Therapeutic targeting of Wnt & Metabolism in Colon Cancer

项目总结/摘要 在这个国家，大多数散发性结肠癌都是由基因突变引起的， 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：化疗方案（如FOLFOX用于III、IV期）和附加抗- 血管生成剂如贝伐单抗或阿瓦斯汀。附加治疗可延长寿命约5个月，但 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 转移性结肠癌的临床试验显然，有必要增加替代治疗。我们最近的工作 WNT信号传导已确定Wnt/β-连环蛋白途径指导癌症代谢，促进有氧代谢 糖酵解和血管生成通过上调糖酵解基因表达，包括丙酮酸 脱氢酶激酶1（PDK 1）和SLC 16 A1/MCT-1。我们还发现，Wnt信号传导活性和 糖酵解在异种移植物和患者肿瘤中是异质的。肿瘤异质性是公认的标志 它被认为来自代谢和信号网络的细胞内重编程， 这有助于癌细胞适应波动的营养供应和压力环境。所述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细胞簇阵列（5-7 细胞，约30µm中心到中心）在肿瘤的增殖，非坏死周边。我们也观察肿瘤 使用单细胞测序分析，在逐个细胞的基础上检测异质性。一系列有规律的 异种移植肿瘤中的细胞簇表明异质性是模式化的和受调节的。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 是这个模型的核心。我们已经测试了关于代谢模式和基因的几个模型预测 表达式，结果支持了模型的有效性。然而，一个重要的预测并没有被 测试-一个最相关的开发癌症治疗的新想法。肿瘤生长的计算机模拟 提示抗Wnt和抗PDK（糖酵解）药物的组合应协同作用， 肿瘤生长因此，这里提出的项目重点是测试这一预测。如果这个预测是 正确的，它将指出一个新的致命弱点，靶向肿瘤。在Aim I中，我们将测试抗- Wnt/抗PDK组合疗法将使用以下药物协同抑制结肠癌异种移植物生长， 目前正在进行临床试验。在目标II中，我们将评估肿瘤和小鼠间质如何对单一和 使用单细胞RNA测序（scRNAseq）、批量RNAseq和 免疫组化这个短期的、高度可行的项目是迈向更大范围的必要步骤 研究集中于人PDX和原发性患者组织的癌症代谢。

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.