第一部分：

研究目的：颈动脉体瘤（CBT）是一种罕见的副神经节瘤，手术切除是唯一有效的治疗方法。由于它们靠近颈动脉、颈静脉和颅神经，手术极其困难，出血和神经血管损伤的风险很高。Shamblin分型常用于CBT的临床评估；然而，它们之间差异的分子机制尚不清楚。本研究的目的是探讨CBT发病机制和不同Shamblin分型之间的致病机制和分子差异。

研究方法：使用基于数据非依赖性采集技术（DIA）的蛋白质组学来鉴定Shamblin I、II和III型CBT的肿瘤组织中的差异表达蛋白（DEPs）。免疫组化用于验证DEPs。

研究结果：三种Shamblin分型的蛋白质组学图谱存在显著差异。生物信息学分析显示，肾上腺髓质素信号通路、间隙连接信号通路、蛋白激酶A信号通路、肾上腺素受体信号通路、IL-1信号通路、肌动蛋白细胞骨架信号通路、VEGF信号通路、内皮素-1信号通路、血管生成素信号通路、PPAR信号通路、BMP信号通路、HIF1α信号通路和IL-6信号通路显著富集。此外，60种差异表达蛋白（DEP）随着肿瘤进展而发生显著变化。免疫组织化学验证了重要的DEPs，包括AOX1、MED22、CPT1A和HSF1。

研究结论：这是首次在CBT肿瘤组织中使用蛋白质组学进行研究。我们的研究结果将加深对CBT相关发病机制的理解，并有助于确定CBT的治疗靶点。

第二部分：

研究目的：颈动脉体瘤（CBT）是一种罕见的位于颈动脉分叉处的肿瘤。临床中常用Shamblin分型进行评估，其中Shamblin III型的CBT特点是侵袭性强，与颈动脉血管关系复杂，需要手术切除，并与神经血管并发症的高发病率有关。本文研究了利用蛋白质组学技术识别颈动脉体瘤靶点，将手术与靶向治疗相结合，以减少术后并发症。

研究方法：本研究包括12例颈动脉体瘤患者（6例Shamblin II型和6例Shamblin III型）。440蛋白芯片用于筛选Shamblin II/III型颈动脉体瘤的新的基于组织的生物标志物。使用生物信息学分析两组之间差异表达蛋白，并使用定制蛋白芯片对22例颈动脉体瘤患者（11例Shamblin II型和11例Shamblin III型）进行了验证。
研究结果：使用定制蛋白芯片选择了15个靶点进行验证，其中12个生物标志物：TARC、CD40、6ckine、ENA-78、Tie-2、MCSF、IL-15R、TRAIL-R3、IL-16、LRIG3、Shh-N和IL-2-Rg与先前研究中报道的一致。
研究结论：我们的研究发现ENA-78是一种很有前景的生物标志物或靶点，可以区分II型和III型颈动脉体瘤（曲线下面积0.950）或治疗靶点。此外，TARC、CD40、6ckine、Tie-2、MCSF、白介素-15R、TRAIL-R3、LRIG3、Shh-N和IL-2-Rg代表了颈动脉体瘤的一组有前途的诊断或辅助治疗靶点。
 

Part 1

Objective: Carotid body tumors (CBTs) are a rare type of paraganglioma, with surgical resection as the only effective treatment. Because of their proximity to the carotid artery, jugular vein, and cranial nerve, surgery is extremely difficult. with high risks of hemorrhage and neurovascular injury. The Shamblin classification is used for the clinical evaluation of CBTs; however, the molecular mechanisms underlying their differences remain unclear. The purpose of this study was to explore the pathogenic mechanisms and molecular differences between CBT types.

Methods: Direct data-independent acquisition (DIA)-based proteomics was used to identify differentially expressed proteins (DEPs) in Shamblin I, II, and III tumors. Immunohistochemistry was used to validate the DEPs.

Results: Proteomics profiling of three Shamblin subtypes differed significantly. Bioinformatics analysis showed that adrenomedullin signaling pathway, gap-junction signaling, protein kinase A signaling, ephrin receptor signaling, IL-1 signaling, actin cytoskeleton signaling, VEGF signaling, endothelin-1 signaling, Angiopoietin signaling， PPAR signaling, BMP signaling pathway, HIF1α signaling, and IL-6 signaling pathways were significantly enriched. In addition, 60 differentially expressed proteins (DEPs) changed significantly with tumor progression. Immunohistochemistry validated several important DEPs, including AOX1, MED22, CPT1A, and HSF1.

Conclusions: To our knowledge, this is the first application of proteomics quantification in CBT. Our results will deepen the understanding of CBT-related pathogenesis and aid in identifying therapeutic targets for the treatment of CBT.

Part 2

Objective: Carotid body tumors are rare neoplasms located at the carotid bifurcation. Shamblin type III lesions are characterized by aggressive invasion and a complex relationship with carotid vessels, for which surgical excision is required and associated with high incidence of neurovascular complications. Herein, the use of proteomics technology was investigated for identifying carotid body tumor targets to combine surgery with targeted treatment to reduce postoperative complications.

Methods: This study included 12 patients with carotid body tumors (six type II and six type III). Protein array 440 was used to screen novel tissue-based biomarkers for Shamblin type II/III carotid body tumors. Proteins differentially expressed between the two groups were analyzed using bioinformatics and subjected to a customized array.

Results: Fifteen targets were selected for validation using a customized array, and 12 biomarkers were consistent with those reported in previous studies.

Conclusions: ENA-78 was found to be a promising biomarker or target to distinguish between type II and III carotid body tumors (area under the curve 0.950) or cure targets. Further, TARC, CD40, 6ckine, Tie-2, MCSF, interleukin-15R, TRAIL-R3, leucine-rich repeats and immunoglobulin-like domain 3, Sonic hedgehog-N, and interleukin 2Rg represent a panel of promising diagnostic or adjuvant therapy targets for carotid body tumors.