Therapeutic targeting of Wnt & Metabolism in Colon Cancer

项目摘要/摘要 在这个国家，大多数散发性结肠癌是由基因突变引起的，这些基因突变激活了典型的 WNT/β-连环蛋白信号通路。然而，在临床上，结肠癌患者的治疗方式与他们 已经有几十年了：一种化疗方案(例如，FOLFOX用于III、IV期)和一种附加的抗- 血管生成剂，如贝伐单抗或阿瓦斯丁。附加治疗可延长约5个月的生命，但它们 几乎总是失败，因为肿瘤产生了抗药性。只有一种Wnt抑制剂进入了II期临床 转移性结肠癌试验。显然，有必要增加治疗替代方案。我们最近在以下方面的工作 Wnt信号已确定Wnt/β-连环蛋白通路引导癌症代谢，促进有氧代谢 包括丙酮酸在内的糖酵解基因上调表达与血管生成 脱氢酶激酶1(PDK1)和SLC16A1/MCT-1。我们还发现，Wnt信号活性和 糖酵解在异种移植物和患者肿瘤中是不同的。肿瘤的异质性是公认的标志 它被认为源于新陈代谢和信号网络的细胞内重新编程，一 帮助癌细胞适应不断变化的营养供应和压力环境的灵活性。WNT 我们在异种移植的结肠肿瘤中观察到的糖酵解异质性表现为一系列细胞簇(5-7 细胞，约30微米中心到中心)在肿瘤的增殖性、非坏死性的外围。我们还观察了肿瘤 使用单细胞测序分析逐个细胞的异质性。出现了一系列规则的 异种移植瘤中的细胞团表明，异质性是有模式的和受调控的。因此，我们 开发了一个数学模型，在肿瘤和Wnt信号的前沿重现了这种模式 是这一模式的核心。我们已经测试了几个关于代谢模式和基因的模型预测。 表达式和结果支持了该模型的有效性。然而，一个重要的预测并没有 经过测试--这是与开发癌症治疗新想法最相关的一个。在肿瘤生长的电子计算机模型中 提示联合应用抗WNT和抗PDK(糖酵解)药物应协同作用以抑制 肿瘤生长。因此，这里提出的项目重点是验证这一预测。如果这个预测是 正确，它将指向一种新的针对肿瘤的阿喀琉斯鞋跟。在AIM I中，我们将测试反 WNT/抗PDK联合治疗将使用以下药物协同抑制结肠癌移植瘤生长 目前正在进行临床试验。在AIM II中，我们将评估肿瘤和小鼠间质对单一和 使用单细胞RNA测序(ScRNAseq)、批量RNAseq和 免疫组织化学。这个短期的、高度可行的项目是迈向更大范围的必要步骤。 研究集中在人PDX和原发患者组织中的癌症代谢。

Project Summary/Abstract Most of the sporadic colon cancer in this country arises from genetic mutations that activate the canonical WNT/beta-catenin signaling pathway. Yet in the clinic, colon cancer patients are treated the same way they have been for decades: a chemotherapy regimen (e.g. FOLFOX for stages III, IV) and an add-on anti- angiogenic agent such as Bevacizumab or Avastin. Add-on treatments extend life for ~5 months, but they almost always fail because tumors develop resistance. Only one Wnt inhibitor has made it to a Phase II clinical trial for metastatic colon cancer. Clearly there is a need to increase treatment alternatives. Our recent work on WNT signaling has determined that the Wnt/β-catenin pathway directs cancer metabolism, promoting aerobic glycolysis and angiogenesis through upregulated expression of glycolysis genes including Pyruvate Dehydrogenase Kinase 1 (PDK1) and SLC16A1/MCT-1. We also discovered that Wnt signaling activity and glycolysis are heterogeneous in xenografts and patient tumors. Tumor heterogeneity is a recognized hallmark of cancer, and it is thought to derive from intracellular reprogramming of metabolic and signaling networks, a flexibility that helps cancer cells adapt to fluctuating nutrient availability and a stressful environment. The Wnt and glycolytic heterogeneity we observe in xenografted colon tumors presents as an array of cell clusters (5-7 cells, ~30µm center-to-center) at the proliferating, non-necrotic periphery of tumors. We also observe tumor heterogeneity on a cell-by-cell basis using single cell sequencing analysis. The emergence of a regular array of cell clusters in xenografted tumors suggests that the heterogeneity is patterned and regulated. We therefore developed a mathematical model that recreates this pattern at the leading edge of tumors and Wnt signaling lies at the core of this model. We have tested several model predictions about metabolic patterning and gene expression and the results support the validity of the model. However, an important prediction has not been tested - the one most relevant to developing new ideas for cancer therapy. In silico modeling of tumor growth suggests that a combination of anti-Wnt and anti-PDK (glycolysis) drugs should act synergistically to repress tumor growth. Therefore, the project proposed here is focused on testing this prediction. If this prediction is correct, it would point to a new Achilles Heel for targeting in tumors. In Aim I we will test whether anti- Wnt/anti-PDK combination therapy will synergistically suppress colon cancer xenograft growth using drugs that are currently in clinical trials. In Aim II we will assess how tumor and mouse stroma respond to the single and combination therapy regimen using single cell RNA sequencing (scRNAseq), bulk RNAseq and immunohistochemistry. This short-term, highly feasible project is a necessary step towards a larger scope study focused on cancer metabolism with human PDX and primary patient tissue.