Investigate heterogeneous neutrophils in NSCLC

在基因工程小鼠肺腺癌模型中，我们发现SiglecF+中性粒细胞是一个独特的中性粒细胞亚群。SiglecF+中性粒细胞在肿瘤微环境中丰富，但其功能尚不清楚。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分析。我们的生物信息学分析预测这组中性粒细胞中线粒体功能障碍和中性粒细胞受损。为了进一步了解SiglecF+中性粒细胞的功能，我们正在建立一个条件基因敲除SiglecF的小鼠模型(SiglecF Flox/Flox小鼠)，并计划通过将这个新的中性粒细胞缺乏SiglecF的小鼠模型与另一个具有中性粒细胞特异性Cre的小鼠品系(Catchup小鼠)杂交来创建一个小鼠模型。第一批SiglecF FLOX/FLOX小鼠诞生了，我们正在鉴定过程中。为了分析SiglecF的表达是如何控制的，我们进行了RNA测序，并鉴定了几个调节SiglecF转录的潜在转录因子。我们正在使用一个小鼠模型来探索是否有任何候选转录因子对SiglecF的表达是重要的，在该模型中，我们可以通过CRISPR特异性地敲除或过度表达中性粒细胞中的靶标。除了研究小鼠中不同种类的中性粒细胞外，我们还修改了我们的临床方案，允许我们收集肺癌患者的骨髓样本和肺癌样本。我们计划使用先进的测序技术来研究骨髓中的异质性中性粒细胞和肿瘤微环境。我们已经成功地建立了我们先进的测序平台，并已经进行了第一批测序实验。

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.