Investigate heterogeneous neutrophils in NSCLC

研究 NSCLC 中的异质中性粒细胞

• 批准号: 10262613
• 负责人: Chen Zhao
• 金额: $ 34.61万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10262613
• 关键词: Address; Anesthesia procedures; Bioinformatics; Blood; Bone Marrow; Bone Marrow Aspiration; Bone Marrow Neoplasms; Cells; Clinical Protocols; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Development; Epitopes; Excision; Expression Profiling; Flow Cytometry; Genetic Transcription; Genetically Engineered Mouse; Harvest; Hematopoiesis; Impairment; Knock-out; LoxP-flanked allele; Lung Adenocarcinoma; Lung Neoplasms; Malignant neoplasm of lung; Mitochondria; Mouse Strains; Mus; Myeloid Cells; Non-Small-Cell Lung Carcinoma; Operative Surgical Procedures; Pathway Analysis; Patients; Play; Population; Process; Role; Sampling; Site; Subgroup; Techniques; Tumor-Derived; conditional knockout; experimental study; healthy volunteer; indexing; mouse model; neutrophil; overexpression; peripheral blood; sequencing platform; single cell analysis; single cell sequencing; single-cell RNA sequencing; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; tumor; tumor microenvironment

We identified SiglecF+ neutrophils as a unique neutrophil subgroup in a genetically engineered mouse model of lung adenocarcinoma. SiglecF+ neutrophils are enriched in the tumor microenvironment, but their function is poorly understood. We first confirmed the existence of this group of neutrophils and then did comprehensive transcriptomic analysis. Our bioinformatic analysis predicts mitochondria disfunction and impaired neutrophil swarming in this group of neutrophils. To further understand the function of SiglecF+ neutrophils, we are generating a conditional knockout mouse model of SiglecF (SiglecF flox/flox mouse), and plan to create a mouse model specifically lacking SiglecF in neutrophils by crossing this new floxed strain to another mouse strain with neutrophil specific Cre (Catchup mice). The first batch of SiglecF flox/flox mice were born and we are in the process of characterization. To analyze how SiglecF expression is controlled, we performed RNA sequencing and identified several potential transcription factors regulating SiglecF transcription. We are exploring whether any of the candidate transcription factors are important for SiglecF expression using a mouse model in which we can specifically knock out or overexpress targets in neutrophils via CRISPR. In addition to studying heterogenous neutrophil populations in mice, we amended our clinical protocol to allow us to collect bone marrow samples and lung tumor samples from patients with lung cancers. We plan to use advanced sequencing techniques to study the heterogeneous neutrophils in bone marrow and the tumor microenvironment. We have successfully established our advanced sequencing platform and already performed the first batch of sequencing experiments.

我们将SiglecF+中性粒细胞鉴定为肺腺癌基因工程小鼠模型中独特的中性粒细胞亚群。SiglecF+中性粒细胞在肿瘤微环境中富集，但其功能知之甚少。我们首先确认了这组中性粒细胞的存在，然后进行了全面的转录组学分析。我们的生物信息学分析预测了这组中性粒细胞中线粒体功能障碍和受损的中性粒细胞群集。为了进一步了解SiglecF+嗜中性粒细胞的功能，我们正在生成SiglecF的条件性敲除小鼠模型（SiglecF flox/flox小鼠），并计划通过将这种新的floxed品系与具有嗜中性粒细胞特异性Cre的另一种小鼠品系（Catchup小鼠）杂交来创建嗜中性粒细胞中特异性缺乏SiglecF的小鼠模型。第一批SiglecF flox/flox小鼠诞生，我们正在进行表征。为了分析SiglecF表达是如何被控制的，我们进行了RNA测序并鉴定了几种调节SiglecF转录的潜在转录因子。我们正在探索是否有任何候选转录因子对SiglecF表达很重要，使用小鼠模型，我们可以通过CRISPR特异性敲除或过表达中性粒细胞中的靶点。除了研究小鼠中的异源中性粒细胞群体外，我们修改了我们的临床方案，允许我们从肺癌患者中收集骨髓样本和肺肿瘤样本。我们计划使用先进的测序技术来研究骨髓和肿瘤微环境中的异质性中性粒细胞。我们已经成功建立了先进的测序平台，并已经进行了第一批测序实验。