Project 3: Identifying transcriptional driver genes and targeting transcription in TSC

项目3：摘要 在这个项目中，我们将研究驱动肿瘤发展的转录因子和表达途径 结节性硬化症（TSC）。我们已经发表了，转录是一个关键的依赖， TSC肿瘤，MITF是血管平滑肌脂肪瘤（AMLK）发展的驱动转录因子。此外，本发明还提供了一种方法， 我们最近通过诱导TSC 2-/-人的肾分化产生了一种新的TSC肾AML模型， 诱导多能干细胞（hIPSC）以产生肾类器官。TSC 2-/-肾的主要细胞亚群 与对照肾分化的hIPSC相比，类器官具有AML表达表型。分开的 研究中，我们使用单细胞RNA测序（scRNA-Seq）来表征AML的组成， 高分辨率，并证实了MITF驱动的转录在AML细胞中的重要性。在当前 我们将研究TSC肿瘤发展中的JUN-AXL通路，这导致了一种新的激酶 抑制剂敏感性使用果蝇模型，我们将进一步详细分析mTORC 1如何调节Mitf 活性，然后将关键发现转化为哺乳动物细胞。最后，我们将使用TSC 2-/- hiPSC肾 类器官模型，以确定AML发展所需的转录电路，使用scRNA-Seq， H3 K27 ac和MITF的scATAC-Seq和scChIP-Seq。我们还将确定整个小组的转录 使用应用于TSC 2-/- hiPSC肾类器官的Perturb-Seq的AML细胞发育所需的因子 模型预期的健康相关结局包括识别多种潜在治疗方法 对于TSC患者和具有TSC 2/TSC 1突变的癌症。

Project 3: Abstract In this project, we will examine the transcription factors and expression pathways that drive tumor development in tuberous sclerosis complex (TSC). We have published already that transcription is a key dependence for TSC tumors, and that MITF is a driver transcription factor for angiomyolipoma (AMLK) development. In addition, we have recently generated a new model of TSC renal AML by inducing renal differentiation in TSC2-/- human induced pleuripotent stem cells (hIPSCs) to generate renal organoids. A major cell subset of TSC2-/- renal organoids have an AML expression phenotype in contrast to control renal-differentiated hIPSCs. In separate studies, we have used single cell RNA sequencing (scRNA-Seq) to characterize the composition of AML at high resolution, and have confirmed the importance of MITF-driven transcription in AML cells. In the current proposal, we will examine the JUN-AXL pathway in TSC tumor development, which leads to a novel kinase inhibitor sensitivity. Using a Drosophila model, we will dissect in further detail how mTORC1 regulates Mitf activity, and then translate key findings to mammalian cells. Last, we will use the TSC2-/- hiPSC renal organoid model to identify the transcriptional circuitry required for AML development, using scRNA-Seq, scATAC-Seq, and scChIP-Seq for H3K27ac and MITF. We will also determine the entire panel of transcription factors required for AML cell development using Perturb-Seq applied to the TSC2-/- hiPSC renal organoid model. Expected health-related outcomes include identification of multiple potential therapeutic approaches for both TSC patients and cancers with TSC2/TSC1 mutations.