimann）和我的顾问团队的指导，以及 我的协作团队的技术培训，包括轻度显微镜，SCRNA-SEQ实验 设计和心血管表型分析。我的职业发展计划将进一步培训 智力发展与协作，科学写作和沟通以及指导和教学， 在R00阶段促进我向独立研究者的过渡。发展部分 Cu Anschutz医疗校园的儿科系内的生物学是一个非常支持的 环境，提供广泛的资源，将有助于我持续的学术成功和 专业发展，促进我的职业目标的实现。