Aberrant Glycogen in Lung Adenocarcinoma Tumorigenesis

摘要： 肺腺癌(LUAD)是肺癌的主要组织亚型，也是肺癌的主要病因。 世界范围内与癌症相关的死亡率。对于相当数量的LUAD患者来说，唯一的治疗选择 可用的是传统的多药剂化疗结合手术和/或放射治疗。对于这些患者来说， 令人失望的是，5年存活率仍然很低。驱动LUAD扩散的其他分子机制 和肿瘤发生仍然是肺癌研究中的一个关键知识缺口，也是阻碍 个性化治疗的发展。最近的报告，包括我们自己的报告，揭示了糖原在 肺部肿瘤进展。在这些基础研究的基础上，我们开发了一种健壮和精确的 利用质谱学成像技术以50微米的空间分辨率原位显示糖原 与以前的方法相比，灵敏度提高了1,000倍。使用这项技术，我们定义了 在肯塔基大学国家癌症研究所指定的癌症中心治疗的122名非小细胞肺癌患者的糖原水平。 我们的初步数据表明：1)LUAD患者的糖原显著升高，而LUAD患者的糖原未见显著升高 正常肺组织。2)糖原升高是LUAD的组织特异性标志，在肺中未观察到 鳞状细胞癌。3)LUAD-糖原在结构上是独特的，具有更高的磷酸化和 分支。4)这种LUAD表型与糖原磷酸酶显著的蛋白降低有关 拉福林。引人注目的是，在模型肺细胞系和KrasG12D/P53-/-LUAD小鼠模型中，Laforin基因敲除驱动：1) 糖原过度磷酸化，2)增加与AMP激活的主要代谢调节因子的亲和力 蛋白激酶(AMPK)，3)AMPK活性降低，4)促进肿瘤增殖和进展。 我们假设结构上独特的LUAD-糖原是LUAD代谢的关键成分， 扩散和进展。本研究的总体目标是明确LUAD-糖原的病因学。 癌症新陈代谢和肿瘤进展。为了实现这一点，我们将：定义LUAD-糖原临床 课程及其与AMPK的相互作用(目标1)。然后，我们将确定LUAD-糖原在体内的信号转导作用 通过AMPK的细胞代谢(目标2)。最后，我们将确定LUAD-糖原在肿瘤中的作用 体内进展和早期转化(Aim3)。这项建议建立在令人兴奋和严谨的初步基础上 数据，并提出了一种综合的方法，以定义这一独特的LUAD特征的过量糖原利用 强大、互补和最先进的方法，如质谱学成像、蛋白质和 糖原生物化学和靶向代谢组学。这项提案的显著发现将显著 推进糖原在LUAD生物学和进展中的作用的知识库，并推动 发现可用于仅符合以下条件的LUAD人群的个性化疗法 常规化疗。

Abstract: Lung adenocarcinoma (LUAD) is the major histological subtype of lung cancer and the leading cause of cancer-related mortalities worldwide. For a substantial number of LUAD patients, the only treatment options available are traditional multi-agent chemotherapy coupled with surgery and/or radiation. For these patients, the 5-year survival remains disappointingly low. Additional molecular mechanisms driving LUAD proliferation and tumorigenesis remain a critical knowledge gap in lung cancer research, and a major barrier for the development of personalized therapies. Recent reports, including our own, reveal critical roles for glycogen in lung tumor progression. Building on these foundational studies, we developed a robust and precision technology to visualize glycogen in situ with 50 µm spatial resolution using mass spectrometry imaging that provides 1,000x increased sensitivity compared to previous methods. Using this technology, we defined glycogen levels in 122 NSCLC patients treated at the University of Kentucky’s NCI Designated Cancer Center. Our preliminary data demonstrate that: 1) significantly elevated glycogen is observed in LUAD and not in normal lung tissue. 2) Elevated glycogen is a LUAD tissue-specific hallmark and is not observed in lung squamous cell carcinoma. 3) LUAD-glycogen is structurally unique with increased phosphorylation and branching. 4) This LUAD phenotype correlated with marked protein decreases in the glycogen phosphatase laforin. Strikingly, laforin knockout in model lung cell lines and the KrasG12D/p53-/- LUAD mouse model drives: 1) glycogen hyper-phosphorylation, 2) increased affinity with the master metabolic regulator AMP-activated protein kinase (AMPK), 3) decreased AMPK activity, and 4) accelerated tumor proliferation and progression. We hypothesize that the structurally unique LUAD-glycogen is a critical component of LUAD metabolism, proliferation, and progression. The overall objective of this study is to define the etiology of LUAD-glycogen on both cancer metabolism and tumor progression. To achieve this, we will: Define the LUAD-glycogen clinical course and its interaction with AMPK (Aim 1). Then, we will define the signaling role of LUAD-glycogen in cellular metabolism through AMPK (Aim 2). Finally, we will establish the role of LUAD-glycogen in tumor progression and early transformation in vivo (Aim3). This proposal builds on exciting and rigorous preliminary data and presents an integrated approach to define this unique LUAD hallmark of excess glycogen utilizing robust, complementary, and state-of-the-art methodologies such as mass spectrometry imaging, protein and glycogen biochemistry, and targeted metabolomics. The salient findings from this proposal will significantly advance the knowledge base regarding the roles of glycogen in LUAD biology and progression and drive the discovery of personalized therapies that can be leveraged for the LUAD population that only qualify for conventional chemotherapy.