Investigating the role of RUNX2 activation across cancer evolution in lung adenocarcinoma

项目摘要 肺腺癌(LUAD)是癌症相关死亡的主要原因之一；然而，只有 基于某些致癌驱动因素突变的存在，患者有资格接受靶向治疗。在……里面 此外，许多治疗方法针对的是致癌事件，但肿瘤细胞获得了替代方案 肿瘤进展过程中的依赖性。在这项提议中，我的目标是询问一种遗传学上的肿瘤进展 工程化小鼠(GEM)肺腺癌模型称为KP(KrasG12D/；p53f/f)模型。 忠实地概括了人类疾病的特征。在KP肿瘤中发生转录失调 进展，并不能用获得体细胞突变来解释，这表明肿瘤的进化是 主要由表观遗传机制驱动。在之前利用单细胞染色质图谱的工作中，我 确定KP肿瘤具有显著的表观遗传学肿瘤内异质性。有趣的是，我 发现晚期肿瘤细胞群体表现出RUNX2转录的渐进性激活 活动。此外，RUNX2激活的细胞增加了相关基因周围染色质的可及性 然而，RUNX2改变染色质状态和基因的机制(S) 表达程序并不完善。基于这个和其他数据，我的中心假设是 Runx2在晚期肿瘤进展中作为主要调节转录因子发挥作用 异常地启动基因程序，服务于：1)重塑局部肿瘤微环境；2)促进 不同的间质/免疫成分，以及iii)增加了肿瘤内的异质性。我的第一次 AIM将询问全局染色质变化，并确定RUNX2-中与RUNX2相关的辅因子 激活的细胞。第二个目标将特别关注RUNX2激活在血管重构中的作用。 局部肿瘤微环境。第三个目标将更广泛地评估 晚期癌细胞中的异质性基因表达程序。这项建议寻求在成果的基础上再接再厉 在我博士后培训期间产生的，并将作为我的独立研究小组的初始重点。 我在研究生院接受的癌症生物学和表观遗传学背景，加上我广泛的 在我博士后培训中获得的宝石模型和表观基因组学技术方面的专业知识，使我能够在 成功地过渡到独立。作为这笔赠款的一部分，我将在我的机构组建一个顾问团队 支持我在本提案中概述的过渡和职业发展培训活动。我要买下 利用我博士后机构的各种资源以及我获得 具有独立的职位，能够发展与指导、沟通和研究道德相关的技能。总体而言， 这份提案中概述的培训将极大地支持我的继续发展，目标是使 在表观遗传学和癌症生物学领域做出了重大贡献。

Project Summary Lung adenocarcinoma (LUAD) is one of the leading causes of cancer-related death; however, only a subset of patients are eligible for targeted therapies based on the presence of mutations in certain oncogenic drivers. In addition, many therapeutic approaches target cancer-initiating events, yet tumor cells acquire alternative dependencies during tumor progression. In this proposal, I aim to interrogate tumor progression in a genetically engineered mouse (GEM) model of lung adenocarcinoma termed the KP (KrasG12D/+; p53f/f) model, which faithfully recapitulates features of human disease. Transcriptional dysregulation occurs across KP tumor progression and is not explained by the acquisition of somatic mutations, suggesting that tumor evolution is driven predominantly by epigenetic mechanisms. In previous work leveraging single-cell chromatin profiling, I determined that KP tumors were marked by substantial epigenetic intratumoral heterogeneity. Interestingly, I found that late-stage populations of tumor cells exhibited progressive activation of RUNX2 transcriptional activity. In addition, RUNX2 activated cells have increased chromatin accessibility surrounding genes involved in extracellular secretion; however, the mechanism(s) by which RUNX2 alters chromatin state and gene expression programs is not well-established. Based on this and other data, my central hypothesis is that RUNX2 functions as a master regulatory transcription factor during late-stage tumor progression that aberrantly initiates gene programs that serve to i) reshape the local tumor microenvironment, ii) facilitate differential stromal/immune composition, and iii) contribute to increased intratumoral heterogeneity. My first aim will interrogate the global chromatin changes and identify RUNX2-associated cofactors in RUNX2- activated cells. The second aim will specifically focus on the role of RUNX2 activation on the remodeling of the local tumor microenvironment. The third aim will more broadly assess the spatial localization of heterogeneous gene expression programs in late-stage cancer cells. This proposal seeks to build on results generated during my postdoctoral training and will serve as the initial focus of my independent research group. My background in cancer biology and epigenetics from my graduate school training, paired with my extensive expertise in GEM models and epigenomic technologies from my postdoctoral training, positions me for a successful transition to independence. As part of this grant, I will assemble an advisory team at my institution to support my transition and career development training activities outlined in this proposal. I will take advantage of the varied resources at my postdoctoral institutions and the institution where I secure an independent position to develop skills related to mentorship, communication, and research ethics. Overall, the outlined training plain in this proposal will greatly support my continued development, with the goal of making substantial contributions in the fields of epigenetics and cancer biology.