Clariom D Pico gene array of microdissected GBM areas or PBMCs.. Clariom D Pico gene array of microdissected GBM areas or PBMCs.

Glioblastoma multiforme (GBM) presents a formidable clinical challenge due to its complex microenvironment. Here, we introduce lipid droplet (LD)-loaded macrophages, or tumor-associated foam cells (TAFs), as a previously unidentified immune cell population in GBM. Through extensive analyses of patient tumors, together with in vitro and in vivo investigations, we reveal that TAFs exhibit distinct pro-tumorigenic characteristics related to hypoxia, mesenchymal transition, angiogenesis, and impaired phagocytosis. Moreover, TAF presence correlates with worse patient outcome. Our mechanistic investigations demonstrate that TAF formation is facilitated by lipid cargo from extracellular vesicles released by GBM cells. Importantly, we demonstrate that targeting key enzymes involved in LD formation, such as DGAT1 or ACSL, effectively disrupts TAF functionality. This study establishes TAFs as a prominent immune cell entity in GBM and provides valuable insights into their interplay within the microenvironment. Disruptin Overall design: 13 samples, out of which: 4 are from microdissected regions high in macrophage marker intensity (2 with high lipid droplet (LD+) and 2 with low lipid droplet (LD-)); and 9 samples are from patient PBMCs (4 monocytes (HC_t0 and P_t0), 2 macrophages (HC_CTRL), and 2 of macrophages plus extracellular vesicles (HC_EVs)). The LD- and HC_CTRL samples are reference/control samples. The brain samples were microdissected from glioblastoma tissues, underwent RNA extraction with AllPrep DNA/RNA Micro Kit for RNA, and hybridization on Affymetrix Clariom Human D Pico microarray cartrige. The PBMCs samples were isolated by CD14+ microbeads, and underwent RNA extraction with AllPrep DNA/RNA Micro Kit for RNA, and hybridization on Affymetrix Clariom Human D Pico microarray cartrige

多形性胶质母细胞瘤（Glioblastoma multiforme, GBM）因其复杂的肿瘤微环境，成为极具挑战性的临床难题。本研究首次报道了负载脂滴（lipid droplet, LD）的巨噬细胞——肿瘤相关泡沫细胞（tumor-associated foam cells, TAFs）——作为GBM中此前未被识别的免疫细胞亚群。通过对患者肿瘤组织的大规模分析，结合体外与体内实验研究，我们揭示TAFs展现出与缺氧、间质转化、血管生成及吞噬功能受损相关的独特促肿瘤特性。此外，TAFs的存在与患者不良预后显著相关。机制研究证实，TAFs的形成可由GBM细胞释放的细胞外囊泡所携带的脂质货物所促进。重要的是，我们发现靶向参与LD形成的关键酶（如DGAT1或ACSL），可有效破坏TAFs的功能。本研究确立TAFs作为GBM中一类重要的免疫细胞实体，并为其在肿瘤微环境中的相互作用提供了宝贵见解。整体实验设计：本数据集共包含13例样本，其中4例来自巨噬细胞标志物表达水平较高的显微切割区域（2例为高脂滴阳性（LD+）样本，2例为低脂滴阳性（LD-）样本）；剩余9例来自患者外周血单个核细胞（peripheral blood mononuclear cell, PBMCs）：包括4例单核细胞（HC_t0与P_t0）、2例巨噬细胞（HC_CTRL）以及2例添加细胞外囊泡的巨噬细胞（HC_EVs）。其中LD-样本与HC_CTRL样本作为参考对照样本。脑组织样本均从胶质母细胞瘤组织中显微切割获取，采用AllPrep DNA/RNA Micro Kit进行RNA提取，随后在Affymetrix Clariom Human D Pico微阵列芯片上完成杂交。外周血单个核细胞样本采用CD14+磁珠进行分离，同样使用AllPrep DNA/RNA Micro Kit进行RNA提取，并在Affymetrix Clariom Human D Pico微阵列芯片上完成杂交。