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，而是呈现出一系列 合并症。常见的例子包括一组心脏、四肢和肺相连的先天性疾病。 缺陷。以斑马鱼为模型，我的研究目标是揭示单个致病基因是如何驱动 心脏、四肢和肺的多器官并存。了解潜在的机制将有助于 对跨疾病组的治疗方法的发展和改善患者的预测性诊断 复杂的多脏器疾病。在成人体内，心脏、四肢和肺的血管系统似乎在 结构水平，但在胚胎中有着密切的发育起源。这些结构都源于 未固定的侧板中胚层，含有许多器官系统的祖细胞。我最重要的是 假说认为心脏、四肢和肺血管的谱系(心咽血管) 起源于侧板中胚层内的共同祖细胞群体，并形成相互连接的 通过TBX-成纤维细胞生长因子和谱系特异性信号输入的作用方式。 在目标1中，我将测试心脏、胸鳍和心咽血管之间的血统联系 使用转基因谱系标记和基于CRISPR-Cas9的条码实验。我的工作将直接考验我的 假设心脏、四肢和肺缺陷由受影响细胞的共同发育起源联系在一起 类型。在目标2中，我将从功能上测试tbx-成纤维细胞生长因子信号通路的作用，以及谱系特异性。 关节心脏、四肢和心咽血管谱系模式的输入。这些实验将测试 共有的和谱系特有的信号扰动对心脏、胸鳍、 和心咽部血管缺陷。在完成我的目标后，我将测试如何分享血统 共同的信号通路和谱系特异性输入的起源和扰动导致人类疾病 心脏、四肢和肺部都有缺陷。 总之，我提出的研究目标、我的技术培训和我的职业发展计划将提供 我的职业目标是成为一家领先研究机构的独立调查员。在.期间 在K99阶段，我将接受我的导师Christian Mosimann博士和我的顾问团队的指导，以及 来自我的协作团队的技术培训，包括光片显微镜、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加强我在 智力开发和协作、科学写作和交流、导师和教学， 帮助我在R00阶段转变为独立调查员。《发展篇》 CU Anschutz医学院儿科的生物学是一个例外的支持 环境，提供了广泛的资源，这将有助于我继续学业成功和 专业发展，促进我职业目标的实现。