Mapping and prediction of quantitative transcription factor dosage effects to understand variation in craniofacial morphology and disease

项目总结 越来越多的证据表明转录调控的健壮性，然而人类基因研究表明 复杂性状和罕见疾病的表型敏感性对转录因子(TF)剂量的影响 组织因子剂量效应的定量研究。这些特征以人类的发展为例证 面部，在一系列具有典型特征的证候中经常被扰乱。这项提议的目的是 为了开发和应用工具来精确地调节颅面神经嵴细胞中TFS的剂量， 面部的祖细胞，并评估多个生物水平的影响-分子、细胞和 形态上的。该提案最初侧重于SOX9，使用不同的方法来理解规则 决定了哪些调控元件和基因对SOX9剂量敏感(目标1)，以及它们的 表型后果(目标2)。然后，这样的规则将被推广到另外五个颅面TF (AIM 3)，并用于剖析与颅面部相关的遗传变异的潜在机制 形态或疾病(目标4)。这些研究将揭示颅面形态和疾病的驱动因素 变异，提供对转录调控的一般见解，并帮助转变我们的方法 研究TF函数，从本质上从二进制到定量。 这项拟议的研究，以及对我职业发展的全面规划，结合了我的科学 功能基因组学、基因调控和人类遗传学的背景和博士后培训 在机器学习、老鼠遗传学和形态分析方面的额外培训机会。这就做 在充满活力的科学环境中得到乔安娜·威索卡和乔纳森·普里查德博士的指导(斯坦福大学 化学和系统生物学)，并从我的顾问委员会詹姆斯博士那里接受额外的培训 费雷尔、安舒尔·昆达杰和贝尼迪克特·哈尔格里姆松。 参加科学训练和非科学训练将促进我向独立的过渡 和研讨会，以及在科学会议上展示我的工作的经验。我也会收获 有指导学生和技术人员的经验，并将磨练我的赠款管理和写作技能。 总的来说，我的导师已经指导了他们的25名博士后进行独立的学术研究 在我找工作的过程中，他们会给我提供实用的建议和反馈。 我的长期职业目标是指导一个独立的研究项目，旨在了解 头面部形态和疾病风险的数量变异的分子基础。到目前为止，我已经 通过我的研究经验、出版物和 与更广泛的科学界接触。然而，我坚信，K99指导阶段 将通过提供访问关键资源和培训来最大化我的成功机会，这将是 否则我的博士后经验就不够丰富了。

PROJECT SUMMARY Accumulating evidence suggests robustness of transcriptional regulation, yet human genetic studies indicate phenotypic sensitivity to transcription factor (TF) dosage in both complex traits and rare disorders, motivating quantitative studies of TF dosage effects. These features are exemplified by the development of the human face, which is frequently disrupted in a range of syndromes with typical features. The aim of this proposal is to develop and apply tools to precisely modulate dosage of craniofacial TFs in cranial neural crest cells, the progenitors of the face, and assess effects across multiple biological levels – molecular, cellular, and morphological. The proposal initially focuses on SOX9, using diverse approaches to understand the rules that determine which regulatory elements and genes are sensitive to SOX9 dosage (Aim 1), as well as their phenotypic consequences (Aim 2). Then, such rules will be generalized to five additional craniofacial TFs (Aim 3) and used to dissect mechanisms underlying genetic variants associated with craniofacial morphology or disease (Aim 4). These studies will reveal drivers of craniofacial morphological and disease variation, provide general insights into transcriptional regulation, and help transform our approach to studying TF function, from binary to quantitative in nature. The proposed research, along with a comprehensive plan for my career development, combines my scientific background and postdoctoral training in functional genomics, gene regulation, and human genetics with opportunities for additional training in machine learning, mouse genetics, and morphological analysis. I will be mentored by Drs. Joanna Wysocka and Jonathan Pritchard in a vibrant scientific environment (Stanford Chemical and Systems Biology) and receive additional training from my advisory committee of Drs. James Ferrell, Anshul Kundaje, and Benedikt Hallgrimsson. My transition to independence will be facilitated by participation in both scientific and non-scientific trainings and workshops, as well as experience in presenting my work at scientific conferences. I will also gain experience mentoring students and technicians, and will hone my grant management and writing skills. Collectively, my mentors have guided over 25 of their former postdocs to independent academic research positions, and they will provide practical advice and feedback during my faculty job search. My long-term career goal is to direct an independent research program aimed at understanding the molecular underpinnings of quantitative variation in craniofacial morphology and disease risk. So far I have achieved significant progress towards this goal through my research experience, publications, and engagement with the broader scientific community. I firmly believe, however, that a K99 mentored phase will maximize my chances for success by providing access to key resources and training that would be otherwise lacking from my postdoctoral experience.