Characterising the surfaceome of hypoxic myeloid cells infiltrating the tumour microenvironment.

肿瘤相关巨噬细胞(TAMs)是肿瘤微环境中含量最丰富的细胞之一，与肿瘤的进展密切相关。巨噬细胞的行为受到环境刺激的影响，包括缺氧(缺氧)，以进一步促进治疗抵抗和癌症复发。虽然人们对靶向TAMs以提高抗肿瘤作用感兴趣，但在开发治疗方法之前，还需要进一步研究以确定缺氧性TAMs的标志物。一个电子表面组数据集(Bausch-Fluck等人)研究发现，在低氧模拟物二甲氧甘氨酸(DMOG)处理的人单核细胞中，有100多个编码细胞表面分子的基因上调。为了确定候选的标记，然后根据其已知的表达谱筛选基因，并进行文献综述，以确定9个感兴趣的基因以供进一步探索。单核细胞被招募到肿瘤微环境中，在那里它们分化为TAMs。其中，在缺氧性人单核细胞来源的巨噬细胞(MDM)中，TNFRSF1B、TREM1、HCAR3和PLAUR表达增加。这些基因的产物随后通过流式细胞术在不同治疗的MDM中进行评估。细胞表面CD87(由PLAUR编码)、TREM1和TNFR2(TNFRSF1B)在低氧条件下以及在偏向促炎细胞因子的MDM中表达上调，代表了相关的低氧敏感标记，有待进一步研究。为了研究这些标记物在更多临床相关模型中的表达，用癌细胞系条件培养液(CM)产生的样巨噬细胞对它们的表达进行了评估。在常氧条件下，可能表达低氧相关蛋白的细胞系的CM显著上调了候选标志物。为了阐明它们的表达调控因素，癌细胞释放的细胞因子、趋化因子和外切体将通过酶联免疫吸附试验、聚合酶链式反应和超速离心法进行评估。这些标记物将通过免疫组织化学在原发人类肿瘤样本中进一步评估。在随后的验证之后，将获得或产生能够调节受体的单抗，以探索它们调节巨噬细胞行为的能力。

Tumour-associated macrophages (TAMs) are one of the most abundant populations in the tumour microenvironment and can contribute to cancer progression. The behaviour of macrophages is influenced by environmental stimuli, including hypoxia (oxygen deprivation), to further promote therapy resistance and cancer relapse. Whilst there is an interest in targeting TAMs to improve anti-tumour efforts, further study is needed to identify markers for hypoxic TAMs before developing therapies. An in silico surfaceome dataset (Bausch-Fluck et al.) revealed over 100 genes encoding cell surface molecules were upregulated in human monocytes treated with dimethyloxalylglycine (DMOG), a hypoxia mimetic. To identify candidate TAM markers, genes were then filtered based on their known expression profile and a literature review was conducted to identify 9 genes of interest to explore further. Monocytes are recruited to the tumour microenvironment where they differentiate into TAMs. Of these 9, TNFRSF1B, TREM1, HCAR3, and PLAUR expression was increased in hypoxic human monocyte-derived macrophages (MDMs). The product of these genes was then assessed by flow cytometry in differentially treated MDMs. Cell surface CD87 (encoded by PLAUR), TREM1, and TNFR2 (TNFRSF1B) was upregulated in the presence of hypoxia and also in MDMs skewed with pro-inflammatory cytokines, representing relevant hypoxia-sensitive markers to investigate further. To investigate the expression of these markers in more clinically relevant models, their expression was assessed on TAM-like macrophages generated using cancer cell line conditioned media (CM). CM from cell lines that may constitutively express hypoxia-related proteins considerably upregulated the candidate markers at normoxia. To elucidate the factors regulating their expression, cytokines, chemokines, and exosomes released by cancer cells will be assessed by ELISA, PCR, and ultracentrifugation. These markers will be further assessed by immunohistochemistry in primary human tumour samples. Following subsequent validation, monoclonal antibodies capable of modulating the receptor will be sourced or generated to explore their ability to modulate macrophage behaviour.