Effect of Cyclical Intermittent Hypoxia on Lung Cancer Progression

目的：肺癌的预后仍然很差，退伍军人的肺癌发病率更高。 阻塞性睡眠呼吸暂停（OSA）与肺癌和肺癌的预后有关。 癌症发病率2 -5和死亡率2，6-8更高，退伍军人的OSA患病率更高， 9我们的首要目标是确定周期性间歇性缺氧 (CIH)是OSA的主要潜在病理，促进肺癌进展。我们假设CIH 增加未成熟免疫细胞向抑制性免疫细胞的分化，然后，无意中， 保护癌症。我们已经发表了CIH加速原发性肺癌的进展， 转基因KrasG 12 D +; p53 fl/fl;肉豆蔻化p110 fl/fl ROSA-gfp（TT-Kpp）小鼠（由VA CPPF资助）。这 下一步是探索CIH如何促进癌症进展的机制。研究设计： 这项资助的技术创新之处在于，我们将首先衡量轻度、中度和 严重CIH对TT-Kpp小鼠中癌症进展的影响（目的1）。这项资助的概念创新在于， 将是第一个使用这种TT-Kpp模型来探索系统性CIH是否会增加产量、分化 以及抑制性免疫细胞对原发性肺癌的募集。方法学：目标1：TT-Kpp小鼠 注射Ad 5CC 10 Cre病毒的小鼠将暴露于4种条件之一：假手术（室内空气），轻度CIH（CIH 15， （FiO 2下降0.21至FiO 2 0.15 [SaO 2最低值为82%]）中度CIH（CIH 10，（FiO 2下降0.21至FiO 2 0.15 [SaO2最低点为61%]）或严重CIH（CIH 5; FiO 2下降0.21至FiO 2 0.05 [SaO 2最低点为37%]）。初级 结果将是定量肿瘤体积（每月microCT扫描）;次要结果将是生存率。目的 2：评估CIH 5（相对于假手术）对抑制性免疫细胞的产生、分化和募集的影响。 在患癌和未患癌的小鼠中的细胞。主要结局将是定量骨中的M-MDSC/TAM/TdR 早期（2周）和晚期（4个月）时间点的骨髓、脾脏、胸腺、左肺和右肺;次要 结果将是抑制性免疫细胞的功能测定。探索目标3：开发克隆癌细胞 TT-Kpp小鼠的肺癌。此外，评估CIH 5（相对于假手术）调节（1）趋化因子/细胞因子的作用 肺癌细胞内的基因（通过早期和晚期时间点的PCR组）和（2）其他基因（通过早期和晚期时间点的RNA-seq）。 一个时间点）。PCR和RNA-seq信息将一起用于未来的资助， CRISPR编辑克隆细胞内的重要基因，并将其注射到野生型小鼠（同基因小鼠）中。 原位模型）以进一步理解CIH对癌症进展的作用机制。调查结果： 成绩.临床关系：我们可以提供证据表明，CIH，阻塞性睡眠的主要病理 呼吸暂停是一个可改变的风险因素，可以提高肺癌的生存率。影响/意义：气道正压通气 压力，治疗OSA没有副作用，可能是一个强大的辅助治疗，以防止肺 癌症进展

Objective: Lung cancer continues to have a very poor prognosis, with Veterans having a higher incidence of lung cancer and worse outcome compared to civilians.1 Obstructive sleep apnea (OSA) has been associated with higher cancer incidence2-5 and mortality,2, 6-8 and again, Veterans have a higher prevalence of OSA that is more severe compared to civilians.9 Our overarching objective is to determine how cyclical intermittent hypoxia (CIH), a major underlying pathology of OSA, promotes lung cancer progression. Our hypothesis is that CIH increases the differentiation of immature immune cells to suppressor immune cells which then, inadvertently, protects the cancer. We have published that CIH accelerates primary lung cancer progression in Triple Transgenic KrasG12D+; p53fl/fl; myristolated p110fl/fl ROSA-gfp (TT-Kpp) mice (funded by VA CPPF grant). This grant is the next step - to explore mechanisms of HOW CIH promotes cancer progression. Research Design: The technical innovation of this grant is that we will be the first to measure the effect of mild, moderate and severe CIH on cancer progression in TT-Kpp mice (Aim 1). The conceptual innovation of this grant is that we will be the first to use this TT-Kpp model to explore whether systemic CIH increases production, differentiation and recruitment of suppressor immune cells to primary lung cancer. Methodology: Aim 1: TT-Kpp mice injected with Ad5CC10Cre virus will be exposed to one of 4 conditions: sham (room air), mild CIH (CIH15, (drops in FiO2 0.21 to FiO2 0.15 [SaO2 nadir of 82%]) moderate CIH (CIH10, (drops in FiO2 0.21 to FiO2 0.15 [SaO2 nadir of 61%]) or severe CIH (CIH5; drops in FiO2 0.21 to FiO2 0.05 [SaO2 nadir of 37%]). Primary outcome will be quantitative tumor volumes (monthly microCT scans); secondary outcome will be survival. Aim 2: Assess the effect of CIH5 (vs Sham) on production, differentiation and recruitment of suppressor immune cells in mice with and without cancer. Primary outcome will be quantitative M-MDSC/TAM/Tregs in bone marrow, spleen, thymus, left lung and right lung at early (2 weeks) and late (4 months) time points; secondary outcome will be functional assays of suppressor immune cells. Exploratory Aim 3: Develop a clone cancer cell from lung cancer of TT-Kpp mice. Also, assess the effect of CIH5 (vs Sham) regulating (1) chemokine/cytokine genes within lung cancer cell (via PCR panel at early and late time point) and (2) other genes (via RNA-seq at one time point). Together, the PCR and RNA-seq information will be utilized in a future grant where we will use CRISPR editing of significant genes within clone cells and inject them into a wild type mouse (syngeneic orthotopic model) to further understand mechanism of effect of CIH on cancer progression. Findings: First submission. Clinical Relationships: We may provide evidence that CIH, a major pathology of obstructive sleep apnea, is a modifiable risk factor that can improve lung cancer survival. Impact/Significance: Positive airway pressure, a treatment for OSA has no side effects and may be a powerful adjunct therapy to prevent lung cancer progression.